研究者詳細 - 二神　泰基

Enkang Zhu, Kentaro Hiramatsu, Taiga Inoue, Kazuki Mori, Kosuke Tashiro, Kiyotaka Fujita, Takefumi Karashima, Hideharu Takashita, Kayu Okutsu, Yumiko Yoshizaki, Kazunori Takamine, Hisanori Tamaki, Taiki Futagami . Deficiency of β-xylosidase activity in Aspergillus luchuensis mut. kawachii IFO 4308. . Bioscience, biotechnology, and biochemistry 2024年4月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）

In this study, we investigated a deleterious mutation in the β-xylosidase gene, xylA (AkxylA), in Aspergillus luchuensis mut. kawachii IFO 4308 by constructing an AkxylA disruptant and complementation strains of AkxylA and xylA derived from A. luchuensis RIB2604 (AlxylA), which does not harbor the mutation in xylA. Only the AlxylA complementation strain exhibited significantly higher growth and substantial β-xylosidase activity in medium containing xylan, accompanied by an increase in XylA expression. This resulted in lower xylobiose and higher xylose concentrations in the mash of barley shochu. These findings suggest that the mutation in xylA affects xylose level during the fermentation process. Because the mutation in xylA was identified not only in the genome of strain IFO 4308 but also the genomes of other industrial strains of A. luchuensis and A. luchuensis mut. kawachii, these findings enhance our understanding of the genetic factors that affect the fermentation characteristics.

DOI： 10.1093/bbb/zbae046

Atsushi Nishitani, Kentaro Hiramatsu, Chihiro Kadooka, Kyoka Hiroshima, Kazutaka Sawada, Kayu Okutsu, Yumiko Yoshizaki, Kazunori Takamine, Masatoshi Goto, Hisanori Tamaki, Taiki Futagami . Overexpression of the DHA1 family, ChlH and ChlK, leads to enhanced dicarboxylic acids production in koji fungi, Aspergillus luchuensis mut. kawachii and Aspergillus oryzae. . Journal of bioscience and bioengineering 2024年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

The white koji fungus Aspergillus luchuensis mut. kawachii secretes substantial amounts of citric acid through the expression of the citric acid exporter CexA, a member of the DHA1 family. In this study, we aimed to characterize 11 CexA homologs (Chl proteins) encoded in the genome of A. luchuensis mut. kawachii to identify novel transporters useful for organic acid production. We constructed overexpression strains of chl genes using a cexA disruptant of the A. luchuensis mut. kawachii as the host strain, which prevented excessive secretion of citric acid into the culture supernatant. Subsequently, we evaluated the effects of overexpression of chl on producing organic acids by analyzing the culture supernatant. All overexpression strains did not exhibit significant citric acid accumulation in the culture supernatant, indicating that Chl proteins are not responsible for citric acid export. Furthermore, the ChlH overexpression strain displayed an accumulation of 2-oxoglutaric and fumaric acids in the culture supernatant, while the ChlK overexpression strain exhibited the accumulation of 2-oxoglutaric, malic and succinic acids. Notably, the ChlH and ChlK overexpression led to a substantial increase in the production of 2-oxoglutaric acid, reaching approximately 25 mM and 50 mM, respectively. Furthermore, ChlH and ChlK overexpression also significantly increased the secretory production of dicarboxylic acids, including 2-oxoglutaric acid, in the yellow koji fungus, Aspergillus oryzae. Our study demonstrates that overexpression of DHA1 family gene results in enhanced secretion of organic acids in koji fungi of the genus Aspergillus.

DOI： 10.1016/j.jbiosc.2024.01.010

Chihiro Kadooka, Shun Yakabe, Daisuke Hira, Taiki Futagami, Masatoshi Goto, Takuji Oka . Functional redundancy and divergence of UDP-glucose 4-epimerases in galactose metabolism and cell wall biosynthesis in Aspergillus nidulans. . Fungal genetics and biology : FG & B 103972 - 103972 2025年2月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）

Galactose-containing polysaccharides in the cell walls of filamentous fungi are vital for hyphal formation, mycelial aggregation, and adhesion. Uridine diphosphate (UDP)-glucose 4-epimerase, an enzyme capable of reversibly converting UDP-glucose to UDP-galactose, plays a key role in galactose metabolism. This study investigates the functional specialization and overlapping roles of UDP-glucose 4-epimerases, UgeA and UgeB, in Aspergillus nidulans. Enzyme activity assays revealed that UgeA catalyzes the interconversion of UDP-glucose and UDP-galactose, while UgeB facilitates both UDP-glucose/UDP-galactose and UDP-N-acetylglucosamine/UDP-N-acetylgalactosamine interconversions. Both UgeA and UgeB successfully restored growth in a yeast gal10 disruptant, indicating their involvement in galactose metabolism in vivo. Additionally, the ugeB disruptant of A. nidulans exhibited growth retardation during galactose metabolism, a defect that was alleviated by complementation with ugeB or multiple-copy expression of ugeA. These findings elucidate the complex interplay between sugar metabolism and cell wall synthesis in filamentous fungi and offer insights for the development of novel antifungal therapies.

DOI： 10.1016/j.fgb.2025.103972

Yuki Kobashi, Eri Nakayama, Naoki Fukumori, Ayane Shimojima, Manami Tabira, Yuki Nishimura, Manami Mukae, Ai Muto, Naoto Nakashima, Kayu Okutsu, Yumiko Yoshizaki, Taiki Futagami, Kazunori Takamine, Hisanori Tamaki . Homozygous gene disruption in diploid yeast through a single transformation. . Journal of bioscience and bioengineering137 ( 1 ) 31 - 37 2024年1月

記述言語：英語掲載種別：研究論文（学術雑誌）

As industrial shochu yeast is a diploid strain, obtaining a strain with mutations in both allelic genes was considered difficult. We investigated a method for disrupting two copies of a homozygous gene with a single transformation. We designed a disruption cassette containing an intact LYS5 flanked by nonfunctional ura3 gene fragments divided into the 5'- and 3'-regions. These fragments had overlapping sequences that enabled LYS5 removal as well as URA3 regeneration through loop-out. Furthermore, both ends of the disruption cassette had an additional repeat sequence that allowed the cassette to be removed from the chromosome through loop-out. First, 45 bases of 5'- and 3'-regions of target gene sequences were added on both ends of this cassette using polymerase chain reaction; the resultant disruption cassette was introduced into a shochu yeast strain (ura3/ura3 lys5/lys5); then, single allele disrupted strains were selected on Lys drop-out plates; and after cultivation in YPD medium, double-disrupted strains, in which replacement of another allelic gene with disruption cassette by loss of heterozygosity and regeneration of URA3 in one of the cassettes by loop-out, were obtained by selection on Ura and Lys drop-out plates. The disruption cassettes were removed from the double-disrupted strain via loop-out between repeat sequences in the disruption cassette. The strains that lost either URA3 or LYS5 were counter-selected on 5-fluoroorotic acid or α-amino adipic acid plates, respectively. Using this method, we obtained leu2/leu2 and leu2/leu2 his3/his3 strains in shochu yeast, demonstrating the effectiveness and repeatability of this gene disruption technique in diploid yeast Saccharomyces cerevisiae.

DOI： 10.1016/j.jbiosc.2023.11.003

Yuki Kobashi, Yumiko Yoshizaki, Kayu Okutsu, Taiki Futagami, Hisanori Tamaki, Kazunori Takamine . THI3 contributes to isoamyl alcohol biosynthesis through thiamine diphosphate homeostasis. . Journal of bioscience and bioengineering 2023年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Isoamyl alcohol is a precursor of isoamyl acetate, an aromatic compound that imparts the ginjo aroma to sake. The isoamyl alcohol biosynthesis pathway in yeasts involves the genes PDC1, PDC5, PDC6, ARO10, and THI3 encoding enzymes that decarboxylate α-ketoisocaproic acid to isovaleraldehyde. Among these genes, THI3 is the main gene involved in isoamyl alcohol biosynthesis. Decreased production of isoamyl alcohol has been reported in yeast strains with disrupted THI3 (Δthi3). However, it has also been reported that high THI3 expression did not enhance decarboxylase activity. Therefore, the involvement of THI3 in isoamyl alcohol biosynthesis remains unclear. In this study, we investigated the role of THI3 in isoamyl alcohol biosynthesis. While reproducing previous reports of reduced isoamyl alcohol production by the Δthi3 strain, we observed that the decrease in isoamyl alcohol production occurred only at low yeast nitrogen base concentrations in the medium. Upon investigating individual yeast nitrogen base components, we found that the isoamyl alcohol production by the Δthi3 strain reduced when thiamine concentrations in the medium were low. Under low-thiamine conditions, both thiamine and thiamine diphosphate (TPP) levels decreased in Δthi3 cells. We also found that the decarboxylase activity of cell-free extracts of the Δthi3 strain cultured in a low-thiamine medium was lower than that of the wild-type strain, but was restored to the level of the wild-type strain when TPP was added. These results indicate that the loss of THI3 lowers the supply of TPP, a cofactor for decarboxylases, resulting in decreased isoamyl alcohol production.

DOI： 10.1016/j.jbiosc.2023.11.006

Nishitani A, Hiramatsu K, Kadooka C, Mori K, Okutsu K, Yoshizaki Y, Takamine K, Tashiro K, Goto M, Tamaki H, Futagami T . Expression of heterochromatin protein 1 affects citric acid production in Aspergillus luchuensis mut. kawachii. . Journal of bioscience and bioengineering 2023年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Bioscience and Bioengineering

A putative methyltransferase, LaeA, controls citric acid production through epigenetic regulation of the citrate exporter gene, cexA, in the white koji fungus Aspergillus luchuensis mut. kawachii. In this study, we investigated the role of another epigenetic regulator, heterochromatin protein 1, HepA, in citric acid production. The ΔhepA strain exhibited reduced citric acid production in liquid culture, although to a lesser extent compared to the ΔlaeA strain. In addition, the ΔlaeA ΔhepA strain showed citric acid production similar to the ΔlaeA strain, indicating that HepA plays a role in citric acid production, albeit with a less-significant regulatory effect than LaeA. RNA-seq analysis revealed that the transcriptomic profiles of the ΔhepA and ΔlaeA strains were similar, and the expression level of cexA was reduced in both strains. These findings suggest that the genes regulated by HepA are similar to those regulated by LaeA in A. luchuensis mut. kawachii. However, the reductions in citric acid production and cexA expression observed in the disruptants were mitigated in rice koji, a solid-state culture. Thus, the mechanism by which citric acid production is regulated differs between liquid and solid cultivation. Further investigation is thus needed to understand the regulatory mechanism in koji.

DOI： 10.1016/j.jbiosc.2023.09.004

Okutsu K, Yamamoto Y, Matsuo F, Yoshizaki Y, Futagami T, Tamaki H, Maeda G, Tsuchida E, Takamine K . Characterization of aroma profiles of kokuto-shochu prepared from three different cultivars of sugarcane. . Journal of bioscience and bioengineering135 ( 6 ) 458 - 465 2023年6月

記述言語：英語出版者・発行元：Journal of Bioscience and Bioengineering

Kokuto-shochu is a traditional Japanese spirit prepared from kokuto, obtained by evaporating water from sugarcane (Saccharum officinarum L.) juice. To clarify the effects of sugarcane cultivars on the sensory quality of kokuto-shochu, we investigated the flavor characteristics and composition of volatiles in kokuto-shochu prepared from kokuto using three different sugarcane cultivars, NiF8, Ni15, and RK97-14. Furthermore, experiments were conducted by using the cultivars collected between 2018 and 2020 to observe annual variations in their properties. The amino acid content of the three kokuto varieties did not differ significantly, but the amino acid content of NiF8 was two to five times higher than that of RK97-14, which was the same for all samples collected in the selected years. The browning degrees of kokuto were also higher in NiF8, and they were positively correlated to the amino acid contents of kokuto. The kokuto-like aroma of shochu made from Ni15 was stronger than that of shochu made from RK97-14. The concentration of ethyl lactate in shochu made from Ni15 was higher, however, the concentration of guaiacol was the lowest in the three cultivars’ products. Shochu made from NiF8 had the highest levels of Maillard reaction products (MRPs; pyrazines and furans), β-damascenone, and guaiacol amounts. In contrast, shochu made from RK97-14 tended to have a fruity flavor, and lower MRP levels than those made from NiF8. Thus, it was shown that sugarcane cultivars affect the sensory characteristics and volatiles in kokuto-shochu.

DOI： 10.1016/j.jbiosc.2023.03.010

Sobol MS, Hoshino T, Delgado V, Futagami T, Kadooka C, Inagaki F, Kiel Reese B . Genome characterization of two novel deep-sea sediment fungi, Penicillium pacificagyrus sp. nov. and Penicillium pacificasedimenti sp. nov., from South Pacific Gyre subseafloor sediments, highlights survivability. . BMC genomics24 ( 1 ) 249 - 249 2023年5月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：BMC Genomics

Background: Marine deep subsurface sediments were once thought to be devoid of eukaryotic life, but advances in molecular technology have unlocked the presence and activity of well-known closely related terrestrial and marine fungi. Commonly detected fungi in deep marine sediment environments includes Penicillium, Aspergillus, Cladosporium, Fusarium, and Schizophyllum, which could have important implications in carbon and nitrogen cycling in this isolated environment. In order to determine the diversity and unknown metabolic capabilities of fungi in deep-sea sediments, their genomes need to be fully analyzed. In this study, two Penicillium species were isolated from South Pacific Gyre sediment enrichments during Integrated Ocean Drilling Program Expedition 329. The inner gyre has very limited productivity, organic carbon, and nutrients. Results: Here, we present high-quality genomes of two proposed novel Penicillium species using Illumina HiSeq and PacBio sequencing technologies. Single-copy homologues within the genomes were compared to other closely related genomes using OrthoMCL and maximum-likelihood estimation, which showed that these genomes were novel species within the genus Penicillium. We propose to name isolate SPG-F1 as Penicillium pacificasedimenti sp. nov. and SPG-F15 as Penicillium pacificagyrus sp. nov. The resulting genome sizes were 32.6 Mbp and 36.4 Mbp, respectively, and both genomes were greater than 98% complete as determined by the presence of complete single-copy orthologs. The transposable elements for each genome were 4.87% for P. pacificasedimenti and 10.68% for P. pacificagyrus. A total of 12,271 genes were predicted in the P. pacificasedimenti genome and 12,568 genes in P. pacificagyrus. Both isolates contained genes known to be involved in the degradation of recalcitrant carbon, amino acids, and lignin-derived carbon. Conclusions: Our results provide the first constructed genomes of novel Penicillium isolates from deep marine sediments, which will be useful for future studies of marine subsurface fungal diversity and function. Furthermore, these genomes shed light on the potential impact fungi in marine sediments and the subseafloor could have on global carbon and nitrogen biogeochemical cycles and how they may be persisting in the most energy-limited sedimentary biosphere.

DOI： 10.1186/s12864-023-09320-6

Hiramatsu K, Nishitani A, Okutsu K, Yoshizaki Y, Takamine K, Tamaki H, Futagami T . Efficient gene targeting in Aspergillus chevalieri used to produce katsuobushi. . Bioscience, biotechnology, and biochemistry87 ( 6 ) 672 - 682 2023年3月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioscience, Biotechnology and Biochemistry

In this study, we developed an efficient gene targeting system for the osmophilic fungus Aspergillus chevalieri, which is commonly used in the production of a dried bonito, katsuobushi. Specifically, we utilized the clustered regularly interspaced short palindromic repeats/Cas9 system to disrupt the ATP sulfurylase encoding sC gene. This results in methionine auxotroph and selenate-resistance. Additionally, we disrupted the DNA ligase IV encoding ligD gene, which is required for nonhomologous end joining. Using the sC marker and selenate-resistance as a selection pressure, we were able to rescue the sC marker and generate a ligD sC strain. We determined that the gene targeting efficiency of the ligD sC strain was significantly higher than that of the parental sC strain, which indicates that this strain provides efficient genetic recombination for the genetic analysis of A. chevalieri.

DOI： 10.1093/bbb/zbad033

Tiantian Wang, Isao Hanashiro, Yumiko Yoshizaki, Yuki Kobashi, Suzuka Noda, Kayu Okutsu, Taiki Futagami, Hisanori Tamaki, Kazunori Takamine . Shochu Koji Microstructure and Starch Structure during Preparation. . Journal of applied glycoscience70 ( 4 ) 109 - 117 2023年

記述言語：英語掲載種別：研究論文（学術雑誌）

In this study, we investigated the changes in composition, microstructure, and starch molecular structure of shochu koji during preparation. We observed that the gelatinized and outer part of starch was decomposed in priority during the early and middle preparation stages. The gap between the starch granules increased with the delayed time. Finally, the koji microstructure became spongy. Shochu koji mold produced two α-amylases in different expression manners. Acid-labile α-amylase was produced in the early and middle preparation stages. Acid-stable α-amylase and saccharification power were produced in the middle and late stages. Throughout the koji preparation, reducing sugars content reached approximately 13-20 % of the total sugar content, with glucose representing over 70 % of the reducing sugars. α-Glucan fragments with C chains of degree of polymerization (DP) 4-73 were observed in the early and middle stages (<23 h), indicating the degradation of amylopectin at long B chains. In the latter stage, the amount of C chains of DP 6-30 decreased, while the longer C chains (DP 30<) did not change. These results showed that acid-labile α-amylase, acid-stable α-amylase, and saccharification enzymes including glucoamylase and α-glucosidase work preferentially on the amorphous regions of starch granules, and cooperative action of these enzymes during koji preparation contributes to the formation of the observed microstructure. Our study is the first report on the decomposition schemes of starch and the microstructure forming process in shochu koji.

DOI： 10.5458/jag.jag.JAG-2023_0006

Li B, Yoshizaki Y, Okutsu K, Haruyama Y, Sato Y, Futagami T, Tamaki H, Takamine K . Factors that cause replacement of shochu yeast by wild yeast upon Sashi-moto repetition. . Journal of bioscience and bioengineering134 ( 4 ) 326 - 330 2022年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Bioscience and Bioengineering

In shochu-making, a small amount of fermenting moromi is added to a koji/water mixture instead of yeast culture to initiate fermentation. This is a characteristic process called Sashi-moto. It is known that shochu yeast is replaced by wild yeast upon repetition of Sashi-moto. The shochu yeast strains Kagoshima No. 2 (K2), Kagoshima No. 4 (C4), and Kagoshima No. 5 (H5), but not Kagoshima No. 6 (A6), were replaced by wild yeast (strain No. S5-g). K2 and C4 were easily replaced compared to H5, and the specific growth rates of K2 and C4 were lower than that of S5-g under higher osmotic pressure. Although the specific growth rate of H5 was higher than that of S5-g, its yeast population at the stationary phase was smaller than S5-g. On the other hand, both the specific growth rate and yeast population of A6 were higher than those of S5-g. The specific growth rate of yeast would be affected by osmotic tolerance and specific characters of the yeast strain.

DOI： 10.1016/j.jbiosc.2022.07.011

劉根僑, 吉﨑由美子, 奥津果優, 二神泰基, 玉置尚徳, 髙峯和則 . サツマイモ固体糖化後仕込みによる芋焼酎製造の可能性 . 日本醸造協会誌117 ( 10 ) 712 - 723 2022年10月

曾伝濤, 劉夢迪, 吉﨑由美子, 奥津果優, 二神泰基, 玉置尚徳, 髙峯和則 . サトイモ焼酎の香味特性と焼酎粕の機能性 . 日本醸造協会誌117 ( 10 ) 701 - 711 2022年10月

Futagami T . The white koji fungus Aspergillus luchuensis mut. kawachii. . Bioscience, biotechnology, and biochemistry86 ( 5 ) 574 - 584 2022年4月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Bioscience, Biotechnology and Biochemistry

The white koji fungus, Aspergillus luchuensis mut. kawachii, is used in the production of shochu, a traditional Japanese distilled spirit. White koji fungus plays an important role in the shochu production process by supplying amylolytic enzymes such as α-amylase and glucoamylase. These enzymes convert starch contained in primary ingredients such as rice, barley, buckwheat, and sweet potato into glucose, which is subsequently utilized by the yeast Saccharomyces cerevisiae to produce ethanol. White koji fungus also secretes large amounts of citric acid, which lowers the pH of the shochu mash, thereby preventing the growth of undesired microbes and enabling stable production of shochu in relatively warm regions of Japan. This review describes the historical background, research tools, and recent advances in studies of the mechanism of citric acid production by white koji fungus.

DOI： 10.1093/bbb/zbac033

Ketkaeo S, Nagano Y, Baba S, Kimura K, Futagami T, Sanpamongkolchai W, Kobayashi G, Goto M . Development of Monascus purpureus monacolin K-hyperproducing mutant strains by synchrotron light irradiation and their comparative genome analysis. . Journal of bioscience and bioengineering133 ( 4 ) 362 - 368 2022年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Bioscience and Bioengineering

Monascus purpureus have been used for making koji and other fermented foods and supplements. M. purpureus characteristically produces monacolin K (MK), a secondary metabolite that competitively inhibits cholesterol synthesis. Synchrotron light irradiation was applied to induce mutation in the strain KUPM5 to improve the MK-producing ability of M. purpureus strain KUPM5. Screening by a bioassay utilizing sensitivities to yeast Saccharomyces cerevisiae from 936 colonies allows isolating three mutant strains: SC01, SC02, and SC03. These mutant strains and the parental strain KUPM5 were subjected to make koji using rice, and their metabolites were compared. All strains SC01, SC02, and SC03 in koji showed higher production of MK than the strain KUPM5. Particularly, the SC02 strain produced MK threefold higher than KUPM5 and maintained the production capabilities of other metabolites, including red, yellow, and orange pigments, mycelial contents, and α-amylase activity comparable to those of the strain KUPM5. Comparative genome analysis among strain KUPM5 and the mutants revealed that synchrotron light irradiation introduced mutations in approximately 90% of the total genes, including SNV, MNV, and indel mutations. The frequencies of SNV substitution in the whole genome occupied 68.96% of all mutations, of which 92.38% were transversions and 7.62% were transitions. This study, therefore, proved the synchrotron light irradiation was highly efficient for the strain improvement of a filamentous fungus, M. purpureus, and provided insights into the properties of mutation in the fungus by this mutagen.

DOI： 10.1016/j.jbiosc.2021.11.011

Mori K, Kadooka C, Oda K, Okutsu K, Yoshizaki Y, Takamine K, Tashiro K, Goto M, Tamaki H, Futagami T . Chromosome-level genome sequence data and analysis of the white koji fungus, Aspergillus luchuensis mut. kawachii IFO 4308. . Data in brief41 107888 - 107888 2022年4月国際誌

Oussou-Azo FA, Futagami T, Vestergaard MCM . Immuno-Dipstick for Colletotrichum gloeosporioides Detection: Towards On-Farm Application. . Biosensors12 ( 2 ) 49 - 49 2022年1月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Biosensors

Early and quick detection of pathogens are crucial for managing the spread of infections in the biomedical, biosafety, food, and agricultural fields. While molecular diagnostics can offer the specificity and reliability in acute infectious diseases, detection of pathogens is often slowed down by the current benchtop molecular diagnoses, which are time consuming, labor intensive, and lack the mobility for application at the point-of-need. In this work, we developed a complete on-farm use detection protocol for the plant-devastating anthracnose agent: Colletotrichum gloeosporioides. Our methods combined a simplified DNA extraction on paper that is compatible with loop-mediated isothermal amplification (LAMP), coupled with paper-based immunoassay lateral flow sensing. Our results offer simple, quick, easy, and a minimally instrumented toolkit for Colletotrichum gloeosporioides detection. This scalable and adaptable platform is a valuable alternative to traditional sensing systems towards on-the-go pathogen detection in food and agriculture, biomedical, and other fields.

DOI： 10.3390/bios12020049

Futagami T, Goto M . Insights regarding sirtuin-dependent gene regulation during white koji production. . Communicative & integrative biology15 ( 1 ) 92 - 95 2022年査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Communicative and Integrative Biology

White koji, a solid-state culture of Aspergillus luchuensis mut. kawachii using grains such as rice and barley, is used as a source of amylolytic enzymes and citric acid for the production of shochu, a traditional Japanese distilled spirit. We previously characterized changes in gene expression that affect the properties of white koji during the shochu production process; however, the underlying regulatory mechanisms were not determined. We then characterized the NAD+-dependent histone deacetylase sirtuin, an epigenetic regulator of various biological phenomena, in A. l. mut. kawachii and found that sirtuin SirD is involved in expression of α-amylase activity and citric acid accumulation. In this addendum study, we measured the NAD+/NADH redox state and found that the NAD+ level and NAD+/NADH ratio decrease during koji production, indicating that sirtuin activity declines in the late stages of koji culture. By comparing these results with transcriptomic data obtained in our previous studies, we estimate that approximately 35% of the gene expression changes during white koji production are SirD dependent. This study provides clues to the mechanism of gene expression regulation in A. l. mut. kawachii during the production of white koji.

DOI： 10.1080/19420889.2022.2051844

Zhao Z., Sugimachi M., Yoshizaki Y., Yin X., Han X.L., Okutsu K., Futagami T., Tamaki H., Takamine K. . Correlation between key aroma and manufacturing processes of rice-flavor baijiu and awamori, Chinese and Japanese traditional liquors . Food Bioscience44 2021年12月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：Food Bioscience

In this study, we compared the flavor profiles Chinese rice-flavor baijiu and Japanese awamori—traditional liquors with similar manufacturing processes. Citric acid and lactic acid were the major acid constituents in the mash of awamori and rice-flavor baijiu, respectively. The amino acid content of awamori mash was greater than that of rice-flavor baijiu owing to the higher proteolytic activities in the former. The high amino acid content also induced yeast growth at the early stage of fermentation in awamori mash, thus increasing the fermentation rate. Sensory evaluation revealed that rice-flavor baijiu had a significantly stronger alcoholic, floral, and fruity aroma, whereas the awamori had a strong sour, koji-like, oily, and cereal-like aromas. We determined the aroma threshold of (+)-ethyl D-lactate (15,400 μg/L) and (−)-ethyl L-lactate (12,500 μg/L). Both compounds have a common sweet, fruity, and green leaves-like aroma and a different nuance; (+)-ethyl D-lactate has a peach- and citrus-like aromas, and (−)-ethyl L-lactate has an apple-like and milky aromas. In rice-flavor baijiu, (−)-ethyl L-lactate constitutes the major isoform, as evident from the higher concentration of L (+)-lactic acid in the mash. Higher alcohols, (−)-ethyl L-lactate, and isoamyl acetate contents were greater in rice-flavor baijiu, while 1-octen-3-ol and acetic acid were greater in awamori. These results showed that solid-state saccharification contributed to the production of ethyl lactate by supplying in the mash a large amount of lactic acid produced by Rhizopus oryzae and koji contributed to the production of 1-octen-3-ol via the high fatty acid oxygenase activity produced by Aspergillus luchuensis.

DOI： 10.1016/j.fbio.2021.101375

Web of Science

Zitai Wang, Kayu Okutsu, Yumiko Yoshizaki, Taiki Futagami, Hisanori Tamaki, Takuro Maruyama, Kazufumi Toume, Katsuko Komatsu, Fumio Hashimoto, Kazunori Takamine . Effects of microbial fermentation on enzyme activity and volatile properties of Massa Medicata Fermentata . Traditional & Kampo Medicine9 ( 1 ) 10 - 17 2021年11月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Aim: Massa Medicata Fementata (MMF) is a crude drug used in East Asia to treat anorexia and dyspepsia. It is prepared from wheat and several herbs through microbial fermentation using Aspergillus sp. and Rhizopus sp. There is great difference in the quality of commercial MMF, and the microbes of MMF are suggested to affect its quality. We investigated the effects of microbial fermentation on the quality of MMF. Methods: Raw materials of MMF were mixed according to the ratio listed in the National Standard for Chinese Patent Drugs, and MMF was prepared using pure cultures of Aspergillus oryzae or Rhizopus oryzae. Digestive enzyme activities (α-amylase, protease, and lipase) and volatile compounds were measured using an analytical kit and GC–MS, respectively. Results: Enzyme activity increased in MMF. MMF prepared with A. oryzae (MMF-A) showed higher α-amylase and lipase activities than that prepared with R. oryzae (MMF-R). Protease activity was marginally higher in MMF-R than in MMF-A. GC–MS analysis revealed that terpenoids decreased with fermentation; however, 2,3-butanediol, acetoin, and guaiacol were detected in MMF only. C8 compounds such as 1-octen-3-ol were higher in MMF-A than MMF-R; however, aromatic compounds such as 4-vinylguaiacol and pyrazines were higher in MMF-R than MMF-A. Conclusion: Microbial fermentation contributes to increased enzyme activity and changes in MMF volatiles. These properties of MMF were considerably affected by the microbes used, and it is proposed in this study that it is important to have microbial control in the production of commercial MMF.

DOI： 10.1002/tkm2.1303

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/tkm2.1303

Hirose J, Watanabe T, Futagami T, Fujihara H, Kimura N, Suenaga H, Goto M, Suyama A, Furukawa K . A New ICEclc Subfamily Integrative and Conjugative Element Responsible for Horizontal Transfer of Biphenyl and Salicylic Acid Catabolic Pathway in the PCB-Degrading Strain Pseudomonas stutzeri KF716. . Microorganisms9 ( 12 ) 2021年11月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Microorganisms

Integrative and conjugative elements (ICEs) are chromosomally integrated self-transmissible mobile genetic elements. Although some ICEs are known to carry genes for the degradation o-f aromatic compounds, information on their genetic features is limited. We identified a new member of the ICEclc family carrying biphenyl catabolic bph genes and salicylic acid catabolic sal genes from the PCB-degrading strain Pseudomonas stutzeri KF716. The 117-kb ICEbph-sal KF716 contains common core regions exhibiting homology with those of degradative ICEclc from P. knackmussii B13 and ICEXTD from Azoarcus sp. CIB. A comparison of the gene loci collected from the public database revealed that several putative ICEs from P. putida B6-2, P, alcaliphila JAB1, P. stutzeri AN10, and P. stutzeri 2A20 had highly conserved core regions with those of ICEbph-sal KF716, along with the variable region that encodes the catabolic genes for biphenyl, naphthalene, toluene, or phenol. These data indicate that this type of ICE subfamily is ubiquitously distributed within aromatic compound-degrading bacteria. ICEbph-sal KF716 was transferred from P. stutzeri KF716 to P. aeruginosa PAO1 via a circular extrachromosomal intermediate form. In this study, we describe the structure and genetic features of ICEbph-sal KF716 compared to other catabolic ICEs.

DOI： 10.3390/microorganisms9122462

Zhao Z, Sugimachi M, Yoshizaki Y, Yin X, Han XL, Okutsu K, Futagami T, Tamaki H, Takamine K . Impact of solid-state saccharification on the flavor of rice-flavor baijiu. . Journal of food science86 ( 11 ) 4958 - 4968 2021年11月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Rice-flavor baijiu is a traditional Chinese liquor. The flavor profile and volatiles presented with or without the solid-state saccharification (SSS) were investigated to reveal the effects of SSS process on the quality of rice-flavor baijiu. The liquor prepared with SSS had a sweet flavor. It contained significantly higher contents of β-phenylethyl alcohol, β-phenylethyl acetate, and ethyl lactate with odor active value of >1. The liquor prepared without SSS had a cheese-like flavor. It was confirmed that the cheese-like flavor derived from butanoic acid was only detected in the liquor prepared without SSS. SSS facilitated the biosynthesis of β-phenylethyl alcohol and ethyl lactate by supplying a large amount of phenylalanine and lactic acid at the initial stage of fermentation, and it prevented contamination. These results indicated that the SSS process contributed to produce the characteristic flavor compounds of rice-flavor baijiu. Practical Application: Solid-state saccharification (SSS) process of rice-flavor baijiu contributes not only in brewing, but also in the production of the characteristic flavor compounds and the repression of the off-flavor derived from the contamination. Therefore, SSS is a critical process to control the flavor of rice-flavor baijiu.

DOI： 10.1111/1750-3841.15935

Kadooka C, Mori K, Okutsu K, Yoshizaki Y, Takamine K, Tashiro K, Tamaki H, Futagami T . Chromosome-Level Genome Sequence of Aspergillus chevalieri M1, Isolated from Katsuobushi. . Microbiology resource announcements10 ( 37 ) e0038521 2021年9月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Microbiology Resource Announcements

In this study, we report the chromosome-level genome sequence of the osmophilic filamentous fungus Aspergillus chevalieri M1, which was isolated from a dried bonito, katsuobushi. This fungus plays a significant role in the fermentation and ripening process. Thus, elucidating the sequence data for this fungus will aid in subsequent genomic research on the fungi involved in katsuobushi production.

DOI： 10.1128/MRA.00385-21

Futagami T, Mori K, Kadooka C, Niihara H, Tashiro K, Tamaki H, Tanaka T . Chromosome-Level Genome Sequence of Aspergillus puulaauensis MK2, a Fungus Isolated from a Dead Hard Tick. . Microbiology resource announcements10 ( 36 ) e0037221 2021年9月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Society for Microbiology

<jats:p>
<jats:named-content content-type="genus-species">Aspergillus puulaauensis</jats:named-content>
strain MK2 was isolated from a dead hard tick (
<jats:named-content content-type="genus-species">Haemaphysalis longicornis</jats:named-content>
). Here, we determined the chromosome-level genome sequence of
<jats:named-content content-type="genus-species">A. puulaauensis</jats:named-content>
MK2.
</jats:p>

DOI： 10.1128/MRA.00372-21

Mori K, Kadooka C, Nishitani A, Okutsu K, Yoshizaki Y, Takamine K, Tashiro K, Goto M, Tamaki H, Futagami T . Chromosome-Level Genome Sequence of the Black Koji Fungus Aspergillus luchuensis RIB2601. . Microbiology resource announcements10 ( 29 ) e0038421 2021年7月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Society for Microbiology

<jats:p>
<jats:named-content content-type="genus-species">Aspergillus luchuensis</jats:named-content>
is used for the production of awamori and shochu, which are traditional Japanese distilled alcoholic beverages. Here, we determined the chromosome-level genome sequence of
<jats:italic>A. luchuensis</jats:italic>
RIB2601.
</jats:p>

DOI： 10.1128/MRA.00384-21

Hayashi K, Kajiwara Y, Futagami T, Goto M, Takashita H . Making Traditional Japanese Distilled Liquor, Shochu and Awamori, and the Contribution of White and Black Koji Fungi. . Journal of fungi (Basel, Switzerland)7 ( 7 ) 2021年6月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Fungi

The traditional Japanese single distilled liquor, which uses koji and yeast with designated ingredients, is called “honkaku shochu.” It is made using local agricultural products and has several types, including barley shochu, sweet potato shochu, rice shochu, and buckwheat shochu. In the case of honkaku shochu, black koji fungus (Aspergillus luchuensis) or white koji fungus (Aspergillus luchuensis mut. kawachii) is used to (1) saccharify the starch contained in the ingredients, (2) produce citric acid to prevent microbial spoilage, and (3) give the liquor its unique flavor. In order to make delicious shochu, when cultivating koji fungus during the shochu production process, we use a unique temperature control method to ensure that these three important elements, which greatly affect the taste of the produced liquor, are balanced without any excess or deficiency. This review describes in detail the production method of honkaku shochu, a distilled spirit unique to Japan and whose market is expected to expand worldwide, with special attention paid to the koji fungi cultivation step. Furthermore, we describe the history of the koji fungi used today in the production of shochu, and we provide a thorough explanation of the characteristics of each koji fungi. We also report the latest research progress on this topic.

DOI： 10.3390/jof7070517

二神泰基, 門岡千尋, 後藤正利, 玉置尚徳 . 白麹菌のクエン酸高生産機構 . 化学と生物59 ( 5 ) 241 - 246 2021年5月

記述言語：日本語出版者・発行元：公益社団法人日本農芸化学会

清酒，味噌，醤油などの製造には一般的に黄麹菌が用いられるが，焼酎の製造には白麹菌あるいは黒麹菌が主に用いられる．焼酎造りに用いられる麹菌は，穀類原料の糖化のほかにクエン酸を高分泌生産して雑菌汚染を防ぐ役割を有している．クエン酸の分泌機構については長年不明であったが，近年，クエン酸輸送体についての研究が進展した．見いだされたクエン酸輸送体を黄麹菌にて発現させるとクエン酸の高分泌生産能力を獲得することから，本輸送体がクエン酸の排出において中心的役割を担っていると考えられる．本稿では，白麹菌において見いだされたクエン酸輸送体による排出プロセスの重要性について解説したい．

DOI： 10.1271/kagakutoseibutsu.59.241

Zeng C, Yoshizaki Y, Yin X, Wang Z, Okutsu K, Futagami T, Tamaki H, Takamine K . Additional moisture during koji preparation contributes to the pigment production of red koji (Monascus-fermented rice) by influencing gene expression. . Journal of food science86 ( 3 ) 969 - 976 2021年3月国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Food Science

Abstract: Additional moisture in preparing red koji, Monascus-fermented rice, is a characteristic production process. To determine how additional moisture affects red koji preparation as per quality, we compared the growth of Monascus purpureus, enzyme and pigment production, and related gene expressions using our findings. We considered two kinds of red koji: one prepared with additional moisture at the middle part of the preparation and the other prepared without additional moisture. Our results showed that additional moisture did not promote the growth of M. purpureus, but it was significantly increased the pigment (red and yellow) and tended to increase the α-amylase level and saccharification power. Although adding a high amount of moisture (approximately 60% moisture content) promoted pigment production, it slightly repressed enzyme production. In contrast, adding approximately 50% moisture content promoted enzyme production. These findings showed that the additional moisture can affect the quality of red koji on the purpose. The expression of 10 pigment biosynthetic gene clusters and two glycohydrolase genes in red koji after adding moisture was analyzed through real-time qPCR. Eight genes were upregulated within 1 hr after adding water, with mppR2 being the first upregulated gene within 30 min. The expression of genes as per pigment production quickly responded to additional moisture during solid-state fermentation. Moreover, acetyl-CoA, which is a starting substrate for pigment content in red koji was increased within 3 hr after adding water. This study first described the relationship between additional moisture and expression of pigment biosynthetic genes by Monascus spp. during red koji preparation.

DOI： 10.1111/1750-3841.15610

Zeng C. . The expression profiles of acid-stable α-amylase and acid-labile α-amylase of Aspergillus luchuensis mut. Kawachii effect on the microstructure of koji and alcohol fermentation . LWT139 2021年3月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：LWT

We investigated the contribution of the complex expressions of two α-amylases, namely acid-stable α-amylase (AS-amylase) and acid-labile α-amylase (AL-amylase), from Aspergillus kawachii to the microstructure of koji and the brewing. AL-amylase was found to be the primary contributor to the decomposition of starch in the early stage of koji making. From the middle stage, both α-amylases decomposed the starch in a coordinated manner, and at the final stage, acid-stable α-amylase and glucoamylase decomposed the starch granules. Characterization of koji prepared by the single disruptions of AS-amylase or AL-amylase genes, double disruption of both α-amylase genes, and triple disruption of two α-amylase and glucoamylase genes in A. kawachii cells indicated that both α-amylases can work in a synergistic manner to decompose starches during koji making. AL-amylase was found, for the first time, to play an important role in starch decomposition in koji. The speed of alcohol fermentation and ester contents of the fermented mash were higher the mash prepared the control strain, followed by single, double, and triple disruptants. These results indicate that the microstructure of koji plays a role in promoting alcohol fermentation and flavor development.

DOI： 10.1016/j.lwt.2020.110580

Web of Science

Nakamura E, Kadooka C, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . Citrate exporter enhances both extracellular and intracellular citric acid accumulation in the koji fungi Aspergillus luchuensis mut. kawachii and Aspergillus oryzae. . Journal of bioscience and bioengineering131 ( 1 ) 68 - 76 2021年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Bioscience and Bioengineering

Citrate exporter CexA plays a key role in the production of citric acid in fungi; however, its role in intracellular metabolism has remained unclear. In this study, we comparably characterized homologous cexA genes in the white koji fungus Aspergillus luchuensis mut. kawachii and the yellow koji fungus Aspergillus oryzae, which exhibit high and low abilities, respectively, to produce citric acid. Disruption of cexA caused a significant decline of both extracellular and intracellular citric acid accumulation in Aspergillus kawachii, while overexpression of the A. kawachii cexA gene (AkcexA) into A. oryzae significantly enhanced both extracellular and intracellular citric acid accumulation in A. oryzae to a level comparable to that of A. kawachii. In addition, overexpression of two intrinsic cexA homologs (AocexA and AocexB) in A. oryzae also enhanced its extracellular and intracellular citric acid accumulation. Comprehensive analysis of intracellular metabolites from an AkcexA-overexpressing strain of A. oryzae compared with its control strain identified metabolic changes associated with intracellular citric acid accumulation via the glycolytic pathway, pentose phosphate pathway, and tricarboxylic acid cycle. Our results indicate that citric acid export enhances not only extracellular citric acid accumulation but also intracellular metabolic fluxes to generate citric acid.

DOI： 10.1016/j.jbiosc.2020.09.002

Genqiao Liu, Juna Serikawa, Kayu Okutsu, Yumiko Yoshizaki, Taiki Futagami, Hisanori Tamaki, Kazunori Takamine . Impact of fermentation temperature on the quality and sensory characteristics of imo‐shochu . Journal of the Institute of Brewing127 ( 4 ) 417 - 423 2021年1月

Wakamatsu T, Mizobuchi S, Mori F, Futagami T, Terada T, Morono Y . Construction of Aerobic/Anaerobic-Substrate-Induced Gene Expression Procedure for Exploration of Metagenomes From Subseafloor Sediments. . Frontiers in microbiology12 726024 - 726024 2021年国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

Substrate-induced gene expression (SIGEX) is a high-throughput promoter-trap method. It is a function-based metagenomic screening tool that relies on transcriptional activation of a reporter gene green fluorescence protein (gfp) by a metagenomic DNA library upon induction with a substrate. However, its use is limited because of the relatively small size of metagenomic DNA libraries and incompatibility with screening metagenomes from anaerobic environments. In this study, these limitations of SIGEX were addressed by fine-tuning metagenome DNA library construction protocol and by using Evoglow, a green fluorescent protein that forms a chromophore even under anaerobic conditions. Two metagenomic libraries were constructed for subseafloor sediments offshore Shimokita Peninsula (Pacific Ocean) and offshore Joetsu (Japan Sea). The library construction protocol was improved by (a) eliminating short DNA fragments, (b) applying topoisomerase-based high-efficiency ligation, (c) optimizing insert DNA concentration, and (d) column-based DNA enrichment. This led to a successful construction of metagenome DNA libraries of approximately 6 Gbp for both samples. SIGEX screening using five aromatic compounds (benzoate, 3-chlorobenzoate, 3-hydroxybenzoate, phenol, and 2,4-dichlorophenol) under aerobic and anaerobic conditions revealed significant differences in the inducible clone ratios under these conditions. 3-Chlorobenzoate and 2,4-dichlorophenol led to a higher induction ratio than that for the other non-chlorinated aromatic compounds under both aerobic and anaerobic conditions. After the further screening of induced clones, a clone induced by 3-chlorobenzoate only under anaerobic conditions was isolated and characterized. The clone harbors a DNA insert that encodes putative open reading frames of unknown function. Previous aerobic SIGEX attempts succeeded in the isolation of gene fragments from anaerobes. This study demonstrated that some gene fragments require a strict in vivo reducing environment to function and may be potentially missed when screened by aerobic induction. The newly developed anaerobic SIGEX scheme will facilitate functional exploration of metagenomes from the anaerobic biosphere.

DOI： 10.3389/fmicb.2021.726024

白石洋平, 竹浦澪, 奥津果優, 吉崎由美子, 二神泰基, 玉置尚徳, 和久豊, 高峯和則 . 麹菌菌種の違いが芋焼酎の香味形成に及ぼす影響(第1報)酵素活性と芋焼酎醪の差異 . 日本醸造協会誌116 ( 1 ) 2021年

白石洋平, 奥津果優, 吉崎由美子, 二神泰基, 玉置尚徳, 和久豊, 高峯和則 . 麹菌菌種の違いが芋焼酎の香味形成に及ぼす影響(第2報)香気成分と官能評価での差異 . 日本醸造協会誌116 ( 1 ) 2021年

Kadooka C, Yamaguchi M, Okutsu K, Yoshizaki Y, Takamine K, Katayama T, Maruyama JI, Tamaki H, Futagami T . A CRISPR/Cas9-mediated gene knockout system in Aspergillus luchuensis mut. kawachii . Biosci Biotechnol Biochem.84 ( 10 ) 2179 - 2183 2020年10月査読

Iwasaki F, Sunao M, Okutsu K, Yoshizaki Y, Futagami T, Tamaki H, Takamine K, Sameshima Y . Effects of liming on the flavor of kokuto-shochu, a spirit made from non-centrifugal sugar . J Biosci Bioeng130 ( 4 ) 360 - 366 2020年10月査読

Iwasaki F, Sunao M, Okutsu K, Yoshizaki Y, Futagami T, Tamaki H, Takamine K, Sameshima Y . Effects of liming on the flavor of kokuto-shochu, a spirit made from non-centrifugal sugar. . Journal of bioscience and bioengineering130 ( 4 ) 360 - 366 2020年10月

Kadooka C, Yamaguchi M, Okutsu K, Yoshizaki Y, Takamine K, Katayama T, Maruyama JI, Tamaki H, Futagami T . A CRISPR/Cas9-mediated gene knockout system in Aspergillus luchuensis mut. kawachii. . Bioscience, biotechnology, and biochemistry84 ( 10 ) 2179 - 2183 2020年10月

Kadooka C, Nakamura E, Kubo S, Okutsu K, Yoshizaki Y, Takamine K, Tamaki H, Futagami T . Analysis of the fungal population involved in Katsuobushi production. . The Journal of general and applied microbiology66 ( 4 ) 239 - 243 2020年9月招待査読

Nakamura E, Kadooka C, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T. . Citrate exporter enhances both extracellular and intracellular citric acid accumulation in the koji fungi Aspergillus luchuensis mut. kawachii and Aspergillus oryzae . J Biosci Bioeng 2020年9月査読

Wang Z, Okutsu K, Futagami T, Yoshizaki Y, Tamaki H, Maruyama T, Toume K, Komatsu K, Hashimoto F, Takamine K . Microbial Community Structure and Chemical Constituents in Shinkiku, a Fermented Crude Drug Used in Kampo Medicine . Front Nutr7 ( 115 ) 2020年7月査読

門岡千尋、合原翔平、久保臣悟、尾上昌平、奥津果優、吉﨑由美子、髙峯和則、後藤正利、玉置尚徳、二神泰基 . 白麹菌Aspergillus luchuensis mut. kawachii NBRC 4308のアルビノ表現型の原因遺伝子の解析 . 日本醸造協会誌116 ( 6 ) 369 - 377 2020年6月査読

Oussou-Azo AF, Nakama T, Nakamura M, Futagami T, Vestergaard MCM . Antifungal Potential of Nanostructured Crystalline Copper and Its Oxide Forms . Nanomaterials (Basel)10 ( 5 ) 1003 2020年5月

Oussou-Azo AF, Nakama T, Nakamura M, Futagami T, Vestergaard MCM . Antifungal Potential of Nanostructured Crystalline Copper and Its Oxide Forms. . Nanomaterials (Basel, Switzerland)10 ( 5 ) 2020年5月

Miyamoto A, Kadooka C, Mori K, Tagawa Y, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . Sirtuin SirD is involved in α-amylase activity and citric acid production in Aspergillus luchuensis mut. kawachii during a solid-state fermentation process. . Journal of bioscience and bioengineering129 ( 4 ) 454 - 466 2020年4月招待査読

瀬戸口翔, 章超, 飯島遼平, 二神泰基, 河野邦晃, 岩井謙一, 玉置尚徳 . 焼酎醸造において発酵遅延を引き起こす黄変黒麹から見出された抗菌物質の抗菌効果 . 日本醸造学会誌115 151 - 158 2020年3月査読

Xuan Yin, Yumiko Yoshizaki, Shugo Kurazono, Mina Sugimachi, Haruka Takeuchi, Xing-Lin Han, Kayu Okutsu, Taiki Futagami, Hisanori Tamaki, Kazunori Takamine . Characterization of Flavor Compounds in Rice-flavor baijiu, a Traditional Chinese Distilled Liquor, Compared with Japanese Distilled Liquors, awamori and kome-shochu. . Food Science and Technology Research26 ( 3 ) 411 - 422 2020年3月査読

Yin X, Yoshizaki Y, Ikenaga M, Han XL, Okutsu K, Futagami T, Tamaki H, Takamine K . Manufactural impact of the solid-state saccharification process in rice-flavor baijiu production. . Journal of bioscience and bioengineering129 ( 3 ) 315 - 321 2020年3月招待査読

Kadooka C, Nakamura E, Mori K, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . LaeA Controls Citric Acid Production through Regulation of the Citrate Exporter-Encoding cexA Gene in Aspergillus luchuensis mut. kawachii. . Applied and environmental microbiology86 ( 5 ) 1 - 17 2020年2月

Kadooka C, Nakamura E, Mori K, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . LaeA Controls Citric Acid Production through Regulation of the Citrate Exporter-Encoding cexA Gene in Aspergillus luchuensis mut. kawachii . Appl Environ Microbiol86 ( 5 ) e01950-19 2020年2月査読

瀬戸口翔, 益留綾菜, 大岩達郎, 二神泰基, 河野邦晃, 岩井謙一, 玉置尚徳 . 酸性プロテアーゼを高生産する黒麹菌の育種と芋焼酎醸造試験 . 日本醸造学会誌115 54 - 61 2020年1月査読

Wang Z, Okutsu K, Futagami T, Yoshizaki Y, Tamaki H, Maruyama T, Toume K, Komatsu K, Hashimoto F, Takamine K . Microbial Community Structure and Chemical Constituents in Shinkiku, a Fermented Crude Drug Used in Kampo Medicine. . Frontiers in nutrition7 115 2020年

Setoguchi S, Mizutani O, Yamada O, Futagami T, Iwai K, Takase Y, Tamaki H. . Effect of pepA deletion and overexpression in Aspergillus luchuensis on sweet potato shochu brewing . J. Biosci. Bioeng128 ( 4 ) 456 - 462 2019年10月査読

Setoguchi S, Mizutani O, Yamada O, Futagami T, Iwai K, Takase Y, Tamaki H . Effect of pepA deletion and overexpression in Aspergillus luchuensis on sweet potato shochu brewing. . Journal of bioscience and bioengineering128 ( 4 ) 456 - 462 2019年10月

Kurokawa Y, Konishi R, Yoshida A, Tomioku K, Futagami T, Tamaki H, Tanabe K, Fujita A. . Essential and distinct roles of phosphatidylinositol 4-kinases, Pik1p and Stt4p, in yeast autophagy. . Biochim Biophys Acta Mol Cell Biol Lipids1864 ( 9 ) 1214 - 1225 2019年9月査読

Kurokawa Y, Konishi R, Yoshida A, Tomioku K, Futagami T, Tamaki H, Tanabe K, Fujita A . Essential and distinct roles of phosphatidylinositol 4-kinases, Pik1p and Stt4p, in yeast autophagy. . Biochimica et biophysica acta. Molecular and cell biology of lipids1864 ( 9 ) 1214 - 1225 2019年9月

Hirose J, Fujihara H, Watanabe T, Kimura N, Suenaga H, Futagami T, Goto M, Suyama A, Furukawa K . Biphenyl/PCB Degrading bph Genes of Ten Bacterial Strains Isolated from Biphenyl-Contaminated Soil in Kitakyushu, Japan: Comparative and Dynamic Features as Integrative Conjugative Elements (ICEs). . Genes10 ( 5 ) 2019年5月

Hirose J, Fujihara H, Watanabe T, Kimura N, Suenaga H, Futagami T, Goto M, Suyama A, Furukawa K. . Biphenyl/PCB Degrading bph Genes of Ten Bacterial Strains Isolated from Biphenyl-Contaminated Soil in Kitakyushu, Japan: Comparative and Dynamic Features as Integrative Conjugative Elements (ICEs). . Genes (Basel)10 ( 5 ) E404 2019年5月査読

Kadooka C, Izumitsu K, Onoue M, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . Mitochondrial Citrate Transporters CtpA and YhmA Are Required for Extracellular Citric Acid Accumulation and Contribute to Cytosolic Acetyl Coenzyme A Generation in Aspergillus luchuensis mut. kawachii. . Applied and environmental microbiology85 ( 8 ) 2019年4月

Kojo T, Kawai M, Shiraishi Y, Kurazono S, Kadooka C, Okutsu K, Yoshizaki Y, Ikenaga M, Futagami T, Takamine K, Tamaki H. . Effect of maturation time on koji-like smell and volatile compounds of barley miso (Japanese soybean paste) during fermentation . Food Sci. Technol. Res.25 313 - 319 2019年4月査読

Kadooka C, Izumitsu K, Onoue M, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . Mitochondrial citrate transporters CtpA and YhmA are required for extracellular citric acid accumulation and contribute to cytosolic acetyl-CoA generation in Aspergillus luchuensis mut. kawachii . Appl Environ Microbiol. AEM.03136-18 2019年2月査読

Sobol MS, Hoshino T, Futagami T, Inagaki F, Kiel Reese B. . Draft Genome Sequences of Penicillium spp. from Deeply Buried Oligotrophic Marine Sediments . Microbiol Resour Announc.8 e01613-18 2019年2月査読

Sobol MS, Hoshino T, Futagami T, Inagaki F, Kiel Reese B . Draft Genome Sequences of Penicillium spp. from Deeply Buried Oligotrophic Marine Sediments. . Microbiology resource announcements8 ( 6 ) 2019年2月

Kimoto D, Kadooka C, Saenrungrot P, Okutsu K, Yoshizaki Y, Takamine K, Goto M, Tamaki H, Futagami T . Pex16 is involved in peroxisome and Woronin body formation in the white koji fungus, Aspergillus luchuensis mut. kawachii. . Journal of bioscience and bioengineering127 ( 1 ) 85 - 92 2019年1月

Kojo T. . Effect of Maturation Time on Koji-like Smell and Volatile Compounds of Barley MISO (Japanese Soybean Paste) during Fermentation . Food Science and Technology Research25 ( 2 ) 313 - 319 2019年

Wakamatsu T. . Metal-ion-induced expression of gene fragments from subseafloor micro-organisms in the Kumano forearc basin, Nankai Trough . Journal of Applied Microbiology125 ( 5 ) 1396 - 1407 2018年11月

Kimura N, Watanabe T, Suenaga H, Fujihara H, Futagami T, Goto M, Hanada S, Hirose J. . Pseudomonas furukawaii sp. nov., a polychlorinated biphenyldegrading bacterium isolated from biphenyl-contaminated soil in Japan . Int J Syst Evol Microbiol68 ( 5 ) 1429 - 1435 2018年5月査読

Kan-na Tomioku, Mikiko Shigekuni, Hiroki Hayashi, Akane Yoshida, Taiki Futagami, Hisanori Tamaki, Kenji Tanabe, Akikazu Fujita . Nanoscale domain formation of phosphatidylinositol 4-phosphate in the plasma and vacuolar membranes of living yeast cells . European Journal of Cell Biology 2018年5月査読

Kimura N, Watanabe T, Suenaga H, Fujihara H, Futagami T, Goto M, Hanada S, Hirose J . Pseudomonas furukawaii sp. nov., a polychlorinated biphenyl-degrading bacterium isolated from biphenyl-contaminated soil in Japan. . International journal of systematic and evolutionary microbiology68 ( 5 ) 1429 - 1435 2018年5月

Tomioku KN, Shigekuni M, Hayashi H, Yoshida A, Futagami T, Tamaki H, Tanabe K, Fujita A . Nanoscale domain formation of phosphatidylinositol 4-phosphate in the plasma and vacuolar membranes of living yeast cells. . European journal of cell biology97 ( 4 ) 269 - 278 2018年5月

吉﨑由美子、奥津果優、二神泰基、玉置尚徳、鮫島吉廣、髙峯和則 . プロテアーゼまたは細胞壁分解酵素製剤添加による紅麹焼酎製造法の改良 . 日本醸造教会誌113 ( 4 ) 265 - 272 2018年4月査読

Kojo T, Kadooka C, Komohara M, Onitsuka S, Tanimura M, Muroi Y, Kurazono S, Shiraishi Y, Oda K, Iwashita K, Onoue M, Okutsu K, Yoshizaki Y, Takamine K, Futagami T, Mori K, Tamaki H . Characterization of amylolytic enzyme overproducing mutant of Aspergillus luchuensis obtained by ion beam mutagenesis. . The Journal of general and applied microbiology63 ( 6 ) 339 - 346 2018年1月

Mori K, Kadooka C, Masuda C, Muto A, Okutsu K, Yoshizaki Y, Takamine K, Futagami T, Tamaki H . Genome Sequence of Saccharomyces cerevisiae Strain Kagoshima No. 2, Used for Brewing the Japanese Distilled Spirit Shōchū. . Genome announcements5 ( 41 ) 2017年10月

Futagami T, Kadooka C, Ando Y, Okutsu K, Yoshizaki Y, Setoguchi S, Takamine K, Kawai M, Tamaki H . Multi-gene phylogenetic analysis reveals that shochu-fermenting Saccharomyces cerevisiae strains form a distinct sub-clade of the Japanese sake cluster. . Yeast (Chichester, England)34 ( 10 ) 407 - 415 2017年10月

Suenaga H, Fujihara H, Kimura N, Hirose J, Watanabe T, Futagami T, Goto M, Shimodaira J, Furukawa K . Insights into the genomic plasticity of Pseudomonas putida KF715, a strain with unique biphenyl-utilizing activity and genome instability properties. . Environmental microbiology reports9 ( 5 ) 589 - 598 2017年10月

白石洋平, 安藤有加, 奥津果優, 吉﨑由美子, 二神泰基, 玉置尚徳, 和久豊, 髙峯和則 . 芋焼酎の香気形成に及ぼすアミノ酸の影響 . 日本醸造協会誌112 ( 8 ) 563 - 568 2017年8月査読

Suenaga H, Fujihara H, Kimura N, Hirose J, Watanabe T, Futagami T, Goto M, Shimodaira J, Furukawa K . Insights into the genomic plasticity of Pseudomonas putida KF715, a strain with unique biphenyl-utilizing activity and genome instability properties . Environmental Microbiology Reports 2017年7月査読

白石洋平、安藤有加、奥津果優、吉﨑由美子、二神泰基、玉置尚徳、和久豊、髙峯和則 . 芋焼酎醪へのプロテアーゼ剤添加による揮発成分と官能評価への影響 . 日本醸造協会誌112 ( 7 ) 517 - 523 2017年7月査読

Futagami T, Kadooka C, Ando Y, Okutsu K, Yoshizaki Y, Setoguchi S, Takamine K, Kawai M, Tamaki H. . Multi-gene phylogenetic analysis reveals that shochu-fermenting Saccharomyces cerevisiae strains form a distinct sub-clade of the Japanese sake cluster . Yeast 2017年7月査読

Rahayu YY, Yoshizaki Y, Yamaguchi K, Okutsu K, Futagami T, Tamaki H, Sameshima Y, Takamine K. . Key volatile compounds in red koji-shochu, a Monascus-fermented product, and their formation steps during fermentation . Food Chemistry 2017年6月査読

Rahayu YY, Yoshizaki Y, Yamaguchi K, Okutsu K, Futagami T, Tamaki H, Sameshima Y, Takamine K . Key volatile compounds in red koji-shochu, a Monascus-fermented product, and their formation steps during fermentation. . Food chemistry224 398 - 406 2017年6月

Suenaga H, Yamazoe A, Hosoyama A, Kimura N, Hirose J, Watanabe T, Fujihara H, Futagami T, Goto M, Furukawa K . Complete Genome Sequence of the Polychlorinated Biphenyl-Degrading Bacterium Pseudomonas putida KF715 (NBRC 110667) Isolated from Biphenyl-Contaminated Soil. . Genome announcements5 ( 7 ) 2017年2月

Suenaga H, Yamazoe A, Hosoyama A, Kimura N, Hirose J, Watanabe T, Fujihara H, Futagami T, Goto M, Furukawa K. . Complete genome sequence of the polychlorinated biphenyl-degrading bacterium Pseudomonas putida KF715 (NBRC 110667) isolated from biphenyl-contaminated soil . Genome Announc. 2017年2月

奥津果優、門岡千尋、小城章裕、吉﨑由美子、二神泰基、玉置尚徳、髙峯和則. . 中国及び韓国の市場品「神麹」における菌叢と含有成分の実態調査 . 生薬学雑誌 2017年2月

奥津果優, 門岡千尋, 小城章裕, 吉崎由美子, 二神泰基, 玉置尚徳, 高峯和則 . 中国及び韓国の市場品「神麹」における菌叢と含有成分の実態調査 . 生薬学雑誌71 ( 1 ) 41 - 48 2017年2月

出版者・発行元：(一社)日本生薬学会

中国および韓国で入手した市場品の神麹15種をサンプルとし、菌叢、酵素活性(アミラーゼ、リパーゼ、プロテアーゼ)およびフェルラ酸含量について調査した。Denaturing gradient gel electrophoresis(DGGE)解析の結果、神麹には乳酸菌や枯草菌など複数の微生物が存在し、製品によって微生物の種類が大きく異なることが判明した。また、半数以上のサンプルでアミラーゼ活性およびリパーゼ活性を認め、全サンプルでフェルラ酸の含有を認めた。色調により暗褐色系、白色系、黄褐色系の3群に分類して主成分分析した結果、酵素活性が検出されなかったのは全て黄褐色群で、明度が低いほどフェルラ酸含量が高い傾向であった。以上より、神麹は、色調によって酵素活性およびフェルラ酸含量の予測が可能であると考えられた。

Kojo T, Kadooka C, Komohara M, Onitsuka S, Tanimura M, Muroi Y, Kurazono S, Shiraishi Y, Oda K, Iwashita K, Onoue M, Okutsu K, Yoshizaki Y, Takamine K, Futagami T, Mori K, Tamaki H . Characterization of amylolytic enzyme overproducing mutant of Aspergillus luchuensis obtained by ion beam mutagenesis . The Journal of General and Applied Microbiology 2017年査読

奥津果優, 門岡千尋, 小城章裕, 吉﨑由美子, 二神泰基, 玉置尚徳, 髙峯和則 . 中国及び韓国の市場品「神麹」における菌叢と含有成分の実態調査 . 生薬学雑誌71 ( 1 ) 41 - 48 2017年

出版者・発行元：一般社団法人日本生薬学会

"Shinkiku" is a traditional digestive drug prepared by the fermentation of wheat and some herbs with fermentative microbes. Shinkiku is manufactured in China and Korea, and also used in Japanese Kampo medicine as a component of Hangebyakujutsutemmato. However, there are currently no quality standards for shinkiku, and thus, the quality of shinkiku has considerable variation depending on its manufacturer. Although these variations would be partially derived from the differences in fermentative microbes, there are no studies about microbial diversities or chemical constituents in commercial shinkiku. Thus, we investigated the microbial diversity and chemical constituents of 15 commercial shinkiku samples to standardize its quality. PCR-denaturing gradient gel electrophoresis of 16S rDNA and ITS1 sequences revealed that different microbes such as Lactobacillus sp. and Candida sp. were present in each shinkiku sample. On the other hand, most shinkiku samples showed amylase (12/15 samples) and lipase activities (9/15 samples) that behave as digestants. In addition, all samples commonly contained ferulic acid (>10 nmol/g), which has anti-inflammatory and anti-oxidant activities. Thus, enzyme activities and ferulic acid were suggested to be one of the candidates for use as reference standards for the quality control of shinkiku. Exceptional shinkiku samples without enzyme activities showed a baked brown color, and ferulic acid content was inversely related with the brightness color of shinkiku (R2=0.47). Therefore, it seems that color indices would be effective to predict the quality of shinkiku such as enzyme activities and ferulic acid.

DOI： 10.24684/jspharm.71.1_41