Improvement of Saccharomyces cerevisiae for bioethanol production by modified genome shuffling

• Project/Area Number: 25450100
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Osaka University
• Principal Investigator: Sugiyama Minetaka 大阪大学, 工学(系)研究科(研究院), 准教授 (80379130)
• Co-Investigator(Kenkyū-buntansha): HARASHIMA Satosi 崇城大学, 生物生命学部, 教授 (70116086) ; 笹野 佑 大阪大学, 大学院工学研究科, 助教 (90640194)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: Saccharomyces cerevisiae / Thermotolerance / Bioethanol / Thermoresistance / Genome shuffling / Xylose fermentation

Outline of Final Research Achievements

In this study, we tried to improve thermotolerance of S.cerevisiae by modified genome shuffling. In the first round of genome shuffling, S. cerevisiae TJ14 and C3867 strains showing thermotolerance up to 41°C with high ethanol productivity were used as recipient strains. Non-conventional yeast Hansenula polymorpha showing robust thermotolerance up to 49°C was chosen as donor strains. After transformation of recipient strains with genomic DNA from donor strain, 3 transformants were isolated at 42°C and they showed pretty better growth and 7 times higher ethanol production than their recipient strains at 42°C. In the second round of genome shuffling, 2 transformants were isolated at 42.5°C and they showed significantly better growth and 2 times higher ethanol production than their recipient strains at 42.5°C. These strains will contribute to providing better platforms for further development of thermotolerant yeast for bioethanol production.

Research Products

• [Journal Article] Overexpression of ESBP6 improves lactic acid resistance and production in Saccharomyces cerevisiae (2016)
  • Author(s): Sugiyama, M., Akase, S., Nakanishi, R., Kaneko, Y., Harashima, S.
  • Journal Title: J. Biosci. Bioeng. Volume: 印刷中
  • NAID: 40020966107
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Nuclear localization of Haa1, which is linked to its phosphorylation status, mediates lactic acid tolerance in Saccharomyces cerevisiae. (2014)
  • Author(s): Sugiyama, M., Akase, SP., Nakanishi, R., Horie, H., Kaneko, Y., Harashima, S.
  • Journal Title: Appl. Environ. Microbiol Volume: 80 Issue: 11 Pages: 3488-3495
  • DOI: 10.1128/aem.04241-13
  • Peer Reviewed
• [Presentation] Improvement of thermotolerance of Saccharomyces cerevisiae for bioethanol production by genome shuffling (2015)
  • Author(s): Sugiyama, M., Wu, J., Sasano, Y., Kaneko, Y., Boonchird, C., Harashima S.
  • Organizer: 27th Annual Meeting of the Thai Society for Biotechnology and International Conference
  • Place of Presentation: Bangkok (Thailand)
  • Year and Date: 2015-11-18
  • Int'l Joint Research / Invited
• [Presentation] バイオエタノール生産のためのゲノムシャフリングによる高温耐性酵母の開発 (2015)
  • Author(s): 杉山峰崇, 呉 俊元, 笹野 佑, 金子嘉信, 原島 俊
  • Organizer: 第67回日本生物工学会大会
  • Place of Presentation: 城山観光ホテル（鹿児島）
  • Year and Date: 2015-10-26
• [Presentation] ゲノムシャフリングによるキシロース発酵性酵母Spathaspora passalidarumの高温耐性 (2015)
  • Author(s): 笹野 佑, 林 真帆, 藤原悠人, 杉山峰崇、原島 俊
  • Organizer: 第67回日本生物工学会大会
  • Place of Presentation: 城山観光ホテル（鹿児島）
  • Year and Date: 2015-10-26
• [Presentation] Improvement of thermotolerance of Saccharomyces cerevisiae for bioethanol production by genome shuffling (2015)
  • Author(s): Sugiyama, M., Wu, J., Sasano, Y., Kaneko, Y., Boonchird, C., Harashima S.
  • Organizer: 32nd International Specialized Symposium on Yeasts
  • Place of Presentation: Perugia (Italy)
  • Year and Date: 2015-09-04
  • Int'l Joint Research
• [Presentation] バイオエタノール生産のためのゲノムシャフリングによる高温耐性酵母の開発 (2015)
  • Author(s): 杉山峰崇, 呉 俊元, 笹野 佑, 金子嘉信, 原島 俊
  • Organizer: 酵母遺伝学フォーラム第48回研究報告会
  • Place of Presentation: 広島大学（広島）
  • Year and Date: 2015-08-31
• [Presentation] ゲノムシャフリングによる出芽酵母の高温耐性の改良と高温アルコール発酵 (2014)
  • Author(s): 杉山 峰崇 , 呉 俊元 , Badua MarkAnthonyA., 笹野 佑 , 金子 嘉信 , 原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-09-09
• [Presentation] 過剰発現で乳酸耐性を付与する出芽酵母転写活性化因子 Haa1 の機能解析 (2014)
  • Author(s): 中西良太、堀江仁志、松田祐典、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 第47会酵母遺伝学フォーラム研究報告会
  • Place of Presentation: 東京大学弥生講堂
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] 高温耐性出芽酵母 C3723 株における CDC19-C 遺伝子による高温耐性化 (2014)
  • Author(s): 木村駿太、王 棍、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 第47会酵母遺伝学フォーラム研究報告会
  • Place of Presentation: 東京大学弥生講堂
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] ゲノムシャフリングによる出芽酵母高温耐性の改良と高温アルコール発酵 (2014)
  • Author(s): 杉山峰崇、呉 俊元、Badua Mark Anthony A.、笹野 佑、金子嘉信、原島 俊
  • Organizer: 第47会酵母遺伝学フォーラム研究報告会
  • Place of Presentation: 東京大学弥生講堂
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] 出芽酵母のPKC1とGAS1の2重過剰発現による高温・酸ストレス条件下での高効率エタノール生産
  • Author(s): 大原佑介、荒木泰治、則木奈都子、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第65回大会
  • Place of Presentation: 広島国際会議場（広島）
• [Presentation] 高温・酸ストレス条件下でのPKC1 GAS1過剰発現出芽酵母による高効率エタノール発酵
  • Author(s): 杉山峰崇、大原佑介、荒木泰治、則木奈都子、笹野 佑、原島 俊
  • Organizer: 酵母遺伝学フォーラム第46回研究報告会
  • Place of Presentation: 東北学院大学押川記念ホール（仙台）
• [Book] Stress Biology of Yeasts and Fungi: Application for Industrial Brewing and Fermentation (Mechanism of yeast adaptation to weak organic acid stress) (2015)
  • Author(s): Sugiyama M, Sasano Y, Harashima S.
  • Total Pages: 218
  • Publisher: Springer Japan
• [Book] 酵母の生命科学と生物工学：第12章 pHストレス（酸・アルカリ） (2013)
  • Author(s): 杉山峰崇、西沢正文、原島 俊
  • Total Pages: 14
  • Publisher: 化学同人