KINDAI UNIVERSITY


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KAJIYAMA Shinichiro

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FacultyDepartment of Biotechnological Science / Graduate School of Biology-Oriented Science and Technology
PositionProfessor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/142-kajiyama-shinichirou.html
URLhttp://www.waka.kindai.ac.jp/tea/biotech/labs/labs.html
Mail
Last Updated :2020/11/25

Education and Career

Education

  •  - 1989 , Kyoto University, Faculty of Agriculture
  •   1985 04  - 1989 03 , Kyoto University, Faculty of Agriculture
  •   1989 04  - 1991 03 , Kyoto University

Academic & Professional Experience

  •   2011 04 ,  - 現在, Professor, School of Bio-oriented Science and technology, Kinki Univ.
  •   2008 ,  - 2011 03 , Associate professor, School of Bio-oriented Science and technology, Kinki Univ.
  •   2005 ,  - 2008 , Invited Associate Professor, Grad. school of Engineering, Osaka Univ.
  •   1995 ,  - 2005 , Assintant Professor, Faculty of Engineering, Osaka Univ.
  •   1991 ,  - 1995 , Research Assistant, Faculty of Agriculture, Okayama Univ.

Research Activities

Research Areas

  • Life sciences, Bioorganic chemistry
  • Manufacturing technology (mechanical, electrical/electronic, chemical engineering), Applied biofunctional and bioprocess engineering
  • Nanotechnology/Materials, Biochemistry
  • Nanotechnology/Materials, Green/sustainable/environmental chemistry
  • Nanotechnology/Materials, Analytical chemistry
  • Environmental science/Agricultural science, Environmental materials/recycling technology
  • Environmental science/Agricultural science, Environmental load reduction/restoration technology
  • Environmental science/Agricultural science, Environmental policy and society
  • Environmental science/Agricultural science, Environmental impact assessment

Research Interests

  • post genome science, metabolite analysis, non-linear photonics, laser cell manipulation, Cell enginerring

Published Papers

  • Promotion of lipopeptide antibiotic production by Bacillus sp. IA in the presence of rice husk biochar, Pesticide Science, 44(1), 33 - 40, Sep. 2019 , Refereed
  • Antimicrobial activities by the floral scents of the Japanese bamboo lily Lilium japonicum, the symbolic flowers offered to the deity at the Saikusa Festival, 花﨑 直史, 岩佐 郁, 藤澤 瑞希, 高橋 康樹, 柳川 綾, 梶山 慎一郎, 瀧川 義浩, 松川 哲也, 荒井 滋, 秋野 順治, Aroma research = アロマリサーチ : journal of aroma science technology and safety, Aroma research = アロマリサーチ : journal of aroma science technology and safety, 20(2), 137 - 143, May 28 2019 , Refereed
  • Antimicrobial activities by floral scents of the Japanese bamboo lily Lilium japonicum, the symbolic flowers offered the deity at the Saikusa Festival., Naoshi Hanasaki, Iku Iwasa, Yasuki, Takahashi, Aya Yanagawa, Shiichirou Kajiwara, AROMA RESEARCH, AROMA RESEARCH, 78(20), 41 - 47, May 2019 , Refereed
  • A Questionnaire-based Investigation Indicated Mental Relaxation Effects in the Scent of the Sacred Flower of the Saikusa Festival, Lilium japonicum, 藤澤 瑞希, 花崎 直史, 瀧川 義浩, 柳川 綾, 松川 哲也, 秋野 順治, 梶山 慎一郎, 荒井 滋, 生薬学雑誌, 生薬学雑誌, 73(1), 1 - 6, Feb. 20 2019 , Refereed
  • Past and Current trends in research of Lilium japnicum, sasayuri, 瀧川義浩, 田中普祟, 藤澤瑞希, 松川哲也, 梶山慎一郎, 近畿大学先端技術総合研究所紀要, 近畿大学先端技術総合研究所紀要, (23), 11‐22, Mar. 31 2018
  • Biological Control Potential of Streptomyces sp. AR10 Producing Albocycline Isolated from Soil around Ant Nest, Tatsuya Ohike, Tetsuya Matsukawa, Masahiro Okanami, Shin’ichiro Kajiyama, Takashi Ano, Journal of Agricultural Science, Journal of Agricultural Science, 10(3), 54 - 61, Jan. 2018 , Refereed
  • In vitro and in vivo Assay for Assessment of the Biological Control Poterntial of Streptomyces sp. KT, Tatsuya Ohike, Minori Maeda, Tetsuya Matsukawa, Masahiro Okanami, Shin’ichiro Kajiyama, Takashi Ano, Journal of Plant Studies, Journal of Plant Studies, 7(1), 10 - 18, Jan. 2018 , Refereed
  • Inhibition of Advanced Glycation End Products Formation by Mangifera indica Leaf Extract, Kimihisa Itoh, Kazuya Murata, Nao Sakaguchi, Kohei Akai, Tomoka Yamaji, Kohsuke Shimizu, Kaoru Isaki, Tetsuya Matsukawa, Shin'ichiro Kajiyama, Masahiko Fumuro, Morio Iijima, Hideaki Matsuda, Journal of Plant Studies, Journal of Plant Studies, 6(2), 102 - 107, Jul. 2017 , Refereed
  • Protection and current situation of rare plant "Sasa yuri" in satoyama, 瀧川 義浩, 松川 哲也, 宮下 実, 梶山 慎一郎, Memoirs of Institute of Advanced Technology, Kinki University, Memoirs of Institute of Advanced Technology, Kinki University, (22), 35 - 40, Mar. 31 2017
    Summary:本論では里山におけるササユリの自生環境、個体数の減少の要因および近畿大学 先端技術総合研究所(研究所)と生物理工学部で実施している活動や研究について記載した。Sasa yuri(Lilium japnicum)is a wild lily in japan and also known as an index plant for evaluating a natural environment of satoyama. In the present paper, we described current situation and protection of the rare plant "Sasa yuri ".
  • A Pancreatic Lipase Inhibitory Activity by Mango (Mangifera indica) Leaf Methanolic Extract, Kimihisa Itoh, Kazuya Murata, Yuta Nakagaki, Ayaka Shimizu, Yusuke Takata, Kohsuke Shimizu, Tetsuya Matsukawa, Shin'ichiro Kajiyama, Masahiko Fumuro, Morio Iijima, Hideaki Matsuda, Journal of Plant Studies, Journal of Plant Studies, 5(2), 72 - 78, Jun. 2016 , Refereed
  • Lansioside D, a new triterpenoid glycoside antibiotic, Eufrocinio C. Marfori, Shin Ichiro Kajiyama, Ei-Ichiro Fukusaki and, Journal of Pharmacognosy and Phytochemistry, Journal of Pharmacognosy and Phytochemistry, 13(5), 258 - 261, May 2015 , Refereed
  • バイオディーゼル原料植物Jatropha Curcasの毒性代謝産物とその制御, MATSUKAWA TETSUYA, KAJIYAMA SHIN'ICHIRO, スマートプロセス学会誌, スマートプロセス学会誌, 3(5), 258 - 261, Sep. 20 2014
  • マウス二段階発がん試験とEBV早期抗原誘導活性を指標としたJatropha curcas種子抽出物の発がんプロモーション活性評価, MATSUKAWA TETSUYA, SHODA MASAHIRO, TAMAKI KEIKO, MURAKAMI HIKARU, KAJIYAMA SHIN'ICHIRO, Mem Fac Biol Oriented Sci Technol Kinki Univ, Mem Fac Biol Oriented Sci Technol Kinki Univ, 33(33), 9 - 16, Mar. 31 2014
    Summary:[要旨]トウダイグサ科の落葉低木であるJatropha curcasは, 種子に豊富な中性脂質を含み, 荒廃地でも生育可能であるため次世代バイオディーゼル原料植物として有望視されている。しかし一方で, 本植物の種子には発がんプロモーターであるphorbol ester類を含んでおり, 生産者や消費者の健康に与える負の影響が懸念されている。本研究ではJ. curcas 種子抽出物の発がんプロモーション活性を, マウスを用いた二段階発がん試験およびEBV早期抗原誘導活性を指標としたアッセイ系により評価した。その結果, J. curcas種子抽出物の発がんプロモーション活性は, 発がんプロモーション活性を有する代表的なホルポールエステルであるphorbol-12-myristate-13-acetate(PMA)より低いことが見いだされた。また,種子抽出物はPMAの発がん活性を抑制することが示された。これらの結果から, J. curcas種子抽出物には化学発がんを抑制する物質が含まれることが示唆された。 [Abstract] Jatr•opha eurcas, a tropical plant belonging to the family of Euphorbiaceac, has recently been attracted the considerable attention as a potential source for bio-diesel due to the high adaptability to semi-arid areas and the high oil content in seed kernel. The seed, however, contains phorbol esters which exhibit tumor-promoting activity, and thus negative impact of these compounds on health of producers or consumers should be concerned_ In this study, tumor-promoting activity of I cur eas seed extracts was evaluated by the two-stage mouse skin tumorgenesis assay and Epstein-Barr virus early antigen induction assay. Our results showed that tumor-promoting activity of J. curcas seed extracts was significantly lower than phorbol-l2-myristate-1 3-acetate (PMA), which is well known as a potent tumor promotor. In addition, J. cur eas seed extracts were revealed to suppress the tumor-promoting activity ofPMA. These results suggest that./ curcas seed extracts contain some suppressor of tumorgenesis.
  • 〈Original Papers〉 Evaluation of tumor promoting activity of Jatropha curcas seed extracts by the two-stage mouse skin tumorgenesis assay and Epstein-Barr virus early antigen induction assay., 松川 哲也, 正田 将大, 玉置 恵子, 村上 ひかる, 梶山 慎一, 近畿大学生物理工学部紀要 = Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University, 近畿大学生物理工学部紀要 = Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University, 33, 9 - 16, Mar. 01 2014
    Summary:[要旨]トウダイグサ科の落葉低木であるJatropha curcasは, 種子に豊富な中性脂質を含み, 荒廃地でも生育可能であるため次世代バイオディーゼル原料植物として有望視されている。しかし一方で, 本植物の種子には発がんプロモーターであるphorbol ester類を含んでおり, 生産者や消費者の健康に与える負の影響が懸念されている。本研究ではJ. curcas 種子抽出物の発がんプロモーション活性を, マウスを用いた二段階発がん試験およびEBV早期抗原誘導活性を指標としたアッセイ系により評価した。その結果, J. curcas種子抽出物の発がんプロモーション活性は, 発がんプロモーション活性を有する代表的なホルポールエステルであるphorbol-12-myristate-13-acetate(PMA)より低いことが見いだされた。また,種子抽出物はPMAの発がん活性を抑制することが示された。これらの結果から, J. curcas種子抽出物には化学発がんを抑制する物質が含まれることが示唆された。 [Abstract] Jatr•opha eurcas, a tropical plant belonging to the family of Euphorbiaceac, has recently been attracted the considerable attention as a potential source for bio-diesel due to the high adaptability to semi-arid areas and the high oil content in seed kernel. The seed, however, contains phorbol esters which exhibit tumor-promoting activity, and thus negative impact of these compounds on health of producers or consumers should be concerned_ In this study, tumor-promoting activity of I cur eas seed extracts was evaluated by the two-stage mouse skin tumorgenesis assay and Epstein-Barr virus early antigen induction assay. Our results showed that tumor-promoting activity of J. curcas seed extracts was significantly lower than phorbol-l2-myristate-1 3-acetate (PMA), which is well known as a potent tumor promotor. In addition, J. cur eas seed extracts were revealed to suppress the tumor-promoting activity ofPMA. These results suggest that./ curcas seed extracts contain some suppressor of tumorgenesis.
  • Plant metabolite analysis with laser optics., KAJIYAMA Shinichiro, Regulation of Plant Growth & Development, Regulation of Plant Growth & Development, 48(2), 163 - 168, Dec. 20 2013
    Summary:In recent decades, laser optics has been strenuously adapted to biological and biotechnological applications. Laser has high monochromaticity, directivity, and coherency and these properties are considered to be suitable for manipulating and/or observing intact cells with high special resolution. In this technical note, several laser-based microsurgery and non-linear optical microscopy techniques for plant metabolite analysis was discussed.
  • Distributional Analysis of Phorbol Esters in J. curcas and J. integerrima., 大池 達矢, 松川 哲也, 岡南 政宏, 阿野 貴司, 梶山 慎一郎, Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University, Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University, (27), 29 - 37, Mar. 2011
    Summary:[要旨] トウダイグサ科の落葉低木であるJatropha curcasは、種子の仁に40-60%の中性脂質を含んでおり、乾燥地域や貧栄養土壌などの荒廃地でも生育可能であるため、現在、東南アジア諸国で使用され、バイオディーゼル燃料(BDF)となる資源として注目されている。しかし、J. curcasの種子中からは、発がんプロモーション活性物質であるフォルボールエステル類が検出されており、BDF生産者の健康や環境への影響が懸念されている。その一方で、Jatrophaは、観賞用植物としても親しまれており、街路樹に利用されるなど現在でも多くの種が流通している。しかしながら、観賞用のJatrophaは、BDF用のJatrophaに比べると商業的な価値が低いため、種子や各器官に関する基礎的な知見が乏しい。そこで本研究では、Jatropha integerrimaにおけるフォルボールエステル類の分布ををJatropha curcasを元に、種子油の脂肪酸組成分析を行なった。その結果、Jatropha integerrimaの種子および葉においてフォルボールエステル類の存在が見出され、種子油に含まれる脂肪酸の組成は、両植物では大きく異なることが分かった。本研究の結果は、観賞用Jatrophaにおけるフォルボールエステル類のリスク評価と、BDF植物の優良品種開発に寄与すると期待される。 [Abstract]Jatropha curcas, a member of Euphorbiaceae family, has recently been attracted the considerable attention as a potential source of bio-diesel due to the high adaptbility to arid and oligotrophic soil and oil concern (40-60%) in seed kernel. The seed, however, contains toxic compounds including phorbol esters, well known as a tumor promoting agent, and thus influence of these compounds on environment and people engaged in the production should be concerned. In contrast, I integerrima, widely distributed for ornamental use, has not been focused, and little is know about phorbol esters contained in this species. In this study, we performed phorbol esters analysis and comparison of fatty acid composition between J. curcas and I integerrima. Phorbol esters were detected in J. integerrima seeds and leaves as well as J. curcas, while profiles of fatty acids were different among these species.
  • Distributional Analysis of Phorbol Esters in J. curcas and J. integerrima., 大池 達矢, 松川 哲也, 岡南 政宏, 阿野 貴司, 梶山 慎一郎, Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University = 近畿大学 生物理工学部 紀要, Memoirs of the Faculty of Biology-Oriented Science and Technology of Kinki University = 近畿大学 生物理工学部 紀要, (27), 29 - 37, Mar. 01 2011
    Summary:[要旨] トウダイグサ科の落葉低木であるJatropha curcasは、種子の仁に40-60%の中性脂質を含んでおり、乾燥地域や貧栄養土壌などの荒廃地でも生育可能であるため、現在、東南アジア諸国で使用され、バイオディーゼル燃料(BDF)となる資源として注目されている。しかし、J. curcasの種子中からは、発がんプロモーション活性物質であるフォルボールエステル類が検出されており、BDF生産者の健康や環境への影響が懸念されている。その一方で、Jatrophaは、観賞用植物としても親しまれており、街路樹に利用されるなど現在でも多くの種が流通している。しかしながら、観賞用のJatrophaは、BDF用のJatrophaに比べると商業的な価値が低いため、種子や各器官に関する基礎的な知見が乏しい。そこで本研究では、Jatropha integerrimaにおけるフォルボールエステル類の分布ををJatropha curcasを元に、種子油の脂肪酸組成分析を行なった。その結果、Jatropha integerrimaの種子および葉においてフォルボールエステル類の存在が見出され、種子油に含まれる脂肪酸の組成は、両植物では大きく異なることが分かった。本研究の結果は、観賞用Jatrophaにおけるフォルボールエステル類のリスク評価と、BDF植物の優良品種開発に寄与すると期待される。 [Abstract]Jatropha curcas, a member of Euphorbiaceae family, has recently been attracted the considerable attention as a potential source of bio-diesel due to the high adaptbility to arid and oligotrophic soil and oil concern (40-60%) in seed kernel. The seed, however, contains toxic compounds including phorbol esters, well known as a tumor promoting agent, and thus influence of these compounds on environment and people engaged in the production should be concerned. In contrast, I integerrima, widely distributed for ornamental use, has not been focused, and little is know about phorbol esters contained in this species. In this study, we performed phorbol esters analysis and comparison of fatty acid composition between J. curcas and I integerrima. Phorbol esters were detected in J. integerrima seeds and leaves as well as J. curcas, while profiles of fatty acids were different among these species.
  • Enzymatic Production of (-)-Indolactam V by LtxB, a Cytochrome P450 Monooxygenase, Minh U. Huynh, Matthew C. Elston, Nick M. Hernandez, David B. Ball, Shin-ichiro Kajiyama, Kazuhiro Irie, William H. Gerwick, Daniel J. Edwards, JOURNAL OF NATURAL PRODUCTS, JOURNAL OF NATURAL PRODUCTS, 73(1), 71 - 74, Jan. 2010 , Refereed
    Summary:The P450 cytochrome monooxygenase gene, ltxB, was cloned and overexpressed in Escherichia coli as a 6xHis-taoged protein. The resulting recombinant LtxB was purified by Ni-NTA affinity chromatography and characterized biochemically. Purified LtxB demonstrated typical cytochrome P450 spectroscopic properties including substrate-induced transition from a low-spin (lambda(max) = 414 nm) to high-spin state (lambda(max) = 386 nm) upon incubation with N-methyl-L-valyl-L-tryptophanol. The catalytic activity of LtxB was verified by demonstrating the oxidation/cyclization of N-methyl-L-valyl-L-tryptophanol to (-)-indolactam V. LtxB shows a relaxed specificity for analogue substrates in which the valyl group is substituted for other aliphatic groups. The relaxed substrate specificity of LtxB, along with the relaxed specificity of the prenyltransferase, LtxC, allowed for the enzymatic production of a series of (-)-indolactam V and lyngbyatoxin analogues.
  • Development of an enzyme activity screening system for beta-glucosidase-displaying yeasts using calcium alginate micro-beads and flow sorting, Yan Zhou, Shin'ichiro Kajiyama, Kouichi Itoh, Takanori Tanino, Nobuo Fukuda, Tsutomu Tanaka, Akihiko Kondo, Kiichi Fukui, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 84(2), 375 - 382, Aug. 2009
    Summary:Recent reports on high-speed affinity screening systems for yeast cells using flow cytometry have not been adapted to screening yeast cells that display hydrolyzing enzymes, since the fluorescent molecules which are released from fluoresceinated substrate diffuse into solution after enzymatic reaction. In this research, yeast cells displaying beta-glycosidase were individually captured in micro-sized calcium alginate beads by using the newly developed reverse micelle method to prevent diffusion of hydrolyzed fluorescent substrates. By adopting flow sorting to these captured cells, active cells were successfully enriched about 82-fold from a mixed suspension with negative controls. This system should be a useful method for high-speed screening of yeast cells that display various hydrolyzing enzymes and has potential application to screening randomized libraries of enzyme-displayed yeast cells with higher activities.
  • A new size and shape controlling method for producing calcium alginate beads with immobilized proteins, J.Biomed. Sci Eng., J.Biomed. Sci Eng., 2(5), 287 - 293, 2009
  • Single cell sampling and analysis by ultra short pulse laser, Kajiyama Shinichiro, Sakakura Masaaki, Ueda Mitsuyoshi, Itoh Kazuyoshi, Fukui Kiichi, PROGRESS ON POST-GENOME TECHNOLOGIES, PROGRESS ON POST-GENOME TECHNOLOGIES, 25 - 28, 2007 , Refereed
  • JST Innovation Plaza KYOTO UEDA Project(Gravure & Interview,Precision Engineering for Visualization of Biological Functions), YABUKI Akihiko, Journal of the Japan Society of Precision Engineering, Journal of the Japan Society of Precision Engineering, 72(11), 1307 - 1310, Nov. 05 2006
  • JST Innovation Plaza KYOTO UEDA Project, UEDA Mitsuyoshi, KAJIYAMA Shin'ichiro, YABUKI Akihiko, 精密工学会誌 = Journal of the Japan Society of Precision Engineering, 精密工学会誌 = Journal of the Japan Society of Precision Engineering, 72(11), 1307 - 1310, Nov. 05 2006
  • Plant metabolite analysis using laser single cell sampling technique, S Kajiyama, JOURNAL OF PESTICIDE SCIENCE, JOURNAL OF PESTICIDE SCIENCE, 30(1), 57 - 60, 2005
  • The LASER Micro-Processing of Living Cells and Its Application for Plant Biotechnology, KOBAYASHI Akio, KAJIYAMA Shin'ichiro, 化学と生物, 化学と生物, 42(9), 596 - 603, Sep. 25 2004
  • A Novel Gene Delivery System in Plants with Calcium Alginate, 曽根 岳史, 長森 英二, 池内 智彦, 水上 温司, 高倉 友紀子, 梶山 慎一郎, 福崎 英一郎, 原島 俊, 小林 昭雄, 福井 希一, Seibutsu-kogaku Kaishi, Seibutsu-kogaku Kaishi, 82(3), Mar. 25 2004
  • The LASER Micro-Processing of Living Cells and Its Application for Plant Biotechnology, KOBAYASHI Akio, KAJIYAMA Shin'ichiro, KAGAKU TO SEIBUTSU, KAGAKU TO SEIBUTSU, 42(9), 596 - 603, 2004
  • Chemical and Biochemical Aspects of Desiccation Tolerant Cyanobacterium Nostoc commune (The Seminar, "Stresses and Responses and Adaptation of Living Organisms : Molecular and Cellular Mechanisms and its Application"), KAJIYAMA Shinichiro, KOBAYASHI Akio, Cryobiology and cryotechnology, Cryobiology and cryotechnology, 49(1), 37 - 42, Aug. 30 2003
    Summary:Nostoc commune, a nitrogen-fixing filamentous cyanobacterium, produces copious amounts of the EPS (extracellular polysaccharide) around the cells and has remarkable desiccation tolerance. Recently, biochemical experiment revealed that the EPS can serve to stabilize cyanobacterial cells during desiccation and subsequent rehydration. The EPS also seems to act as the physical and/or chemical barrier to the surroundings. Despite marked features of EPS of this cyanobacteria, little is known about its chemical structure, due largely to structural complexity. In this paper, the chemical and biochemical aspects of this cyanobacterial EPS are discussed.
  • SELEX for tubulin affords specific T-rich DNA aptamers, FUKUSAKI Ei-ichiro, HASUNMA Tomohisa, KAJIYAMA Shin-ichiro, OKAZAWA Atsushi, ITOH J Tomohiko, KOBAYASHI Akio, Bioorganic & Medicinal Chemistry Letters, Bioorganic & Medicinal Chemistry Letters, Vol. 11, No. 22, pp. 2927-2930, Nov. 2001 , Refereed
  • Quantitative structure-activity studies on indole alkaloid tumor promoter indolactam congeners, K. Irie, S. Okuno, S. Kajiyama, K. Koshimizu, H. Nishino, A. Iwashima, Carcinogenesis, 12/10, 1883-1886, Carcinogenesis, 12/10, 1883-1886, 1991 , Refereed
  • Biosynthesis of indole alkaloid-tumor promoters teleocidins (I): possible biosynthetic pathway of the monoterpenoid moieties of teleocidins, K. Irie, S. Kajiyama, A. Funaki, K. Koshimizu, H.Hayashi, M. Arai, Tetrahedron, 46/8, 2773-2788, Tetrahedron, 46/8, 2773-2788, 1990 , Refereed
  • Pancreatic Lipase Inhibitory Activity of Citrus unshiu Leaf Extract, Kimihisa Itoh, Tetsuya Matsukawa, Kazuya Murata, Ryota Nishitani, Momona Yamagami, Norimichi Tomohiro, Shin'ichiro Kajiyama, Masahiko Fumuro, Morio Iijima, Shigeru Shigeoka, Yuichi Endo, Hideaki Matsuda, Natural Product Communications, Natural Product Communications, 14(9), 1934578X1987343 - 1934578X1987343, Sep. 2019 , Refereed
    Summary:In the cultivation of Citrus fruit, pruning in late winter after harvest is important to obtain excellent flower buds in the next spring and a rich harvest of superior fruit in the following winter; a large number of pruned leaves are usually discarded. The purpose of this study was to determine the utility value of pruned leaves of Citrus unshiu. A methanol extract of pruned C. unshiu leaves showed a porcine pancreatic lipase inhibitory activity. Fractionation of C. unshiu leaf extract (CUL-ext) followed by bibliographic and chromatographic analyses revealed that a part of the pancreatic lipase inhibitory activity of CUL-ext was attributable to nobiletin (1), rutin (2), and hesperidin (3). Thus, pruned C. unshiu leaves may be a reasonable natural resource for the preparation of ingredients with lipase inhibitory activity.
  • Metabolomic analysis of primary metabolites in citrus leaf during defense responses, Tomonori Asai, Tetsuya Matsukawa, Shin'ichiro Kajiyama, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 123(3), 376 - 381, Mar. 2017 , Refereed
    Summary:Mechanical damage is one of the unavoidable environmental stresses to plant growth and development. Plants induce a variety of reactions which defend against natural enemies and/or heal the wounded sites. Jasmonic acid (JA) and salicylic acid (SA), defense-related plant hormones, are well known to be involved in induction of defense reactions and play important roles as signal molecules. However, defense related metabolites are so numerous and diverse that roles of individual compounds are still to be elucidated. In this report, we carried out a comprehensive analysis of metabolic changes during wound response in citrus plants which are one of the most commercially important fruit tree families. Changes in amino acid, sugar, and organic acid profiles in leaves were surveyed after wounding, JA and SA treatments using gas chromatography mass spectrometry (GC/MS) in seven citrus species, Citrus sinensis, Citrus limon, Citrus paradisi, Citrus unshiu, Citrus kinokuni, Citrus grandis, and Citrus hassaku. GC/MS data were applied to multivariate analyses including hierarchical cluster analysis (HCA), primary component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) to extract stress-related compounds. HCA showed the amino acid cluster including phenylalanine and tryptophan, suggesting that amino acids in this cluster are concertedly regulated during responses against treatments. OPLS-DA exhibited that tryptophan was accumulated after wounding and JA treatments in all species tested, while serine was down regulated. Our results suggest that tryptophan and serine are common biomarker candidates in citrus plants for wound stress. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.
  • Isolation and characterization of wound-induced compounds from the leaves of Citrus hassaku, Tomonori Asai, Tetsuya Matsukawa, Atsushi Ishihara, Shin'ichiro Kajiyama, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 122(2), 208 - 212, Aug. 2016 , Refereed
    Summary:Citrus plants are world widely cultivated as horticultural tree crops, and nowadays their pharmacological activities have been well studied. Since research of defense responses in citrus plants have been mainly focused on the post-harvested fruits because of their commercial importance, defense mechanisms during their developmental stages have not been well understood. In the present study, two wound-induced compounds were isolated from leaves of Citrus hassalcu, and their structures were elucidated by high-resolution electron spray ionization mass spectra (HRESIMS) and nuclear magnetic resonance (NMR) analyses. One of these compounds was identified as a known flavanone, hesperetin. The other was characterized as a novel furofuran lignan, and was named `biscitrusnin-A'. Their antimicrobial activities were also evaluated. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.
  • Metabolic changes in Citrus leaf volatiles in response to environmental stress, Tomonori Asai, Tetsuya Matsukawa, Shin'ichiro Kajiyamal, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 121(2), 235 - 241, Feb. 2016 , Refereed
    Summary:Citrus plants are well known as a rich source of VOCs, and several have important roles in defense responses. However, how VOCs are regulated in response to environmental stress is not yet well understood. In this study, we investigated dynamic changes of VOCs present in leaves of seven Citrus species (Citrus sinensis, C. limon, C. paradisi, C. unshiu, C. kinokuni, C grandis, and C hassaku) in response to mechanical wounding, jasmonic acid (JA), and salicylic acid (SA) as determined by gas chromatography/mass spectrometric analysis followed by multivariate analysis (principal component analysis, PCA, and orthogonal partial least squares-discriminant analysis, OPLS-DA). PCA and OPLS-DA suggested that changes in VOC profiles against stress stimuli were much diverse among Citrus species. OPLS-DA showed that C6 volatiles, such as hexanal and trans-2-hexenal, were induced in response to JA and SA stimuli in C sinensis and C. grandis, while the other VOCs were decreased under all tested stress conditions. alpha-Farnesene was induced in all species except C. hassaku after wounding or JA treatment. In addition, alpha-farnesene was also induced in response to SA stimuli in C. unshiu and C kinokuni. Therefore these volatiles can be candidates of the common stress biomarkers in Citrus. Our results will give a new insight into defense mechanisms in Citrus species. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.
  • Potential application of oligotrophic bacteria as biocontrol agents, Tatsuya Ohike, Tetsuya Matsukawa, Masahiro Okanami, Shin'ichiro Kajiyama, Takashi Ano, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 248, Aug. 2014 , Refereed
  • Visualization of oil body distribution in Jatropha curcas L. by four-wave mixing microscopy, Makiko Ishii, Susumu Uchiyama, Yasuyuki Ozeki, Sin'ichiro Kajiyama, Kazuyoshi Itoh, Kiichi Fukui, Japanese Journal of Applied Physics, Japanese Journal of Applied Physics, 52(6), 062403, Jun. 2013 , Refereed
    Summary:Jatropha curcas L. (jatropha) is a superior oil crop for biofuel production. To improve the oil yield of jatropha by breeding, the development of effective and reliable tools to evaluate the oil production efficiency is essential. The characteristics of the jatropha kernel, which contains a large amount of oil, are not fully understood yet. Here, we demonstrate the application of four-wave mixing (FWM) microscopy to visualize the distribution of oil bodies in a jatropha kernel without staining. FWM microscopy enables us to visualize the size and morphology of oil bodies and to determine the oil content in the kernel to be 33.2%. The signal obtained from FWM microscopy comprises both of stimulated parametric emission (SPE) and coherent anti-Stokes Raman scattering (CARS) signals. In the present situation, where a very short pump pulse is employed, the SPE signal is believed to dominate the FWM signal. © 2013 The Japan Society of Applied Physics.
  • A Novel Gene Delivery System in Plants with Calcium Alginate Micro-Beads, Naoki Wada, Shin'ichiro Kajiyama, Naruemon Khemkladngoen, Kiichi Fukui, Plant Transformation Technologies, Plant Transformation Technologies, 73 - 81, Mar. 16 2011 , Refereed
  • Sequence Analysis of the Genome of an Oil-Bearing Tree, Jatropha curcas L., Shusei Sato, Hideki Hirakawa, Sachiko Isobe, Elgo Fukai, Akiko Watanabe, Midori Kato, Kumiko Kawashima, Chiharu Minami, Akiko Muraki, Naomi Nakazaki, Chika Takahashi, Shinobu Nakayama, Yoshie Kishida, Mitsuyo Kohara, Manabu Yamada, Hisano Tsuruoka, Shigemi Sasamoto, Satoshi Tabata, Tomoyuki Aizu, Atsushi Toyoda, Tadasu Shin-i, Yohei Minakuchi, Yuji Kohara, Asao Fujiyama, Suguru Tsuchimoto, Shin'ichiro Kajiyama, Eri Makigano, Nobuko Ohmido, Nakako Shibagaki, Joyce A. Cartagena, Naoki Wada, Tsutomu Kohinata, Alipour Atefeh, Shota Yuasa, Sachihiro Matsunaga, Kiichi Fukui, DNA RESEARCH, DNA RESEARCH, 18(1), 65 - 76, Feb. 2011
    Summary:The whole genome of Jatropha curcas was sequenced, using a combination of the conventional Sanger method and new-generation multiplex sequencing methods. Total length of the non-redundant sequences thus obtained was 285 858 490 bp consisting of 120 586 contigs and 29 831 singlets. They accounted for,similar to 95% of the gene-containing regions with the average G + C content was 34.3%. A total of 40 929 complete and partial structures of protein encoding genes have been deduced. Comparison with genes of other plant species indicated that 11529 (4%) of the putative protein-encoding genes are specific to the Euphorbiaceae family. A high degree of microsynteny was observed with the genome of castor bean and, to a lesser extent, with those of soybean and Arabidopsis thaliana. In parallel with genome sequencing, cDNAs derived from leaf and callus tissues were subjected to pyrosequencing, and a total of 21 225 unigene data have been generated. Polymorphism analysis using microsatellite markers developed from the genomic sequence data obtained was performed with 12 J. curcas lines collected from various parts of the world to estimate their genetic diversity. The genomic sequence and accompanying information presented here are expected to serve as valuable resources for the acceleration of fundamental and applied research with J. curcas, especially in the fields of environment-related research such as biofuel production. Further information on the genomic sequences and DNA markers is available at http://www.kazusa.or.jp/jatropha/.
  • Sequence Analysis of the Genome of an Oil-Bearing Tree, Jatropha curcas L., Shusei Sato, Hideki Hirakawa, Sachiko Isobe, Elgo Fukai, Akiko Watanabe, Midori Kato, Kumiko Kawashima, Chiharu Minami, Akiko Muraki, Naomi Nakazaki, Chika Takahashi, Shinobu Nakayama, Yoshie Kishida, Mitsuyo Kohara, Manabu Yamada, Hisano Tsuruoka, Shigemi Sasamoto, Satoshi Tabata, Tomoyuki Aizu, Atsushi Toyoda, Tadasu Shin-i, Yohei Minakuchi, Yuji Kohara, Asao Fujiyama, Suguru Tsuchimoto, Shin'ichiro Kajiyama, Eri Makigano, Nobuko Ohmido, Nakako Shibagaki, Joyce A. Cartagena, Naoki Wada, Tsutomu Kohinata, Alipour Atefeh, Shota Yuasa, Sachihiro Matsunaga, Kiichi Fukui, DNA RESEARCH, DNA RESEARCH, 18(1), 65 - 76, Feb. 2011 , Refereed
    Summary:The whole genome of Jatropha curcas was sequenced, using a combination of the conventional Sanger method and new-generation multiplex sequencing methods. Total length of the non-redundant sequences thus obtained was 285 858 490 bp consisting of 120 586 contigs and 29 831 singlets. They accounted for,similar to 95% of the gene-containing regions with the average G + C content was 34.3%. A total of 40 929 complete and partial structures of protein encoding genes have been deduced. Comparison with genes of other plant species indicated that 11529 (4%) of the putative protein-encoding genes are specific to the Euphorbiaceae family. A high degree of microsynteny was observed with the genome of castor bean and, to a lesser extent, with those of soybean and Arabidopsis thaliana. In parallel with genome sequencing, cDNAs derived from leaf and callus tissues were subjected to pyrosequencing, and a total of 21 225 unigene data have been generated. Polymorphism analysis using microsatellite markers developed from the genomic sequence data obtained was performed with 12 J. curcas lines collected from various parts of the world to estimate their genetic diversity. The genomic sequence and accompanying information presented here are expected to serve as valuable resources for the acceleration of fundamental and applied research with J. curcas, especially in the fields of environment-related research such as biofuel production. Further information on the genomic sequences and DNA markers is available at http://www.kazusa.or.jp/jatropha/.
  • Stimulated Raman scattering microscope with shot noise limited sensitivity using subharmonically synchronized laser pulses, Yasuyuki Ozeki, Yuma Kitagawa, Kazuhiko Sumimura, Norihiko Nishizawa, Wataru Umemura, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, OPTICS EXPRESS, OPTICS EXPRESS, 18(13), 13708 - 13719, Jun. 2010 , Refereed
    Summary:We propose and demonstrate the use of subharmonically synchronized laser pulses for low-noise lock-in detection in stimulated Raman scattering (SRS) microscopy. In the experiment, Yb-fiber laser pulses at a repetition rate of 38 MHz are successfully synchronized to Ti:sapphire laser pulses at a repetition rate of 76 MHz with a jitter of <8 fs by a two-photon detector and an intra-cavity electro-optic modulator. By using these pulses, high-frequency lock-in detection of SRS signal is accomplished without high-speed optical modulation. The noise level of the lock-in signal is found to be higher than the shot noise limit only by 1.6 dB. We also demonstrate high-contrast, 3D imaging of unlabeled living cells. (C) 2010 Optical Society of America
  • Structure and Functional Characterization of Vibrio parahaemolyticus Thermostable Direct Hemolysin, Itaru Yanagihara, Kumiko Nakahira, Tsutomu Yamane, Shuji Kaieda, Kouta Mayanagi, Daizo Hamada, Takashi Fukui, Kiyouhisa Ohnishi, Shin'ichiro Kajiyama, Toshiyuki Shimizu, Mamoru Sato, Takahisa Ikegami, Mitsunori Ikeguchi, Takeshi Honda, Hiroshi Hashimoto, JOURNAL OF BIOLOGICAL CHEMISTRY, JOURNAL OF BIOLOGICAL CHEMISTRY, 285(21), 16267 - 16274, May 2010 , Refereed
    Summary:Thermostable direct hemolysin (TDH) is a major virulence factor of Vibrio parahaemolyticus that causes pandemic foodborne enterocolitis mediated by seafood. TDH exists as a tetramer in solution, and it possesses extreme hemolytic activity. Here, we present the crystal structure of the TDH tetramer at 1.5 angstrom resolution. The TDH tetramer forms a central pore with dimensions of 23 angstrom in diameter and similar to 50 angstrom in depth. pi-Cation interactions between protomers comprising the tetramer were indispensable for hemolytic activity of TDH. The N-terminal region was intrinsically disordered outside of the pore. Molecular dynamic simulations suggested that water molecules permeate freely through the central and side channel pores. Electron micrographs showed that tetrameric TDH attached to liposomes, and some of the tetramer associated with liposome via one protomer. These findings imply a novel membrane attachment mechanism by a soluble tetrameric pore-forming toxin.
  • The effects of puroindoline b on the ultrastructure of endosperm cells and physicochemical properties of transgenic rice plant, Naoki Wada, Shin'ichiro Kajiyama, Joyce A. Cartagena, Linyen Lin, Yukio Akiyama, Motoyasu Otani, Go Suzuki, Yasuhiko Mukai, Noriaki Aoki, Kiichi Fukui, JOURNAL OF CEREAL SCIENCE, JOURNAL OF CEREAL SCIENCE, 51(2), 182 - 188, Mar. 2010
    Summary:Endosperm texture is an important factor governing the end-product quality of cereals. The texture of wheat (Triticum aestivum L) endosperm is controlled by puroindoline a and b genes which are both absent in rice (Oryza sativa L). It has been reported that the endosperm texture of rice can be modified by puroindoline genes. The mechanism, however, by which puroindolines affect the ultrastructure of rice endosperm cells remains to be investigated. In this study, we observed the ultrastructure of endosperm cells and the morphology of isolated starch granules of the transgenic rice expressing the puroindoline b gene. SEM and TEM observations indicated that compound starch granules were embedded within the matrix material in non-transgenic rice, Nipponbare, whereas they were surrounded by spaces in the transgenic rice. The morphology and size of each starch granule were not different between nontransgenic and the transgenic rice. However, the transgenic rice flour showed smaller particle size, higher starch damage, and lower viscosity during gelatinization than that of non-transgenic rice. These results confirm that puroindoline b reduces the grain hardness in rice. Moreover, the results also suggest that puroindoline b functions at the surface of compound starch granules, and not on polygonal starch granules in rice endosperm. (C) 2009 Elsevier Ltd. All rights reserved.
  • Biological applications of novel nonlinear optical microscopy, Shin'ichiro Kajiyama, Yasuyuki Ozeki, Kiichi Fukui, Kazuyoshi Itoh, 2009 INTERNATIONAL WORKSHOP ON INFORMATION OPTICS, 2009 INTERNATIONAL WORKSHOP ON INFORMATION OPTICS, 206, 2010
    Summary:Two types of newly developed nonlinear optical microscopes namely stimulated parametric emission (SPE) microscope and stimulated Raman scattering (SRS) microscope were presented together with their biological applications.
  • Dual-Band Stimulated Parametric Emission Microscopy, Masatomo Yamagiwa, Gen Omura, Yasuyuki Ozeki, Makiko Ishii, Hieu Min Dang, Shin'ichiro Kajiyama, Takeshi Suzuki, Kiichi Fukui, Kazuyoshi Itoh, JAPANESE JOURNAL OF APPLIED PHYSICS, JAPANESE JOURNAL OF APPLIED PHYSICS, 49(1), 16603 - 16606, 2010
    Summary:We demonstrate that two distinct images can be simultaneously acquired using the dual-band configuration of stimulated parametric emission (SPE) microscopy, which is based on the four-wave mixing process enhanced by two-photon electronic resonance. The dual-band SPE microscopy utilizes two SPE signals simultaneously generated at different wavelengths. In the experiment, we image stained and unstained biological tissues, and obtain two different images for each sample. Additionally, we propose an axial differential imaging technique to improve the contrast of SPE images, and verify its applicability to biological imaging. (C) 2010 The Japan Society of Applied Physics
  • Biological applications of stimulated parametric emission microscopy and stimulated raman scattering microscopy, Shin'Ichiro Kajiyama, Yasuyuki Ozeki, Kiichi Fukui, Kazuyoshi Itoh, Information Optics and Photonics: Algorithms, Systems, and Applications, Information Optics and Photonics: Algorithms, Systems, and Applications, 89 - 98, 2010 , Refereed
    Summary:Two types of newly developed nonlinear-optical microscopy are presented, namely, stimulated parametric emission (SPE) microscopy and stimulated Raman scattering (SRS) microscopy, together with their biological applications. These techniques allow high-resolution 3D visualization of unstained living cells. SPE microscopy is also applied to time-lapse observation of dividing cells as well as simultaneous observation of fluorescence emitted from labeled organelles and SPE signal. © 2010 Springer Science+Business Media, LLC.
  • Subharmonic Synchronization of Two-Color Laser Pulses for Stimulated Raman Scattering Microscopy, Yasuyuki Ozeki, Yuma Kitagawa, Kazuhiko Sumimura, Norihiko Nishizawa, Wataru Umemura, Makiko Ishii, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, 2010 CONFERENCE ON LASERS AND ELECTRO-OPTICS (CLEO) AND QUANTUM ELECTRONICS AND LASER SCIENCE CONFERENCE (QELS), 2010 CONFERENCE ON LASERS AND ELECTRO-OPTICS (CLEO) AND QUANTUM ELECTRONICS AND LASER SCIENCE CONFERENCE (QELS), 2010 , Refereed
    Summary:We successfully demonstrate low-jitter synchronization of a 38-MHz Yb-fiber oscillator to a 76-MHz Ti:sapphire oscillator with a two-photon detector and an intra-cavity electro-optic modulator for sensitive lock-in detection of stimulated Raman signal in biological microscopy. (C) 2010 Optical Society of America
  • Three-Dimensional Unstained Live-Cell Imaging Using Stimulated Parametric Emission Microscopy, Hieu M. Dang, Takehito Kawasumi, Gen Omura, Toshiyuki Umano, Shin'ichiro Kajiyama, Yasuyuki Ozeki, Kazuyoshi Itoh, Kiichi Fukui, JAPANESE JOURNAL OF APPLIED PHYSICS, JAPANESE JOURNAL OF APPLIED PHYSICS, 48(9), 97007, Sep. 2009
    Summary:The ability to perform high-resolution unstained live imaging is very important to in vivo study of cell structures and functions. Stimulated parametric emission (SPE) microscopy is a nonlinear-optical microscopy based on ultra-fast electronic nonlinear-optical responses. For the first time, we have successfully applied this technique to archive three-dimensional (3D) images of unstained sub-cellular structures, such as, microtubules, nuclei, nucleoli, etc. in live cells. Observation of a complete cell division confirms the ability of SPE microscopy for long time-scale imaging. (C) 2009 The Japan Society of Applied Physics
  • Development of an enzyme activity screening system for beta-glucosidase-displaying yeasts using calcium alginate micro-beads and flow sorting, Yan Zhou, Shin'ichiro Kajiyama, Kouichi Itoh, Takanori Tanino, Nobuo Fukuda, Tsutomu Tanaka, Akihiko Kondo, Kiichi Fukui, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 84(2), 375 - 382, Aug. 2009 , Refereed
    Summary:Recent reports on high-speed affinity screening systems for yeast cells using flow cytometry have not been adapted to screening yeast cells that display hydrolyzing enzymes, since the fluorescent molecules which are released from fluoresceinated substrate diffuse into solution after enzymatic reaction. In this research, yeast cells displaying beta-glycosidase were individually captured in micro-sized calcium alginate beads by using the newly developed reverse micelle method to prevent diffusion of hydrolyzed fluorescent substrates. By adopting flow sorting to these captured cells, active cells were successfully enriched about 82-fold from a mixed suspension with negative controls. This system should be a useful method for high-speed screening of yeast cells that display various hydrolyzing enzymes and has potential application to screening randomized libraries of enzyme-displayed yeast cells with higher activities.
  • Label-free imaging by stimulated parametric emission microscopy reveals a difference in hemoglobin distribution between live and fixed erythrocytes, Hieu M. Dang, Gen Omura, Toshiyuki Umano, Masatomo Yamagiwa, Shin'ichiro Kajiyama, Yasuyuki Ozeki, Kazuyoshi Itoh, Kiichi Fukui, JOURNAL OF BIOMEDICAL OPTICS, JOURNAL OF BIOMEDICAL OPTICS, 14(4), 040506, Jul. 2009 , Refereed
    Summary:We demonstrate that stimulated parametric emission (SPE) microscopy enables label-free, 3-D visualization of internal hemoglobin distribution of live mouse and chicken erythrocytes with high sensitivity. Change in hemoglobin distribution in chicken erythrocytes before and after ethanol fixation is clearly visualized. (C) 2009 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3207151]
  • Physical Mapping of Ribosomal DNA Genes on Jatropha curcas Chromosomes by Multicolor FISH, Magdalena Witkowska, Nobuko Ohmido, Joyce Cartagena, Nakako Shibagaki, Shin'ichiro Kajiyama, Kiichi Fukui, CYTOLOGIA, CYTOLOGIA, 74(2), 133 - 139, Jun. 2009
    Summary:Jatropha curcas L. belonging to the Euphorbiaceae is becoming a significant crop in tropical areas nowadays. The genetic diversity among 1 African and 5 Asian varieties were analyzed by multicolor fluorescence in situ hybridization (McFISH) using 5S and 45S ribosomal RNA genes (rDNAs). One locus of 5S rDNA and 2 loci for 45S rDNA were detected at specific regions of the chromosomes in all the materials. Also, telomeric repeats (TTTAGGG)(n) were localized on the terminal regions of all chromosomes. The present results confirm the stability of these major repeated sequences among Jatropha lines.
  • Nonlinear Phase Imaging Using Two-Beam Interferometry in Stimulated Parametric Emission Microscopy, Masatomo Yamagiwa, Yasuyuki Ozeki, Gen Omura, Takeshi Suzuki, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, JAPANESE JOURNAL OF APPLIED PHYSICS, JAPANESE JOURNAL OF APPLIED PHYSICS, 48(6), 062501, Jun. 2009
    Summary:We demonstrate that two-beam interferometry is applicable to phase detection of nonlinear susceptibility of biological samples in the stimulated parametric emission (SPE) microscopy, which is one of the coherent nonlinear optical microscopy techniques. We experimentally observed stained mouse kidney cells and successfully confirmed nonlinear phase shift associated with an electronic resonance at stained nuclei. In addition, phase blurring was found to be almost completely eliminated in a nonlinear differential interferometric contrast image, which can be produced through a simple post-processing procedure. (C) 2009 The Japan Society of Applied Physics
  • Bioactive beads-mediated transformation of rice with large DNA fragments containing Aegilops tauschii genes, Naoki Wada, Shin'ichiro Kajiyama, Yukio Akiyama, Shigeki Kawakami, Daisuke No, Susumu Uchiyama, Motoyasu Otani, Takiko Shimada, Naoko Nose, Go Suzuki, Yasuhiko Mukai, Kiichi Fukui, PLANT CELL REPORTS, PLANT CELL REPORTS, 28(5), 759 - 768, May 2009 , Refereed
    Summary:Transformation with large DNA molecules enables multiple genes to be introduced into plants simultaneously to produce transgenic plants with complex phenotypes. In this study, a large DNA fragment (ca. 100 kb) containing a set of Aegilops tauschii hardness genes was introduced into rice plants using a novel transformation method, called bioactive beads-mediated transformation. Nine transgenic rice plants were obtained and the presence of transgenes in the rice genome was confirmed by PCR and FISH analyses. The results suggested that multiple transgenes were successfully integrated in all transgenic plants. The expression of one of the transgenes, puroindoline b, was confirmed at the mRNA and protein levels in the T(2) generation. Our study clearly demonstrates that the bioactive bead method is capable of producing transgenic rice plants carrying large DNA fragments. This method will facilitate the production of useful transgenic plants by introducing multiple genes simultaneously.
  • High-resolution spatial and temporal analysis of phytoalexin production in oats, Yoshihiro Izumi, Shin'ichiro Kajiyama, Ryosuke Nakamura, Atsushi Ishihara, Atsushi Okazawa, Eiichiro Fukusaki, Yasuo Kanematsu, Akio Kobayashi, PLANTA, PLANTA, 229(4), 931 - 943, Mar. 2009 , Refereed
    Summary:The production of oat (Avena sativa L.) phytoalexins, avenanthramides, occurs in response to elicitor treatment with oligo-N-acetylchitooligosaccharides. In this study, avenanthramides production was investigated by techniques that provide high spatial and temporal resolution in order to clarify the process of phytoalexin production at the cellular level. The amount of avenanthramides accumulation in a single mesophyll cell was quantified by a combination of laser micro-sampling and low-diffuse nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) techniques. Avenanthramides, NAD(P)H and chlorophyll were also visualized in elicitor-treated mesophyll cells using line-scanning fluorescence microscopy. We found that elicitor-treated mesophyll cells could be categorized into three characteristic cell phases, which occurred serially over time. Phase 0 indicated the normal cell state before metabolic or morphological change in response to elicitor, in which the cells contained abundant NAD(P)H. In phase 1, rapid NAD(P)H oxidation and marked movement of chloroplasts occurred, and this phase was the early stage of avenanthramides biosynthesis. In phase 2, avenanthramides accumulation was maximized, and chloroplasts were degraded. Avenanthramides appear to be synthesized in the chloroplast, because a fluorescence signal originating from avenanthramides was localized to the chloroplasts. Moreover, our results indicated that avenanthramides biosynthesis and the hypersensitive response (HR) occurred in identical cells. Thus, the avenanthramides production may be one of sequential events programmed in HR leading to cell death. Furthermore, the phase of the defense response was different among mesophyll cells simultaneously treated with elicitor. These results suggest that individual cells may have different susceptibility to the elicitor.
  • Analysis and experimental assessment of the sensitivity of stimulated Raman scattering microscopy, Yasuyuki Ozeki, Fumihiro Dake, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, OPTICS EXPRESS, OPTICS EXPRESS, 17(5), 3651 - 3658, Mar. 2009 , Refereed
    Summary:We theoretically show that the shot-noise-limited sensitivity of stimulated Raman scattering (SRS) microscopy, which enables high-contrast vibrational imaging, is similar to that of coherent anti-Stokes Raman scattering microscopy. We experimentally confirm that the sensitivity of our SRS microscope is lower than the shot-noise limit only by <15 dB, which indicates that the high-sensitivity of SRS microscopy is readily available. (C) 2009 Optical Society of America
  • Background-Free, 3D Vibrational Imaging by Stimulated Raman Scattering Microscopy, Yasuyuki Ozeki, Fumihiro Dake, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, 2009 CONFERENCE ON LASERS AND ELECTRO-OPTICS AND QUANTUM ELECTRONICS AND LASER SCIENCE CONFERENCE (CLEO/QELS 2009), VOLS 1-5, 2009 CONFERENCE ON LASERS AND ELECTRO-OPTICS AND QUANTUM ELECTRONICS AND LASER SCIENCE CONFERENCE (CLEO/QELS 2009), VOLS 1-5, 143 - +, 2009 , Refereed
    Summary:We propose and experimentally show that stimulated Raman scattering can realize novel 3D nonlinear-optical Raman microscopy that is immune to nonresonant background signals. We demonstrate high-contrast molecular imaging of polystyrene beads and plant cells. (C) 2009 Optical Society of America
  • Highly Sensitive Signal Detection in Stimulated Parametric Emission Microscopy Based on Two-Beam Interferometry, Masatomo Yamagiwa, Yasuyuki Ozeki, Takehito Kawasumi, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, JAPANESE JOURNAL OF APPLIED PHYSICS, JAPANESE JOURNAL OF APPLIED PHYSICS, 47(12), 8820 - 8824, Dec. 2008
    Summary:We present a simple and highly sensitive optical detection method based on two-beam interferometry for application to coherent nonlinear optical microscopy (CNOM). The theoretical sensitivity of this method is higher than that of conventional spectral interferometry (SI). We experimentally applied this technique to stimulated parametric emission (SPE) microscopy and achieved a high sensitivity that is only 4 or 5 dB lower than that of a theoretical shot noise limit. In order to validate the practical applicability of this technique, we demonstrated a noise reduction experiment in the observation of a plant cell with an SPE microscope. [DOI: 10.1143/JJAP.47.8820]
  • Transient Gene Expression in Guard Cell Chloroplasts of Tobacco Using ArF Excimer Laser Microablation, Shin'ichiro Kajiyama, Benesh Joseph, Fumihide Inoue, Masaki Shimamura, Eiichiro Fukusaki, Kenichi Tomizawa, Akio Kobayashi, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 106(2), 194 - 198, Aug. 2008
    Summary:In this paper, we report a novel method for delivering genes into chloroplasts of tobacco cells using laser microablation. The plasmid pLD200-GFP was introduced into chloroplasts of Nicotiana tabacum cv. Xanthi guard cells and transient GFP expression was detected in the chloroplasts after 2-3 d of incubation. The technique uses an argon fluoride (ArF) excimer laser to perforate the cell surface in a 4 mu m(2) area in the presence of plasmid coated gold microparticles. Pretreatment of guard cells to promote stomatal closure prior to laser ablation resulted in a significant increase in the survival rate of cells and a transient expression rate of 2-3% in trial number basis was archived. Our method has unique advantages such as avoiding laborious pretreatments that adversely affect cell viability and specific delivery of transgenes into a desired cell in complex leaf tissue. This technique is a potential tool for cell specific transient gene expression studies for elucidation of gene regulation and expression.
  • Supergiant ampholytic sugar chains with imbalanced charge ratio form saline ultra-absorbent hydrogels, Maiko K. Okajima, Takeshi Bamba, Yasuhiro Kaneso, Kazumasa Hirata, Eiichiro Fukusaki, Shin'ichiro Kajiyama, Tatsuo Kaneko, MACROMOLECULES, MACROMOLECULES, 41(12), 4061 - 4064, Jun. 2008
  • Line-Scanning Microscopy for Time-Gated and Spectrally Resolved Fluorescence Imaging, Ryosuke Nakamura, Yoshihiro Izumi, Shin'ichiro Kajiyama, Akio Kobayashi, Yasuo Kanematsu, JOURNAL OF BIOLOGICAL PHYSICS, JOURNAL OF BIOLOGICAL PHYSICS, 34(1-2), 51 - 62, Apr. 2008
    Summary:Laser-scanning fluorescence microscopy for efficient acquisition of time-gated and spectrally resolved fluorescence images was developed based on line illumination of the laser beam and detection of the fluorescence image through a slit. In this optical arrangement, the fluorescence image was obtained by scanning only one axis perpendicular to the excitation line, and the acquisition time was significantly reduced compared with conventional laser-scanning confocal microscopy. A multidimensional fluorescence dataset consisting of fluorescence intensities as a function of x-position, y-position, fluorescence wavelength, and delay time after photoexcitation was analyzed and decomposed based on the parallel factor analysis model. The performance of the line-scanning microscopy was examined by applying it to the analysis of one of the plant defense responses, accumulation of antimicrobial compounds of phytoalexin in oat (Avena sativa), induced by the elicitor treatment.
  • Improvement of transformation efficiency by bioactive-beads-mediated gene transfer using DNA-lipofectin complex as entrapped genetic material, Tomoko Murakawa, Shin'ichiro Kajiyama, Tomohiko Ikeuchi, Shigeki Kawakami, Kiichi Fukui, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 105(1), 77 - 80, Jan. 2008
    Summary:The efficiency of bioactive-beads-mediated plant transformation was improved using DNA-lipofectin complex as the entrapped genetic material instead of naked DNA used in the conventional method. In the improved method, beads aggregated and formed clusters around the protoplasts resulting in a 4-fold higher transformation efficiency than that by the conventional method.
  • Improvement of bioactive bead-mediated transformation by concomitant application of electroporation, Tomoko Murakawa, Shin'ichiro Kajiyama, Kiichi Fukui, PLANT BIOTECHNOLOGY, PLANT BIOTECHNOLOGY, 25(4), 387 - 390, 2008
    Summary:Techniques of exogenous gene introduction in plant cells have been actively applied for producing transgenic cultured cells that produce high levels of useful biological substances and are used for the molecular breeding of plants. A novel gene transfer technique has been developed using calcium alginate micro-beads (bioactive beads) to entrap genetic material. Although this technique has several advantages such as high transformation efficiency and the ability to introduce large DNA molecules, the precise optimal application conditions remain to be determined. Here, the optimal conditions for the concomitant use of bioactive beads and electroporation, such as polyethylene glycol (PEG) concentration, alginate concentration, electric field intensity, pulse duration, and pulse numbers, were determined for improved gene transfer efficiency of bioactive bead-mediated plant transformation. Consequently, highly efficient transient transgene expression (up to 4.7%) was achieved by applying electrical pulses (0.75 kV cm(-1), 30 mu s, 3 x) to a protoplast suspension in 12% polyethylene glycol (PEG).
  • Femtosecond pulsed laser as a microscalpel for microdissection and isolation of specific sections from biological samples, Masaaki Sakakura, Shinichiro Kajiyama, Masafumi Tsutsumi, Jinhai Si, Eiichiro Fukusaki, Yutaka Tamaru, Shin-Ichi Akiyama, Kiyotaka Miura, Kazuyuki Hirao, Mitsuyoshi Ueda, JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS, JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS, 46(9A), 5859 - 5864, Sep. 2007
    Summary:The fermosecond laser is a candidate tool for dissecting and obtaining specific tissues or cells from biological samples. In this paper, we report on the dissection of flammable (heat-sensitive) plant tissues (Eucommia ulmoides stem) and that of a transparent zebrafish embryo placed in water. During laser dissection in the plant tissues, we observed that a low repetition rate of the irradiations is crucial for reducing the risk of thermal damage. During laser dissection of the zebrafish embryo, the laser-induced bubble formation at the tissue surface disturbed the laser-induced dissection. We concluded that the "interior laser dissection method", in which the dissection is performed by moving the laser spot inside a biological sample, is an efficient way to dissect a sample that is placed in water. In view of our findings, the advantages of fs laser dissection are discussed on the basis of the physical characteristics of fs laser machining.
  • Novel plant transformation system by gene-coated gold particle introduction into specific cell using ArF excimer laser, Shin'ichiro Kajiyama, Fumihide Inoue, Yoshinobu Yoshikawa, Takeshi Shoji, Eiichiro Fukusaki, Akio Kobayashi, PLANT BIOTECHNOLOGY, PLANT BIOTECHNOLOGY, 24(3), 315 - 320, Jun. 2007
    Summary:Novel methods for foreign-gene introduction into specific cells are required for the functional analysis of specific cells. We report a new transfection method for intact plant cells that uses ArF excimer laser-induced shock waves to introduce foreign gene-coated gold particles into plant cells through the cell wall. Foreign gene (sGFP)-coated gold particles were randomly scattered on the surface of Torenia tissue samples; the laser was irradiated toward the gold particles using a hollow optical fiber and a quartz light-condensing tip to introduce the particles into the target cells. Treated plant tissue samples were then cultured for several weeks on the selection medium, and transformation was assessed by fluorescence microscopy and PCR. Irradiation-induced damage to the cells and coated DNA was insignificant, and the transformation efficiency was similar to that of biolistic transformation based on the particle number. Although this method requires a suitable laser setup and is time-consuming, it could be applied in transformation experiments on specific target cells. Moreover, this method can possibly be used to introduce not only DNA but also biologically active substances such as protein regulators or inhibitors, whose introduction into cells is difficult through the cell wall.
  • Single cell-based analysis of Torenia petal pigments by a combination of ArF excimer laser micro sampling and nano-high performance liquid chromatography (HPLC)-mass spectrometry, Shin'ichiro Kajiyama, Kazuo Harada, Eiichiro Fukusaki, Akio Kobayashi, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 102(6), 575 - 578, Dec. 2006
    Summary:The molecular constituents of the petal pigments of the Torenia plant (Torenia hybrida) were analyzed on a single-cell basis by a combination of newly developed laser-microsampling and nano-flow liquid chromatography-electro spray ionization mass spectrometry (LC-ESIMS) techniques. Our method should provide a facile method for obtaining precise metabolic profiles of each cell in a single plant tissue.
  • A novel microsurgery method for intact plant tissue at the single cell level using ArF excimer laser microprojection, S Kajiyama, T Shoji, S Okuda, Y Izumi, E Fukusaki, A Kobayashi, BIOTECHNOLOGY AND BIOENGINEERING, BIOTECHNOLOGY AND BIOENGINEERING, 93(2), 325 - 331, Feb. 2006
    Summary:A novel microsurgery technique for the partial removal of rigid cell-walls in intact plant tissue is established. Using a size-variable slit, an ArF excimer laser was microprojected on the surface of the targeted cell, and this method enabled the area- and depth-controllable processing of the cortical structure of plant cells including the cuticle and cell wall layer. In epidermal cells of all tested plants, viabilities of more than 90% were retained 24 h after irradiation. Scanning electron microscope (SEM) observation revealed that the cuticle layer of the irradiated region was completely ablated, and the cellulose microfibrils of the secondary cell wall were partially removed; furthermore, 4 days after laser treatment, the regeneration of cell wall fibrils was observed. As a model experiment, the transient expression of synthetic green fluorescent protein (sGFP) was performed by the microinjection of cauliflower mosaic virus (CMV) 35S promoter-derived sGFP gene through an "aperture" in the treated cell surface. Moreover, micron-sized fluorescent beads were successfully introduced by the same method into the onion cells indicating that this method can be used to introduce foreign materials as large as organelles. (c) 2005 Wiley Periodicals, Inc.
  • Highly sensitive spectral interferometric fourwave mixing microscopy near the shot noise limit and its combination with two-photon excited fluorescence microscopy, Keisuke Isobe, Yasuyuki Ozeki, Takehito Kawasumi, Shogo Kataoka, Shin'ichiro Kajiyama, Kiichi Fukui, Kazuyoshi Itoh, Optics Express, Optics Express, 14(23), 11204 - 11214, 2006
    Summary:We present spectral interferometric four-wave mixing (FWM) microscopy with a nearly shot-noise limited sensitivity and with the capability of separating FWM signals from fluorescence signals. We analyze the requirements for obtaining the shot-noise limited sensitivity and experimentally achieve the sensitivity that is only 4-dB lower than the shotnoise limit. Moreover, we show that only FWM signals can be extracted through the Fourier filtering even when the FWM spectrum is overlapped and overwhelmed by the fluorescence spectrum. We demonstrate simultaneous acquisition of FWM and two-photon excited fluorescence images of fluorescent monodispersed polystyrene microspheres. © 2006 Optical Society of America.
  • Syntheses and screening tests of new chlorin derivatives as photosensitizer, Y Nakae, E Fukusaki, S Kajiyama, A Kobayashi, S Nakajima, Sakata, I, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, 174(3), 187 - 193, Sep. 2005
    Summary:New chlorin derivatives were synthesized from protoporphyrin IX dimethyl ester (1) as a starting material. The tumor-localizing abilities were determined by bathochromic and hyperchromic shift test using albumin and the photosensitivities were evaluated by photooxidation test using dansyl-L-methionine. NOH-P-H (3) was the most excellent in the both tests. NOH-P-H (3) was conjugated with aspartic acid to evaluate the biodistribution with the time progress by nitrogen pulsed laser spectrofluorometry. The concentration of NOH-P-Asp (15) in cancer tissues showed the highest concentration at 6 h after the administration and corresponded to five to seven times in other organs. The concentration in cancer tissues showed a rapid decrease at 24 h after the administration. PDT effect of NOH-P-Asp (15) was evaluated using tumor-bearing mice. Remarkable anti-tumor effect was observed. NOH-P-Asp (15) was expected to be applicable to a new photosensitizer for PDT. (c) 2005 Elsevier B.V. All rights reserved.
  • The convenient screening method using albumin for the tumor localizing property of Ga-porphyrin complexes, Y Nakae, EI Fukusaki, S Kajiyama, A Kobayashi, S Nakajima, Sakata, I, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, 172(1), 55 - 61, May 2005
    Summary:Eleven Ga-phorphyrin complexes bearing various lengths of carbon-chain were synthesized and the UV-vis spectra shift patterns in various concentrations of albumin solutions were evaluated. The distributions of these derivatives into the organs were also determined by surface fluorescence using nitrogen-pulsed laser spectrophotometry. The shift patterns between 0 and 0.9% albumin concentrations (albumin test) were correlated to their tumor localizing property. The albumin test was proved to be useful for evaluation of tumor localizing properties of porphyrin derivatives. The tumor localizing ability was increased along with increasing their lipophilicity and the derivative bearing decyl groups as side chains was a maximum. The porphyrin-albumin complex might have key mission for the passive accumulation focusing on the tumor. © 2004 Elsevier B.V. All rights reserved.
  • Convenient screening methods for the photosensitivity, Y Nakae, E Fukusaki, S Kajiyama, A Kobayashi, Sakata, I, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY, 171(1), 91 - 95, Apr. 2005
    Summary:Two kinds of convenient photosensitivity screening methods (Methods A and B) were established to find new photosensitizers for photodynamic therapy (PDT). Method A is based on oxidation mechanism of cholesterol and types 1 and 11 can be specified. Method B is performed using dansyl-L-methionine and offers the benefits of fast response time. Various porphyrin, chlorin and phthalocyanine derivatives were synthesized and subjected to the screening methods. The assays have yielded similar results. The results were also identical to the photosensitivity (Ga > Zn > Mn-complex) of water-soluble metalloporphyrins bearing the same side chains. Therefore, the Methods A and B were thought to be appropriate as screening methods to find lead compounds of photosensitizers for PDT. (c) 2004 Elsevier B.V. All rights reserved.
  • Isolation and characterization of a cDNA encoding phytochrome a in the non-photosynthetic parasitic plant, Orobanche minor sm., C Trakulnaleamsai, A Okazawa, CI An, S Kajiyama, E Fukusaki, K Yoneyama, Y Takeuchi, A Kobayashi, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 69(1), 71 - 78, Jan. 2005
    Summary:In this study, the isolation and characterization of a phytochrome A (PHYA) homologous cDNA (OmPHYA) in the non-photosynthetic holoparasitic plant Orobanche minor are described. The present findings provide the first report of the presence of a PHYA homolog in the holoparasite. This study found that OmPHYA is of similar size to the other PHYAs of green plants and shows 72, 77, and 77% amino acid sequence identity with PHYA in Arabidopsis, potato, and tobacco respectively. The OmPHYA contains a conserved chromophore attachment cysteine at position 323. Although OmPHYA shows high sequence identity with other PHYAs in green plants, 13 amino acid substitutions located in both the N and C-terminal domains are observed (a total of 26 amino acids). OmPHYA is encoded by a single gene within the O. minor genome. The abundance of the OmPHYA transcript as well as nuclear translocation of OmphyA occurs in a light-dependent manner.
  • Biosynthetic pathway for the C45 polyprenol, solanesol, in tobacco, E Fukusaki, S Takeno, T Bamba, H Okumoto, H Katto, S Kajiyama, A Kobayashi, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68(9), 1988 - 1990, Sep. 2004
    Summary:Feeding experiments were independently performed with [1-C-13]deoxy-D-xylulose triacetate and (RS)-[2-C-13]mevalonolactone in the tobacco plant. The labeling pattern for solanesol was elucidated to reveal that the isoprene moiety of solanesol would be derived from deoxy-xylulose. The result strongly suggests that tobacco solanesol is biosynthesized via the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway.
  • Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference, E Fukusaki, K Kawasaki, S Kajiyama, CI An, K Suzuki, Y Tanaka, A Kobayashi, JOURNAL OF BIOTECHNOLOGY, JOURNAL OF BIOTECHNOLOGY, 111(3), 229 - 240, Aug. 2004
    Summary:Suppression of biosynthetic genes involved in flower color formation is an important approach for obtaining target flower colors. Here we report that flower color of the garden plant Torenia hybrida was successfully modulated by RNA interference (RNAi) against a gene of chalcone synthase (CHS), a key enzyme for anthocyanin and flavonoid biosynthesis. By using each of the coding region and the 3'-untranslated region of the CHS mRNA as an RNAi target, exhaustive and gene-specific gene silencing were successfully induced, and the original blue flower color was modulated to white and pale colors, respectively. Our results indicate that RNAi is quite useful for modulations of flower colors of commercially important garden plants. (C) 2004 Elsevier B.V. All rights reserved.
  • A chitin-oligomer binding peptide obtained by screening of a phage display random peptide library and its affinity modulation corresponding to oxidation-reduction state, E Fukusaki, K Ogawa, A Okazawa, SI Kajiyama, A Kobayashi, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 28(4-6), 181 - 184, Jun. 2004
    Summary:Screening of a phage display random 9-mer peptide library, in which cysteine residues were at the both terminals of the 7-mer random region, was performed to obtain an oligopeptide that recognizes a chitin-oligomer. Affinity of the obtained peptide (Cys-Ser-Arg-Thr-Thr-Arg-Thr-Arg-Cys) to chitotriose was modulated by its oxidation-reduction state. Only the oxidized form exhibited specific binding to the target molecule, chitotriose. This is the first report of reversible affinity modulation of a synthetic oligopeptide which can recognize a neutral saccharide. (C) 2004 Elsevier B.V. All rights reserved.
  • Transformation of yeast using calcium alginate microbeads with surface-immobilized chromosomal DNA, A Mizukami, E Nagamori, Y Takakura, S Matsunaga, Y Kaneko, S Kajiyama, S Harashima, A Kobayashi, K Fukui, BIOTECHNIQUES, BIOTECHNIQUES, 35(4), 734 - +, Oct. 2003
    Summary:Yeast artificial chromosomes (YACs) are useful cloning vectors that have the capacity to carry large DNA inserts. The largest barrier to using such large DNA molecules in transformation experiments has been their physical instability in solution. We developed a new method of transforming yeast using chromosome-sized DNA. The method uses calcium alginate microbeads to immobilize high-density yeast chromosomal DNA. Chromosomal DNA immobilized on microbeads is physically stabilized compared with naked chromosomal DNA. The microbead-mediated transformation performed well, not only with respect to the transformation frequency with large DNA molecules (> 100 kb) but also in successful tran formation using split chromosome DNA that exceeded 450 kb.
  • Phytotoxicity of the tetramic acid metabolite trichosetin, EC Marfori, SI Kajiyama, EI Fukusaki, A Kobayashi, PHYTOCHEMISTRY, PHYTOCHEMISTRY, 62(5), 715 - 721, Mar. 2003
    Summary:Trichosetin, a tetramic acid-containing metabolite produced in the dual culture of Trichoderma harzianum and Catharanthus roseus (L.) G. Don callus, was subjected to phytotoxicity assays. In seedling growth assays, trichosetin inhibited root and shoot growth of all five plant species tested by damaging the cell membrane, as evidenced by the dose-dependent increase in electrolyte leakage and lipid peroxidation. Vital staining of trichosetin-treated Nicotiana tabacum BY-2 cells, with rhodamine 123, showed a weaker green fluorescence compared to controls indicating damaging effects on mitochondria. FDA-PI staining, to determine cell viability, indicated that cells of the trichosetin-treated roots were mostly dead. (C) 2003 Elsevier Science Ltd. All rights reserved.
  • Purification and properties of an enzyme capable of degrading the polysaccharide of the cyanobacterium, Nostoc commune, Dasman, S Kajiyama, A Okazawa, E Fukusaki, A Kobayashi, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 57(11-12), 1042 - 1046, Nov. 2002
    Summary:A novel Nostoc commune-polysaccharide (NPS)-degrading enzyme with a molecular mass of 128.5 kDa was purified from Paenibacillus glycanilyticus DS-1. The optimum pH and temperature of the enzyme activity were 5.5 and 35degreesC, respectively. The enzyme completely degraded NPS to oligosaccharides, ranging from tetra to hexasaccharides and could degrade the xylan weakly whereas xanthan, gellan, cellulose, curdlan and p-nitrophenyl-beta-D-xylopyra-noside were not degraded. Homology analysis of the N-terminal amino acid sequence of the NPS-degrading enzyme against the PIR and SWISS-PROT databases indicated that the sequence was not homologous to any other polysaccharide-degrading enzyme.
  • Paenibacillus glycanilyticus sp nov., a novel species that degrades heteropolysaccharide produced by the cyanobacterium Nostoc commune, Dasman, S Kajiyama, H Kawasaki, M Yagi, T Seki, E Fukusaki, A Kobayashi, INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 52(5), 1669 - 1674, Sep. 2002
    Summary:A novel bacterial strain, DS-1(T), was isolated that degrades heteropolysaccharide produced by the cyanobacterium Nostoc commune. The isolate was identified by a combination of phenotypic characterization, cellular fatty acid analysis, DNA base composition, DNA-DNA hybridization and 16S rRNA gene sequence analysis. Phylogenetic analysis placed strain DS-1(T) within the Paenibacillus cluster on a phylogenetic tree and the phenotypic characteristics of this strain appear to be similar to those of Paenibacillus curdlanolyticus IFO 15724(T) and Paenibacillus kobensis IFO 15729(T). The strain was distinguished from A curdlanolyticus IFO 15724(T) and A kobensis IFO 15729(T) by its ability to degrade the polysaccharide of Nostoc commune, by assimilation of rhamnose, inositol and L-fucose and by its wide range of optimal growth temperature (28-37degreesC). Like other Paenibacillus species, this strain contains anteiso-C-15:0 as a major cellular fatty acid, and it has a DNA G+C content of 50.5 mol %. Based on these results, it is concluded that this isolate should be placed within a novel species of Paenibacillus, Paenibacillus glycanilyticus sp. nov., with the type strain DS-1(T)(=IFO 16618(T)=JCM 11221(T)=NRRL B-23455(T)).
  • Biosynthetic studies of the tetramic acid antibiotic trichosetin, EC Marfori, T Bamba, S Kajiyama, E Fukusaki, A Kobayashi, TETRAHEDRON, TETRAHEDRON, 58(33), 6655 - 6658, Aug. 2002
    Summary:A feeding experiment with C-13-labeled precursors was conducted in the dual cultures of Trichoderma harzianum H14 and Catharanthus roseus callus to determine the biosynthetic origin of the carbon atoms in trichosetin. Results showed that trichosetin originated from two separate biogenetic units. One unit is an octaketide intermediate directly derived from eight intact acetate units joined in a head-to-tail fashion of a polyketide pathway. The other unit is serine. (C) 2002 Elsevier Science Ltd. All rights reserved.
  • A novel gene delivery system in plants with calcium alginate micro-beads, T Sone, E Nagamori, T Ikeuchi, A Mizukami, Y Takakura, SI Kajiyama, EI Fukusaki, S Harashima, A Kobayashi, K Fukui, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 94(1), 87 - 91, Jul. 2002
    Summary:We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFA With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 It incubation compared to that with the conventional method using a naked plasmid solution. Transfection was up to 0.22% efficient. These results indicate that bio-beads have a possibility for efficient transformation in plants.
  • Trichosetin, a novel tetramic acid antibiotic produced in dual culture of Trichoderma harzianum and Catharanthus roseus Callus, EC Marfori, S Kajiyama, E Fukusaki, A Kobayashi, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 57(5-6), 465 - 470, May 2002
    Summary:The dual culture of Trichoderma harzianum and Catharanthus roseus callus produced an antimicrobial compound with a remarkable activity against the Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis. Structural elucidation revealed that this compound, which we have named trichosetin, is a novel tetramic acid (2,4-pyrrolidinedione) antibiotic and a homolog of the fungal metabolite equisetin. This compound however, was not produced in the individual culture of T. harzianum or C roseus callus.
  • Identification of genes induced by taxol application using a combination of differential display RT-PCR and DNA microarray analysis, E Fukusaki, T Oishi, H Tanaka, S Kajiyama, A Kobayashi, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 56(9-10), 814 - 819, Sep. 2001 , Refereed
    Summary:The differential display reverse transcriptional polymerase chain reaction (DD-RT-PCR) was used to hunt for cDNA fragments specifically expressed by taxol treatment of HeLa cells. Forty-eight cDNA clones were differentially displayed through the experiments. The cDNA fragments obtained were separately spotted onto glass slides to prepare a tailor-made DNA chip. The gene expression pattern of differentially displayed cDNA fragments were checked by DNA microarray analysis.
  • The occurrence of geometric polyprenol isomers in the rubber-producing plant, Eucommia ulmoides Oliver, T Bamba, E Fukusaki, S Kajiyama, K Ute, T Kitayama, A Kobayashi, LIPIDS, LIPIDS, 36(7), 727 - 732, Jul. 2001 , Refereed
    Summary:The chain length and geometric isomerism of polyprenols from Eucommia ulmoides Oliver were analyzed using supercritical fluid chromatography. After intensive effort to establish separation conditions for geometric isomers, a phenyl-bonded silica gel-packed column was found that cleanly separated poly-trans and -cis prenols. The presence of long-chain poly-trans prenols (> 9 mers) was confirmed for the first time in plants. Trans isomers were found in the leaf, seed coat, and root, but not in the bark and seed. Poly-trans prenols in this plant may act as intermediates for trans-polyisoprene biosynthesis.
  • High-resolution analysis of polyprenols by supercritical fluid chromatography, T Bamba, E Fukusaki, S Kajiyama, K Ute, T Kitayama, A Kobayashi, JOURNAL OF CHROMATOGRAPHY A, JOURNAL OF CHROMATOGRAPHY A, 911(1), 113 - 117, Mar. 2001 , Refereed
    Summary:A high-resolution analysis of polyprenol mixtures was achieved by supercritical fluid chromatography (SFC). The separation of polyprenols was examined on an octadecylsilane-packed column with liquid carbon dioxide as the mobile phase and ethanol as modifier. Using this chromatography system, the resolution of separation (R-s) between octadecaprenol (prenol 18) and nonadecaprenol (prenol 19) was two times higher than that using conventional reversed-phase highperformance liquid chromatography. Our SFC technique allows the advantage of baseline separation of polyprenol samples containing hydrophobic components such as terpenes or fatty acids that are unfavorable for good separation. This method is very useful for the analysis of structurally close polyprenol analogues of rubber plant metabolites. (C) 2001 Elsevier Science B.V. All rights reserved.
  • Taxezopidines J, K, and L, new taxoids from Taxus cuspidata inhibiting Ca2+-induced depolymerization of microtubules, H Shigemori, CA Sakurai, H Hosoyama, A Kobayashi, S Kajiyama, J Kobayashi, TETRAHEDRON, TETRAHEDRON, 55(9), 2553 - 2558, Feb. 1999
    Summary:Three new taxoids, taxezopidines J (1), K (2), and L (3), have been isolated from seeds of the Japanese yew Taxus cuspidata Sieb. et Zucc. and the structures were elucidated on the basis of spectroscopic data and chemical means. Taxezopidines K (2) and L (3) markedly inhibited Ca2+-induced depolymerization of microtubules, while taxuspine D (4), a known taxoid from the yew, induced unusual shape change of mitotic spindles like taxol. (C) 1999 Elsevier Science Ltd. Ail rights reserved.
  • Nostofungicidine, an antifungal lipopeptide from the field-grown terrestrial blue-green alga Nostoc commune, S Kajiyama, H Kanzaki, K Kawazu, A Kobayashi, TETRAHEDRON LETTERS, TETRAHEDRON LETTERS, 39(22), 3737 - 3740, May 1998
    Summary:In the course of our screening program for bioactive compounds, a novel lipopeptide, nostofungicidine (1), was isolated from the methanolic extract of a field-grown terrestrial blue-green alga, Nostoc commune. The structure of nostofungicidine was elucidated by chemical degradation and extensive NMR measurements including DQF-COSY, HOHAHA, HMBC, and ROESY techniques. Nostofungicidine contains a novel beta-amino acid 3-amino-6-hydroxy stearic acid (Ahs) in its structure. (C) 1998 Elsevier Science Ltd. All rights reserved.
  • Secondary metabolites of cyanobacteria Nostoc sp., Chin. J. Oceanol. Limnol., Chin. J. Oceanol. Limnol., 16(Suppliment), 109 - 117, 1998
  • Antimicrobial constituents of Angelica dahurica roots, YS Kwon, A Kobayashi, SI Kajiyama, K Kawazu, H Kanzaki, CM Kim, PHYTOCHEMISTRY, PHYTOCHEMISTRY, 44(5), 887 - 889, Mar. 1997
    Summary:One novel coumarin from Angelica dahurica roots was elucidated to be 5,8-di(2,3-dihydroxy-3-methylbutoxy)-psoralen. It occurs together with six other known coumarins and ferulic acid. The antimicrobial activity of the coumarins and ferulic acid were compared. Copyright (C) 1997 Elsevier Science Ltd.
  • Tectona grandis Callus Produces Antibacterial Triterpene Acids not Detected in the Intact plant, Natural Product Sciences, Natural Product Sciences, 3(1), 75 - 80, 1997
  • N-linked sugar chain of 55-kDa royal jelly glycoprotein, Y Kimura, S Kajiyama, J Kanaeda, T Izukawa, M Yonekura, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 60(12), 2099 - 2102, Dec. 1996
    Summary:An N-linked sugar chain from 55-kDa royal jelly glycoportein (RJGP), which maintains the high viability of rat liver primary cultured cell and is a different molecular species from 350-kDa RJGP [Kimura et al., Biosci, Biotech. Biochem., 59, 507-509 (1995)], has been identified, The sugar chains mere released by hydrazinolysis followed by N-acetylation and pyridylamination. The structural analysis of the pyridylaminated sugar chain was done by a combination of sequential eso-mannosidase digestions, MALDI-TOF MS, and 500 MHz H-1-NMR. For the carbohydrate moiety of 55-kDa RJGP, only one N-linked sugar chain has been detected. The structure has been found to be Man alpha 1-->2Man alpha 1-->6(Man alpha 1-->2Man alpha 1-->3)Man alpha 1-->6(Man alpha 1-->2Man alpha 1-->2Man alpha 1-->3)Man beta 1-->4GlcNAc beta 1-->4GlcNAc, which is a non-processed high mannose type structure.
  • A cytotoxic principle of Tamarindus indica, di-n-butyl malate and the structure-activity relationship of its analogues, A Kobayashi, MI Adenan, SI Kajiyama, H Kanzaki, K Kawazu, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 51(3-4), 233 - 242, Mar. 1996
    Summary:Bioassay-guided fractionation of the methanolic extract of Tamarindus indica fruits led to the isolation of L-(-)-di-n-butyl malate which exhibited a pronounced cytotoxic activity against sea urchin embryo cells. In order to study structure-activity relationships, close-structure relatives of di-n-butyl malate were synthesized using D-(+)- and L-(-)-malic acid as starting materials, and their cytotoxic activities were examined for the sea urchin embryo assay. L-(-)-Di-n-pentyl malate was the most effective inhibitor to the development of the fertilized eggs. Significant inhibitory activity was not seen in the esters of D-(-)-isomer.
  • ANTIFUNGAL COMPOUNDS INDUCED IN THE DUAL CULTURE WITH PHYTOLACCA-AMERICANA CALLUS AND BOTRYTIS-FABAE, A KOBAYASHI, K HAGIHARA, S KAJIYAMA, H KANZAKI, K KAWAZU, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 50(5-6), 398 - 402, May 1995
    Summary:In order to investigate new metabolites which are only induced in a plant callus infected by a pathogenic fungus, dual cultures with combinations of 10 species of fungi and 6 plant cell lines from different species were established. Among the combinations tested, the methanolic extract of a dual culture consisting of a plant cell line, Phytolacca americana and a fungus, Botrytis fabae showed a marked antifungal activity to Cladosporium herbarum. The main active constituent of this extract was identified to be phytolaccoside B (Pls B) by the spectroscopic analyses.
  • ANTIFEEDANT AND ANTIOVIPOSITION ACTIVITIES OF THE FRACTIONS OF EXTRACT FROM A TROPICAL PLANT, ANDROGRAPHIS-PANICULATA (ACANTHACEAE), AGAINST THE DIAMONDBACK MOTH, PLUTELLA-XYLOSTELLA (LEPIDOPTERA, YPONOMEUTIDAE), W HERMAWAN, S KAJIYAMA, R TSUKUDA, K FUJISAKI, A KOBAYASHI, F NAKASUJI, APPLIED ENTOMOLOGY AND ZOOLOGY, APPLIED ENTOMOLOGY AND ZOOLOGY, 29(4), 533 - 538, Nov. 1994
    Summary:Antifeedant activity of extracts from Andrographis paniculata against the 1st and the 4th-instar larvae of the diamondback moth (DBM), Plutella xylostella, was tested by using leaf disc of cabbage. Furthermore, the antioviposition effect was also observed on the caged cabbage plant treated with the fractions. The active fractions from the acetone extract suppressed feeding by the 1st and 4th-instar larvae at the concentrations of 60 ppm and 500 ppm, respectively. The antioviposition activity against DBM females was also found at the concentration of 125 ppm.
  • A NEW-TYPE OF ANTIMICROBIAL PHENOLICS PRODUCED BY PLANT PEROXIDASE AND ITS POSSIBLE ROLE IN THE CHEMICAL DEFENSE SYSTEMS AGAINST PLANT-PATHOGENS, A KOBAYASHI, Y KOGUCHI, H KANZAKI, SI KAJIYAMA, K KAWAZU, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 49(7-8), 411 - 414, Jul. 1994
    Summary:Syringaldehyde readily reacted in the horse-radish peroxidase (HRPOD) system. The ethyl acetate extract of the reaction mixture showed a marked antimicrobial activity against bacteria and fungi. After repeated column chromatography three potential antimicrobial compounds were obtained from the extract. The structural elucidation of active compounds was achieved by a combination of spectroscopic techniques and chemical modification.
  • NOSTODIONE-A, A NOVEL MITOTIC SPINDLE POISON FROM A BLUE-GREEN-ALGA NOSTOC-COMMUNE, A KOBAYASHI, SI KAJIYAMA, K INAWAKA, H KANZAKI, K KAWAZU, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES, 49(7-8), 464 - 470, Jul. 1994
    Summary:A novel antimitotic compound named Nostodione A (Nd A) was isolated from a terrestrial blue-green alga Nostoc commune. Nostodione A disturbed the mitotic spindle formation of sea-urchin eggs and gave small spindles with low birefringence density. Nostodione A, however, had no phytotoxicity on the germination of a dicotyledonous plant Medicago sativa. Based on the spectral analysis and chemical degradation, the structure of Nostodione A was elucidated.
  • NEW TELEOCIDIN-RELATED METABOLITES, (-)-7-GERANYLINDOLACTAM-V AND BLASTMYCETIN-F, FROM STREPTOVERTICILLIUM-BLASTMYCETICUM, K IRIE, SI KAJIYAMA, S OKUNO, M KONDO, K KOSHIMIZU, H HAYASHI, M ARAI, H NISHINO, A IWASHIMA, JOURNAL OF NATURAL PRODUCTS, JOURNAL OF NATURAL PRODUCTS, 57(3), 363 - 368, Mar. 1994
    Summary:Two new teleocidin-related metabolites, (-)-7-geranylindolactam-V [2] and blastmycetin F [3], were isolated from fermentation broths of the actinomycete Streptoverticillium blastmyceticum NA34-17, and their structures were determined by spectroscopic methods. Compound 2 bound strongly to phorbol ester receptors in a mouse epidermal particulate fraction, suggesting that it is a potent in vivo rumor promoter comparable to teleocidins A-1 [4] and B-4 [5].
  • PRODUCTION OF A NEW-TYPE OF BIOACTIVE PHENOLIC COMPOUND, A KOBAYASHI, Y KOGUCHI, H KANZAKI, S KAJIYAMA, K KAWAZU, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 58(1), 133 - 134, Jan. 1994
    Summary:Peroxidase (POD) catalyzed the polymerization of naturally occurring phenolics to give a variety of bioactive products. The guaiacol readily reacted in the POD system and afforded potential anti-microbial compounds. Three active phenols, 1, 2, and 3, were isolated from the reaction mixture. Spectroscopic analyses elucidated that 1 and 2 were a dimer and a trimer respectively originating from guaiacol. Compound 3 was readily oxidized to 4 by exposure to air. The structure of 4 was determined by a combination of chemical modification and spectroscopic analyses, and 3 was elucidated to be the hydroxyquinone form of 4.
  • SYNTHESIS OF NEW INDOLACTAM ANALOGS BY MICROBIAL CONVERSION, S KAJIYAMA, K IRIE, T KIDO, K KOSHIMIZU, H HAYASHI, M ARAI, TETRAHEDRON, TETRAHEDRON, 47(29), 5453 - 5462, Jul. 1991
    Summary:Ten indolactam congeners with L-Ala, Abu, gamma, delta-DELTA-Nva, Nva, Nle, tert-Leu, Leu, Ile, allo-Ile, allo-Ile, Phg instead of L-Val in (-)-indolactam-Val, were synthesized from their seco-compounds (N-methyl-L-amino acidyl-L-tryptophanol) by microbial conversion.
  • ISOLATION AND BIOSYNTHESIS OF (-)-INDOLACTAM-I, A NEW CONGENER OF INDOLE ALKALOID TUMOR PROMOTER TELEOCIDINS, K IRIE, S KAJIYAMA, K KOSHIMIZU, H HAYASHI, M ARAI, TETRAHEDRON LETTERS, TETRAHEDRON LETTERS, 31(50), 7337 - 7340, 1990
  • BIOSYNTHESIS OF (-)-INDOLACTAM-V, THE BASIC RING-STRUCTURE OF TUMOR PROMOTERS TELEOCIDINS, K IRIE, S KAJIYAMA, A FUNAKI, K KOSHIMIZU, H HAYASHI, M ARAI, TETRAHEDRON LETTERS, TETRAHEDRON LETTERS, 31(1), 101 - 104, 1990

Books etc

  • Citrus Pathology, etsuya Matsukawa, Tomonori Asai and, Joint author, Chapter 3:Metabolic Changes during Defense Responses against, InTech,   2017 04 , 9789535130727
  • Transgenic Plants - Advances and Limitations, Naruemon Khemkladngoen, Naoki Wada, Suguru Tsuchimoto, Joyce A. Cartagena, Shin-ichiro Kajiyama, Kiichi Fukui, Joint author, Chapter 5 Bioactive Beads-Mediated Transformation of Rice with Large DNA Fragments Containing Aegilops tauschii Genes, with Special Reference to Bead-Production Methodology, InTech,   2012 09 , 9789535101819
  • Plant Transformation Technologies, N. Wada, S. Kajiyama, N. Khemkladngoen, K. Fukui, Joint author, Chapter 4 A Novel Gene Delivery System in Plants with Calcium Alginate Micro-Beads, Wiley-Blackwell,   2011 08 , 0813821959
  • Quantitative structure-activity studies on indole alkaloid tumor promoter indolactam congeners.(共著), Carcinogenesis,   1991
  • Biosynthesis of indole alkaloid tumor promoters teleocidins((]G0001[))Possible biosynthetic pathway of the monoterpenoid moieties of teleocidins. (共著), Tetrahedron,   1990

Conference Activities & Talks

  • Isolation of phorbol ester degrading microbes for Jatropha bio-diesel production., International Union of Microbial Societies 2011,   2011 09 , International Union of Microbial Societies 2011
  • Stimulated parametric emission microscopy for high-resolution 3-D imaging of unstained live-cell,   2008
  • Development of high speed screening system for enzyme surface displaying yeast by flow sorting, The Society for Biotechnology Japan, Annual Meeting 2008,   2008 , The Society for Biotechnology Japan, Annual Meeting 2008
  • A novel gene delivery system in plants with calcium alginate micro-beads, International Conference on Plant Transformation Technologies,   2007 , International Conference on Plant Transformation Technologies
  • Cell imaging by spectral-interferometric stimulated parametric emission microscopy with enhanced sensitivity and fluorescence separation capability, 3rd Asian and Pacific Rim Conference on Biophotonics,   2007 , 3rd Asian and Pacific Rim Conference on Biophotonics
  • Single cell sampling and analysis by ultra short pulse laser, 5th International forum of post genome technologies,   2007 , 5th International forum of post genome technologies
  • Dynamic analysis of plant defence responses based on single-cell metabolite mapping, International Symposium on Molecular Nano Dynamics,   2006 , International Symposium on Molecular Nano Dynamics
  • Single cell based metabolite analysis upon defense response of oat by laser micro sampling and nano flow LC-MS, The 1st Workshop on Approaches to Single-Cell Analysis,   2006 , The 1st Workshop on Approaches to Single-Cell Analysis
  • Single-cell based metabolite analyses by laser microsampling and nano-flow LC-MS, The 4th International forum on post-genome technologies,   2006 , The 4th International forum on post-genome technologies

Misc

  • Jatropha curcas種子に含まれる二次代謝物質の網羅的解析, 松久保康輔, 松川哲也, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.3E7p05 (WEB ONLY),   2019 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201902225254891126
  • 花香分析とRAPD解析によるササユリ(Lilium japonicum)系統間の多様性解析, 藤澤瑞希, 荒井滋, 松川哲也, 瀧川義浩, 秋野順治, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.4D2a03 (WEB ONLY),   2019 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201902242811915482
  • ゼニゴケ(Marchantia polymorpha)の精子誘引物質の探索, 山崎由美子, 竹村美保, 松川哲也, 大和勝幸, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.4D5a10 (WEB ONLY),   2019 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201902255879211422
  • ササユリ花香成分のポリネーターに対する誘引, 花崎直史, 秋野順治, 荒井滋, 松川哲也, 瀧川義浩, 梶山慎一郎, 日本生態学会大会講演要旨(Web), 66th, ROMBUNNO.P1‐322 (WEB ONLY),   2019 02 09 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201902215145693030
  • Promotion of lipopeptide antibiotic production by Bacillus sp. IA in the presence of rice husk biochar, Ebe Shohei, Ohike Tatsuya, Matsukawa Tetsuya, Okanami Masahiro, Kajiyama Shin'ichiro, Ano Takashi, Journal of Pesticide Science, 44, 1, 33, 40,   2019 , http://ci.nii.ac.jp/naid/130007602790
    Summary:<p>The purpose of this study is to isolate the beneficial microorganisms whose growth is promoted in the presence of charcoal materials. We successfully isolated strain IA, whose growth is promoted on an agar plate with charcoal materials, and identified it as a novel strain of the <i>Bacillus</i> sp. The growth of strain IA in the liquid medium was promoted by the addition of both activated charcoal (AC) and rice husk biochar (RHB). Moreover, the sporulation of strain IA in the RHB medium and the antifungal activity of the culture supernatant of the RHB medium were much higher than those with AC. HPLC and MS analyses revealed that strain IA produced an antifungal lipopeptide iturin A, and the yield of iturin A in the RHB medium was 8 times higher than that in the medium without RHB. This is the first paper to describe the positive effect of RHB on microbial metabolisms.</p>
  • ササユリ(Lilium japonicum)系統間および類縁種間の花香組成を指標とした化学分類の試み, 藤澤瑞希, 松川哲也, 瀧川義浩, 荒井滋, 秋野順治, 柳川綾, 梶山慎一郎, 日本生物工学会大会講演要旨集, 70th, 163,   2018 08 07 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802240538984081
  • 放線菌由来揮発性有機化合物が植物病原菌に与える影響, 前田みのり, 大池達矢, 岡南政宏, 松川哲也, 梶山慎一郎, 阿野貴司, 日本生物工学会大会講演要旨集, 70th, 165,   2018 08 07 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802274262418205
  • 在来カンキツのストレス応答反応における二次代謝物質のプロファイリング, 伊佐木芳, 松川哲也, 松川哲也, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2018, ROMBUNNO.2A29a03 (WEB ONLY),   2018 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802253973729387
  • CRISPR/Cas9を用いたJatropha curcas L.のゲノム編集の試み, 藤波友樹, 児玉岳, 十川太輔, 松川哲也, 大和勝幸, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2018, ROMBUNNO.3A28a13 (WEB ONLY),   2018 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802229878554746
  • ササユリ(Lilium japonicum)の二次代謝産物分析, 藤澤瑞希, 松川哲也, 瀧川義浩, 荒井滋, 柳川綾, 秋野順治, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2018, ROMBUNNO.2A17a10 (WEB ONLY),   2018 03 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802278153950332
  • 未利用農産資源の機能性素材としての開発研究―ウンシュウミカン花のリパーゼ阻害作用―, 伊藤仁久, 村田和也, 山上桃奈, 松川哲也, 松川哲也, 梶山慎一郎, 文室政彦, 飯嶋盛雄, 飯嶋盛雄, 松田秀秋, 松田秀秋, 日本薬学会年会要旨集(CD-ROM), 138th, ROMBUNNO.27PA‐pm242,   2018 , http://jglobal.jst.go.jp/public/201802244390124894
  • Jatropha curcasの産生するホルボールエステル類の生理活性評価, 柳内知哉, 小田垣瑞穂, 高田果歩, 松川哲也, 西海史子, 名倉由起子, 柳原格, 梶山慎一郎, 日本生物工学会大会講演要旨集, 69th, 258,   2017 08 08 , http://jglobal.jst.go.jp/public/201702251066789837
  • 種々のストレス状況下におけるカンキツ類の二次代謝物質のプロファイリング, 伊佐木芳, 松川哲也, 松川哲也, 梶山慎一郎, 日本生物工学会大会講演要旨集, 69th, 258,   2017 08 08 , http://jglobal.jst.go.jp/public/201702271391796595
  • 放線菌由来生理活性物質が植物および植物病原菌に与える影響, 前田みのり, 大池達矢, 岡南政宏, 松川哲也, 梶山慎一郎, 阿野貴司, 日本生物工学会大会講演要旨集, 69th, 321,   2017 08 08 , http://jglobal.jst.go.jp/public/201702278589648865
  • ブンタン葉のストレス応答反応における二次代謝物質の変動解析, 伊佐木芳, 松川哲也, 松川哲也, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2017, ROMBUNNO.2J31a01 (WEB ONLY),   2017 03 05 , http://jglobal.jst.go.jp/public/201702279545198307
  • 微生物が生産する揮発性有機化合物の農業利用に向けた基礎的研究, 米田奈央, 江邉正平, 大池達矢, 松川哲也, 岡南政宏, 梶山慎一郎, 阿野貴司, 日本微生物生態学会大会(Web), 2017, ROMBUNNO.P‐154 (WEB ONLY),   2017 , http://jglobal.jst.go.jp/public/201702211854426369
  • 三枝祭で奉られるササユリの花香が示す抗菌性, 花崎直史, 秋野順治, 柳川綾, 梶山慎一郎, 瀧川義浩, 松川哲也, 荒井滋, 日本生態学会大会講演要旨(Web), 64th, ROMBUNNO.P1‐B‐084 (WEB ONLY),   2017 , http://jglobal.jst.go.jp/public/201702268230327293
  • 未利用農産資源の機能性素材としての開発研究―マンゴー葉のリパーゼ阻害作用―, 伊藤仁久, 村田和也, 中垣友太, 清水彩加, 高田雄輔, 志水恒介, 松川哲也, 松川哲也, 梶山慎一郎, 文室政彦, 飯嶋盛雄, 飯嶋盛雄, 松田秀秋, 松田秀秋, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.26PA‐am026,   2017 , http://jglobal.jst.go.jp/public/201702282783195829
  • アルボサイクリン生産放線菌を用いた微生物農薬の開発, 大池達矢, 前田みのり, 松川哲也, 松川哲也, 岡南政宏, 岡南政宏, 梶山慎一郎, 梶山慎一郎, 阿野貴司, 阿野貴司, 日本生物工学会大会講演要旨集, 68th, 243,   2016 08 25 , http://jglobal.jst.go.jp/public/201602229232187172
  • ハッサク葉のストレス応答反応における二次代謝物質の変動解析, 伊佐木芳, 松川哲也, 松川哲也, 梶山慎一郎, 日本生物工学会大会講演要旨集, 68th, 300,   2016 08 25 , http://jglobal.jst.go.jp/public/201602257881755960
  • Streptomyces sp.KT株の微生物農薬としての可能性, 前田みのり, 大池達矢, 岡南政宏, 岡南政宏, 松川哲也, 松川哲也, 梶山慎一郎, 梶山慎一郎, 阿野貴司, 阿野貴司, 日本生物工学会大会講演要旨集, 68th, 243,   2016 08 25 , http://jglobal.jst.go.jp/public/201602290677061238
  • 灌漑用水条件の異なるエジプト産ホホバ種子中の代謝物の変動解析, 藤波友樹, 松川哲也, HEGAZY Adel, 梶山慎一郎, 日本生物工学会大会講演要旨集, 68th, 123,   2016 08 25 , http://jglobal.jst.go.jp/public/201602291058485906
  • 御神花ササユリの葉には抗菌性がある, 岩佐郁, 秋野順治, 梶山慎一郎, 瀧川義浩, 松川哲也, 柳川綾, 荒井滋, 日本応用動物昆虫学会大会講演要旨, 60th, 187,   2016 03 10 , http://jglobal.jst.go.jp/public/201602214943615702
  • ウメの品種間代謝物プロファイリング, 松川哲也, 平山幸千, 北村祐人, 竹中正好, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2016, 2H006 (WEB ONLY),   2016 03 05 , http://jglobal.jst.go.jp/public/201602221422922756
  • カンキツ類の傷害応答反応における二次代謝物質の変動解析, 伊佐木芳, 松川哲也, 浅井智紀, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2016, 2H001 (WEB ONLY),   2016 03 05 , http://jglobal.jst.go.jp/public/201602257193741750
  • カンキツ葉中の一次代謝物質の環境ストレス応答における代謝変動解析, 浅井智紀, 松川哲也, 梶山慎一郎, 日本農芸化学会大会講演要旨集(Web), 2016, 2H002 (WEB ONLY),   2016 03 05 , http://jglobal.jst.go.jp/public/201602258892791631
  • 三輪明神摂社三枝祭の御神花ササユリの花香とリーフワックスの機能, 岩佐郁, 近藤陽香, 秋野順治, 柳川綾, 梶山慎一郎, 瀧川義浩, 荒井滋, 日本生態学会大会講演要旨(Web), 63th, P2‐190 (WEB ONLY),   2016 , http://jglobal.jst.go.jp/public/201602250493738649
  • 放線菌由来抗菌活性物質の探索, UETSUKI YOSHIHIRO, OIKE TATSUYA, MATSUKAWA TETSUYA, OKANAMI MASAHIRO, KAJIYAMA SHIN'ICHIRO, ANO TAKASHI, 日本生物工学会大会講演要旨集, 67th, 213,   2015 09 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201502209849337129
  • ササユリ(Lilium japonicum)香気成分の分析とその抗菌活性, TAKAHASHI KOKI, MATSUKAWA TETSUYA, TAKIGAWA YOSHIHIRO, ARAI SHIGERU, AKINO TOSHIHARU, KAJIYAMA SHIN'ICHIRO, 日本生物工学会大会講演要旨集, 67th, 339,   2015 09 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201502219206740657
  • 未利用農産資源の機能性素材としての開発研究―マンゴー葉の美白作用―, 伊藤仁久, 志水恒介, 堀川勇次, 友廣教道, 文室政彦, 宇都宮直樹, 飯嶋盛雄, 高田雄輔, 松川哲也, 松川哲也, 梶山慎一郎, 赤井康平, 中垣友太, 村田和也, 松田秀秋, 松田秀秋, 日本生薬学会年会講演要旨集, 62nd, 102,   2015 08 31 , http://jglobal.jst.go.jp/public/201602254709799901
  • ウンシュウミカン(Citrus unshiu)葉のリパーゼ阻害活性物質, TAKATA YUSUKE, MATSUKAWA TETSUYA, ITO KIMIHISA, SHIMIZU KOSUKE, HORIKAWA YUJI, TOMOHIRO NORIMICHI, TSUMOTO TERUYOSHI, HASHIZUME JUNJI, FUJITA TAKUYA, FUMIMURO MASAHIKO, UTSUNOMIYA NAOKI, NAKAGAKI YUTA, MURATA KAZUYA, MATSUDA HIDEAKI, KAJIYAMA SHIN'ICHIRO, 日本農芸化学会大会講演要旨集(Web), 2015, 3F37A15 (WEB ONLY),   2015 03 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201502283903737507
  • 2P-155 Isolation of antifungal active substance from actinomycetes, Uetsuki Yoshihiro, Ohike Tatsuya, Matsukawa Tetsuya, Okanami Masahiro, Kajiyama Shin'ichiro, Ano Takashi, 日本生物工学会大会講演要旨集, 67,   2015 , http://ci.nii.ac.jp/naid/110010009242
  • 3P-274 Volatile flavor components of Lillium japonicum and their antimicrobial activity, Takahashi Koki, Matsukawa Tetsuya, Takikawa Yoshihiro, Arai Shigeru, Akino Toshiharu, Kajiyama Shin'ichiro, 日本生物工学会大会講演要旨集, 67,   2015 , http://ci.nii.ac.jp/naid/110010009689
  • カンキツ植物の生活習慣病に対する有効性に関する研究―ウンシュウミカン(Citrus unshiu)葉の抗肥満作用について, ITO KIMIHISA, SHIMIZU KOSUKE, HORIKAWA YUJI, TOMOHIRO NORIMICHI, TSUMOTO MITSUTAKA, HASHIZUME JUNJI, FUJITA TAKUYA, FUMURO MASAHIKO, UTSUNOMIYA NAOKI, TAKADA YUSUKE, MATSUKAWA TETSUYA, MATSUKAWA TETSUYA, KAJIYAMA SHIN'ICHIRO, NAKAGAKI TOMOTA, MURATA KAZUYA, MATSUDA HIDEAKI, MATSUDA HIDEAKI, 日本薬学会年会要旨集(CD-ROM), 135th, ROMBUNNO.27PB-AM159,   2015 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201502295752084393
  • 続・生物工学基礎講座 バイオよもやま話 天然物由来成分に騙されるな(天然物は本当に安全なの?), MATSUKAWA TETSUYA, KAJIYAMA SHIN'ICHIRO, 生物工学会誌, 92, 10, 556, 559,   2014 10 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402219060833510
  • 3P-101 Evaluation of Trichoderma atroviride HNT-01 as biocontrol agent, Ohike Tatsuya, Matsukawa Tetsuya, Okanami Masahiro, Kajiyama Shin'ichiro, Yamamoto Shigeki, Hirose Yoichiro, Ano Takashi, 日本生物工学会大会講演要旨集, 66,   2014 08 05 , http://ci.nii.ac.jp/naid/110009906783
  • 3P-100 Development of biocontrol agents from actinomycetes, Uetsuki Yoshihiro, Ohike Tatsuya, Matsukawa Tetsuya, Okanami Masahiro, Kajiyama Shin'ichiro, Ano Takashi, 日本生物工学会大会講演要旨集, 66,   2014 08 05 , http://ci.nii.ac.jp/naid/110009906782
  • 2P-232 Analysis of metabolic changes in volatile compounds in Citrus leaves against environmental stress, Asai Tomonori, Matsukawa Tetsuya, Kajiyama Shin'ichiro, 日本生物工学会大会講演要旨集, 66,   2014 08 05 , http://ci.nii.ac.jp/naid/110009906616
  • カンキツ葉中の香気成分の環境応答反応における代謝変動解析, ASAI TOMONORI, MATSUKAWA TETSUYA, KAJIYAMA SHIN'ICHIRO, 日本生物工学会大会講演要旨集, 66th, 164,   2014 08 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402236070091327
  • 放線菌を用いた微生物農薬の開発, UETSUKI YOSHIHIRO, OIKE TATSUYA, MATSUKAWA TETSUYA, OKANAMI MASAHIRO, KAJIYAMA SHIN'ICHIRO, ANO TAKASHI, 日本生物工学会大会講演要旨集, 66th, 219,   2014 08 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402242826402429
  • SRS顕微法によるジャトロファ種子油分の非破壊解析, SASAI TOMOHIRO, TSUCHIMOTO SUGURU, SAKAI TOSHIE, WADA NAOKI, OZEKI YASUYUKI, KAJIYAMA SHIN'ICHIRO, ITO KAZUYOSHI, FUKUI KIICHI, 日本生物工学会大会講演要旨集, 66th, 141,   2014 08 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402263902163550
  • 十和田石粉末から単離したTrichoderma atroviride HNT‐01の微生物農薬としての評価, OIKE TATSUYA, MATSUKAWA TETSUYA, OKANAMI MASAHIRO, KAJIYAMA SHIN'ICHIRO, YAMAMOTO SHIGEKI, HIROSE YOICHIRO, ANO TAKASHI, 日本生物工学会大会講演要旨集, 66th, 220,   2014 08 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402269255755561
  • 2P-139 Non-destructive analysis of jatropha seed oil by SRS microscopy, Sasai Tomohiro, Tsuchimoto Suguru, Sakai Toshie, Wada Naoki, Ozeki Yasuyuki, Kajiyama Shin'ichiro, Ito Kazuyoshi, Fukui Kiichi, 日本生物工学会大会講演要旨集, 66,   2014 08 05 , http://ci.nii.ac.jp/naid/110009906523
  • Jatropha curcas種子採油残渣を用いた微生物農薬の培養, 石田結子, 大池達矢, 松川哲也, 岡南政宏, 梶山慎一郎, 阿野貴司, 日本農芸化学会大会講演要旨集(Web), 2014, 2A14P15 (WEB ONLY),   2014 03 05 , http://jglobal.jst.go.jp/public/201402235221867865
  • 低栄養性細菌を用いた微生物農薬候補株の探索, OIKE TATSUYA, MATSUKAWA TETSUYA, OKANAMI MASAHIRO, KAJIYAMA SHIN'ICHIRO, ANO TAKASHI, 日本農芸化学会大会講演要旨集(Web), 2014, 2A14P14 (WEB ONLY),   2014 03 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402236415833097
  • 日本在来藍のインディカン生産に対するストレスホルモンおよび紫外線の影響, TAKATA YUSUKE, MATSUKAWA TETSUYA, HORIBATA AKIRA, KAJIYAMA SHIN'ICHIRO, 日本農芸化学会大会講演要旨集(Web), 2014, 2C05A13 (WEB ONLY),   2014 03 05 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402293447865733
  • ナンヨウアブラギリ由来フォルボールエステル粗画分の発がんプロモーション活性, 正田将大, 玉置恵子, 村上ひかる, 松川哲也, 梶山慎一郎, 日本生物工学会大会講演要旨集, 65th, 161,   2013 08 25 , http://jglobal.jst.go.jp/public/201302290763590812
  • 2P-229 Tumor promoting activity of crude phorbol ester fraction from Jatropha curcas, Shoda Masahiro, Tamaki Keiko, Murakami Hikaru, Matsukawa Tetsuya, Kajiyama Shin'ichiro, 日本生物工学会大会講演要旨集, 65,   2013 08 25 , http://ci.nii.ac.jp/naid/110009737895
  • 1P-206 Production of antifungal lipopeptide iturin A by glycerol as BDF by-product, Ohike Tatsuya, Hashigami Kohei, Ishida Yuko, Matsukawa Tetsuya, Okanami Masahiro, Kajiyama Shin'ichiro, Ano Takashi, 日本生物工学会大会講演要旨集, 65,   2013 08 25 , http://ci.nii.ac.jp/naid/110009737586
  • 2Bp10 Isolation and characterization of wound-induced compounds in Citrus hassaku leaves, Asai Tomonori, Matsukawa Tetsuya, Ishihara Atsushi, Kajiyama Shin'ichirou, 日本生物工学会大会講演要旨集, 63,   2011 08 25 , http://ci.nii.ac.jp/naid/110008911296
  • 2Hp02 Quantitative analysis of phorbol esters in the wastes from Jatropha bio-diesel production, Suzuki Haruka, Matsukawa Tetsuya, Kajiyama Shin'ichiro, 日本生物工学会大会講演要旨集, 21,   2009 08 25 , http://ci.nii.ac.jp/naid/110007500611
  • 3S12a01 Single cell based analysis of oat difence responces, KAJIYAMA Shin'ichiro, Izumi Yashihiro, Nakamura Ryosuke, Ishihara Atsushi, Okazawa Atsushi, Fukusaki Eiichiro, Kanematsu Yasuo, Kobayashi Akio, 日本生物工学会大会講演要旨集, 21,   2009 08 25 , http://ci.nii.ac.jp/naid/110007500959
  • 2Aa01 Development of dispersion method for hydrophobic bioactive substances in water using laser ablation, FUKADA Yosuke, KAJIYAMA Shin'ichiro, ASAHI Tsuyoshi, YASUKUNI Ryohei, FUKUI Kiichi, 日本生物工学会大会講演要旨集, 20,   2008 07 11 , http://ci.nii.ac.jp/naid/110007323577
  • 1Cp14 A novel breeding method for yeast producing a high level of gamma-aminobutyric acid by femtosecond laser., TSUTSUMI Hiroko, MIZUMOTO Makiko, INDO Masaharu, SAKAKURA Masaaki, HASHIMOTO Takuya, KAJIYAMA Shinichiro, FUKUSAKI Eiichiro, UEDA Mitsuyoshi, HIRAO Kazuyuki, HATA Yoji, 日本生物工学会大会講演要旨集, 20,   2008 07 11 , http://ci.nii.ac.jp/naid/110007323702
  • How to use production potential of photosynthetic bacteria and plants for global warning, 小林 昭雄, 梶山 慎一郎, 岡澤 敦司, Lichenology, 6, 2, 154, 157,   2007 , http://ci.nii.ac.jp/naid/40015808716
  • 62.Development of a novel quantification system of transcripts using mass spectrometer, Hirozane Yoshihiko, Fukusaki Eiichiro, Kajiyama Shinichiro, Kobayashi Akio, 植物化学調節学会研究発表記録集, 41, 41,   2006 10 04 , http://ci.nii.ac.jp/naid/110006168824
  • 63.High sensitive nano flow LC-ESI-MS analysis using packed nano capillary column integrated with nano-sprayer for plant metabolite anaylysis, Izumi Yoshihiro, Fukusaki Eiichiro, Kajiyama Shinichiro, Kobayashi Akio, 植物化学調節学会研究発表記録集, 41, 41,   2006 10 04 , http://ci.nii.ac.jp/naid/110006168825
  • 2H12-3 Expression analysis of error-prone DNA polymerases in human cultured cells under stress condition, HIRANO Miki, KURIHARA Sachiko, AKIYAMA Yukio, KAJIYAMA Shin'ichiro, FUKUI Kiichi, 日本生物工学会大会講演要旨集, 18,   2006 08 03 , http://ci.nii.ac.jp/naid/110007322021
  • 3B09-4 Development of a novel quantification system of transcripts using mass spectrometer, HIROZANE Yoshihiko, FUKUSAKI Eiichiro, KAJIYAMA Shin'ichiro, KOBAYASHI Akio, 日本生物工学会大会講演要旨集, 18,   2006 08 03 , http://ci.nii.ac.jp/naid/110007322411
  • 3A-AM5 Laser technology for single cell sampling and analysis, KAJIYAMA Shin'ichiro, KOBAYASHI Akio, FUKUI Kiichi, ITOH Kazuyoshi, 日本生物工学会大会講演要旨集, 18,   2006 08 03 , http://ci.nii.ac.jp/naid/110007322470
  • 2H12-1 Analysis of transcripts using ESI-MS(I) : investigation of ionization condition, KAJIWARA Shin'ichi, RIHARA Yoshiyuki, IZUMI Yoshihiro, HARADA Kazuo, FUKUSAKI Ei-ichiro, KOBAYASHI Akio, 日本生物工学会大会講演要旨集, 17,   2005 09 25 , http://ci.nii.ac.jp/naid/110007321274
  • 3S2-AM6 Application of plused UV laser micro-processing for plant cells, KAJIYAMA Shin'ichiro, KOBAYASHI Akio, 日本生物工学会大会講演要旨集, 16,   2004 08 25 , http://ci.nii.ac.jp/naid/110007321130
  • 47(P-9) Single cell analysis of plant metabolites using LASER micro sampling, Kobayashi A, Kajiyama S, Watabe T, Harada K, Fukusaki E, 天然有機化合物討論会講演要旨集, 45, 275, 280,   2003 09 01 , http://ci.nii.ac.jp/naid/110006682306
    Summary:The analysis of bulk tissue samples gives the informations about solute and metabolite concentrations of all compartments and different cell types, and this do not reflect the fine scale patterns between adjacent cells or provide information about processes, that are regulated on the single cell level. To understand the physiology of a whole organism, it is necessary to understand the behavior of a single cell as its fundamental unit. Such investigations essentially require single cell or even subcellular analysis. Along with the progress of plant physiology, the reliable method for single cell sampling is sought-after to investigate the precise metabolic profiling of different cell types. Research on single cells of higher plants is relatively difficult, mainly because of their small size and rigid cell walls. Single cell sampling of plant cell is usually performed by a fine oil-filled glass microcapillary, mounted on a micromanipulator. However, this method is time consuming and rather needs high skill due to the hard and thick cell wall barrier. We developed a unique method of laser-assisted single cell sampling and applied this to metabolite analysis at the resolution of the individual cell. Some pico-litter cell content of GFP expressed transgenic torenia (Torenia hybrida.) were picked up from a single cell (epidermal and petal cell) using this method. The trans gene (GFP) mRNA was detected by RT-PCR, and the petal pigments i.e. peonidin-3,5-diglucoside and malvidin-3-glucoside-5-(p-coumaroyl)-glucoside were also identified by using nano flow LC-ESI MS/MS and MALDI TOF MS. This method would be one of the most powerful tools for studying metabolic profiling of individual plant cells.
  • Application of UV pulsed laser micro beam to plant cells (IV) : Acquisition of stable transgenic plants by the laser gene introduction, Inoue Fumihide, Kajiyama Shinichiro, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002955917
  • Application of UV pulsed laser micro beam to plant cells (V) : An approach to huge-sized DNA introduction to plant cells, Tabata Atsushi, Kajiyama Shinichiro, Fukusaki Eiichiro, Harashima Satoshi, Fukui Kiichi, Kobayashi Akio, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002955918
  • Application of UV pulsed laser micro beam to plant cells (VI) : One cell analysis of petal pigments using laser microsampling, Kajiyama Shinichiro, Watabe Tatsuya, Harada Kazuo, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002955919
  • Analysis of molecular mechanism of autumnal tints : Localization analysis of Ginkgo biloba chlorophyllase, Okazawa Atsushi, Itoh Yoshiko, Tang Lei, Kajiyama Shinichiro, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002956100
  • Transformation of tobacco SR-1 using bio-active beads, Murakawa Tomoko, Kawabe Akira, Liu Haibo, Yanagisawa Masanobu, Kajiyama Shinichiro, Kobayashi Akio, Harashima Satoshi, Matsunaga Sachihiro, Kiichi, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002956133
  • Expression of Aspergillus niger pectin methylesterase in transgenic tobacco (Nicotiana tabacum) leads to alterations on the level of methanol and a dwarf phenotype, Hasunuma Tomohisa, Fukusaki Eiichiro, Okazawa Atsushi, Kajiyama Shinichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 15,   2003 08 25 , http://ci.nii.ac.jp/naid/110002956134
  • 47(P-9) Single cell analysis of plant metabolites using LASER micro sampling, Kobayashi A, Kajiyama S, Watabe T, Harada K, Fukusaki E, Symposium on the Chemistry of Natural Products, symposium papers, 45, 0, 275, 280,   2003 , http://ci.nii.ac.jp/naid/130005978963
    Summary:The analysis of bulk tissue samples gives the informations about solute and metabolite concentrations of all compartments and different cell types, and this do not reflect the fine scale patterns between adjacent cells or provide information about processes, that are regulated on the single cell level. To understand the physiology of a whole organism, it is necessary to understand the behavior of a single cell as its fundamental unit. Such investigations essentially require single cell or even subcellular analysis. Along with the progress of plant physiology, the reliable method for single cell sampling is sought-after to investigate the precise metabolic profiling of different cell types. Research on single cells of higher plants is relatively difficult, mainly because of their small size and rigid cell walls. Single cell sampling of plant cell is usually performed by a fine oil-filled glass microcapillary, mounted on a micromanipulator. However, this method is time consuming and rather needs high skill due to the hard and thick cell wall barrier. We developed a unique method of laser-assisted single cell sampling and applied this to metabolite analysis at the resolution of the individual cell. Some pico-litter cell content of GFP expressed transgenic torenia (Torenia hybrida.) were picked up from a single cell (epidermal and petal cell) using this method. The trans gene (GFP) mRNA was detected by RT-PCR, and the petal pigments i.e. peonidin-3,5-diglucoside and malvidin-3-glucoside-5-(p-coumaroyl)-glucoside were also identified by using nano flow LC-ESI MS/MS and MALDI TOF MS. This method would be one of the most powerful tools for studying metabolic profiling of individual plant cells.
  • 951 Application of UV plused laser microbeam technique to plant cells (III) : Analysis of plant wounding responses to plused UV laser, Shoji Takeshi, Kajiyama Shinichiro, Kawaoka Akiyoshi, Yoshida Kazuya, Shinmyo Atsuhiko, Yamaguchi Yube, Sano Hiroshi, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 14,   2002 09 25 , http://ci.nii.ac.jp/naid/110007319756
  • 950 Application of UV pulsed laser microbeam technique to plant cells (II) : Sophisticated approach to one cell metabolites analysis, Kajiyama Shinichiro, Yoshikawa Norihiro, Watabe Tatsuya, Shoji Takeshi, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 14,   2002 09 25 , http://ci.nii.ac.jp/naid/110007319757
  • 949 Application of UV pulsed laser microbeam technique to plant cells (I) : Laser microprocessing and its application, Yoshikawa Norihiro, Shoji Takeshi, Kajiyama Shinichiro, Fukusaki Eiichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 14,   2002 09 25 , http://ci.nii.ac.jp/naid/110007319758
  • 537 Elucidation of a molecular recognition mechanism of the chitin binding penticles, Fukusaki Eiichiro, Ogawa Koichi, Kajiyajma Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 日本生物工学会大会講演要旨集, 14,   2002 09 25 , http://ci.nii.ac.jp/naid/110007320327
  • 33 Structure of Hetero Polysaccharide from Desiccation-tolerant Terrestrial Cyanobacterium Nostoc commune, Kajiyama Shin'ichiro, Yagi Masahiro, Kurihara Tatsuyuki, Okazawa Atsushi, Fukusaki Ei-ichiro, Kobayashi Akio, 天然有機化合物討論会講演要旨集, 43, 193, 198,   2001 09 01 , http://ci.nii.ac.jp/naid/110006682078
    Summary:Cyanobacteria (blue-green algae) are photoautotrophic prokaryotes which include a large variety of species with varied morphological, physiological, and biochemical properties. It is well known that some kinds of cyanobacteria produce large amounts of exopolysaccharides. Although the quantity and biochemical properties are diverse depending on the species, these polysaccharides are thought to play important roles in desiccation tolerance, UV tolerance, nutrient uptake, and interactions with other organisms in the establishment of symbiosis. Nostoc commune, a nitrogen-fixing filamentous cyanobacterium, also produces copious amounts of polysaccharide around the cells and has remarkable desiccation tolerance. Despite conspicuous features of polysaccharide of Nostoc commune, little is known about its chemical structure due largely to its structural complexity. In general, polysaccharides of cyanobacteria are more complex both in their sugar composition and in their linkage types than those of other bacteria or macroalgae, and at this time only a few structures of cyanobacterial polysaccharides have been proposed. In this study, we investigated the chemical structure of water-soluble polysaccharide from desiccation-tolerant cyanobacterium Nostoc commune. The water-soluble polysaccharide (glycan) was extracted and purified from field-grown Nostoc commune. Sugar compositional analysis revealed that this polysaccharide contained D-glucose, D-galactose, D-xylose, and D-glucuronic acid in the molecular ratio of 8: 4: 4: 1 together with unknown uronic acids and trace amount of ribose. One of the unknown uronic acid was identified as 2,3-O-(1-carboxyethylidene)-D-glucuronic acid by NMR analysis of the methanolysate of the polysaccharide. Partial hydrolysis of the polysaccharide by acid (0.2 N TFA, 90℃, 5h) or by crude enzyme which was obtained from culture brose of Paenibacillus sp. afforded 5 oligosaccharide fragments. After pyridylamination, these oligosaccharides were isolated by reverse phase column chromatography, and structures were elucidated by 2D NMR. By comparison with the structures of these oligosaccharides, we concluded that the polysaccharide had a 5-membered repeating unit viz.→4)-β-D-Glcp-(1→4)-β-D-Xylp-(1→4)-[β-D-GlcAp-(1→6)]-β-D-Glcp-(1→4)-β-D-Galp-(1→, and residues in the repeating unit were partially substituted by 3-O-(1-carboxyethyl)-D-glucuronic acid or 2,3-O-(1-carboxyethylidene)-D-glucuronic acid in the GlcA moiety and by 2-O-Me-Man in the branched Glc moiety as shown in Figure 5.
  • Structure elucidation of a tetramic acid having an antimicrobial activity, produced in callus and fungus co-culture system., Marfori E. C, Kajiyama Shin'ichiro, Okazawa Atsushi, Fukusaki Ei-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 13,   2001 08 24 , http://ci.nii.ac.jp/naid/110002922853
  • Analysis of yellowing mechanism of Ginkgo biloba leaves : Cloning and expression of chlorophyllase, Okazawa Atsushi, Itoh Yoshiko, Tang Lei, Fukusaki Ei-ichiro, Kajiyama Shin-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 13,   2001 08 24 , http://ci.nii.ac.jp/naid/110002922893
  • 33 Structure of Hetero Polysaccharide from Desiccation-tolerant Terrestrial Cyanobacterium Nostoc commune, Kajiyama Shin'ichiro, Yagi Masahiro, Kurihara Tatsuyuki, Okazawa Atsushi, Fukusaki Ei-ichiro, Kobayashi Akio, Symposium on the Chemistry of Natural Products, symposium papers, 43, 0, 193, 198,   2001 , http://ci.nii.ac.jp/naid/130005978735
    Summary:Cyanobacteria (blue-green algae) are photoautotrophic prokaryotes which include a large variety of species with varied morphological, physiological, and biochemical properties. It is well known that some kinds of cyanobacteria produce large amounts of exopolysaccharides. Although the quantity and biochemical properties are diverse depending on the species, these polysaccharides are thought to play important roles in desiccation tolerance, UV tolerance, nutrient uptake, and interactions with other organisms in the establishment of symbiosis. Nostoc commune, a nitrogen-fixing filamentous cyanobacterium, also produces copious amounts of polysaccharide around the cells and has remarkable desiccation tolerance. Despite conspicuous features of polysaccharide of Nostoc commune, little is known about its chemical structure due largely to its structural complexity. In general, polysaccharides of cyanobacteria are more complex both in their sugar composition and in their linkage types than those of other bacteria or macroalgae, and at this time only a few structures of cyanobacterial polysaccharides have been proposed. In this study, we investigated the chemical structure of water-soluble polysaccharide from desiccation-tolerant cyanobacterium Nostoc commune. The water-soluble polysaccharide (glycan) was extracted and purified from field-grown Nostoc commune. Sugar compositional analysis revealed that this polysaccharide contained D-glucose, D-galactose, D-xylose, and D-glucuronic acid in the molecular ratio of 8: 4: 4: 1 together with unknown uronic acids and trace amount of ribose. One of the unknown uronic acid was identified as 2,3-O-(1-carboxyethylidene)-D-glucuronic acid by NMR analysis of the methanolysate of the polysaccharide. Partial hydrolysis of the polysaccharide by acid (0.2 N TFA, 90℃, 5h) or by crude enzyme which was obtained from culture brose of Paenibacillus sp. afforded 5 oligosaccharide fragments. After pyridylamination, these oligosaccharides were isolated by reverse phase column chromatography, and structures were elucidated by 2D NMR. By comparison with the structures of these oligosaccharides, we concluded that the polysaccharide had a 5-membered repeating unit viz.→4)-β-D-Glcp-(1→4)-β-D-Xylp-(1→4)-[β-D-GlcAp-(1→6)]-β-D-Glcp-(1→4)-β-D-Galp-(1→, and residues in the repeating unit were partially substituted by 3-O-(1-carboxyethyl)-D-glucuronic acid or 2,3-O-(1-carboxyethylidene)-D-glucuronic acid in the GlcA moiety and by 2-O-Me-Man in the branched Glc moiety as shown in Figure 5.
  • Peptide-degrading capacity of pitcher fluid from the carnivorous plant Nepenthes alata., AN Chung-I, FUKUSAKI Ei-ichiro, TAKEKAWA Shoji, OKAZAWA Atsushi, KAJIYAMA Shin-ichiro, KOBAYASHI Akio, Chemical Regulation of Plants, 35, 2,   2000 12 25 , http://ci.nii.ac.jp/naid/110001855794
  • Effects of C1 compounds on plant metabolisms (part7) : Analysis of the proteins responding to methylamine treatment in rice plant, FUKUSAKI Ei-ichiro, KAWAMOTO Yuri, SHIRAISHI Takehiko, KAJIYAMA Shinichiro, OKAZAWA Atsushi, KOBAYASI Akio, Chemical Regulation of Plants, 35, 2,   2000 12 25 , http://ci.nii.ac.jp/naid/110001855802
  • Effects of C1 compounds on plant metabolisms (part 6) : Analysis of the genes responding to methylamine treatment in rice plant, SHIRAISHI Takehiko, WATANABE Tetsuya, FUKUSAKI Ei-ichiro, KAJIYAMA Shinichiro, OKAZAWA Atsushi, KOBAYASHI Akio, Chemical Regulation of Plants, 35, 2, 226, 227,   2000 12 25 , http://ci.nii.ac.jp/naid/110001855803
  • Peptide-degrading capacity of pitcher fluid from the carnivorous plant Nepenthes alata., An Chung-Il, Fukusaki Ei-ichiro, Takekawa Shoji, Okazawa Atsushi, Kajiyama Shin-ichiro, Kobayashi Akio, 植物化学調節学会研究発表記録集, 35, 29, 30,   2000 11 02 , http://ci.nii.ac.jp/naid/110001853515
    Summary:Nepenthes is a unique genus of carnivorous plants that can capture insects in trapping organs called pitchers and digest them in pitcher fluid. The pitcher fluid includes digestive enzymes and is strongly acidic. Previous reports revealed that the pitcher fluid has strong proteolytic activity. The activity is thought to be essential for degradation of insect proteins. To elucidate the degradation process of proteins in the pitcher fluid, we evaluated peptide-degrading capacity of the fluid with B chain of oxidized insulin as a model substrate. LC-MS analysis demonstrated that the substrate was rapidly cleaved around hydrophobic amino acid residues. These cleavage sites were close to those by porcine pepsin A.
  • Effects of C1 compounds on plant metabolisms (part 7) : Analysis of the proteins responding to methylamine treatment in rice plant, Fukusaki Ei-ichiro, Kawamoto Yuri, Shiraishi Takehiko, Kajiyama Shinichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 35, 45, 46,   2000 11 02 , http://ci.nii.ac.jp/naid/110001853523
    Summary:The metabolism of C1 compounds in higher plants is an attractive research target. Rice plant growth test revealed that methylamine treatment inhibited the aerial part but stimulated the growth of root part. Proteins from methylamine treated rice and untreated rice were separated by two-dimensional gel electrophoresis (2D-PAGE). Eleven variable spots were observed by the overlap between comparisons of the methylamine treated and untreated rice plants. Peptide sequencing analysis of the variable spots revealed following fashion: 1) The protein level of glyceraldehyde-3-phosphate dehydrogenase are increased in both aerial part and root; 2) one of germin-like protein was decreased in root; 3) one of malate dehydrogenases and voltage-dependent anion channel proteins were increased in the aerial part.
  • Effect of C1 compounds on plant metabolisms (part 6) : Analysis of the genes responding to methylamine treatment in rice plant, Shiraishi Takehiko, Watanabe Tetsuya, Fukusaki Ei-ichiro, Kajiyama Shinichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 35, 47, 48,   2000 11 02 , http://ci.nii.ac.jp/naid/110001853524
    Summary:The metabolism of C1 compounds in higher plants in an attractive research target. Rice plant growth test revealed that methylamine treatment inhibited the aerial part by contrast stimulated the growth of root part. Homologues of seven independent genes, accumulated by methylamine treatment were isolated by the DD (Differential Display) method. Increase of mRNA level of two genes (MS1, cytosolic phosphoglycerate kinase homologue; MR1, chloroplast 23S ribosomal RNA homologue) was confirmed by Northern blot analysis. Further Northern blot analysis of the genes related to glycolytic pathway also indicated that stimulation of glycolysis system in methylamine treated plant.
  • The analysis of viscous colony where Escherichia coli and Dictyostelium discoidium are coexisting., Oki S, Todoriki M, Kajiyama S, Fukuzaki E, Kobayash A, Urabe I, Yomo T, Biophysics, 40, 1,   2000 08 05 , http://ci.nii.ac.jp/naid/110001154312
  • In vitro selection of DNA aptamers that bind specifically to tubulin, Hasunuma Tomohisa, Fukusaki Ei-ichiro, Okazawa Atsushi, Kajiyama Shin-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 12,   2000 07 10 , 10.1016/S0960-894X(00)00013-5, http://ci.nii.ac.jp/naid/110002922262
  • Structure elucidation of exocellular polysaccharide produced by desiccation tolerant cyanobacterium Nostoc sp., Kajiyama Shin'ichiro, Yagi Masahiro, Fukusaki Eiichiro, Okazawa Atsushi, Kobayashi Akio, 日本生物工学会大会講演要旨集, 12,   2000 07 10 , http://ci.nii.ac.jp/naid/110002922430
  • Effects of C1 compounds on plant metabolisms (part4) : protecting effects against photoinhibition., SHIRAISHI Takehiko, FUKUSAKI Ei-ichiro, MIYAKE Chikahiro, YOKOTA Akiho, KAJIYAMA Shin-ichiro, OKAZAWA Atsushi, KOBAYASHI Akio, Chemical Regulation of Plants, 34, 2, 351, 352,   1999 12 25 , http://ci.nii.ac.jp/naid/110001846932
  • Effects of C1 compounds on plant metabolisms (part5) : C1 compounds metabolism in Arabidopsis thaliana., FUKUSAKI Ei-ichiro, IKEDA Tatsuhiko, NISHIKAWA Takashi, SHIRAISHI Takehiko, KAJIYAMA Shin-ichiro, OKAZAWA Atsushi, KOBAYASHI Akio, Chemical Regulation of Plants, 34, 2,   1999 12 25 , http://ci.nii.ac.jp/naid/110001846933
  • Analyses of the expression pattern of the genes stimulated by elicitor in French bean plant., FUKUSAKI Ei-ichiro, MATSUNAWA Ayako, WATANABE Tetsuya, KAJIYAMA Shin-ichiro, OKAZAWA Atsushi, KOBAYASHI Akio, Chemical Regulation of Plants, 34, 2,   1999 12 25 , http://ci.nii.ac.jp/naid/110001846956
  • cDNA cloning of plasma membrane proton pump of carnivorous plant Nepenthes hybrida., FUKUSAKI Ei-ichiro, AN Tadaichi, KAJIMYAMA Shin-ichiro, OKAZAWA Atsushi, KOBAYASHI Akio, Chemical Regulation of Plants, 34, 2,   1999 12 25 , http://ci.nii.ac.jp/naid/110001846957
  • 61 Effects of C1 compounds on plant metabolisms (part4)(protecting effects against photoinhibition), Shiraishi Takehiko, Fukusaki Ei-ichiro, Miyake Chikahiro, Yokota Akiho, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 34, 121, 122,   1999 10 01 , http://ci.nii.ac.jp/naid/110001853471
    Summary:In plants, environmental adversity often leads to the formation of highly reactive oxy gen species. Even though light is the source of energy for photosynthesis, it can also be harmful to plants. Light-induced damage is targeted mainly to photosystem II and leads to the inactivation of electron transport systems and subsequent oxidative destruction of the reaction centers, in particular to the deactivation of PS II center proteins. We showed that incubation of rice leaf under the high light with CO_2-depleted air causes serious demage on photosynthesis, however the application of potassium formate or methanol could protect rice photosynthesis form photoinhibition.
  • 62 Effects of C1 compounds on plant metabolisms (part5)(C1 compounds metabolism in Arabidopsis thaliana), Fukuzaki Ei-ichiro, Ikeda Tatsuhiko, Nishikawa Takashi, Shiraishi Takehiko, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 34, 123, 124,   1999 10 01 , http://ci.nii.ac.jp/naid/110001853472
    Summary:Our experiments showed that methanol and formate are oxidized to CO_2 in Arabidopsis thaliana. Full length of FDH(formate dehydrogenase) gene, which catalyzed the conversion of formate to CO_2, was cloned by the RT-PCR and 3'-and 5'-RACE.
  • 85 Analyses of the expression pattern of the genes stimulated by elicitor in French bean plant, Fukusaki Ei-ichiro, Matsunawa Ayako, Watanabe Tetsuya, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 34, 163, 164,   1999 10 01 , http://ci.nii.ac.jp/naid/110001853495
    Summary:Plants respond activily to infection and other environmental stresses and these responses are known to be induced by compounds, elicitors. We have focused on specific gene expressions induced by elicitors. To obtain desired specific genes, we performed differential display RT-PCR method with elicitor-treated plant. We will present the result of expression analysis of the gene family in French bean(Phaseolus vulgaris) induced by a carbohydrate elicitor, laminaran.
  • 86 cDNA cloning of plasma membrane proton pump of carnivorous plant Nepenthes hybrida, Fukusaki Ei-ichiro, An Tadaichi, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 34, 165, 166,   1999 10 01 , http://ci.nii.ac.jp/naid/110001853496
    Summary:Carnivorous plants inhabit nutrient-poor environment and they capture insects to supplement their nutrient demand. Carnivorous plant Nepenthes hybrida has a special organ to capture and digest insects. Inside of this trapping organ(pitcher), acidic fluid which contains digestive enzymes is secreted. To elucidate the role of acidification of pitcher fluid in nutrient acquisition process, we cloned cDNA fragments which encode putative plasma membrane H^+-ATPase homologues. Northern analysis showed that one of the cloned products was expressed in roots, stems, petioles and pitchers. Among these tissues, the highest expression was observed in lower part of opened pitcher with natural prey.
  • Structure elucidation of exocellular polysaccharide produced by desiccation tolerant terrestrial cyanobacterium Nostoc sp., Kajiyama Shin'ichiro, Yagi Masahiro, Fukusaki Eiichiro, Okazawa Atsushi, Kobayashi Akio, 日本生物工学会大会講演要旨集, 11,   1999 08 16 , http://ci.nii.ac.jp/naid/110002920819
  • Chlorophyll catabolism in yellowing of Ginkgo green leaves, Okazawa Atsushi, Tang Lei, Fukusaki Ei-ichiro, Kajiyama Shin-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 11,   1999 08 16 , http://ci.nii.ac.jp/naid/110002920820
  • In vitro selection of DNA aptamers that bind specifically to hematoporphyrin, Okazawa Atsushi, Maeda Hiroshi, Fukusaki Ei-ichiro, Kajiyama Shin-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 11,   1999 08 16 , http://ci.nii.ac.jp/naid/110002920821
  • A-10 Effect of C1 compounds on the plant metabolism : Growth stimulation by formate in rice plant, FUKUZAKI Eiichiro, SHIRAISHI Takehiko, KAJIYAMA Shin-ichiro, OKAZAWA Atsushi, KOBAYASHI Akio, 植物化学調節学会研究発表記録集, 33, 29, 30,   1998 10 01 , http://ci.nii.ac.jp/naid/110001853358
    Summary:The metabolism of C1 compounds in higher plants is an attractive research target. As a first step, we investigated the effect of formate on the growth of rice plant.Growth stimulation of the aerial part and increase of fresh weight were observed by the (Formate Dehydrogenase)specific activity was increased. These findings suggest the existence of formate metabolism in rice plant. We obtained FDH homologue sequence based on potato FDH sequence by RT-PCR method. The cDNA contains 1230 bp open reading frame. The deduced amino acid sequence shows high homology to other plant FDHs, so far reported.
  • Radical coupling reaction of hydroxystilbene by Vitacea cell culture., Fukusaki Eiichiro, Kobayashi Yoshitaka, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 日本生物工学会大会講演要旨集, 10,   1998 08 31 , http://ci.nii.ac.jp/naid/110002921534
  • 76(P55) Microbial flora controlling substances in the jelly clumps of Nostoc commune, Kajiyama Shin-ichiro, Nakao Motonao, Kobayashi Akio, Sasaki Katsuharu, Kawazu Kazuyoshi, 天然有機化合物討論会講演要旨集, 40, 449, 454,   1998 08 31 , http://ci.nii.ac.jp/naid/110006679682
    Summary:Recently blue-green algae (cyanobacteria) attract much attention because of their secondary metabolites with potent biological activities and unusual structures. Blue-green algae have also been checked for their metabolism influenced by symbiotic or parasitic microorganisms. For example, lichens, symbiotic organisms consisting of algae and fungi, produce peculiar secondary metabolites called lichen substances. Nostoc commune is a blue-green alga which often grows on wet ground surface in jelly clumps consisting of polysaccharides. Our preliminary experiment showed that many microorganisms including bacteria, fungi, and actinomycetes were residing in the jelly clumps of field grown Nostoc commune. On the other hand the methanol extract of this alga exhibited antifungal activity against 7 fungi (Aspergillus cardidus, Cladosporium herbarum, Botrytis fabae, Glomerella cingulata, Sclerotium rolfsii, Rhizoctoniasolani, Cercospora beticala) suggesting that the antimicrobial substance(s) may control microbial flora of the jelly clumps. As the first step to clarify the relationship between the alga and co-residing microorganisms, we isolated one of the antimicrobial principles-named nostofungicidine (1) from field-grown algal sample. The structure of nostofungicidine was elucidated mainly by chemical degradation and by extensive NMR measurements including DQF-COSY, HOHAHA, HMBC, and ROES Y techniques. Nostofungicidine was a novel lipopeptide containing a long chain beta amino acid, 3-amino-6-hydroxy stearic acid (Ahs) in its structure. Nostofungicidine was not found in the axenic culture of this alga and in the individual cultures of each tested microbial strain isolated from jelly clumps. On the other hand, one of the co-residing strain (tentatively identified as an actinomycete) produced a known antibiotic, echinomycin. Echinomycin had no antialgal effect toward axenic Nostoc commune at the concentration of 100 micro-g/ml. These results suggest that co-residing microorganisms may play an important role in the accumulation of microbial flora controlling substances not only through production of antimicrobial compound(s) but also through stimulation of the algal secondary metabolism.
  • 76(P55) Microbial flora controlling substances in the jelly clumps of Nostoc commune, Kajiyama Shin-ichiro, Nakao Motonao, Kobayashi Akio, Sasaki Katsuharu, Kawazu Kazuyoshi, Symposium on the Chemistry of Natural Products, symposium papers, 40, 0, 449, 454,   1998 , http://ci.nii.ac.jp/naid/130005978424
    Summary:Recently blue-green algae (cyanobacteria) attract much attention because of their secondary metabolites with potent biological activities and unusual structures. Blue-green algae have also been checked for their metabolism influenced by symbiotic or parasitic microorganisms. For example, lichens, symbiotic organisms consisting of algae and fungi, produce peculiar secondary metabolites called lichen substances. Nostoc commune is a blue-green alga which often grows on wet ground surface in jelly clumps consisting of polysaccharides. Our preliminary experiment showed that many microorganisms including bacteria, fungi, and actinomycetes were residing in the jelly clumps of field grown Nostoc commune. On the other hand the methanol extract of this alga exhibited antifungal activity against 7 fungi (Aspergillus cardidus, Cladosporium herbarum, Botrytis fabae, Glomerella cingulata, Sclerotium rolfsii, Rhizoctoniasolani, Cercospora beticala) suggesting that the antimicrobial substance(s) may control microbial flora of the jelly clumps. As the first step to clarify the relationship between the alga and co-residing microorganisms, we isolated one of the antimicrobial principles-named nostofungicidine (1) from field-grown algal sample. The structure of nostofungicidine was elucidated mainly by chemical degradation and by extensive NMR measurements including DQF-COSY, HOHAHA, HMBC, and ROES Y techniques. Nostofungicidine was a novel lipopeptide containing a long chain beta amino acid, 3-amino-6-hydroxy stearic acid (Ahs) in its structure. Nostofungicidine was not found in the axenic culture of this alga and in the individual cultures of each tested microbial strain isolated from jelly clumps. On the other hand, one of the co-residing strain (tentatively identified as an actinomycete) produced a known antibiotic, echinomycin. Echinomycin had no antialgal effect toward axenic Nostoc commune at the concentration of 100 micro-g/ml. These results suggest that co-residing microorganisms may play an important role in the accumulation of microbial flora controlling substances not only through production of antimicrobial compound(s) but also through stimulation of the algal secondary metabolism.
  • 21 Gene expression of phenylpropanoid biosynthetic enzyme in elicitor treated French bean, Matsunawa Ayako, Watanabe Takashi, Fukusaki Ei-ichiro, Kajiyama Shin-ichiro, Okazawa Atsushi, Kobayashi Akio, 植物化学調節学会研究発表記録集, 32, 47, 48,   1997 11 28 , http://ci.nii.ac.jp/naid/110001854012
  • Isolation and analysis of genes induced by elicitor, Fukusaki Ei-ichirou, Watanabe Takashi, Kajiyama Sin-ichirou, Kobayashi Akio, 日本生物工学会大会講演要旨集, 9,   1997 09 05 , http://ci.nii.ac.jp/naid/110002924565
  • Elicitor activities of chimera oligosaccharides consisting of glucose and glucosamine., Fukusaki Eiichiro, Yabushita Setsuko, Nakanishi Tsuyoshi, Kajiyama Shin-ichiro, Kobayashi Akio, 日本生物工学会大会講演要旨集, 8,   1996 09 05 , http://ci.nii.ac.jp/naid/110002925358
  • Production of bioactive compounds by peroxidase reaction system, Kobayashi Akio, Bamba Takeshi, Kajiyama Shin-ichiro, Kawazu Kazuyoshi, 日本生物工学会大会講演要旨集, 7,   1995 09 15 , http://ci.nii.ac.jp/naid/110002948036
  • Enzymatic synthesis of elicitor-active thiophenyl hetero-N-acetyl-chitooligosaccharides utilizing lysozyme-catalyzed transglycosylation reaction., Kobayashi Akio, Kanzaki Hiroshi, Akiyama Kohki, Kajiyama Shin-ichiro, Kawazu Kazuyoshi, 日本生物工学会大会講演要旨集, 6,   1994 10 10 , http://ci.nii.ac.jp/naid/110002923667
  • 47 THE ROLE OF PEROXIDASE IN THE CHEMICAL DEFENCE OF PLANTS : A NOVEL PEROXIDASE INHIBITOR AND A NEW TYPE OF BIOACTIVE PHENOLICS PRODUCED BY PEROXIDASE, Kobayashi A, Koguchi Y, Kanzaki H, Kajiyama S, Kawazu K, 天然有機化合物討論会講演要旨集, 35, 361, 368,   1993 09 10 , http://ci.nii.ac.jp/naid/110006679211
    Summary:Plants induce varied resistance-response toward microbial challenge. In order to establish an infection, pathogenic fungi must break such defense barriers as lignin, phytoalexins, and hydrolytic enzymes. At the early stage of infection, POD (peroxidase) activity is enhanced dramatically. POD and lignin accumulates at the infected site and then protects from further fungal invasion. The suppression of lignin deposition at the site might be of advantage to susceptible pathogenic fungi. We, therefore, conducted a comprehensive screening to obtain POD inhibitors from fungal cultures and a potent radical scavenger named ST-1 was isolated from a fungus, and its chemical structure was elucidated to be 1,3-dihydro-4,5,6-trihydroxy-1,3-dimethoxy-7-methylisobenzofuran by a combination of chemical modification and spectroscopic techniques. ST-1 effectively scavenged oxygen radicals and also suppressed the phytoalexin accumulation in kidney bean. These results suggested that such a mechanism can be enumerated as one of significant defense systems intervening plants and microbes. On the other hand, the POD, activated upon pathogenic challenge, could convert common plant phenolics into bioactive compounds which interfered with pathogen and insect attacks. This idea prompted us to examine the possibility. Guaiacol and other phenols were chosen as model substrates for the enzyme reaction system, and several products with novel structures were confirmed to have unique biological activities. This paper deals with the chemical structures of a novel radical scavenger (ST-1) and the POD products with anti-microbial activities and phytotoxicities.
  • 47 THE ROLE OF PEROXIDASE IN THE CHEMICAL DEFENCE OF PLANTS : A NOVEL PEROXIDASE INHIBITOR AND A NEW TYPE OF BIOACTIVE PHENOLICS PRODUCED BY PEROXIDASE, Kobayashi A, Koguchi Y, Kanzaki H, Kajiyama S, Kawazu K, Symposium on the Chemistry of Natural Products, symposium papers, 35, 0, 361, 368,   1993 , http://ci.nii.ac.jp/naid/130005977632
    Summary:Plants induce varied resistance-response toward microbial challenge. In order to establish an infection, pathogenic fungi must break such defense barriers as lignin, phytoalexins, and hydrolytic enzymes. At the early stage of infection, POD (peroxidase) activity is enhanced dramatically. POD and lignin accumulates at the infected site and then protects from further fungal invasion. The suppression of lignin deposition at the site might be of advantage to susceptible pathogenic fungi. We, therefore, conducted a comprehensive screening to obtain POD inhibitors from fungal cultures and a potent radical scavenger named ST-1 was isolated from a fungus, and its chemical structure was elucidated to be 1,3-dihydro-4,5,6-trihydroxy-1,3-dimethoxy-7-methylisobenzofuran by a combination of chemical modification and spectroscopic techniques. ST-1 effectively scavenged oxygen radicals and also suppressed the phytoalexin accumulation in kidney bean. These results suggested that such a mechanism can be enumerated as one of significant defense systems intervening plants and microbes. On the other hand, the POD, activated upon pathogenic challenge, could convert common plant phenolics into bioactive compounds which interfered with pathogen and insect attacks. This idea prompted us to examine the possibility. Guaiacol and other phenols were chosen as model substrates for the enzyme reaction system, and several products with novel structures were confirmed to have unique biological activities. This paper deals with the chemical structures of a novel radical scavenger (ST-1) and the POD products with anti-microbial activities and phytotoxicities.
  • 43 SYNTHESIS OF NEW INDOLACTAM ANALOGUES BY MICROBIAL CONVERSION AND THEIR IN VITRO BIOLOGICAL ACTIVITIES RELATED TO TUMOR PROMOTION, Irie K, Kajiyama S, Okuno S, Koshimizu K, Nishino H, Iwashima A, Hayashi H, Arai M, 天然有機化合物討論会講演要旨集, 33, 329, 336,   1991 09 07 , http://ci.nii.ac.jp/naid/110006679011
    Summary:Teleocidins are potent tumor promoters produced by actinomycetes. (-)-Indolactam-Val (ILV), which has the fundamental structure of teleocidins and is the minimum unit for tumor-promoting activity, is useful as a lead compound for structure-activity studies and receptor analysis. Hitherto, total syntheses of ILV and its analogues have been intensively investigated. However, more convenient syntheses are desirable from the view point of synthesizing various indolactam congeners for structure-activity studies. We have demonstrated that ILV was biosynthesized from L-Trp, L-Val and L-Met via N-Me-L-Val-L-Trp-ol using Streptoverticillium blastmyceticum NA34-17. Since N-Me-L-Val-L-Trp-ol can be chemically synthesized without difficulty, utilization of the microbial cyclization enzyme would be a convenient synthetic method of various indolactam analogues for structure-activity studies. We have metabolized twelve synthetic seco-compounds and obtained ten ILV congeners with L-Ala, Abu, γ,δ-Δ-Nva, Nva, Nle, tert-Leu, Leu, Ile, allo-Ile, Phg, instead of L-Val in ILV. These indolactam congeners were examined for two biological tests related to tumor promotion. They were binding ability to the 12-O-tetradecanoylphorbol-13-acetate receptor and stimulation of radioactive inorganic phosphate incorporation into phospholipids of HeLa cells. The results indicated that both the hydrophobicity and the bulkiness of the substituents at position 12 increased these activities, demonstrating the recent hypothesis that the isopropyl group at position 12 of ILV is involved in the hydrophobic interaction on the receptor site.
  • 43 SYNTHESIS OF NEW INDOLACTAM ANALOGUES BY MICROBIAL CONVERSION AND THEIR IN VITRO BIOLOGICAL ACTIVITIES RELATED TO TUMOR PROMOTION, Irie K, Kajiyama S, Okuno S, Koshimizu K, Nishino H, Iwashima A, Hayashi H, Arai M, Symposium on the Chemistry of Natural Products, symposium papers, 33, 0, 329, 336,   1991 , http://ci.nii.ac.jp/naid/130005977497
    Summary:Teleocidins are potent tumor promoters produced by actinomycetes. (-)-Indolactam-Val (ILV), which has the fundamental structure of teleocidins and is the minimum unit for tumor-promoting activity, is useful as a lead compound for structure-activity studies and receptor analysis. Hitherto, total syntheses of ILV and its analogues have been intensively investigated. However, more convenient syntheses are desirable from the view point of synthesizing various indolactam congeners for structure-activity studies. We have demonstrated that ILV was biosynthesized from L-Trp, L-Val and L-Met via N-Me-L-Val-L-Trp-ol using Streptoverticillium blastmyceticum NA34-17. Since N-Me-L-Val-L-Trp-ol can be chemically synthesized without difficulty, utilization of the microbial cyclization enzyme would be a convenient synthetic method of various indolactam analogues for structure-activity studies. We have metabolized twelve synthetic seco-compounds and obtained ten ILV congeners with L-Ala, Abu, γ,δ-Δ-Nva, Nva, Nle, tert-Leu, Leu, Ile, allo-Ile, Phg, instead of L-Val in ILV. These indolactam congeners were examined for two biological tests related to tumor promotion. They were binding ability to the 12-O-tetradecanoylphorbol-13-acetate receptor and stimulation of radioactive inorganic phosphate incorporation into phospholipids of HeLa cells. The results indicated that both the hydrophobicity and the bulkiness of the substituents at position 12 increased these activities, demonstrating the recent hypothesis that the isopropyl group at position 12 of ILV is involved in the hydrophobic interaction on the receptor site.
  • 41 STUDIES ON THE BIOSYNTHESIS OF INDOLE ALKALOID TUMOR PROMOTER TELEOCIDIN, Irie K, Kajiyama S, Funaki A, Koshimizu K, Hayashi H, Arai M, 天然有機化合物討論会講演要旨集, 31, 308, 315,   1989 09 17 , http://ci.nii.ac.jp/naid/110006678826
    Summary:Teleocidins produced by actinomycetes are potent tumor promoters and peculiar indole alkaloids containing a nine-membered lactam ring and a complex monoterpenoid moiety. Among the teleocidin-producing actinomycetes, Streptoverticillium blastmyceticum NA34-17, which we had found to produce the Epstein-Barr virus early antigen inducing indole alkaloids, has a characteristic feature of producing (-)-indolactam V (1), the common biosynthetic intermediate of teleocidins, in quantity. This characteristic would be advantageous to obtain a wide variety of biosynthetic intermediates of teleocidins. Using this microorganism, the possible biosynthetic pathway of teleocidins shown in Fig. 8 was proposed by feeding experiments with several D or ^<13>C-labelled precursors and isolation of new teleocidin-related metabolites named blastmycetin D (8) and E (9). Feeding experiments with several D or ^<13>C-labelled amino acids and possible biosynthetic precursor showed that (-)-indolactam V (1) was biosynthesized from L-tryptophan, L-valine and L-methionine via N-methyl-L-valyl-L-tryptophanol (3). From the isolation of blastmycetin D (8) and E (9), and feeding experiments with D-labelled L-methionine, we proposed the hypothesis that (-)-N^1-nerylindolactam V was a common biosynthetic intermediate of teleocidins, and that the complex monoterpenoid moieties of teleocidins, for example those of olivoretin A, C and E (see Fig. 8), arose by difference of the sequence of the methylation and the aza-Claisen rearrangement, and the position of the methylation.
  • 41 STUDIES ON THE BIOSYNTHESIS OF INDOLE ALKALOID TUMOR PROMOTER TELEOCIDIN, Irie K, Kajiyama S, Funaki A, Koshimizu K, Hayashi H, Arai M, Symposium on the Chemistry of Natural Products, symposium papers, 31, 0, 308, 315,   1989 , http://ci.nii.ac.jp/naid/130005977304
    Summary:Teleocidins produced by actinomycetes are potent tumor promoters and peculiar indole alkaloids containing a nine-membered lactam ring and a complex monoterpenoid moiety. Among the teleocidin-producing actinomycetes, Streptoverticillium blastmyceticum NA34-17, which we had found to produce the Epstein-Barr virus early antigen inducing indole alkaloids, has a characteristic feature of producing (-)-indolactam V (1), the common biosynthetic intermediate of teleocidins, in quantity. This characteristic would be advantageous to obtain a wide variety of biosynthetic intermediates of teleocidins. Using this microorganism, the possible biosynthetic pathway of teleocidins shown in Fig. 8 was proposed by feeding experiments with several D or ^<13>C-labelled precursors and isolation of new teleocidin-related metabolites named blastmycetin D (8) and E (9). Feeding experiments with several D or ^<13>C-labelled amino acids and possible biosynthetic precursor showed that (-)-indolactam V (1) was biosynthesized from L-tryptophan, L-valine and L-methionine via N-methyl-L-valyl-L-tryptophanol (3). From the isolation of blastmycetin D (8) and E (9), and feeding experiments with D-labelled L-methionine, we proposed the hypothesis that (-)-N^1-nerylindolactam V was a common biosynthetic intermediate of teleocidins, and that the complex monoterpenoid moieties of teleocidins, for example those of olivoretin A, C and E (see Fig. 8), arose by difference of the sequence of the methylation and the aza-Claisen rearrangement, and the position of the methylation.
  • Comprehensive analysis of wound induced metabolic change in citrus plants, Tomonori Asai, Tetsuya Matsukawa, Atsushi Ishihara, Shin'ichiro Kajiyama, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 248,   2014 08
  • DNA aptamers that bind to chitin, E Fukusaki, T Kato, H Maeda, N Kawazoe, Y Ito, A Okazawa, S Kajiyama, A Kobayashi, BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 10, 5, 423, 425,   2000 03 , 10.1016/S0960-894X(00)00013-5
    Summary:We have succeeded in the acquisition of DNA aptamers that recognize chitin using in vitro selection. The obtained DNA aptamers have the stem-loop or bulge loop structures with guanine rich loop clusters and the clockwise B-form stems. (C) 2000 Elsevier Science Ltd. All rights reserved.
  • An artificial plastic receptor that discriminates axial asymmetry.(共著), EI-ICHIRO FUKUSAKI, AKIRA SAIGO, SHIN-ICHIRO KAJIYAMA, AKIO KOBAYASHI, ┣DBJ. Biosci. Bioeng.(/)-┫DB, 90, 6, 665, 668,   2000 , 10.1263/jbb.90.665
  • A simple assay for formate dehydrogenase activity by gas chromatography-mass spectrometry, T Shiraishi, E Fukusaki, S Kajiyama, A Kobayashi, JOURNAL OF CHROMATOGRAPHY A, 855, 1, 337, 340,   1999 09 , 10.1016/S0021-9673(99)00653-6
    Summary:A new method using GC-MS was devised for the convenient measurement of formate dehydrogenase (FDH) activity in crude tissue samples. FDH activity was detected by measuring headspace (CO2)-C-13, which was enzymically converted from [C-13]formic acid. This method proved to be sensitive and simple for the estimation of FDH activity without complicated pretreatment. (C) 1999 Published by Elsevier Science B.V. All rights reserved.

Research Grants & Projects

  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(基盤研究(C)), Development of novel screening system for enzyme-displaying yeasts using calcium-alginate based beads., In this study, a novel screening method for enzyme-displaying yeasts were investigated. Enzyme-displayed yeast cells were captured in the micro-sized calcium alginate beads each by each using newly developed reverse micelle method to prevent diffusion of hydrolyzed fluorescent substrates. Adopting flow sorting to these captured cells, active cells were successfully enriched. This system would be a useful method for high speed screening of yeast cells which display various hydrolyzing enzymes with high activity.
  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(特定領域研究), Subcellular dissection and dynamic analysis of intracellular metabolites using non-linear photonics
  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(基盤研究(C)), Investigation on plant intercellular chemical network using laser micro sampling technique, Unlike animal organs controlled by the central nervous system, multicellular plants axe composed of highly independent cells in functional and in metabolic aspect. This means metabolite analysis of whole plant or tissue samples only gives the average information of individual cell functions. For example, upon the infection of pathogens, some plants induce antimicrobial metabolite, phytoalexin. However the way and degree of defense response might not be same among individual cells. In this study, cellular metabolite accumulation and transcription of biosynthetic genes, among individual plant cells are investigated by using the technique of angle cell sampling and the analysis technique i.e. laser assisted single cell sampling, laser micro spectroscopy, and Nano Flow LC-ESI-MS. Through this study, we developed a novel experimental approach for functional cell biology of plants.
  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(基盤研究(C)), Tracking of petal cell maturing process by single-cell based metabolic analysis, In this study, the petal cell maturing process of Torenia plant was investigated by using laser single-cell sampling and analysis technique. In the first year, sample preparation method and laser irradiation condition, and nano-flow LC-MS analysis condition of single-cell origin samples were investigated. Through this investigation, the facile analytical method for petal cell pigment was established. In second year, the quantitative analysis of mRNA concerning petal pigment biosynthesis was investigated. In particular, novel high sensitive quantitative mRNA analysis based on the ESI-MS was developed. In this method, the reporter molecule such as FAM (fluorescein phosphoramidite) was attached to the 5' end of the PCR primer and according to the PCR polymerizing reaction, the reporter molecule and the 5'end nucleotide was released. By quantify the released molecule using ESI-MS, the amount of targeted mRNA in the sample was estimated. The FAM attached nucleotide (FAM-A, FAM-T, FAM-G, FAM-C) was completely separated and quantified by LC-ESI-MS. Using this method, we are succeeded in the estimation of CHS and GADPH quantities in single petal cell of Torenia plant.
  • The Other Research Programs, Development of UV-laser micro processing for plant cells
  • The Other Research Programs, Structure and function of exocellular polysaccharide of cyanobacteria
  • The Other Research Programs, Studies on bioactive compounds of terrestrial blue-green algae