Development of novel screening system for enzyme-displaying yeasts using calcium-alginate based beads.

• Project/Area Number: 21560812
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kinki University
• Principal Investigator: KAJIYAMA Shinichiro 近畿大学, 生物理工学部, 教授 (20243496)
• Co-Investigator(Kenkyū-buntansha): TANAKA Tsutomu 神戸大学, 自然科学系先端融合研究環重点研究部, 准教授 (90436551)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: シングルセル / 酵母表層工学 / 活性スクリーニング / 細胞包摂 / ソーティング

Research Abstract

In this study, a novel screening method for enzyme-displaying yeasts were investigated. Enzyme-displayed yeast cells were captured in the micro-sized calcium alginate beads each by each using newly developed reverse micelle method to prevent diffusion of hydrolyzed fluorescent substrates. Adopting flow sorting to these captured cells, active cells were successfully enriched. This system would be a useful method for high speed screening of yeast cells which display various hydrolyzing enzymes with high activity.

Research Products

• [Journal Article] Direct ethanol production from cellulosic materials using a cellulolytic enzymes expression optimized diploid Saccharomyces cerevisiae strain (2011)
  • Author(s): Yamada, R., Taniguchi, N., Tanaka, T., Ogino, C., Fukuda, H., Kondo, A
  • Journal Title: Biotechnology for Fuels Volume: 4 Issue: 1 Pages: 8-8
  • DOI: 10.1186/1754-6834-4-8
  • Peer Reviewed
• [Journal Article] Ethanol production from cellulosic materials using cellulase expressing yeast (2010)
  • Author(s): Yanase, S., Yamada, R., Kaneko, S., Noda, H., Hasunuma, T., Tanaka, T., Ogino, C., Fukuda, H., Kondo, A
  • Journal Title: Biotechnology Journal Volume: 5 Issue: 5 Pages: 449-455
  • DOI: 10.1002/biot.200900291
  • Peer Reviewed
• [Journal Article] Novel strategy for yeast construction using δ-integration and cell fusion to efficiently produce ethanol from raw starch (2010)
  • Author(s): Yamada, R., et.al.
  • Journal Title: Applied Microbiology and Biotechnology Volume: 85 Pages: 1491-1498
  • Peer Reviewed
• [Journal Article] Development of an enzyme activity screening system for ss-glucosidase-displaying yeasts using calcium alginate micro-beads and flow sorting (2009)
  • Author(s): Yan Zhou, Shin ichiro Kajiyama, Kouichi Itoh, Takanori Tanino, Nobuo Fukuda, Tsutomu Tanaka, Akihiko Kondo and Kiichi Fukui
  • Journal Title: Appl Microbiol Biotechnol Volume: 84(2) Issue: 2 Pages: 375-82
  • DOI: 10.1007/s00253-009-2091-8
  • Year and Date: 2009-06-27
  • Peer Reviewed
• [Journal Article] A new size and shape controlling method for producing calcium alginate beads with immobilized proteins (2009)
  • Author(s): Yan Zhou, Shin ichiro Kajiyama, Hiroshi Masuhara, Yoichiro Hosokawa, Takahiro Kaji, Kiichi Fukui
  • Journal Title: J. Biomed. Sci Eng Volume: Vol.2 No.5 Issue: 07 Pages: 287-293
  • DOI: 10.4236/jbise.2009.25043
  • Peer Reviewed
• [Journal Article] Development of an enzyme activity screening system for β-glucosidase-displaying yeasts using calcium alginate micro-beads and flow sorting (2009)
  • Author(s): Y.Zhou, et.al.
  • Journal Title: Appl Microbiol Biotechnol 84 Pages: 375-382
  • Peer Reviewed
• [Journal Article] A new size and shape controlling method for producing calcium alginate beads with immobilized proteins (2009)
  • Author(s): Y.Zhou, et.al.
  • Journal Title: J.Biomed.Sci Eng. 1.2 Pages: 287-293
  • Peer Reviewed
• [Presentation] セルラーゼ発現バランス最適化酵母を用いたセルロースからのエタノール発酵平成22年度 (2010)
  • Author(s): 山田亮祐, 田中勉, 荻野千秋, 近藤昭彦
  • Organizer: 日本生物工学会
  • Place of Presentation: 日宮崎シーガイアワールドコンベンションセンター
  • Year and Date: 2010-10-28
• [Presentation] セルラーゼ発現バランス最適化酵母を用いたセルロースからのエタノール発酵 (2010)
  • Author(s): 山田亮裕, ら
  • Organizer: 平成22年度日本生物工学会
  • Place of Presentation: 宮崎シーガイアワールドコンベンションセンター
  • Year and Date: 2010-10-28
• [Presentation] Direct fermentation of cellulosic materials to ethanol using yeast strains codisplaying three types of cellulolytic enzyme (2009)
  • Author(s): Suhei Yanase ; Ryosuke Yamada ; Tsutomu Tanaka ; Chiaki Ogino ; Hideki Fukuda ; Akihiko Kondo
  • Organizer: APBIOCHEC 09
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2009-09-24
• [Presentation] Direct fermentation of cellulosic materials to ethanol using yeast strains codisplaying three types of cellulolytic enzyme (2009)
  • Author(s): S.Yanase, et al.
  • Organizer: APBioChEC'09
  • Place of Presentation: 神戸国際会議場(神戸市)
  • Year and Date: 2009-09-24
• [Book] Transgenic Plants - Advances and Limitations. (2012)
  • Author(s): Naruemon Khemkladngoen, Naoki Wada, Suguru Tsuchimoto, Joyce A. Cartagena, Shin-ichiro Kajiyama and Kiichi Fukui
  • Publisher: InTech
• [Book] (2011)
  • Author(s): N. Wada, S. Kajiyama, N. Khemkladngoen, K. Fukui
• [Book] Plant Transformation Technologies Section 2 Chapter 4. (ISBN-10 : 0-8138-2195-9) (2011)
  • Author(s): N.Wada, et.al.
  • Total Pages: 296
  • Publisher: Wiley-Blackwell Publishers