Construction of versatile flocculation system and its application for two-stage production system

• Project/Area Number: 26870363
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied microbiology; Biofunction/Bioprocess
• Research Institution: Kobe University
• Principal Investigator: Tsuge Yota 神戸大学, 自然科学系先端融合研究環, 特命助教 (00647422)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: コリネ型細菌 / 有機酸 / アミノ酸 / フルフラール / 発酵 / コハク酸 / リジン / タンパク質工学 / 凝集 / アミノ酸生産 / 有機酸生産

Outline of Final Research Achievements

We firstly investigated cell growth of amino acid producing Corynebacterium glutamicum under furfural stress. The growth was inhibited by furfural, but furfural was degraded into furfuryl alcohol and 2-furoic acid overtime. Through constructing deletion strains of alcohol dehydrogenase genes, we identified the gene responsible for the conversion of furfural into furfuryl alcohol. Purified protein of this gene converted furfural into furfuryl alcohol in vitro using NADPH as a co-factor.
We moreover constructed a strain by metabolic engineering producing amino acid lysine under aerobic conditions and organic acid succinic acid under anaerobic conditions using oxygen as a metabolic switch.

Research Products

• [Journal Article] Engineering cell factories for producing building block chemicals for bio-polymer synthesis (2016)
  • Author(s): Tsuge Y, Kawaguchi H, Sasaki K, Kondo A
  • Journal Title: Microbial Cell Factories Volume: 15 Issue: 1 Pages: 19-19
  • DOI: 10.1186/s12934-016-0411-0
  • Peer Reviewed / Open Access
• [Journal Article] FudC, a protein primary responsible for furfural detoxification in Corynebacterium glutamicum (2016)
  • Author(s): Tsuge Y, Kudou M, Kawaguchi H, Ishii J, Hasunuma T, Kondo A
  • Journal Title: Applied Microbiology and Biotechnology Volume: 100 Issue: 6 Pages: 2685-2692
  • DOI: 10.1007/s00253-015-7115-y
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Overexpression of the phosphofructokinase encoding gene is crucial for achieving high production of D-lactate in Corynebacterium glutamicum under oxygen deprivation (2015)
  • Author(s): Tsuge Y, Yamamoto S, Kato N, Suda M, Vertès AA, Yukawa H and Inui M
  • Journal Title: Applied Microbiology and Biotechnology Volume: 99 Issue: 11 Pages: 4679-4689
  • DOI: 10.1007/s00253-015-6546-9
  • Peer Reviewed
• [Journal Article] Glucose consumption rate critically depends on redox state in Corynebacterium glutamicum under oxygen deprivation (2015)
  • Author(s): Tsuge Y, Uematsu K, Yamamoto S, Suda M, Yukawa H, Inui M
  • Journal Title: Applied Microbiology and Biotechnology Volume: 99 Issue: 13 Pages: 5573-5582
  • DOI: 10.1007/s00253-015-6540-2
  • Peer Reviewed
• [Journal Article] Recent advances in the metabolic engineering of Corynebacterium glutamicum for the production of lactate and succinate from renewable resources (2015)
  • Author(s): Tsuge Y, Hasunuma T, Kondo A
  • Journal Title: Journal of Industrial Microbiology and Biotechnology Volume: 42 Issue: 3 Pages: 375-389
  • DOI: 10.1007/s10295-014-1538-9
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Detoxification of furfural in Corynebacterium glutamicum under aerobic and anaerobic conditions (2014)
  • Author(s): Tsuge Y, Hori Y, Kudou M, Ishii J, Hasunuma T, Kondo A
  • Journal Title: Applied Microbiology and Biotechnology Volume: 98 Issue: 20 Pages: 8675-8683
  • DOI: 10.1007/s00253-014-5924-z
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Two-step production of D-lactate from mixed sugars by growing and resting cells of metabolically engineered Lactobacillus plantarum (2014)
  • Author(s): Tsuge Y, Kawaguchi H, Sasaki K, Tanaka T, Kondo A
  • Journal Title: Applied Microbiology and Biotechnology Volume: 98 Issue: 11 Pages: 4911-4918
  • DOI: 10.1007/s00253-014-5594-x
  • Peer Reviewed
• [Presentation] コリネ型細菌によるUV吸収性アミノ酸の生産 (2016)
  • Author(s): 柘植陽太、山本省吾、曽田匡洋、荻野千秋、近藤昭彦
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-03-27
• [Presentation] コリネ菌におけるフルフラールの分解 (2014)
  • Author(s): 柘植陽太、堀良美、蓮沼誠久、近藤昭彦
  • Organizer: 日本生物工学会大会
  • 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: 2014-09-09 – 2014-09-11
• [Presentation] コリネ型細菌を用いたD-乳酸生産における解糖系遺伝子高発現の効果 (2014)
  • Author(s): 柘植陽太、山本省吾、加藤直人、須田雅子、湯川英明、乾将行
  • Organizer: 生物工学会若手研究者の集い
  • Place of Presentation: 神戸セミナーハウス（兵庫県・神戸市）
  • Year and Date: 2014-07-12 – 2014-07-13
• [Remarks] 研究室ホームページ
  • URL: http://www2.kobe-u.ac.jp/~akondo/tsuge.html
• [Remarks] Pubmed業績集
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/?term=yota+tsuge