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High-density cultivation method that does not allow lactic acid bacteria to produce lactic acid

Research Project

Project/Area Number 21K05372
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38020:Applied microbiology-related
Research InstitutionKansai University

Principal Investigator

Katakura Yoshio  関西大学, 化学生命工学部, 教授 (50263207)

Co-Investigator(Kenkyū-buntansha) 山崎 思乃  関西大学, 化学生命工学部, 准教授 (50602182)
Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords乳酸菌 / 流加培養 / 呼吸 / 好気代謝 / 比増殖速度 / 酸素ストレス / 高濃度培養 / 高密度培養 / カタボライト抑制 / RNAseq / 好気培養 / ヘム呼吸 / Lactococcus lactis / 好気呼吸 / 酸素比消費速度 / ヘム / 過酸化水素
Outline of Research at the Start

乳酸菌は善玉菌としての需要が高まっているが、その名の通り著量の乳酸を生産し、それが障害となるために高濃度・高収率で培養することができない。当研究室では、糖を少量ずつ添加し、酸素を与えて呼吸させることによって、乳酸菌に乳酸を生産させることなく培養できることを見出した。しかし、乳酸菌の呼吸能力は十分ではなく、酸素障害を受けやすい。そこで本研究では、どうすれば乳酸菌の呼吸能力を高められるか、そして、どうすれば酸素障害を回避できるかを検討する。これによって、従来にはない、革新的な乳酸菌の高効率培養法を確立する。

Outline of Final Research Achievements

A highly efficient culture of lactic acid bacteria, which have various useful functions such as immunostimulation, was investigated. In conventional anaerobic batch culture, in which all nutrients are first provided under anaerobic conditions, lactobacilli produce significant amounts of lactic acid, so that even if the pH is adjusted, the bacteria concentration is limited to 1-3 g-dry-cell/L. In contrast, this study revealed that the production of organic acids such as lactic acid can be suppressed under aerobic condition by fed-batch culture, in which nutrients are gradually added. Furthermore, the concentration of Lactococcus lactis could be increased to about 20 g-dry-cell/L in 15 hours by aerobic fed-batch culture with addition of catalase and hem which reduses oxigen stress and assists respiration, respectively.

Academic Significance and Societal Importance of the Research Achievements

乳酸菌はヒトに対して免疫賦活や成人病予防など様々な有用な効果をもち、プロバイオティクスとして注目されている。プロバイオティクス市場は2030年には1000億ドルを超えるとされ、乳酸菌を効率よく安価に培養する技術が望まれている。本研究では、乳酸菌の好気流加培養について検討した結果、乳酸などの副生を抑制することによって、従来の嫌気回分培養に比べて2倍以上の収率で一桁高い菌体濃度を達成した。この成果は、乳酸の培養コストの大幅な低減を可能にし、プロバイオティクスのさらなる普及によって人々の健康に貢献する。また、カタボライト抑制が解除される流加培養でのみ発現する酸素利用関連遺伝子の存在を明らかにした。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (12 results)

All 2023 2022 2021

All Journal Article (2 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (10 results) (of which Int'l Joint Research: 5 results,  Invited: 3 results)

  • [Journal Article] Analysis of the effects of specific growth rate of Lactococcus lactis MG1363 on aerobic metabolism and its application to high-density culture2023

    • Author(s)
      Ryo Ichinose, Shino Yamasaki-Yashiki and Yoshio Katakura
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 136 Issue: 2 Pages: 129-135

    • DOI

      10.1016/j.jbiosc.2023.05.005

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Aerobic fed-batch culture of lactic acid bacteria2022

    • Author(s)
      片倉 啓雄、一瀬 涼、神原 大樹
    • Journal Title

      Japanese Journal of Lactic Acid Bacteria

      Volume: 33 Issue: 1 Pages: 12-19

    • DOI

      10.4109/jslab.33.12

    • ISSN
      1343-327X, 2186-5833
    • Year and Date
      2022-03-14
    • Related Report
      2022 Research-status Report 2021 Research-status Report
  • [Presentation] High cell density culture of lactic acid bacteria by aerobic fed-batch culture2023

    • Author(s)
      Ryo Ichinose, Daiki Kambara, Takatoshi Nakahara, Shino Yamasaki-Yashiki and Yoshio Katakura
    • Organizer
      The 13th Asian Conference of Lactic Acid Bacteria
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 好気流加培養による乳酸菌の高密度培養2023

    • Author(s)
      一瀬涼, 山崎思乃, 片倉啓雄
    • Organizer
      日本生物工学会第74回大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 好気流加培養における乳酸菌のグリセリン代謝2023

    • Author(s)
      神原大樹, 林田充基, 山崎思乃, 片倉啓雄
    • Organizer
      日本乳酸菌学会2023年度大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 好気的流加培養におけるカタラーゼ添加による酸素障害の回避2023

    • Author(s)
      中原尭駿, 山崎思乃, 片倉啓雄
    • Organizer
      日本乳酸菌学会2023年度大会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Interaction between lactic acid bacteria and carbohydrates2022

    • Author(s)
      Yoshio Katakura
    • Organizer
      The 12th Asian Conference on Lactic Acid Bacteria
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Avoiding oxygen damage in high efficiency culture of lactic acid bacteria2022

    • Author(s)
      Daiki Kambara, Honoka Nagai, Shino Yamasaki-Yashiki, Yoshio Katakura
    • Organizer
      The 15th International Symposium in Science and Technology
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research
  • [Presentation] Highly efficient cultivation of lactic acid bacteria that does not produce lactic acid2022

    • Author(s)
      Ryo Ichinose, Shino Yamasaki-Yashiki, Yoshio Katakura
    • Organizer
      The 15th International Symposium in Science and Technology
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research
  • [Presentation] L. plantarumの好気流加培養における網羅的遺伝子発現解析2022

    • Author(s)
      神原 大樹、山崎 思乃、片倉 啓雄
    • Organizer
      日本乳酸菌2022年度学会秋期セミナー
    • Related Report
      2022 Research-status Report
  • [Presentation] Efficient production of lactic acid bacteria by aerobic fed-batch culture2021

    • Author(s)
      Yoshio Katakura
    • Organizer
      The 11th Asian Conference on Lactic Acid Bacteria
    • Related Report
      2021 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 好気的流加培養による乳酸菌の高効率培養2021

    • Author(s)
      片倉啓雄
    • Organizer
      日本乳酸菌学会設立30周年記念シンポジウム
    • Related Report
      2021 Research-status Report
    • Invited

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Published: 2021-04-28   Modified: 2025-01-30  

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