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• Study on lactic acid bacteria

• Application of Awamori Moromi vinegar

• Analysis of microbes in Awamori Brewing

• Defence Proteins of Tropical Plant

• Relationship Between Structure and Antifungal Activity of Plant Chitinases

Published Papers 【 display / non-display 】

Published Papers 【 display / non-display 】

• Enhancement of the Antifungal Activity of Chitinase by Palmitoylation and the Synergy of Palmitoylated Chitinase with Amphotericin B.

  Santoso P, Minamihata K, Ishimine Y, Taniguchi H, Komada T, Sato R, Goto M, Takashima T, Taira T, Kamiya N

  ACS infectious diseases     2022.04 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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• GH-16 type β-1,3-glucanase from <i>Lysobacter</i> sp. MK9-1 enhances antifungal activity of GH-19 type chitinase, and its glucan-binding domain binds to fungal cell-wall

  Otsuka Yuitsu, Sato Koki, Yano Shigekazu, Kanno Haruki, Suyotha Wasana, Konno Hiroyuki, Makabe Koki, Taira Toki

  Ｊｏｕｒｎａｌ　ｏｆ　Ａｐｐｌｉｅｄ　Ｇｌｙｃｏｓｃｉｅｎｃｅ ( 日本応用糖質科学会 )  advpub ( 0 )   2022.04 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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• Identification of Genes Involved in the Synthesis of the Fungal Cell Wall Component Nigeran and Regulation of Its Polymerization in Aspergillus <i>luchuensis</i>.

  Uechi K, Yaguchi H, Tokashiki J, Taira T, Mizutani O

  Applied and environmental microbiology   87 ( 21 ) e0114421   2021.10 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

  　View Summary

  Certain Aspergillus and Penicillium spp. produce the fungal cell wall component nigeran, an unbranched D-glucan with alternating α-1,3- and α-1,4-glucoside linkages, under nitrogen starvation. The mechanism underlying nigeran biosynthesis and the physiological role of nigeran in fungal survival are not clear. We used RNA-seq to identify genes involved in nigeran synthesis in the filamentous fungus Aspergillus luchuensis when grown under nitrogen-free conditions. agsB, which encodes a putative α-1,3-glucan synthase, and two adjacent genes (agtC and gnsA) were upregulated under conditions of nitrogen starvation. Disruption of agsB in A. luchuensis (ΔagsB) resulted in the complete loss of nigeran synthesis. Furthermore, overexpression of agsB in an Aspergillus oryzae strain that cannot produce nigeran resulted in nigeran synthesis. These results indicated that agsB encodes a nigeran synthase. Therefore, we have renamed the A. luchuensis agsB as nigeran synthase gene (nisA). Nigeran synthesis in an agtC mutant (ΔagtC) increased to 121%; conversely, that in ΔgnsA and ΔagtCgnsA decreased to 64% and 63%, respectively, compared to that in the wild-type strain. Our results revealed that AgtC and GnsA play an important role in regulating not only the quantity of nigeran but also its polymerization. Collectively, our results demonstrated that nisA (agsB) is essential for nigeran synthesis in A. luchuensis, whereas agtC and gnsA contribute to the regulation of nigeran synthesis and its polymerization. This research provides insights into fungal cell wall biosynthesis, specifically the molecular evolution of fungal α-glucan synthase genes and the potential utilization of nigeran as a novel biopolymer. Importance The fungal cell wall is composed mainly of polysaccharides. Under nitrogen-free conditions, some Aspergillus and Penicillium spp. produce significant levels of nigeran, a fungal cell wall polysaccharide composed of alternating α-1,3-/1,4-glucosidic linkages. The mechanisms regulating the biosynthesis and function of nigeran are unknown. Here, we performed RNA sequencing of Aspergillus luchuensis cultured under nitrogen-free or low-nitrogen conditions. A putative α-1,3-glucan synthase gene, whose transcriptional level was upregulated under nitrogen-free conditions, was demonstrated to encode nigeran synthase. Furthermore, two genes encoding an α-glucanotransferase and a hypothetical protein were shown to be involved in controlling nigeran content and molecular weight. This study reveals genes involved in the synthesis of nigeran, a potential biopolymer, and provides a deeper understanding of fungal cell wall biosynthesis.

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• Orthogonal Enzymatic Conjugation Reactions Create Chitin Binding Domain Grafted Chitinase Polymers with Enhanced Antifungal Activity.

  Minamihata K, Tanaka Y, Santoso P, Goto M, Kozome D, Taira T, Kamiya N

  Bioconjugate chemistry ( Bioconjugate Chemistry )  32 ( 8 ) 1688 - 1698   2021.08 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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• cDNA cloning, expression, and antifungal activity of chitinase from Ficus microcarpa latex: difference in antifungal action of chitinase with and without chitin-binding domain.

  Takashima T, Henna H, Kozome D, Kitajima S, Uechi K, Taira T

  Planta ( Planta )  253 ( 6 ) 120 - 120   2021.05 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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  Main conclusion A chitin-binding domain could contribute to the antifungal ability of chitinase through its affinity to the fungal lateral wall by hydrophobic interactions. Complementary DNA encoding the antifungal chitinase of gazyumaru (Ficus microcarpa), designated GlxChiB, was cloned and expressed in Escherichia coli cells. The results of cDNA cloning showed that the precursor of GlxChiB has an N-terminal endoplasmic reticulum targeting signal and C-terminal vacuolar targeting signal, whereas mature GlxChiB is composed of an N-terminal carbohydrate-binding module family-18 domain (CBM18) and a C-terminal glycoside hydrolase family-19 domain (GH19) with a short linker. To clarify the role of the CBM18 domain in the antifungal activity of chitinase, the recombinant GlxChiB (wild type) and its catalytic domain (CatD) were used in quantitative antifungal assays under different ionic strengths and microscopic observations against the fungus Trichoderma viride. The antifungal activity of the wild type was stronger than that of CatD under all ionic strength conditions used in this assay; however, the antifungal activity of CatD became weaker with increasing ionic strength, whereas that of the wild type was maintained. The results at high ionic strength further verified the contribution of the CBM18 domain to the antifungal ability of GlxChiB. The microscopic observations clearly showed that the wild type acted on both the tips and the lateral wall of fungal hyphae, while CatD acted only on the tips. These results suggest that the CBM18 domain could contribute to the antifungal ability of chitinase through its affinity to the fungal lateral wall by hydrophobic interactions.

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Other Papers 【 display / non-display 】

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• Phenolic acid decarboxylase of <i>Aspergillus luchuensis</i> plays a crucial role in 4-vinylguaiacol production during <i>awamori</i> brewing

  Seibutsu-kogaku Kaishi ( The Society for Biotechnology, Japan )  100 ( 2 ) 79 - 79   2022.02

   

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Presentations 【 display / non-display 】

Presentations 【 display / non-display 】

• Development of antifungal system based on chitinases

  Toki Taira, Haruna Tsuhako, Shigekazu Yano, Keiko Uechi

  Workshop on Chitin Biology and Beyond  (大連（中国）)  2019.09  -  2019.09 

• Antifungal activity of Pteris ryukyuensis chitinase-A and its enhancement with LysM domain multimers

  Tomoya Takashima, Ryo Sunagawa, Keiko Uechi, Toki Taira

  Workshop on Chitin Biology and Beyond  (大連（中国）)  2019.09  -  2019.09 

• Antifungal activities of LysM domain multimers and their fusion chitinases

  Tomoya Takashima, Keiko Uechi, Toki Taira

  14th International Chitin and Chitosan Conference  (Osaka)  2018.08  -  2018.08 

• Lysin motifs from Plant Chitinases

  Y. Kitaoku, S. Nishimura, T. Numata, W. Suginta, T. Taira, T. Fukamizo, T. Ohnuma

  14th International Chitin and Chitosan Conference  (Osaka)  2018.08  -  2018.08 

• Antifungal activity of chitinases

  Toki Taira

  14th International Chitin and Chitosan Conference  (Osaka)  2018.08  -  2018.08 

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Grant-in-Aid for Scientific Research 【 display / non-display 】

Grant-in-Aid for Scientific Research 【 display / non-display 】

• Grant-in-Aid for Young Scientists(B)

  Project Year: 2009  -  2010 

  Direct: 2,500,000 (YEN)  Overheads: 750,000 (YEN)  Total: 3,250,000 (YEN)

Preferred joint research theme 【 display / non-display 】

Preferred joint research theme 【 display / non-display 】

• Study on lactic acid bacteria

• Application of Awamori Moromi vinegar

• Analysis of microbes in Awamori Brewing

• Defence Proteins of Tropical Plant

• Relationship Between Structure and Antifungal Activity of Plant Chitinases