研究者詳細 - 二神　泰基

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