Takino Junya
| Faculty of Science | Assistant Professor |
Last Updated :2026/01/07
■Researcher basic information
Mail Address
- juntak
sci.hokudai.ac.jp
Researchmap personal page
Researcher number
- 10979082
J-Global ID
Educational Organization
- Bachelor's degree program, School of Science
- Master's degree program, Graduate School of Chemical Sciences and Engineering
- Doctoral (PhD) degree program, Graduate School of Chemical Sciences and Engineering
■Career
Career
Educational Background
■Research activity information
Papers
- Chain-length preference of trans-acting enoylreductases involved in the biosynthesis of fungal polyhydroxy polyketides
Yuichiro Takekawa, Junya Takino, Shusuke Sato, Hideaki Oikawa, Toyoyuki Ose, Atsushi Minami
Biochemical and Biophysical Research Communications, 761, 151737, 151737, Elsevier BV, May 2025, [Lead author]
Scientific journal - A fungal transcription factor converts a beneficial root endophyte into an anthracnose leaf pathogen
Ren Ujimatsu, Junya Takino, Seishiro Aoki, Masami Nakamura, Hiromi Haba, Atsushi Minami, Kei Hiruma
Current Biology, 35, 9, 1989, 2005.e6, Elsevier BV, May 2025
Scientific journal - Bioinformatics-Guided Reconstitution of Biosynthetic Machineries of Fungal Eremophilane Sesquiterpenes
Yoshiro Sato, Xinge Shi, Ying Ye, Saori Domon, Junya Takino, Taro Ozaki, Chengwei Liu, Hideaki Oikawa, Atsushi Minami
ACS Chemical Biology, 19, 4, 861, 865, American Chemical Society (ACS), 03 Apr. 2024
Scientific journal - A fungal sesquiterpene biosynthesis gene cluster critical for mutualist-pathogen transition in Colletotrichum tofieldiae
Kei Hiruma, Seishiro Aoki, Junya Takino, Takeshi Higa, Yuniar Devi Utami, Akito Shiina, Masanori Okamoto, Masami Nakamura, Nanami Kawamura, Yoshihiro Ohmori, Ryohei Sugita, Keitaro Tanoi, Toyozo Sato, Hideaki Oikawa, Atsushi Minami, Wataru Iwasaki, Yusuke Saijo
Nature Communications, 14, 1, Springer Science and Business Media LLC, 06 Sep. 2023
Scientific journal, Abstract
Plant-associated fungi show diverse lifestyles from pathogenic to mutualistic to the host; however, the principles and mechanisms through which they shift the lifestyles require elucidation. The root fungus Colletotrichum tofieldiae (Ct) promotes Arabidopsis thaliana growth under phosphate limiting conditions. Here we describe a Ct strain, designated Ct3, that severely inhibits plant growth. Ct3 pathogenesis occurs through activation of host abscisic acid pathways via a fungal secondary metabolism gene cluster related to the biosynthesis of sesquiterpene metabolites, including botrydial. Cluster activation during root infection suppresses host nutrient uptake-related genes and changes mineral contents, suggesting a role in manipulating host nutrition state. Conversely, disruption or environmental suppression of the cluster renders Ct3 beneficial for plant growth, in a manner dependent on host phosphate starvation response regulators. Our findings indicate that a fungal metabolism cluster provides a means by which infectious fungi modulate lifestyles along the parasitic–mutualistic continuum in fluctuating environments. - Elucidation of Late-Stage Biosynthesis of Phomoidride: Proposal of Cyclization Mechanism Affording Characteristic Nine-Membered Ring of Fungal Dimeric Anhydride
Shintaro Yamamoto, Taro Matsuyama, Taro Ozaki, Junya Takino, Hajime Sato, Masanobu Uchiyama, Atsushi Minami, Hideaki Oikawa
Journal of the American Chemical Society, 144, 46, 20998, 21004, American Chemical Society (ACS), 14 Nov. 2022
Scientific journal - Heterologous expression of a polyketide synthase ACRTS2 in Aspergillus oryzae produces host-selective ACR toxins: coproduction of minor metabolites
Akari Kotani, Taro Ozaki, Junya Takino, Susumu Mochizuki, Kazuya Akimitsu, Atsushi Minami, Hideaki Oikawa
Bioscience, Biotechnology, and Biochemistry, 86, 3, 287, 293, Informa UK Limited, 11 Dec. 2021
Scientific journal, ABSTRACT
Previously, we succeeded to produce the core structure of the host-selective ACR toxin (1) on brown leaf spot on rough lemon when the polyketide synthase ACRTS2 gene was heterologously expressed in Aspergillus oryzae (AO). To confirm the production of 1 in AO, the detection limit and suppressing decarboxylation were improved, and these efforts led us to conclude the direct production of 1 instead of its decarboxylation product. During this examination, minor ACR-toxin-related metabolites were found. Their structure determination enabled us to propose a decarboxylation mechanism and a novel branching route forming byproducts from the coupling of the dihydropyrone moiety of 1 with the acetaldehyde and kojic acid abundant in AO. The involvement of putative cyclase ACRTS3 in the chain release of linear polyketide was excluded by the coexpression analysis of ACRTS2 and ACRTS3. Taken together, we concluded that the production of 1 in AO is solely responsible for ACRTS2. - Biochemistry‐Guided Prediction of the Absolute Configuration of Fungal Reduced Polyketides
Junya Takino, Akari Kotani, Taro Ozaki, Wenquan Peng, Jie Yu, Yian Guo, Susumu Mochizuki, Kazuya Akimitsu, Masaru Hashimoto, Tao Ye, Atsushi Minami, Hideaki Oikawa
Angewandte Chemie International Edition, 60, 43, 23403, 23411, Wiley, 14 Sep. 2021, [Lead author]
Scientific journal, Abstract
Highly reducing polyketide synthases (HR‐PKSs) produce structurally diverse polyketides (PKs). The PK diversity is constructed by a variety of factors, including the β‐keto processing, chain length, methylation pattern, and relative and absolute configurations of the substituents. We examined the stereochemical course of the PK processing for the synthesis of polyhydroxy PKs such as phialotides, phomenoic acid, and ACR‐toxin. Heterologous expression of a HR‐PKS gene, a trans‐acting enoylreductase gene, and a truncated non‐ribosomal peptide synthetase gene resulted in the formation of a linear PK with multiple stereogenic centers. The absolute configurations of the stereogenic centers were determined by chemical degradation followed by comparison of the degradation products with synthetic standards. A stereochemical rule was proposed to explain the absolute configurations of other reduced PKs and highlights an error in the absolute configurations of a reported structure. The present work demonstrates that focused functional analysis of functionally related HR‐PKSs leads to a better understanding of the stereochemical course. - Elucidation of biosynthetic pathway of a plant hormone abscisic acid in phytopathogenic fungi
Junya Takino, Takuto Kozaki, Taro Ozaki, Chengwei Liu, Atsushi Minami, Hideaki Oikawa
Bioscience, Biotechnology, and Biochemistry, 83, 9, 1642, 1649, Informa UK Limited, 02 Sep. 2019, [Lead author]
Scientific journal, ABSTRACT
Abscisic acid (ABA) is one of the plant hormones that regulates physiological functions in various organisms, including plants, sponges, and humans. The biosynthetic machinery in plants is firmly established, while that in fungi is still unclear. Here, we elucidated the functions of the four biosynthetic genes, bcABA1-bcABA4, found in Botrytis cinerea by performing biotransformation experiments and in vitro enzymatic reactions with putative biosynthetic intermediates. The first-committed step is the cyclization of farnesyl diphosphate to give α-ionylideneethane catalyzed by a novel sesquiterpene synthase, BcABA3, which exhibits low amino acid sequence identities with sesquiterpene synthases. Subsequently, two cytochrome P450s, BcABA1 and BcABA2, mediate oxidative modifications of the cyclized product to afford 1ʹ,4ʹ-trans-dihydroxy-α-ionylideneacetic acid, which undergoes alcohol oxidation to furnish ABA. Our results demonstrated that production of ABA does not depend on the nucleotide sequence of bcABA genes. The present study set the stage to investigate the role of ABA in infections. - Unveiling Biosynthesis of the Phytohormone Abscisic Acid in Fungi: Unprecedented Mechanism of Core Scaffold Formation Catalyzed by an Unusual Sesquiterpene Synthase
Junya Takino, Takuto Kozaki, Yoshiro Sato, Chengwei Liu, Taro Ozaki, Atsushi Minami, Hideaki Oikawa
Journal of the American Chemical Society, 140, 39, 12392, 12395, American Chemical Society (ACS), 18 Sep. 2018, [Lead author]
Scientific journal
Research Themes
- 酵素固定化を基軸とした天然物ストリームライン合成
科学研究費助成事業
31 Aug. 2023 - 31 Mar. 2025
瀧野 純矢
有用な天然資源である天然物は、主に化学反応による化学合成と酵素反応によるバイオ合成によって供給される。しかしながら、化学合成では工程数の多さ、バイオ合成では汎用性の乏しさなど、従来の合成法はそれぞれに課題がある。本研究課題では、酵素を固相担体に担持した固定化酵素を利用し、化学反応と酵素反応を連続的に行うことで、従来法の課題を解決するストリームライン合成法の確立を目指した。
本研究課題では、１）酵素の固定化、２）膜タンパクの固定化、３）フロー法への展開の３段階で計画した。本年度は、このうち１）２）について検討を行なった。１）では、予備検討で利用した還元酵素を、フロー法への利用を見据えた圧力損失の少ない4種の担体に吸着させ、酵素の担持量および酵素活性を測定した。その結果、新たに検討した全ての担体において酵素の吸着は行われるものの、吸着能の高さや担体構成成分などが問題となり、十分な酵素活性が得られなかった。２）では、固定化酵素を利用した反応系の適用範囲を拡張するため、天然物生合成で頻繁に利用される酸化酵素シトクロムP450の固定化検討を行なった。本酵素は生体内において細胞膜と結合している膜結合型タンパクである。そのため、酵素全体から膜結合領域を除いた部分のみをクローニングし、大腸菌を用いた組換タンパクの調製、酵素活性測定を行なった。現在までに、検討したP450について酵素の取得に成功したが、酵素活性は見られていない。
日本学術振興会, 研究活動スタート支援, 北海道大学, 23K19277 - ゲノム編集を基軸とした巨大酵素の機能解析とエンジニアリング
科学研究費助成事業
24 Apr. 2020 - 31 Mar. 2023
瀧野 純矢
本研究課題は、ゲノム編集を基軸とした巨大酵素の機能解析とエンジニアリングを目的として研究を進めている。昨年度までに、ゲノム編集による麹菌ホットスポットへの遺伝子導入によって、繰り返し型ポリケタイド合成酵素 (HR-PKS)の網羅的解析を行い、立体制御の一般的な規則を提唱した。本年度は、HR-PKSによる骨格形成以降の反応に焦点をあて、酸無水物二量体ホモイドライドB生合成を研究対象として、構造多様性創出に寄与する酵素遺伝子の探索を行った。
ホモイドライドBはコレステロール値低下作用を示す糸状菌由来天然物であり、無水マレイン型モノマーが二量化した構造を有する。糸状菌は同様のモノマーによって合成される酸無水物二量体を多数生産するが、ホモイドライドBは他の類縁体には見られない特徴的なカゴ型構造を有している。本年度は、カゴ型構造形成機構を明らかにし、ポリケタイド系天然物の構造多様性創出の鍵となる酵素の探索を行った。以前に構築していた骨格構造生産麹菌に対して、生合成遺伝子クラスター中の機能未解析遺伝子4種の追加導入を行った結果、最終産物ホモイドライドBを観測することに成功した。次いで、ゲノム編集による迅速な機能解析によって、導入遺伝子の組み合わせを変えた形質転換体を作製することで、メチル基転移酵素がカゴ型構造形成の鍵酵素であることを明らかにした。本酵素の反応機構を詳細に解析する為、大腸菌により組換えタンパク質を調製し、補酵素の有無、阻害剤の添加、メチル化副生成物の構造決定によって、SAM依存的メチル化を起点としたアセタール形成によって、カゴ型構造が構築されることを明らかにした。この機構は初の解析例であることに加え、TstEホモログが類縁PKSクラスターでも観測されたことから、同様のカゴ型構造を有する天然物探索にも展開できると考えられる。
日本学術振興会, 特別研究員奨励費, 北海道大学, 20J20880
