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Periplasmic metabolic engineering in acetic acid bacteria

Research Project

Project/Area Number 20H02902
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 38020:Applied microbiology-related
Research InstitutionYamaguchi University

Principal Investigator

Yakushi Toshiharu  山口大学, 大学研究推進機構, 教授 (30324388)

Co-Investigator(Kenkyū-buntansha) 阿野 嘉孝  愛媛大学, 農学研究科, 准教授 (00403642)
Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2022: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2021: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000)
Keywords応用微生物学 / 発酵 / バイオテクノロジー / 酵素 / 酢酸菌 / 代謝工学 / 応用微生物 / イメージング / タンパク質細胞内局在
Outline of Research at the Start

酢酸菌はペリプラズム空間と呼ばれる細胞表層に物質酸化系持つという特徴を持つ。ここに,酸化反応以外の代謝系を移植する「ペリプラズミック代謝工学」を施す。細胞質での代謝に比べて,①細胞質への取り込みや細胞質からの排出に関わる時間とエネルギーの抑制,②毒性物質そのもの,あるいは毒性の中間体を経る代謝経路の構築,③代謝上不安定で短寿命な化合物の生産,に優位性がある。ペリプラズム空間で営まれる代謝の,物質生産における優位性を実験で検証する。これを達成するために,酸化反応に加え,脱水,水和,脱炭酸反応をペリプラズム空間に持ち込む。この実現によって代謝工学分野に新しい可能性を提示する。

Outline of Final Research Achievements

Acetic acid bacteria have a unique oxidation system on the cell surface and therefor, have been used for acetic acid fermentation and also sorbose fermentation in vitamin C production process. Considering that metabolism in the periplasmic space, which is the intracellular compartment of bacteria, has advantages over metabolism in the cytoplasm, this aimed at investigation for efficacy of "periplasmic metabolic engineering" that expands metabolism in the periplasm. The first attempt was to relocate a dehydratase (lyase), which is localized in the cytoplasm in the acetic acid bacteria, to the periplasm. Next, we tried to translocate isomerase, which is naturally localized in the cytoplasm. Both periplasmic metabolic engineering increased the biotransformation rate. Expression of the protein involved in transport to the periplasm also increased the bioconversion rate.

Academic Significance and Societal Importance of the Research Achievements

持続可能な発展的経済活動の必要性や二酸化炭素排出低減が訴えられている中,微生物を用いた物質生産は廃棄物利用や環境負荷の低減という意味で魅力的なソリューションの一つである。本研究は,古くから物質生産に利用されている酢酸菌を用いた,新しい概念の代謝工学の検証を行った。いくつかの良好な結果を得ることができたので,当初期待していた新しい代謝工学の有効性を確認することができた。本菌でのさらなる展開はさることながら,大腸菌などの他の微生物を用いた展開が想定できる。この研究が,広く微生物による代謝工学の考え方を変え,新しい物質生産系の構築という形で波及していくことを期待する。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (25 results)

All 2023 2022 2021 2020 Other

All Int'l Joint Research (6 results) Journal Article (9 results) (of which Int'l Joint Research: 6 results,  Peer Reviewed: 9 results,  Open Access: 4 results) Presentation (8 results) Remarks (2 results)

  • [Int'l Joint Research] ユーリッヒ研究所(ドイツ)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] カセサート大学/ラジャマンガラ工科大学/メイファールアン大学(タイ)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] カセサート大学/ラジャマンガラ工科大学(タイ)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] ユーリッヒ研究所(ドイツ)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] カセサート大学/ラジャマンガラ工科大学(タイ)

    • Related Report
      2020 Annual Research Report
  • [Int'l Joint Research] ユーリッヒ研究所(ドイツ)

    • Related Report
      2020 Annual Research Report
  • [Journal Article] Development of efficient 5-ketogluconate production system by Gluconobacter japonicus2022

    • Author(s)
      Kataoka N, Naoki K, Ano Y, Matsushita K, Yakushi T.
    • Journal Title

      Appl Microbiol Biotechnol.

      Volume: 106 Issue: 23 Pages: 7751-7761

    • DOI

      10.1007/s00253-022-12242-0

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Periplasmic dehydroshikimate dehydratase combined with quinate oxidation in Gluconobacter oxydans for protocatechuate production2022

    • Author(s)
      Nagaki K, Kataoka N, Theeragool G, Matsutani M, Ano Y, Matsushita K, Yakushi T.
    • Journal Title

      Biosci Biotechnol Biochem.

      Volume: 86 Issue: 8 Pages: 1151-1159

    • DOI

      10.1093/bbb/zbac090

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Membrane-bound D-mannose isomerase of acetic acid bacteria: finding, characterization, and application.2022

    • Author(s)
      Osao Adachi, Naoya Kataoka, Kazunobu Matsushita, Yoshihiko Akakabe, Toshihiro Harada, Toshiharu Yakushi.
    • Journal Title

      Bioscience. Biotechnology. Biochem.

      Volume: 86 Issue: 7 Pages: 938-948

    • DOI

      10.1093/bbb/zbac049

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characterization of 3 phylogenetically distinct membrane-bound D-gluconate dehydrogenases of Gluconobacter spp. and their biotechnological application for efficient 2-keto-D-gluconate production2022

    • Author(s)
      Kataoka Naoya、Saichana Natsaran、Matsutani Minenosuke、Toyama Hirohide、Matsushita Kazunobu、Yakushi Toshiharu
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 86 Issue: 5 Pages: 681-690

    • DOI

      10.1093/bbb/zbac024

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Mutations in degP and spoT Genes Mediate Response to Fermentation Stress in Thermally Adapted Strains of Acetic Acid Bacterium Komagataeibacter medellinensis NBRC 32882022

    • Author(s)
      Kataoka N., Matsutani M., Matsumoto N., Oda M., Mizumachi Y., Ito K., Tanaka S., and Kanesaki Y., Yakushi T., and Matsushita K.
    • Journal Title

      Frontiers in Microbiology

      Volume: 13 Pages: 802010-802010

    • DOI

      10.3389/fmicb.2022.802010

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Dissection and Reconstitution Provide Insights into Electron Transport in the Membrane-Bound Aldehyde Dehydrogenase Complex of Gluconacetobacter diazotrophicus2022

    • Author(s)
      Miah Roni、Nina Shun、Murate Takeru、Kataoka Naoya、Matsutani Minenosuke、Ano Yoshitaka、Matsushita Kazunobu、Yakushi Toshiharu
    • Journal Title

      Journal of Bacteriology

      Volume: 204 Issue: 3 Pages: 1-15

    • DOI

      10.1128/jb.00558-21

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Relocation of dehydroquinate dehydratase to the periplasmic space improves dehydroshikimate production with Gluconobacter oxydans strain NBRC32442021

    • Author(s)
      Nakamura Kentaro、Nagaki Kakeru、Matsutani Minenosuke、Adachi Osao、Kataoka Naoya、Ano Yoshitaka、Theeragool Gunjana、Matsushita Kazunobu、Yakushi Toshiharu
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 105 Issue: 14-15 Pages: 5883-5894

    • DOI

      10.1007/s00253-021-11476-8

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Major aldehyde dehydrogenase AldFGH of Gluconacetobacter diazotrophicus is independent of pyrroloquinoline quinone but dependent on molybdopterin for acetic acid fermentation2021

    • Author(s)
      Miah Roni、Nina Shun、Murate Takeru、Kataoka Naoya、Matsutani Minenosuke、Matsushita Kazunobu、Yakushi Toshiharu
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 105 Issue: 6 Pages: 2341-2350

    • DOI

      10.1007/s00253-021-11144-x

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The Auxiliary NADH Dehydrogenase Plays a Crucial Role in Redox Homeostasis of Nicotinamide Cofactors in the Absence of the Periplasmic Oxidation System in Gluconobacter oxydans NBRC32932020

    • Author(s)
      Sriherfyna Feronika Heppy、Matsutani Minenosuke、Hirano Kensuke、Koike Hisashi、Kataoka Naoya、Yamashita Tetsuo、Nakamaru-Ogiso Eiko、Matsushita Kazunobu、Yakushi Toshiharu
    • Journal Title

      Applied and Environmental Microbiology

      Volume: 87 Issue: 2 Pages: 1-14

    • DOI

      10.1128/aem.02155-20

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] 酢酸菌Gluconobacter thailandicus の2つの第二級アルコール脱水素酵素が関与する2,3-ブタンジオール代謝2023

    • Author(s)
      浅野連太朗,小東 陽,後藤 勝,平田花織,吉冨 宙,片岡 尚也,松下 一信,藥師 寿治
    • Organizer
      日本農芸化学会2023年度大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] グルコノバクター属酢酸菌の細胞表層酸化系代謝の細胞増殖における役割2023

    • Author(s)
      吉冨 宙,平田花織,片岡 尚也,松下 一信,藥師 寿治
    • Organizer
      日本農芸化学会2023年度大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Acetobacter pasteurianus の酢酸耐性因子AarCとそのパラログによる酢酸代謝2023

    • Author(s)
      村上 果穂,片岡 尚也,松下 一信,松谷 峰之介,藥師 寿治
    • Organizer
      日本農芸化学会2023年度大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Komagataeibacter 属酢酸菌の遺伝子操作法の開発2022

    • Author(s)
      山下 璃貢,片岡 尚也,松谷 峰之介,Gunjana Theeragool,松下 一信,藥師 寿治
    • Organizer
      日本農芸化学会中四国支部 第62回講演会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 酢酸菌膜結合型アルデヒド脱水素酵素における鉄・硫黄クラスターの関与2022

    • Author(s)
      和泉 理緒,Roni Miah,片岡 尚也,松下 一信,藥師 寿治
    • Organizer
      第95回日本生化学会大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Acinetobacter baylyi 由来ペリプラズム型デヒドロシキミ酸脱水酵素の異種発現とプロトカテク酸生産への利用2022

    • Author(s)
      永木 翔、松谷 峰之介、片岡 尚也、松下 一信、藥師 寿治
    • Organizer
      日本農芸化学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] キナ酸からプロトカテク酸を生産するための酢酸菌のペリプラズミック代謝工学2021

    • Author(s)
      永木翔,松谷峰之介,片岡尚也,松下一信,藥師寿治
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 酵素の細胞内局在を改変することによる酢酸菌のペリプラズミック酸化系の拡張2020

    • Author(s)
      中村謙太郎,永木翔,松谷峰之介,片岡尚也,松下一信,足立収生,藥師寿治
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Remarks] 山口大学応用微生物学研究室

    • URL

      http://ds0n.cc.yamaguchi-u.ac.jp/~obi/index.html

    • Related Report
      2022 Annual Research Report 2021 Annual Research Report 2020 Annual Research Report
  • [Remarks] 山口大学中高温微生物研究センター

    • URL

      https://ds0n.cc.yamaguchi-u.ac.jp/~yurctmr/

    • Related Report
      2022 Annual Research Report 2021 Annual Research Report 2020 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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