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Genetic engineering on increasing NADPH supply for reductive biotransformation by acetic acid bacteria.

Research Project

Project/Area Number 26450095
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Applied microbiology
Research InstitutionYamaguchi University

Principal Investigator

Yakushi Toshiharu  山口大学, 創成科学研究科, 准教授 (30324388)

Co-Investigator(Kenkyū-buntansha) 松下 一信  山口大学, 創成科学研究科, 教授(特命) (50107736)
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords酢酸菌 / 発酵 / 応用微生物 / 酵素 / バイオテクノロジー / 物質生産 / 還元反応 / NADPH / シキミ酸 / トランスヒドロゲナーゼ / 糖代謝
Outline of Final Research Achievements

Gluconobacter sp., acetic acid bacterium, oxidizes glucose on the outer surface of the cytoplasmic membrane with membrane-bound glucose dehydrogenase (GdhM) and accumulates gluconic acid and its derivatives. Thus, only limited parts of glucose are transported into the cytoplasm and metabolized. Because glucose metabolism in the cytoplasm mostly depends on pentose phosphate pathway, it is reasonably assumed that G. oxydans likely produces much NADPH, if high amounts of glucose are metabolized in the cytoplasm. This study aimed to increase NADPH supply from glucose by inactivating GdhM. This study tried to couple NADPH produced in the ΔgdhM cells and the reduction reaction producing shikimate from dehydroshikimate. When cultivated on glucose, biomass yield and amounts of an end metabolite in the ΔgdhM strain were elevated. Taken together with enzyme activity measurements, some changes on glucose metabolism in the ΔgdhM cells suggested increases in the NADPH level.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (22 results)

All 2017 2016 2015 2014 Other

All Int'l Joint Research (2 results) Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 4 results,  Acknowledgement Compliant: 2 results) Presentation (7 results) Book (2 results) Remarks (3 results)

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

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] Forschungszentrum Juelich(ドイツ)

    • Related Report
      2015 Research-status Report
  • [Journal Article] Mem brane-bound glycerol dehydrogenase catalyzes oxidation of D-pentonbates to 4-keto-D-pentonates, D-fructose to 5-keto-D-fructose, and D-psicose to 5-keto-D-psicose.2017

    • Author(s)
      Y. Ano, R.A. Hours, Y. Akakabe, N. Kataoka, T. Yakushi, K. Matsushita, O. Adachi
    • Journal Title

      Biosci. Biotechnol. Biochem

      Volume: 81 Issue: 2 Pages: 411-418

    • DOI

      10.1080/09168451.2016.1254535

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Complete Genome Sequencing and Comparative Genomic Analysis of the Thermotolerant Acetic Acid Bacterium, <i>Acetobacter pasteurianus</i> SKU1108, Provide a New Insight into Thermotolerance2016

    • Author(s)
      Matsutani, M., Hirakawa, H., Hiraoka, E., Theeragool, G., Yakushi, T. and Matsushita, K.
    • Journal Title

      Microbes and Environments

      Volume: 31 Issue: 4 Pages: 395-400

    • DOI

      10.1264/jsme2.ME16023

    • NAID

      130005434990

    • ISSN
      1342-6311, 1347-4405
    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Genomic analyses of thermotolerant microorganisms used for high-temperature fermentations2016

    • Author(s)
      Matsushita K, Azuma Y, Kosaka T, Yakushi T, Hoshida H, Akada R, Yamada M
    • Journal Title

      Biosci Biotechnol Biochem

      Volume: 80(4) Issue: 4 Pages: 655-668

    • DOI

      10.1080/09168451.2015.1104235

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Determination of Dehydrogenase Activities Involved in D-Glucose Oxidation in Gluconobacter and Acetobacter Strains.2016

    • Author(s)
      Sainz, F., Jesus Torija, M., Matsutani, M., Kataoka, N., Yakushi, T., Matsushita, K. and Mas, A.
    • Journal Title

      Frontiers in Microbiology

      Volume: 7 Pages: 1358-1358

    • DOI

      10.3389/fmicb.2016.01358

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] A functionally critical single nucleotide polymorphism in the gene encoding the membrane-bound alcohol dehydrogenase found in ethanol oxidation-deficient Gluconobacter thailandicus2015

    • Author(s)
      P Charoenyingcharoen, M Matsutani, T Yakushi, G Theeragool, P Yukphan, K Matsushita
    • Journal Title

      Gene

      Volume: in press Issue: 2 Pages: 3552-3560

    • DOI

      10.1016/j.gene.2015.04.080

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient production of 2,5-diketo-D-gluconate via heterologous expression of 2-keto-D-gluconate dehydrogenase in Gluconobacter japonicus.2015

    • Author(s)
      Kataoka N, Matsutani M, Yakushi T, Matsushita K.
    • Journal Title

      Appl. Environ. Microbiol.

      Volume: 81(10) Issue: 10 Pages: 3552-3560

    • DOI

      10.1128/aem.04176-14

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Overexpression of a type II 3-dehydroquinate dehydratase enhances the biotransformation of quinate to 3-dehydroshikimate in Gluconobacter oxydans2014

    • Author(s)
      Nishikura-Imamura S, Matsutani M, Insomphun C, Vangnai AS, Toyama H, Yakushi T, Abe T, Adachi O, Matsushita K.
    • Journal Title

      Appl Microbiol Biotechnol

      Volume: 98 Issue: 7 Pages: 2955-2963

    • DOI

      10.1007/s00253-013-5439-z

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Replacement of a terminal cytochrome c oxidase by ubiquinol oxidase during the evolution of acetic acid bacteria2014

    • Author(s)
      Matsutani M, Fukushima K, Kayama C, Arimitsu M, Hirakawa H, Toyama H, Adachi O, Yakushi T, Matsushita K.
    • Journal Title

      Biochim Biophys Acta

      Volume: 1837 Issue: 10 Pages: 1810-1820

    • DOI

      10.1016/j.bbabio.2014.05.355

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Presentation] 酢酸菌Gluconobacter thailandicusにおける ジヒドロキシアセトン・リン酸化酵素の遺伝学的・生化学的解析2017

    • Author(s)
      平田 花織,松谷 峰之介,足立 収生,片岡 尚也,薬師 寿治,松下 一信
    • Organizer
      日本農芸化学会 2017年度大会
    • Place of Presentation
      京都女子大(京都府京都市)
    • Year and Date
      2017-03-18
    • Related Report
      2016 Annual Research Report
  • [Presentation] Gluconobacter属酢酸菌キノプロテイン・グリセロール脱水素酵素における新たな基質の発見とその反応メカニズムの提唱2017

    • Author(s)
      寺田 優花,尾崎 聖士朗,片岡 尚也,足立 収生,赤壁 善彦,薬師 寿治,松下 一信
    • Organizer
      日本農芸化学会 2017年度大会
    • Place of Presentation
      京都女子大(京都府京都市)
    • Year and Date
      2017-03-18
    • Related Report
      2016 Annual Research Report
  • [Presentation] Gluconobacter属酢酸菌由来膜結合型キナ酸脱水素酵素の高発現2016

    • Author(s)
      小松和貴,藥師寿治,松谷峰之介,吉原望,片岡尚也,足立収生,松下一信
    • Organizer
      日本農芸化学会 2016年度中四国支部大会
    • Place of Presentation
      高知大学(高知県高知市)
    • Year and Date
      2016-09-15
    • Related Report
      2016 Annual Research Report
  • [Presentation] 酢酸菌Gluconobacter thailandicusのジヒドロキシアセトン代謝に関する研究:ジヒドロキシアセトンリン酸化酵素2016

    • Author(s)
      平田花織,松谷峰之介,足立収生, 片岡尚也,藥師寿治,松下一信
    • Organizer
      日本農芸化学会大会
    • Place of Presentation
      札幌コンベンションセンター(北海道札幌市)
    • Year and Date
      2016-03-30
    • Related Report
      2015 Research-status Report
  • [Presentation] Gluconobacter oxydansにおける細胞内グルコース代謝の活性化2016

    • Author(s)
      小池尚史,松谷峰之介,片岡尚也,藥師寿治,松下一信
    • Organizer
      日本農芸化学会中四国支部例会
    • Place of Presentation
      岡山県立大学(岡山県総社市)
    • Year and Date
      2016-01-23
    • Related Report
      2015 Research-status Report
  • [Presentation] 酢酸菌Gluconobacter oxydansの膜結合型キナ酸脱水素酵素の高発現2015

    • Author(s)
      小松 和貴, 吉原 望, 今村 志穂美, 片岡 尚也, 薬師 寿治, 足立 収生, 松下 一信
    • Organizer
      日本農芸化学会 2015年度大会
    • Place of Presentation
      岡山大学 津島キャンパス (岡山市)
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 2-ケト-D-グルコン酸脱水素酵素の異種発現によるGluconobacter japonicusでの効率的2,5-ジケト-D-グルコン酸生産2015

    • Author(s)
      片岡 尚也, 松谷 峰之介, 藥師 寿治, 松下 一信
    • Organizer
      日本農芸化学会 2015年度大会
    • Place of Presentation
      岡山大学 津島キャンパス (岡山市)
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Book] Acetic acid bacteria: Ecology and Physiology2016

    • Author(s)
      Edited by Matsushita, K., Toyama, H., Tonouchi, N., and Okamoto-Kainuma, A.
    • Total Pages
      350
    • Publisher
      Springer
    • Related Report
      2016 Annual Research Report
  • [Book] Acetic Acid Bacteria - Ecology and Physiology -2016

    • Author(s)
      Adachi, O. and Yakushi, T.
    • Total Pages
      347
    • Publisher
      Springer
    • Related Report
      2015 Research-status Report
  • [Remarks]

    • URL

      http://web.cc.yamaguchi-u.ac.jp/~oubi/

    • Related Report
      2016 Annual Research Report
  • [Remarks] http://web.cc.yamaguchi-u.ac.jp/~oubi/

    • Related Report
      2015 Research-status Report
  • [Remarks] 山口大学農学部応用微生物学研究室

    • URL

      http://web.cc.yamaguchi-u.ac.jp/~oubi/

    • Related Report
      2014 Research-status Report

URL: 

Published: 2014-04-04   Modified: 2018-03-22  

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