Seriniquinone was isolated as a melanoma-selective anti-cancer agent from a culture broth of the marine-derived bacterium Serinicoccus marinus CNJ927 in 2014. It targets the unique small protein, dermcidin, which affects the drug resistance of cancer cells. Due to its significant activity against cancer cells, particularly melanoma, and its unique target, seriniquinone has been developed as a new pharmacophore. However, it has the disadvantage of poor solubility in drug discovery research, which needs to be resolved. A new seriniquinone glycoside (1) was synthesized by the biological transformation of seriniquinone using the deep sea-derived bacterium Bacillus licheniformis KDM612. Compound 1 exhibited selective anti-cancer activity against melanoma, similar to seriniquinone, and was 50-fold more soluble in DMSO than seriniquinone.

Nectriatide 1a, a naturally occurring cyclic tetrapeptide, has been reported to a potentiator of amphotericin B (AmB) activity. In order to elucidate its structure-activity relationships, we synthesized nectriatide derivatives with different amino acids in solution-phase synthesis and evaluated AmB-potentiating activity against Candida albicans. Among them, C-and N-terminal protected linear peptides were found to show the most potent AmB-potentiating activity.

Habiterpenol is a G2 checkpoint inhibitor isolated from the culture broth of Phytohabitans sp. 3787_5. Here, we report the synthesis of new habiterpenol analogs through the total synthesis process of habiterpenol and evaluating the analogs for G2 checkpoint inhibitory activity. We investigated two different synthetic approaches for total synthesis, with intramolecular conjugate addition and Ti(III)-mediated radical cyclization as key reactions. Although the former was unsuccessful, the latter reaction facilitated stereoselective total synthesis and determination of the absolute configuration of habiterpenol. The extension of these chemistries to a structure-activity relationship (SAR) study gave new habiterpenol analogs, which could not be derived from natural habiterpenol and only be synthesized by applying the total synthesis. Therefore, this study provides important insights into SAR studies of habiterpenol.

A new compound, designated nectriatide (1), was isolated as a potentiator of amphotericin B (AmB) activity against Candida albicans from the culture broth of Nectriaceae sp. BF-0114. This structure was elucidated based on spectroscopic analyses (1D and 2D NMR data), chemical methods, and total synthesis. Compound 1 was a unique cyclotetrapeptide consisting of l-N-methyltyrosine, anthranilic acid, l-alanine, and l-valine. Compound 1 and several synthetic derivatives, including linear peptides, potentiated AmB activity against C. albicans by up to 16-fold (the MIC value of AmB decreased from 0.5 μg/mL to 0.031 μg/mL in combination with test compound).

New indanones, designated celludinones A ((±)-1) and B (2), were isolated from the culture broth of the fungal strain Talaromyces cellulolyticus BF-0307. The structures of celludinones were elucidated by spectroscopic data, including 1D and 2D NMR. Celludinone A was found to be a mixture of racemic isomers ((±)-1), which were isolated by a chiral column. Compounds (+)-1 and (-)-1 inhibited the sterol O-acyltransferase (SOAT) 1 and 2 isozymes in a cell-based assay using SOAT1- and SOAT2-expressing Chinese hamster ovary (CHO) cells, while 2 selectively inhibited the SOAT2 isozyme.

The eŠect of temperature treatment before thawing on the delay of discoloration of frozen skipjack Katsuwonus pelamis meat after thawing was investigated. The NAD hydrolyzing activities of skipjack meat at -6°C and -8°C were 3.66×10-3 mmol/(g min) and 2.78×10-3 mmol/(g min), respectively. After 24 h temperature treatment at -6°C and -8°C, NAD+ concentrations of the meat decreased by up to 3.8%and 5.2%, respectively. After 1-day storage at 5°C, pH of the untreated meat reached around 5.6 however, that of the samples treated at -6°C and -8°C was around 6.3. After 1-day storage at 5°C, metmyoglobin concentrations of meats treated at either temperature with vacuum packaging did not increase however, those of untreated meat and meats treated at the same temperatures with air packaging increased significantly.

A new cytotoxic agent designated isomethoxyneihumicin (1 and 2), a mixture of lactam-lactim tautomers, was isolated along with methoxyneihumicin (3) from the culture broth of the marine Nocardiopsis alba KM6-1. The structures of 1 and 2 were elucidated in spectroscopic analyses (1D and 2D NMR data, and ROESY correlations). Isomethoxyneihumicin (15.0 mu M) and 3 (15.0 mu M) arrested the cell cycle of Jurkat cells at the G2/M phase (66 and 67%) in 12 h. Isomethoxyneihumicin and 3 exhibited cytotoxicity against Jurkat cells with IC50 values of 6.98 and 30.5 mu M in 20 h, respectively. These results strongly suggest that isomethoxyneihumicin and 3 exhibit cytotoxicity against Jurkat cells by inhibiting the cell cycle at the G2/M phase.

The fungus, Aspergillzts nidulans BF0142, was isolated from hot spring-derived soil collected at Hell Valley in Noboribetsu, Hokkaido, Japan. A new furanone compound designated helvafuranone (1) was isolated along with microperfuranone (2), 9-hydroxymicroperfuranone (3), diorcinol (4), emestrin (5), and sterigmatocystin (6) from a culture broth of A. nidulans BF0142. The structure of 1 was elucidated as 5-hydroxy-4-(4-hydroxybenzyl)-3-(4hydroxybenzyl)furanone based on various NMR experiments and chemical modifications.

Eight new thiodiketopiperazines, designated as graphiumins A to H (1-8), were isolated along with bisdethiobis(methylthio)-deacetylaranotin (9) and bisdethiobis(methylthio)-deacetylapoaranotin (10) from the culture broth of the marine-derived fungus Graphium sp. OPMF00224. The structures of the graphiumins were elucidated based on spectroscopic analyses (1D and 2D NMR data, ROESY correlations and CD data) and chemical methods. The absolute configuration of the common (3S)-3-hydroxyoctanoyl acid residue in 1, 3 and 4 was determined by hydrolysis, benzoyl derivatization and HPLC analysis using a chiral column. Five graphiumins moderately inhibited yellow pigment production by methicillin-resistant Staphylococcus aureus.

Marine-derived Streptomyces sp. OPMA00072 was found to produce inhibitors of the synthesis of neutral lipids in a cell-based assay using Chinese hamster ovary (CHO) cells. A new 16-membered macrolide named bafilomycin L (BFL) (1) was isolated along with the known structurally related bafilomycin C-1 (BFC1) (3) from the culture broth of the actinomycete by solvent extraction, octadecylsilyl column chromatography and HPLC. BFL inhibited cholesteryl ester (CE) synthesis in CHO cells with an IC50 value of 0.83 nm and also in mouse peritoneal macrophages with an IC50 of 6.1 nm. In addition, BFL blocked cellular acidification in He La cells by interfering with vacuolar H+-ATPase (V-ATPase) as well as other bafilomycins. These data strongly suggest that BFL disturbed the lysosome function to block cholesterol metabolism, leading to the inhibition of CE accumulation in mammalian cells.

Two new thiodiketopiperazines (TDKPs), designated graphiumins I (1) and J (2), were isolated from the culture broth of the marine-derived fungus Graphium sp. OPMF00224 by solvent extraction, silica gel column chromatography, and HPLC. Their absolute structures were elucidated by spectroscopic analyses (1D and 2D NMR data, ROESY correlations, and CD data) and chemical methods. They were found to be structurally rare TDKPs with a phenylalanine-derived indolin substructure. Compounds 1 and 2 inhibited yellow pigment production by methicillin-resistant Staphylococcus aureus (MRSA) with IC50 values of 63.5 and 76.5 µg/ml, respectively, without inhibiting its growth, even at 250 µg/ml.

Natural products continue to provide vital treatment options for cancer. Although their translation into chemotherapeutics is complex, collaborative programs continue to deliver productive pipelines for cancer chemotherapy. A new natural product, seriniquinone, isolated from a marine bacterium of the genus Serinicoccus, demonstrated potent activity over a select set of tumor cell lines with particular selectivity toward melanoma cell lines. Upon entering the cell, its journey began by localization into the endoplasmic reticulum. Within 3 h, cells treated with seriniquinone underwent cell death marked by activation of autophagocytosis and gradually terminated through a caspase-9 apoptotic pathway. Using an immunoaffinity approach followed by multipoint validation, we identified the target of seriniquinone as the small protein, dermcidin. Combined, these findings revealed a small molecule motif in parallel with its therapeutic target, whose potential in cancer therapy may be significant. This discovery defines a new pharmacophore that displayed selective activity toward a distinct set of cell lines, predominantly melanoma, within the NCI 60 panel. This selectivity, along with the ease in medicinal chemical modification, provides a key opportunity to design and evaluate new treatments for those cancers that rely on dermcidin activity. Further, the use of dermcidin as a patient preselection biomarker may accelerate the development of more effective personalized treatments.

Citridone A (1), originally isolated as a potentiator of antifungal miconazole activity from a fungal culture broth, has a phenyl-R-furopyridone structure. Because of its unique ring structure, 11 derivatives were chemically synthesized and their biological activity was evaluated. Derivatives 17, 20 and 21 potentiated miconazole activity against Candida albicans. Furthermore, 1, 14, 20 and 21 were found to inhibit yellow pigment production in methicillin-resistant Staphylococcus aureus.

A new compound, designated epi-trichosetin (1), was isolated along with the known compound trichosetin (2) from the culture broth of Fusarium oxysporum FKI-4553 by solvent extraction, silica gel column chromatography and reversed-phase HPLC. The structure of 1 was elucidated by comparing various spectral data with those of 2, revealing that 1 was a stereoisomer of 2. Compounds 1 and 2 inhibited the undecaprenyl pyrophosphate synthase activity of Staphylococcus aureus with IC50 values of 83 and 30 mu M, respectively, and showed antimicrobial activity, particularly against Gram-positive bacteria, including methicillin-sensitive and -resistant S. aureus.

Cyanosporasides are marine bacterial natural products containing a chlorinated cyclopenta[a]indene core of suspected enediyne polyketide biosynthetic origin. Herein, we report the isolation and characterization of novel cyanosporasides C-F (3-6) from the marine actinomycetes Salinispora pacifica CNS-143 and Streptomyces sp. CNT-179, highlighted by the unprecedented C-2' N-acetylcysteamine functionalized hexose group of 6. Cloning, sequencing, and mutagenesis of homologous similar to 50 kb cyanosporaside biosynthetic gene clusters from both bacteria afforded the first genetic evidence supporting cyanosporaside's enediyne, and thereby p-benzyne biradical, biosynthetic origin and revealed the molecular basis for nitrile and glycosyl functionalization. This study provides new opportunities for bioengineering of enediyne derivatives and expands the structural diversity afforded by enediyne gene clusters.

Eight new dinapinones, AB1, AB2, AC1, AC2, AD1, AD2, AE1 and AE2, were isolated from the culture broth of Talaromyces pinophilus FKI-3864. The structures of these dinapinones were elucidated by various NMR experiments. All these dinapinones possessed the same biaryl dihydronaphthopyranone skeleton consisting of a heterodimer with one monapinone A and one different monapinone. Dinapinones AB1 and AB2, consisting of monapinones A and B, were atropisomers. Similarly, dinapinones AC1 and AC2, consisting of monapinones A and C, dinapinones AD1 and AD2, consisting of monapinones A and D, and dinapinones AE1 and AE2, consisting of monapinones A and E, were atropisomers. Dinapinone AB2 showed potent inhibition of triacylglycerol (TG) synthesis in intact mammalian cells with an IC50 value of 1.17 μM, whereas the other dinapinones showed weak inhibition of TG synthesis.

Cylindrol A 5, a new ascochlorin congener, was isolated along with 14 known compounds from the culture broth of Cylindrocarpon sp. FKI-4602 by solvent extraction, octadecylsilane column chromatography and HPLC. The structure of cylindrol A 5 was elucidated by spectral analyses, including NMR. The compound has an ascochlorin skeleton consisting of a resorcin aldehyde and a cyclohexanone moieties. Cylindrol A 5 showed moderate antimicrobial activity against Bacillus subtilis, Kocuria rhizophila, Mycobacterium smegmatis and Acholeplasma laidlawii. The biosynthetic pathway to cylindrol A 5 was deduced from the 14 isolated metabolites of the fungal strain. © 2013 Japan Antibiotics Research Association All rights reserved.

Racemates of the diphenolic metabolites (+/-)-tylopilusin A (1) and (+/-)-tylopilusin B (2) were isolated from the fruiting bodies of Tylopilus eximius. Their structures were elucidated on the basis of spectroscopic analyses (1D and 2D NMR data and ROESY correlations) and X-ray crystallography. Each racemate was separated into its individual enantiomers, and electronic circular dichroism calculations were used to assign the absolute configuration of (+)- and (-)-tylopilusin A (1) and (+)- and (-)-tylopilusin B (2).

Two new butenolides, designated trichocyalides A and B, were isolated along with the known compound harzianolide, from the culture broth of Trichoderma sp. FKI-5513 by solvent extraction, ODS column chromatography and HPLC. Their structures were elucidated by several spectral analyses, showing that they have the common skeleton of butenofuranone. Trichocyalides A and B inhibited alkaline phosphatase (ALP) activity, a typical marker enzyme of osteoblastic differentiation (IC50: 83.0 and 187 mu M, respectively), in bone morphogenetic protein (BMP)-stimulated C2C12 myoblasts mutant cells, which stably express BMP receptor activity, whereas harzianolide showed no inhibitory activity against ALP even at 500 mu M. The Journal of Antibiotics (2012) 65, 565-569; doi: 10.1038/ja.2012.70; published online 5 September 2012

Hepatitis C virus core protein (Core) contributes to HCV pathogenicity. Here, we demonstrate that Core impairs growth in budding yeast. We identify HSP90 inhibitors as compounds that reduce intracellular Core protein level and restore yeast growth. Our results suggest that HSC90 (Hsc82) may function in the protection of the nascent Core polypeptide against degradation in yeast and the C-terminal region of Core corresponding to the organelle-interaction domain was responsible for Hsc82-dependent stability. The yeast system may be utilized to select compounds that can direct the C-terminal region to reduce the stability of Core protein. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Most cancer cells have mutations in genes at the Cl checkpoint and repair DNA only in the 62 phase; therefore, the 62 checkpoint is a potential target to develop novel therapy. In the course of screening, a known compound, pycnidione, was isolated from the fungal culture broth of Gloeotinia sp. FKI-3416. Pycnidione irreversibly abrogated bleomycin-induced G2 arrest in Jurkat cells and synergically potentiated the cytotoxicity of bleomycin. To elucidate the mechanism of action, the effect of pycnidione on the signal transduction of the 62 checkpoint was analyzed, showing that the increased phospho-cyclin dependent kinase-1 (CDK1) level caused by bleomycin was abrogated in the presence of pycnidione, indicating that cells did not arrest at the 62 phase. Moreover, under these conditions, Chk1 and Chk2 levels were markedly down-regulated. Thus, we concluded that pycnidione abrogated bleomycin-induced 62 arrest by decreasing Chk1 and Chk2.

Staphyloxanthin, a yellow pigment produced by methicillin-resistant Staphylococcus aureus (M RSA), is a virulent factor escaping from the host immune system. A new screening method for inhibitors of staphyloxanthin production by MRSA was established using paper disks. By this screening method, inhibitors of staphyloxanthin production were selected from the natural product library (ca. 300) and from actinomycete culture broths (ca. 1000). From the natural product library, four known inhibitors of lipid metabolism, cerulenin, dihydrobisvertinol, xanthohumol and zaragozic acid, were found to inhibit staphyloxanthin production; however, typical antibiotics used clinically, including vancomycin, had no effect on staphyloxanthin production. From actinomycete culture broths, two known anthraquinones, 6-deoxy-8-O-methylrabelomycin and tetrangomycin, were found to inhibit staphyloxanthin production by MRSA in the paper disk assay. These results suggested that this screening method is useful and effective to find compounds targeting staphyloxanthin production, leading to a new type of chemotherapeutics against MRSA infection.

Three polyenylpyrone metabolites, pyridinopyrones A to C (1-3), have been isolated from the culture broth of a marine-derived Streptomyces sp., strain CNQ-301. The structures of the pyridinopyrones were assigned on the basis of chemical modification and combined spectroscopic methods, focusing on interpretation of 1D and 2D NMR data. Pyridinopyrones B and C (2, 3), examined as an inseparable mixture of methyl positional isomers, were ultimately defined by hydrogenation and NMR analysis of a saturated derivative. The biosynthesis of these metabolites was defined by the incorporation of stable isotope-labeled precursors, revealing that the biosynthetic starter unit is nicotinic acid, while the polyene chain and pendant methyl groups are acetate- and methionine-derived, respectively.

The first total synthesis of citridone A has been achieved through regioselective intramolecular iodocyclization and regio- and stereoselective Pd(0)-catalyzed coupling as key reactions.

Two new aromatic alkaloids, designated citrinamides A and B, were isolated from the culture broth of Penicillium sp. FKI-1938 by solvent extraction, silica gel column chromatography and HPLC. Their structures were elucidated by spectroscopic analysis, including NMR and amino acid analysis. Citrinamides A and B showed moderate potentiation of miconazole activity against Candida albicans.

The first, concise total synthesis of (+/-)-tensyuic acids B, C, and E, using chemoselective formal S(N)2' type Grignard reactions and selective esterification, is described. In addition, the optical purity of natural (+/-)tensyuic acid B was determined using Chirabite-AR. Synthetic tensyuic acids, together with their intermediate compounds, were found to possess useful bioactive properties, with some of them showing potent activity against Trypanosorna brucei brucei strain GUTat 3.1. (c) 2008 Elsevier Ltd. All rights reserved.

Total synthesis of a fungal cyclic peptide, malformin C, recently rediscovered as a G2 checkpoint inhibitor was completed. Our synthesis involved a convergent approach with respect to a linear pentapeptide, cyclization, and oxidative disulfide formation.

Six new alkylitaconic acids, designated tensyuic acids A to F, were isolated from the culture broth of Aspergillus niger FKI-2342 by solvent extraction, silica gel column chromatography and HPLC. Their structures were elucidated by spectroscopic analysis including UV, NMR, and MS. They are all alkylitaconic acid derivatives. Only tesyuic acid C showed moderate antimicrobial activity against Bacillus subtilis.

A DNA-damaging agent, bleomycin, arrests the cell cycle at the G2 phase of Jurkat cells, which are defective in the G1 checkpoint, while microtubule-disrupting colchicine arrests it at M phase. Fungal cyclopeptides, malformin A 1 and malformin C, were found to abrogate bleomycin-induced G2 arrest (IC50; 0.48 mu M and 0.9 nM, respectively), resulting in a drastic decrease in cells in G2 phase and increase in cells in subG1 phase. On the other hand, malformins showed little effect on the colchicine-induced M phase arrest in Jurkat cells (IC50; 2.7 mu M and 24 nM, respectively). Malformin C (0.026 mu M) also abrogated bleomycin-induced G2 arrest in colon cancer-derived HCT-116 cells. These data strongly suggest that malformin C disrupted the cell cycle at the G2 checkpoint of cancer cells, leading to sensitization of the cancer cells to the anti-cancer reagent.

Citridone D was isolated from the culture broth of Penicillium sp. FKI-1938 by solvent extraction, silica gel column chromatography and HPLC. The structure of citridone D was elucidated by spectroscopic analysis including NMR analysis. Citridone D was found to have a novel phenylfuropyridine skeleton different from those of other citridones. Citridone D potentiated miconazole activity against Candida albicans.

On the basis of a new concept of anti-infective drugs, assay systems were conducted to screen for compounds which potentiate the antifungal activity of an azole compound (miconazole) or the anti-MRSA activity of a β-lactam (imipenem), as a potential approach for the development of anti-infective drugs. Using the conventional assay system, several new compounds, such as beauvericins and phenatic acids were discovered. During continuous screening work, a culture broth of Penicillium sp. FKI-1938, isolated from soil collected at Ishigakijima, was recently selected. Five structurally-related new compounds named citridones were isolated from the culture broth (5L) through EtOAc extract, silicagel column chromatography and preparative HPLC. Finally, citridone A (4.7mg), a mixture of citridones B and B' (59.2mg, purified as a mixture of hemiacetal epimers) and citridone C (11.7mg) were obtained as white needles or amorphous. From another jar fermentation broth (20L), citridone D (16.4mg) was obtained as amorphous. The molecular formulas of citridones B and B' were determined to be C_<19>H_<21>NO_5 on the basis of HRFAB-MS measurement. The structures were elucidated by spectroscopic analyses including various NMR experiments and X-ray crystallography of a tri-O-acetyl derivative prepared from citridone B. Citridones B and B' have a novel cyclic ring system of 7-phenylfuropyridin-2-ol fused with a furan ring. All citridones showed growth inhibition of C. albicans KF-1, but only in combination with miconazole (0.06μM) by the paper disk method. Furthermore, only citridone A potentiated β-lactam imipenem activity against MRSA by decreasing the MIC value from 100μM to 1.6μM.

Two new furopyrrols, designated tensidols A and B, were isolated from the culture broth of Aspergillus niger FKI-2342 by solvent extraction, silica gel column chromatography and HPLC. Their structures were elucidated and shown to have the common skeleton of 6-benzyl-6H-furo[2,3-b]pyrrole. Tensidols A and B potentiated miconazole activity against Candida albicans. Tensidols also showed moderate antimicrobial activity only against Pyricularia oryzae.

New phenylfuropyridinones and related compounds, designated citridones A, B, B' and C, were isolated along with known CJ-16,173, from the culture broth of Penicillium sp. FKI-1938 by solvent extraction, silica gel column chromatography and HPLC. Citridones (75 μ M) potentiate the miconazole activity against Candida albicans, decreasing the IC50 value of miconazole from 14.5 nM to 3.5&SIM; 6.3 nM.

The structures of citridones A, B, B' and C, new potentiators of miconazole activity against Candida albicans produced by Penicillium sp. FKI-1938, were elucidated by various spectroscopic analyses including UV, NMR, and MS and degradation experiments. Although citridones B and B' were isolated as a mixture, each structure was also elucidated, indicating that they exist in equilibrium of hemiacetal epimerization. Citridones A, B and B' have a similar phenylfuropyridone moiety.

Two new phenols, designated phenatic acids A and B, were isolated along with known actiphenol, from the culture of Streptomyces sp. K03-0132 by solvent extraction, silica gel column chromatography and HPLC. Their structures were elucidated by spectroscopic analyses including mainly various NMR experiments. They have a common 1-hydroxy 2, 4-dimethyl benzene ring. These compounds potentiate miconazole activity against Candida albicans. Phenatic acid B also showed moderate antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, Bacteroides fragilis and Acholeplasma laidlawii.

Three new beauvericins, designated beauvericins D, E and F, were isolated along with known beauvericin and beauvericin A, from the culture of Beauveria sp. FKI-1366 by solvent extraction, ODS column chromatography and HPLC. These compounds potentiate miconazole activity against not only wild Candida albicans but also fluconazole resistant C. albicans. Beauvericins D and E decreased the IC50 value of miconazole against fluconazole resistant C. albicans from 1.3 mum to 0.25 and 0.31 mum, respectively.

The structures of beauvericins D, E and F, novel potentiators of miconazole activity against Candida albicans produced by Beauveria sp. FKI 1366, were elucidated by various spectroscopic analyses including UV NMR, and MS and degradation experiments. They have the common skeleton of the 18-membered cyclodepsipeptides.

Three new pentaene macrolides having a 28-membered ring, designated takanawaenes A, B and C, were isolated from the fermentation broth of Streptomyces sp. K99-5278 by solvent extraction, silica-gel column chromatography and HPLC. Takanawaenes showed antifungal activity against Aspergillus niger, Mucor racemosus, Candida albicans and Saccharomyces cerevisiae.

The structures of takanawacnes A, B and C, novel antifungal antibiotics produced by Streptomyces sp. K99-5278, were elucidated by various spectroscopic analyses including UV and NMR, and spectrometric analyses including MS. They have the common skeleton of a 28-membered pentaene macrolide.

A novel phenolic metabolite, ornatipolide, was isolated from the Boletus ornatipes fungus. Its structure was established by a combination of spectroscopic and chemical methods, and by an X-ray crystallographic analysis.

コロナ禍による制限実施もあり、予定していた計画より多少の遅延が認められる。 令和3年度では研究課題対象である抗スキルス胃癌活性を呈する深海・海洋性放線菌32株のうち4株に対して、活性物質の精製法確立および、生産菌ゲノム解析を行なった。4株が生産する活性候補分子は非タンパク質性の有機化合物であることから、C18固相抽出、酢酸エチルを用いた液液分画、C18系Preparative HPLCシステムを用い、ほぼ単一ピークになるまで精製するに至った。活性評価には弊学附属病院で樹立した培養細胞（正常線維芽細胞及びスキルス胃癌細胞株7株）を用い、精製各段階での活性の追跡、および比活性算出に用いた。同活性化合物群を対象とするOrbitrap-MSを用いた精密質量分析では、それらのm/zが500~800の範囲内に収まることから、分析対象とする4活性化合物は低中分子化合物であることが明らかとなった。生産菌ゲノム解析では、long read dataにはONT社MinIONシステム、short read dataにはIllumina社NovaSeqの2プラットフォームを採用し、得られた各データからopen source codeによるバイオインフォマティクス手法でwhole genome配列を構築することに成功した。決定した4株分のゲノム配列はそのゲノムサイズが6Gb~11Gbと幅広く、コードされている16S rRNA遺伝子を指標とした分析からはいずれも新種株に相当することが見出された。解析したゲノム配列は公共データベースであるDDBJに登録申請済みである（データ非公開）。