Proanthocyanidins (PAs) contribute to the quality factors and health benefits of several Rosaceae fruits. Ume, also known as Japanese apricot (Prunus mume Sieb. Et. Zucc), is one of the most popular fruit trees grown in Japan, and its processed products are employed as a folk remedy in eastern Asian countries. However, the information available regarding the PAs present in the fruit remains limited. In this study, we analyzed the PAs and phenolic acid derivatives present in the fruit of ‘Nnanko,’ the most common cultivar, and found that the flesh tissue of the mature fruit contained levels of PAs similar to those of the hydroxycinnamic acids. The concentration of PAs in the endocarp tissue of the mature fruit was approximately 14 times higher than that of the flesh tissue. We also estimated the chemical structures of the PAs from the flesh and the endocarp by HPLC-ESI-MS/MS. Although 13 kinds of PAs, including 2 monomers, 4 B-type dimers, 3 A-type dimers, 2 B-type trimers, 1 A-type trimer, and 1 B-type tetramer were detected in the flesh, only 2 monomers, 1 B-type dimer, and 2 A-type dimers were detected in the endocarp. All PAs detected in the endocarp were also present in the flesh.

Fruits are a major dietary source of phytochemicals with health benefits for humans. In particular, apples are an important source of dietary phytochemicals including procyanidins. Thus, in the present study, we investigated procyanidin concentrations in apples using normal-phase high-performance liquid chromatography with fluorescence detection and hydrophilic oxygen radical absorbance capacity (H-ORAC) values. Apple procyanidin concentrations were significantly correlated with H-ORAC values (r = 0.8284, P < 0.0001) in 30 varieties of apple cultivars (dessert, processing and crab apples). These data suggest that the anti-oxidative activity of apples is mainly due to procyanidins.

Several Japanese citrus fruits, Hassaku, Sweet spring, May pummelo, and Satsuma mandarin, were either peeled by vacuum infusion of a pectinase solution or by hand and then the sections were separated by hand. The citrus segments were then submerged in a cellulase solution to eliminate the segment membranes. There were no significant differences in the microflora, nutrient value (ascorbic acid content and beta-carotene equivalent), physiology (respiration and ethylene production rates), and physicochemical properties (texture, juice leakage, color index, and pH) between enzyme-peeled and hand-peeled segments of Hassaku, Sweet spring, and May pummelo. When enzyme-peeled Satsuma mandarin segments were compared with segments chemically peeled using HCl and NaOH solutions in preparation for a conventional canned product, no differences were found in quality, except that the microbial diversity and color index were less in enzyme-peeled segments, and the respiration rate was less in chemically peeled segments. Enzyme-peeled citrus segments were preferable to hand-or chemically peeled segments for their visual appearance. Enzyme-peeled Satsuma mandarin segments were stored in three types of package films with different O-2 permeability to estimate the shelf-life in a modified atmosphere package (MAP) at 10 degrees C. CO2 approached equilibrium concentrations of 3, 5, and 8% in films of OTR 7000, 1000 and 500 mL m(-2) d(-1) atm(-1), respectively, after 4 d of storage. Microbiological, nutritional, and physicochemical quality of the segments was maintained throughout 6 d of storage regardless of the film package, suggesting that the shelf-life of enzymatically peeled Satsuma mandarin segments as a fresh-cut produce is 6 d in a MAP at 10 degrees C.

Enzymatic peeling of fruit and vegetables is considered superior to conventional peeling methods in terms of minimizing product damage and indeed is expected to improve them. This technology was first proposed for the peeling of citrus fruit and numerous studies on its practical use have been conducted. However, its application to non-citrus fruit and vegetables remains limited. A novel process, including heat treatment prior to enzymatic treatment, is proposed to peel persimmon fruit. Heat treatment induces fine and even cracks on the fruit surface of certain persimmon cultivars, which act as channels via which enzyme solution can enter the peel tissue. Another function of the heat treatment is to inactivate polygalacturonase-inhibiting protein (PGIP), which is inherent in persimmon fruit. Inactivation of PGIP improves the efficiency of enzymatic peeling with exogenous polygalacturonase. Chemical pre-treatments with a food-grade emulsifier solution and a weak basic solution also enhance the permeability of enzyme solution into the peel tissue. These physical and chemical treatments are useful for applying enzymatic peeling to various persimmon cultivars. In future, enzymatic peeling technology is expected to be applied to various agricultural crops as the technology is further refined.

Limonoids in citrus fruits are known to possess multiple biological functions, such as anti-proliferative functions in human cancer cell lines. Therefore, we aimed to investigate the suppressive effect of limonin on intestinal polyp development in Apc-mutant Min mice. Five-week-old female Min mice were fed a basal diet or a diet containing 250 or 500 ppm limonin for 8 weeks. The total number of polyps in mice treated with 500 ppm limonin decreased to 74% of the untreated control value. Neoplastic cell proliferation in the polyp parts was assessed by counting PCNA positive cells, and a tendency of reduction was obtained by limonin treatment. Moreover, expression levels of c-Myc and MCP-1 mRNA in the polyp part were reduced by administration of limonin. We finally confirmed the effects of limonin on beta-catenin signaling, and found limonin significantly inhibited T-cell factor/lymphocyte enhancer factor-dependent transcriptional activity in a dose-dependent manner in the Caco-2 human colon cancer cell line. Our results suggest that limonin might be a candidate chemopreventive agent against intestinal carcinogenesis.

カンキツリモノイドであるリモニン，ノミリンの脂質代謝に与える影響を検討した。Sprague-Dawley系雄性ラットに高脂肪・高コレステロール食を給餌し，リモニンまたはノミリンを50mg/kgBW/day経口投与し，5週間飼育した。摂餌量，体重，肝臓および内臓脂肪重量，血糖値，血中脂質，肝臓中脂質について，ノミリン群でのLDL-Chol上昇を除いて，有意な差は認められなかった。ノミリン投与によりChrebp，Pparα，Fxr，Lxrαの発現量は有意に低下し，Srebflc，Abcalの発現量も低下傾向が示された。脂肪酸生合成（Acaca，Fasn，Scd1，Me1)，β酸化（Cpt1a），コレステロール代謝（Hmgcr，Ldlr，Srebf2，Acat2），胆汁酸代謝（Shp，Lrh-1，Hnf4a，Cyp7a1，Cyp27a1)，トリグリセリド合成（Dgat1，Dgat2）,リポタンパク会合（Mttp），脂質輸送（Abcg5，Abcg8，Abcb11，Cd36）に関連する有意な差は認められなかった。リモニン投与では，いずれの遺伝子発現にも差は認められなかった。

Freshly peeled "baby persimmons", 3-4 cm in diameter and 20-25 g in weight, were prepared by enzymatic peeling of 'Totsutanenashi' cherry persimmon fruit (Diospyros kaki Thunb.), which is a bud mutation and miniature of the full size 'Hiratanenashi' persimmon. The microbiological and visual quality of enzymatically peeled baby persimmons as a fresh-cut fruit was evaluated during storage in air or high CO2 atmospheres (10, 15 and 20%) at 10 degrees C. The flesh of intact baby persimmon fruit showed counts of 2.6 log CFU/g for mesophilic aerobic bacteria and a non-detectable level for coliform groups and fungi. A hot water dipping treatment in the enzymatic peeling process reduced both the counts of mesophiles to levels below the lower limit of detection (2.4 log CFU/g) and the diversity of bacterial flora. Coliforms and fungi remained undetectable throughout the enzymatic peeling process. During storage in air, counts of mesophiles, coliforms, lactic acid bacteria, and fungi of baby persimmons increased to 4-5.5 log CFU/g by day 6, while high CO2 atmospheres reduced the microbial growth, with the greatest reduction observed in 15 or 20% CO2. Diversity of the microflora also decreased in 20% CO2 atmosphere as compared to air on day 6. High CO2 atmospheres inhibited the development of brown discoloration in baby persimmons by day 4, but all samples developed browning to an unmarketable level by day 6. These results indicate that high CO2 atmospheres of 15 to 20% are desirable for enzyme-peeled baby persimmons to extend the shelf life when stored at 10 degrees C.

ウメ由来ポリフェノール抽出物(JAPE)を用いて，ラット小腸二糖類水解酵素への阻害活性ならびに食後血糖値上昇抑制作用を検討した。JAPEは，マルターゼ，グルコアミラーゼ，ラクターゼに対して阻害作用を示し，阻害様式はそれぞれ混合型非拮抗阻害，不拮抗阻害，拮抗阻害であった。スクラーゼ，イソマルターゼ，トレハラーゼに対しては阻害作用を示さなかった。ラットへの経口糖負荷試験では，マルトース，スクロース，デンプン投与時にJAPEによる有意な血糖値上昇抑制作用が認められ，グルコース投与時には，有意な影響は認められなかったことから，JAPEはグルコース吸収には影響せず，血糖値上昇抑制作用は二糖類水解酵素阻害によることが示唆された。AUC 0-120minは変化がなかったことから，JAPEはグルコース吸収量には影響しなかった。JAPEに含まれるヒドロキシ桂皮酸誘導体が二糖類水解酵素阻害にある程度関与していると考えられる。これらの知見より，JAPEは2型糖尿病や肥満などの生活習慣病のリスクを減らす食品素材として有用である可能性が示された。

A large amount of by-product is generated from Satsuma-mandarin (Citrus unshiu Marc.) juice processing, and Satsuma mandarin is a notable source of,beta-cryptoxanthin. Therefore, pulp produced during sieving and centrifugation processes contains large quantities of beta-cryptoxanthin. We developed a process to refine R-cryptoxanthin from raw pulp materials. First, the pulp was subjected to enzymatic degradation, and the resultant precipitate was recovered by centrifugation. The water contained in the precipitate was then replaced with acetone, and the acetone-substituted pulp was directly extracted with hexane. After removal of the hexane, the extract was sequentially separated to soluble and insoluble fractions with hexane, acetone, ethanol, and a hexane/ethanol (4:6 or 3:7) mixture at 30 degrees C. beta-Cryptoxanthin was concentrated in the hexane soluble, acetone soluble, ethanol insoluble, and hexane/ethanol-insoluble fractions. Using alkaline hydrolysis, the hexane/ethanol-insoluble fraction produced non-esterified,beta-cryptoxanthin crystals. In the case of the isolation of beta-cryptoxanthin from Satsuma-mandarin peel, after removal of essential oils in the hexane extract of dried peel under reduced pressure steam distillation, the ethanol-insoluble fraction during the sequential separation was hydrolyzed and subjected to a silica gel flash column chromatography to afford,beta-cryptoxanthin. The above procedure was also successfully applied to carotenoid-containing materials, and Persimmon peel produced a carotenoid mixture in high yield.

In the enzymatic peeling process of persimmon fruit, heat treatment prior to enzyme treatment is indispensable, as heat inactivation of polygalacturonase inhibiting protein (PGIP) greatly enhances the enzymatic degradation of pericarp tissue, and heat treatment produces cracks on the pericarp surface of certain cultivars. These cracks act as channels that introduce enzyme solution into the pericarp tissue. Since the frequency of crack formation affects the quality of peeled fruit, a combination of PGIP heat inactivation and mechanical perforation using metal needles, instead of heat-induced crack formation, has been proposed to improve the enzymatic peeling process of persimmon fruit. However, this process requires specially designed machinery to achieve uniform perforation. In the course of investigating the chemical components of the cuticle membrane of persimmon fruit, we found that the cuticle loses its stretching strength with the removal of wax, one of the main components of the cuticle membrane. Raw fruit treated with chloroform/methanol (1 : 1, v/v) to weaken the cuticle membrane before heat-treatment, showed increases in surface cracks and improved enzymatic peeling efficiency. A practical process applicable to food processing was achieved using a combination of chemical treatment in a detergent solution, for wax extraction, and heat treatment in a weak alkali solution, producing results similar to treatment with perforation. Using this new process, the entire enzymatic peeling process of persimmon fruit can be performed in solutions without mechanical perforation.

The fruit of mume, Japanese apricot (Prunus mume Sieb. et Zucc.), was evaluated for its phenolics content, high performance liquid chromatography (HPLC) pro- file and antioxidative activities. The phenolics content of mume fruit was relatively high, the flesh of fully matured fruit containing up to 1% of phenolics on a dry weight basis. Reflecting such a high content of phenolics, the ORAC (oxygen radical absorbance capacity) value for mume fruit flesh showed high values, ranging from 150 to 320 μmol/g Trolox equivalent, depending upon the stage of maturation. 5-O-Caffeoylqunic acid (chlorogenic acid), 3-O-caffeoylquinic acid and tetra-O-acylated sucrose-related compounds were isolated from the flesh of mume fruit, although many unknown peaks were also apparent in the HPLC chromatogram. An alkali hydrolysate comprised four main phenolic acids, caffeic acid, cis/trans-p-coumaric acid and ferulic acid. No flavonoids were observed in the analysis. These results suggest that the majority of phenolics in mume fruit were hydroxycinnamic acid derivatives.

The microbiological quality and shelf life of enzyme-peeled fresh-cut persimmon slices were evaluated during storage in a high CO2 controlled atmosphere (CA) and active modified atmosphere packaging (MAP) at 10 degrees C. Microbial counts of the enzyme-peeled slices were lower in high CO2 atmospheres (10%, 15%, and 20%) than in air during CA storage for 6 days at 10 degrees C with the 20% CO2 atmosphere being most effective. High CO2 atmospheres did not affect the number of bacterial and fungal species detected in the persimmon slices. The surface color, expressed as C* values, of the peeled side of enzyme-peeled slices was lower in high CO2 than in air after 6 days of CA storage. In contrast, C* values at the cut side were higher for slices stored in 20% CO2 than in air on Day 6. High CO2 atmospheres did not affect other quality of enzyme-peeled slices such as texture, pH, sugar content, and total ascorbic acid content. Based on the optimum 20% CO2 concentration in a CA, enzyme-peeled slices were stored in a MAP flushed with either air or 20% CO2 for 4 days at 10 degrees C. The CO2 concentration approached an equilibrium of either 5% or 10% after 3 days of storage in packages flushed with either air or 20% CO2, respectively, and the O-2 decreased to approximate to 10% in both packages. Adding 20% CO2 to the MAP was effective in reducing the growth of mesophiles and coliforms but not fungi in enzyme-peeled persimmon slices throughout 4 days of storage. The diversity of bacterial and fungal flora was partially similar between packages flushed with air and 20% CO2. Texture, pH, surface color, sugar content, and total ascorbic acid content of enzyme-peeled persimmon slices were unaffected by air or 20% CO2 as the flushing gas, except that C* values of the enzymatically peeled side on Day 4 were lower for slices flushed with 20% CO2 than air. A 20% CO2 atmosphere is recommended for reducing the microbial population of enzyme-peeled persimmon slices stored at 10 degrees C and the shelf life of persimmon slices in an active MAP with 20% CO2 is 4 days at 10 degrees C.

Fruit cuticular membranes (CMs) were isolated from 27 cultivars of Diospyros kaki Thunb., and relationships among their chemical compositions, densities and mechanical properties were analysed. Wide cultivar dependency was observed in both chemical compositions and mechanical properties, however, astringency did not show clear correlation with them. Densities of the CMs showed significant positive correlation with maximum stress and elastic modulus, but negative correlation with maximum strain. Positive correlations were also observed in elastic modulus-polysaccharide content and maximum strain-polysaccharide content, and negative correlation in elastic modulus-cutin content on a weight over unit area (mu g/cm(2)) basis. The integration of densities of fruit CMs and their chemical constituents may explain the mechanical properties which are specific to each cultivar of D. kaki fruit. (C) 2011 Elsevier Ltd. All rights reserved.

Enzymatic peeling of 'Fuyu' and 'Tone-wase' persimmon fruit was conducted for production of fresh-cut slices, and the microbiological and physicochemical quality of enzyme-peeled fresh-cut slices was compared with that of slices manually peeled with a knife. The enzymatic peeling process involved a porous treatment of the peel, heating at 100 degrees C for 45 s, infusion with 3% protopectinase at 37 degrees C for 3 h, and rinsing under running tap water. Initially, the peel of 'Fuyu' persimmon fruit had microbial counts ranging from 3.9 to 4.2 log cfu.g(-1) and a diverse microflora. The heating treatment before the enzymatic peeling process reduced the microbial counts in both the peel and flesh of all fruits to levels below the lower limit of detection. After the enzyme infusion followed by gentle rinsing with tap water, microbial counts of enzyme-peeled fruit were close to or below the level of detection. When microbial contamination of enzyme-peeled and knife-peeled 'Fuyu' and 'Tone-wase' persimmon slices was compared, the bacterial counts and diversity of bacterial and fungal flora were less in enzyme-peeled slices than in knife-peeled slices. With 'Tone-wase' slices, the color index, pH, and texture were unaffected by enzymatic peeling, except for surface lightness, which was lower in enzyme-peeled slices than in knife-peeled slices. These results indicate that enzymatic peeling could be an alternative to knife-peeling of 'Tone-wase' persimmon fruit for fresh-cut production from the point of microbiological and physicochemical quality.

Stone of Prunus mume (P. mume) is a by-product of pickled P. mume industry. Stones of native and pickled P. mume, mainly composed of holocellulose (83.8 +/- 1.8% and 65.1 +/- 0.3%, respectively) and acid-insoluble lignin (25.3 +/- 2.2% and 30.6 +/- 0.9%, respectively), were autohydrolyzed by microwave heating to extract polysaccharides and phenolic compounds. By heating at 200 to 230 degrees C, 48.0% to 60.8% of polysaccharide and 84.1% to 97.9% of phenolic compound were extracted in water along with partial degradation of hemicelluloses and lignin. The extracted liquors showed antioxidant activity against hydroxyl radical and DPPH radical originated from phenolic compounds. The pickled P. mume stone showed higher autohydrolyzability and microwave absorption capacity than the native stone due to absorbed salts and acids during pickling in fruit juice of P. mume with external addition of sodium chloride. Pickling process in salty and weak acidic juice seemed to be a kind of pretreatment for softening the stones prior to autohydrolysis induced by microwave heating.

Okara, a soybean by-product of tofu production, was treated with microwave irradiation in water for solubilization of its components. The solubilization rate increased with increasing temperature and reached higher than 70% around 200 degrees C with a heating time of 7 min. The main solubilized components were neutral carbohydrates consisting of arabinose and galactose, while the residues were mainly composed of cellulose. LV-SEM images confirmed solubilization of cell wall components. Production of new polyphenolic compounds that have antioxidant activity was observed above 180 degrees C. The heating temperature and heating time were optimized for neutral carbohydrate solubilization using the response surface methodology. The optimized condition was heating at 196 degrees C for 2 min, indicating that a short heating time is effective for solubilization of carbohydrates to prevent secondary decomposition.

和歌山特別栽培農産物に認証された青果物を対象に、残留農薬量と微生物数を迅速検出し、それらを基にした情報を加えたトレーサビリティシステムの構築を試みた。HPを作成し、これらの情報がWebサイトで見られるように、デモとして公開した。

市販野菜のバクテリアルフローラを調査した。キュウリ,レタス,ホウレンソウおよびニンジンの一般生菌数と大腸菌群数は,それぞれ4.5〜6.5logCFU/gと3.3〜4.3logCFU/gであるのに対して,トマト,タマネギおよびニンニクの細菌数は検出限界値(2.4logCFU/g)以下であった。キュウリ,レタスおよびニンジンでは,外部組織のほうが内部組織よりも対数値で1〜3程度高い細菌数を示したが,ホウレンソウでは両組織間に差は見られなかった。ニンジンとホウレンソウでは,グラム陰性菌が主体をなしたのに対して,キュウリではグラム陽性菌とグラム陰性菌がほぼ同程度検出された。ニンジン,ホウレンソウおよびキュウリから頻繁に検出された細菌は,グラム陽性菌ではCurtobacterium属菌,Staphylococcus属菌およびEnterococcus属菌で,グラム陰性菌ではPseudomonas属菌,Agrobacterium属菌,Stenotrophomonas属菌,Enterobacter属菌,Pantoea属菌およびKlebsiella属菌であった。なお,いずれの野菜からも食性病原細菌は検出されなかった。

現在わが国で栽培されているカンキツ類の果実8種類を対象にリモノイド配糖体の分析を行った。また, ウンシュウミカンを対象に栽培時の環境因子として日照をとりあげ, 着果位置の違いによる日照条件の差が果実のリモニン配糖体 (LG) 含量に及ぼす影響について検討した。
分析した8種類のカンキツ類果実のリモノイド配糖体濃度は70～260ppmの範囲に分布し, すでに報告されているウンシュウミカン以外にイヨカン, スイートスプリングおよびキヨミなどがリモノイド配糖体含量の多い果実であることが明らかとなった。
キヨミ, シラヌイ, スイートスプリングなどはウンシュウミカンの交雑種であるにもかかわらず, ウンシュウミカンとは大きく異なり, 総リモノイド配糖体含量に占めるLGの割合が小さいという特徴を有していた。早生品種のウンシュウミカンの場合, LGは8月中旬より果実組織に検出されはじめ, その後急速に増加し, 収穫期である11月初旬には100ppm前後に達した。樹冠外側から採取した果実は, 樹冠内側から採取したものに比べて, 常に高いリモノイド配糖体含量を示し, 日照条件を高める栽培方法をとることで, リモノイド配糖体含量を高めた果実を生産できる可能性が示唆された。

合成したフェルラ酸誘導体31種類について, 細菌, カビ, 酵母に対する抗菌活性を評価した。供試した化合物のうち, 12種類の化合物がグラム陽性細菌, カビおよび酵母の増殖を抑制することを見い出した。それらのうちMICの測定に供した化合物のほとんどがフェルラ酸より抗菌力が向上した。フェルラ酸ブチルとフェルラ酸-2-メチル-1-ブチルは, B. subtilis, S.aureus, S. cerevisiaeに対して顕著な増殖抑制を示した。フェルラ酸ヘキシルは, B. subtilis, S.aureusの増殖を顕著に抑制したが, カビ, 酵母に対してはその増殖に影響を与えなかった。一方, フェルラ酸メチル, フェルラ酸エチルはグラム陰性菌を除くいずれの供試菌に対しても増殖抑制作用を有し広い抗菌スペクトルを示した。フェルラ酸エステル類の抗菌活性は, アルキル基鎖の伸長に伴いグラム陽性細菌に対する抗菌力が向上し, 同時に細胞毒性も増すという傾向が認められた。フェルラ酸メチルおよびフェルラ酸-2-メチルー1-ブチルについては, 市販の食品保存料であるソルビン酸や安息香酸と同程度またはそれ以上の抗菌活性を有し, 細胞毒性試験でも100μMの濃度では, マウス由来線維芽細胞株Balb/c3T3A31-1-1の増殖にほとんど影響を与えず, 新たな食品用抗菌剤として検討できるレベルにあることが示唆された。

Novel ferulic acid derivatives in which feruloyl groups were attached to the hydroxyl groups of myo-inositol 1,3,5-orthoformate derivatives were synthesized. These feruloyl-myo-inositols suppressed the cyclooxygenase-2 (COX-2) promoter activity in a concentration-dependent manner. Among the examined compounds, compound 4 showed the highest activity. A treatment with 100 muM of compound 9 for 24 h resulted in a 50% decrease of COX-2 promoter activity without marked cytotoxicity. Both the molecular structure in which two ferulic acid moieties are facing each other and the molecular hydrophobicity may be essential for the suppression of COX-2 promoter activity. (C) 2002 Elsevier Science Ltd. All rights reserved.

An efficient method of hydrolyzing hesperidin to prepare hesperelin 7-glucoside (Hesperetin monoglucoside; HMG) was established. Hesperidin was first converted from its flavanone form to the chalcone form under alkaline conditions, and then hydrolyzed with HCl in the presence of a water-soluble solvent such as isopropyl alcohol (IPA), n-propyl alcohol, ethyl alcohol (EtOH) and methyl alcohol. When the reaction mixture containing optimal concentrations of 6% hesperidin, 20% IPA and 1.5 N HCl was heated for 15 min in a boiling water bath, the maximum amount of HMG were accumulated. HMG can be precipitated and efficiently recovered by dilution and evaporation to lower the IPA concentration. The solubility of crude HMG against distilled water, hot water and EtOH was 42.5, 12.5 and 40.7 times higher than that of reagent grade hesperidin, respectively.

In a study on improving utilization of waste From processing of Satsuma mandarin (Citrus unshiu Marcov.) the limonoid glucosides of fruit, juice, and by-products were measured using HPLC and TLC. All materials had the 17-beta-D-glucopyranoside derivatives of limonoids reported in other commercial citrus fruit. Citrus molasses was a good source for industrial scale extraction of limonoid glucosides. An extraction system using polystyrene divinylbenzene resins, was developed which could be expanded to industrial scale.

In the course of the development of an industrial recovering system of limonoids from citrus processing waste, an efficient method to recover seeds from the citrus processing waste (peel residue) was developed. The residue was first passed on a vibrating sieve (phi 14.5 x 14.5 mm) to remove peel fragments and other contaminants, and then the remaining material was degraded either chemically or enzymatically. The degraded fraction was sieved with mesh screen (phi 4.5 x 50 mm) and 85 similar to 90% of the seeds was recovered in optimal condition. The limonoid content in the seeds was not affected when the first fraction was treated with acid or enzyme. However, the content decreased when it was treated with alkali.

Limonoids and their glucosides in the seeds and barks of Phellondendron amurense (Kihada) were analyzed. The seeds contained limonin (1950 ppm), obakunone (20 ppm), limonin 17-beta-D-glucopyranoside (820 ppm) and obakunnone 17-beta-D-glucopyranoside (1360 ppm). The barks contained limonin (6760 ppm), obakunone (1240 ppm) and nomilin (270 ppm).

Limonoids and their glucosides in the seeds and barks of Phellondendron amurense (Kihada) were analyzed. The seeds contained limonin (1950 ppm), obakunone (20 ppm), limonin 17-β-D-glucopyranoside (820 ppm) and obakunone 17-β-D-glucopyranoside (1360 ppm). The barks contained limonin (6760 ppm), obakunone (1240 ppm) and nomilin (270 ppm).

Limonoids and their glucosides in seeds of sour orange (Citrus aurantium) were measured. The seeds contained neutral limonoids such as ichangin (130 ppm), deacetylnomilin (840 ppm), limonin (2470 ppm), nomilin (178 ppm), and obacunone (32 ppm). They also contained acidic limonoids such as deacetylnomilinic acid (120 ppm), isolimonic acid (110 ppm), and nomilinic acid (20 ppm). Unlike most other species of citrus tested, this species contained very high concentrations of deacetylnomilin and high concentrations of ichanging, deacetylnomilinic acid, and isolimonic acid. The results of thin-layer chromatography suggested that 19-hydroxydeacetylnomilinic acid aglycone was present, which was evidence for the previous suggestion that sour oranges have a biosynthetic pathway for limonoids via 19-hydroxydeacetylnomilinic acid. Sour orange seeds contained very high concentrations of the 17-beta-D-glucopyranosides of deacetylnomilin and deacetylnomilinic acid, accounting for 52.5% and 19.0% of the total limonoid glucosides (7300 ppm) of the seeds, respectively. However, the seeds contained lower concentrations (360 ppm) of nomilin glucoside, which is the most abundant limonoid glucoside in most other citrus seeds tested.

Supercritical carbon dioxide (SC-CO2) extraction of limonoids and limonoid glucosides (LGs) from defatted Natsudaidai seed meals was examined under various conditions. The total amount limonoids extracted by SC-CO2 was less than those extracted by organic solvent. Very little amount of limonin, deacetylnomilin and LGs were extracted by SC-CO2. The results obtained with SC-CO2 extraction with modification of entrainers indicated that acetone was the best entrainers among tested for extracting limonoids. However 70% ethanol was more efficient for LGs. Extraction pressure and the amount of CO2 were important factors in this experiment. The extraction yield of limonids depended upon pressure, but were not affected by temperature. The yield increased with increasing pressure and sharp increase in nomilin extraction was observed between 250 kg/cm2 and 300 kg/cm2. The extraction of deacetylnomilin, nomilin and obacunone was sharply increaed till the 50 g-CO2/g-seed level and remained relatively constant thereafter. The yield of limonin gradually increased with increasing amount of CO2. The optimal extraction ratios of deacetylnomilin, limonin, nomilin and obacunone were 13.9%, 19.1%, 82.1% and 38.9%, respectively. On the other hand, a higher yield (81.0%-99.4%) of LGs was obtained, except for limonin glucoside.

Neutral and acidic limonoids in seeds of Karatachi (Poncirus trifoliata) were quantitatively analyzed. The seeds contained ichangin (20 ppm), deacetylnomilin (230 ppm) and limonin (390 ppm). They also contained acidic limonoids, such as deacetylnomilinic acid (2 ppm), isolimonic acid (2 ppm) and isoobacunoic acid (a trace amount). The concentration of total limonoids was quite low as compared to that of citrus seeds.

Limonoid caused bitterness in citrus juices is a major problem in the citrus industry. Recently limonoids were found to possess anticarcinogenic and insecticidal activities. In this study, we developed an effective method for extracting neutral limonoids from Natsudaidai seeds. Extraction of neutral limonoids from Natsudaidai seeds depended upon the solvents used, time of extraction and methods of sample preparation prior to extraction. Solvent extraction methods were less effective than the enzyme method. This clearly showed that seeds contain two forms of limonoids, dilactones and monolactones. The enzyme methods was the most effective for extracting neutral limonoids from Natsudaidai seeds. This method extracted both dilactones and monolactones, whereas the solvent methods extracted mainly dilactones. Seeds were ground in O.1M Tris buffer at pH 7.0 and incubated for least 20 hr at room temperature. The mixture was then acidified and extracted with dichloromethane.

Ichangensin 17-beta-D-glucopyranoside, a new limonoid glucoside, was isolated from seeds of Yuzu (Citrus junos). This glucoside was also present in seeds of Sudachi (Citrus sudachi) and Kabosu (Citrus sphaerocarpa). The 17-beta-D-glucopyranosides of deacetylnomilin, nomilin, nomilinic acid, obacunone, limonin and deacetylnomilinic acid were also present in seeds of these species of citrus. Like Sudachi and Yuzu, Kabosu also contained ichangensin aglycone. The presence of ichangensin and ichangensin glucoside supports the previous suggestions that these species are related to Ichang Chie (Citrus ichangensis).

The amounts of limonoid glucosides were measured in the seeds of grapefruit (Citrus paradisi), lemon (C. limon), Valencia orange (C. sinensis), tangerine (C. reticulata), Fukuhara (C. sinensis Osbeck Hort.), Hyuganatsu (C. tamurana Hort. ex Tanaka), Shimamikan (C. kinokuni Hort. ex Tanaka) and Sanbokan (C. sulcata Hort. ex Takahashi). All the seeds contained 17-beta-D-glucopyranosides of limonin, nomilin, obacunone, deacetylnomilin, nomilinic acid and deacetylnomilinic acid. The total limonoid glucoside content ranged from 0.31 to 0.87% of the dry weight. The concentration of nomilin glucoside was highest among the glucosides found in the seeds. All the seeds also contained the major neutral limonoid aglycones commonly found in citrus, namely limonin, nomilin, deacetylnomilin, obacunone and ichangin. As previously reported, limonin was the predominant aglycone.

Limonin 17-β-D-glucopyranoside, total limonoid glucosides and limonin in the juice, pericarp and endocarp of hassaku (C. hassaku Hort. ex Tanaka) were determined by HPLC and TLC. Hassaku juice contained 314 ppm of total limonoid glucosides, whereas pericarp and endocarp tissues contained 2300 ppm and 3100 ppm, respectively, on dry basis. Limonin 17-β-D-glucopyranoside was the predominant glucoside derivative in the juice (148ppm, 47% of total limonoid glucosides). However, nomilin 17-β-D-glucopyranoside was the predominant glucoside in the pericarp and endocarp tissues. In addition to limonin and nomilin glucosides, the fruit tissues also contained the glucoside derivatives of typical citrus limonoids, such as nomilinic acid, deacetylnomilinic acid and obacunone.

We found a novel protopectinase that did not catalyze the degradation of polygalacturonic acid in the culture filtrate of Bacillus subtilis IFO 12113. The enzyme was purified by CM-Sephadex C-50 column chromatography, heat treatment at 60℃, and column chromatography on DEAE-Toyopearl 650M and Toyopearl HW-55S. The enzyme was isolated as a homogeneous protein, and was finally isolated as crystals. The molecular weight of the enzyme was estimated to be 30,000 by gel permeation chromatography and by SDS-polyacrylamide gel electrophoresis, and 29,800 by sedimentation analysis. Its isoelectric p...

黒酵母Aureobasidium pullulans由来のβ-グルカンを用いて、p-クマル酸とその類縁体を難水溶性化合物モデルとして、その可溶化とその安定性を検討した。さらに動物試験でのその吸収評価法を検討した。また、β-グルカンの生産性向上についても検討した。その結果、変異株の取得とβ-グルカンの高生産かつ高純度回収法を確立した。アルカリ変性させた変性β-グルカンを用いた場合にｐ-クマル酸の溶解度は約1.5倍に、特にカフェ酸を用いた場合は3倍となった。一方、変性β-グルカンを用いたp-クマル酸分散液を用いた動物経口試験において、その吸収性への阻害影響は見られないことを確認した。