Efficient production of high optical pure hydroxy acid based on the regeneration of coenzyme A

• Project/Area Number: 26420791
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Hokkaido University
• Principal Investigator: Ooi Toshihiko 北海道大学, 工学研究院, 准教授 (40223713)
• Co-Investigator(Kenkyū-buntansha): 松本 謙一郎 北海道大学, 工学研究院, 准教授 (80360642)
• Co-Investigator(Renkei-kenkyūsha): Taguchi Seiichi 北海道大学, 大学院工学研究院, 教授 (70216828)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: キラル化合物 / ヒドロキシ酸 / 3-ヒドロキシ酪酸 / 3-ヒドロキシ酸 / 高光学純度 / キラル / 微生物生産

Outline of Final Research Achievements

Optical purity of building block are essential for the chemical synthesis of bioactive compounds, such as antibiotics and pheromone. Chemical synthesis of optically active compounds are not so easy because of no optical selective synthesis. We developed basic technology of the biosynthesis method for optically active [R]-3-hydroxybutylate (3HB) with high purity using engineered microbiological system.
We have newly developed the quantifying analytical method for intermediate compounds of [R}-3HB biosynthesis and found that the improvement of the flux to Acetyl-CoA is effective to produce [R]-3HB production. On the basis this results Escherichi coli JM109 was selected as a high glucose consumption strains. Using this strain the concentrations of glucose and acetate into the medium was optimized. Productivity of high optically pure [R]-3HB was archived to 15g/l in optimized culture condition. In addition newly cultivation technology was futher developed for the efficient production.

Research Products

• [Journal Article] Xylose-based hydrolysate from eucalyptus extract as feedstock for poly(lactate-co-3-hydroxybutyrate) production in engineered Escherichia coli (2017)
  • Author(s): Takisawa K, Ooi T, Matsumoto K, Kadoya R, Taguchi S
  • Journal Title: Process Biochemistry Volume: 54 Pages: 102-105
  • DOI: 10.1016/j.procbio.2016.12.019
  • Peer Reviewed
• [Journal Article] Molecular weight-dependent degradation of D-lactate-containing polyesters by polyhydroxyalkanoate depolymerases from Variovorax sp. C34 and Alcaligenes faecalis T1 (2015)
  • Author(s): Jian Sun, Ken’ichiro Matsumoto, Yuta Tabata, Ryosuke Kadoya, Toshihiko Ooi, Hideki Abe, Seiichi Taguchi
  • Journal Title: Appl. Microbiol. Biotechnol. Volume: 99 Issue: 22 Pages: 9555-9563
  • DOI: 10.1007/s00253-015-6756-1
  • NAID: 120005855945
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 組換え大腸菌による乳酸ポリマー生産のためのキシロースを炭素源とした高密度培養 (2017)
  • Author(s): 滝沢憲治、Ribordy Greg、山崎貴史、松本謙一郎、Zinn Manfred、田口精一
  • Organizer: 平成29年度日本農芸化学会大会
  • Place of Presentation: 京都女子大学
• [Presentation] Degradation mechanism of poly(lactate-co-3-hydroxybutyrate) by depolymerase from soil bacterium Variovorax sp. C34 (2015)
  • Author(s): Camila Utsunomia, Jian Sun, Ken’ichiro Matsumoto, Toshihiko Ooi, Seiichi Taguchi
  • Organizer: 第67回日本生物工学会大会
  • Place of Presentation: s城山観光ホテル（鹿児島）
  • Year and Date: 2015-10-27
• [Presentation] Mechanism of poly(lactate-co-hydroxybutyrate) degradation by a soil bacterium depolymerase (2015)
  • Author(s): Camila Utsunomia, Jian Sun, Ken’ichiro Matsumoto, Toshihiko Ooi, Seiichi Taguchi
  • Organizer: Japan-Taiwan Bilateral Polymer Symposium
  • Place of Presentation: Hokkaido University、Sapporo
  • Year and Date: 2015-09-04
  • Int'l Joint Research
• [Presentation] 組換え大腸菌を用いた高光学活性(R)-3-ヒドロキシブタン酸の生産におけるCoA転移酵素高活性変異体の効果 (2015)
  • Author(s): 三吉大地、瀬戸望、松本謙一郎、大井俊彦、田口精一
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-03-27
• [Presentation] 組換え大腸菌を利用した高光学純度(S)-3-ヒドロキシブタン酸の生産 (2015)
  • Author(s): 瀬戸望、大井俊彦、松本謙一郎、田口精一
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-03-27