Design of functional bacterionanofibers and creation of fiber-coated microorganisms

• Project/Area Number: 26289313
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Nagoya University
• Principal Investigator: Katsutoshi Hori 名古屋大学, 工学(系)研究科(研究院), 教授 (50302956)
• Co-Investigator(Renkei-kenkyūsha): NAGAMUNE Teruyuki 東京大学, 大学院工学研究科, 教授 (20124373) ; TANAKA Toshiki 名古屋工業大学, 大学院工学研究科, 教授 (70171775) ; SUZUKI Atsuo 名古屋大学, 大学院工学研究科, 准教授 (40196788) ; NAKATANI Hajime 名古屋大学, 大学院工学研究科, 講師 (80456615)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000); Fiscal Year 2016: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2015: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: バクテリオナノファイバー / 機能性ナノファイバー / ファイバータンパク質 / 被毛微生物 / ファイバー提示 / ナノファイバー / 微生物 / 膜タンパク質 / バイオテクノロジー / ナノバイオ / 蛋白質

Outline of Final Research Achievements

The purpose of this research is to design functional nanofibers obtained by fusing functional molecules to the bacteria nanofiber protein AtaA found by the applicant and to create microorganisms covered with functional "hair". I accomplished following thing. ① A protease sensitive fiber with protease recognition / cleavage sequence inserted at the base of the fiber was designed and grown. ② His-tag was inserted into the fiber tip with varying length to make it function at the cell surface. ③ The recognition sequence for sortase was inserted and a modification method after fiber presentation was established . ④ The fiber which was fused to the SNAP tag as a model enzyme was peritrichously grown from bacterial cells. ⑤
Two kinds of homotrimeric fibers were successfully grown from a single cell.

Research Products

• [Int'l Joint Research] マックスプランク研究所(ドイツ)
• [Int'l Joint Research] オスロ大学(ノルウェー)
• [Journal Article] Cell surface protein engineering for high-performance whole-cell catalysts (2017)
  • Author(s): Hajime Nakatani, Katsutoshi Hori
  • Journal Title: Frontiers of Chemical Science and Engineering Volume: 11 Issue: 1 Pages: 46-57
  • DOI: 10.1007/s11705-017-1609-3
  • Peer Reviewed / Open Access
• [Journal Article] バイオフィルム・微生物付着の工学利用 (2017)
  • Author(s): 堀 克敏
  • Journal Title: 膜 Volume: 42 Pages: 54-59
  • NAID: 130005492511
• [Journal Article] An Acinetobacter trimeric autotransporter adhesin reaped from cells exhibits its nonspecific stickiness via a highly stable 3D structure (2016)
  • Author(s): Shogo Yoshimoto, Hajime Nakatani, Keita Iwasaki, Katsutoshi Hori
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 28020-28020
  • DOI: 10.1038/srep28020
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 細胞表層タンパク質の合成生物学 (2016)
  • Author(s): 石川聖人、堀 克敏
  • Journal Title: 生物工学会誌 Volume: 94 Pages: 701-703
• [Journal Article] 細菌接着タンパク質AtaA分子の特性と応用 (2016)
  • Author(s): 堀 克敏
  • Journal Title: バイオサイエンスとインダストリー Volume: 74 Pages: 417-419
  • NAID: 40020960334
• [Journal Article] Secretion of the Intimin Passenger Domain Is Driven by Protein Folding (2016)
  • Author(s): J. C. Leo, P. Oberhettinger, S. Yoshimoto, D. B. R. K. G. Udatha, J. P. Morth, M. Schutz, K. Hori, and D. Linke
  • Journal Title: J. Biol. Chem. Volume: 291 Issue: 38 Pages: 20096-20112
  • DOI: 10.1074/jbc.m116.731497
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 細菌の持つ接着ナノファイバータンパク質AtaA (2016)
  • Author(s): 堀 克敏
  • Journal Title: バイオサイエンスとインダストリー Volume: 74 Pages: 380-381
• [Presentation] AtaA, a new trimeric autotransporter adhesin mediating strong and nonspecific adhesion of bacterial cells (2017)
  • Author(s): K. Hori
  • Organizer: 第90回日本細菌学会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-19
  • Invited
• [Presentation] Innovation of bioprocess through the bacterionanofiber AtaA (2017)
  • Author(s): K .Hori
  • Organizer: 2nd Korea-Japan Smart Biodesign Workshop
  • Place of Presentation: Sendai
  • Year and Date: 2017-02-03
  • Int'l Joint Research / Invited
• [Presentation] Innovation of bioprocess through the bacterionanofiber AtaA (2016)
  • Author(s): K. Hori
  • Organizer: i-BioP2016
  • Place of Presentation: Busan, Corea
  • Year and Date: 2016-12-16
  • Int'l Joint Research / Invited
• [Presentation] 微生物細胞やバクテリオナノファイバーの表面化学 (2016)
  • Author(s): 堀 克敏
  • Organizer: 真空・表面科学合同講演会
  • Place of Presentation: 名古屋
  • Year and Date: 2016-12-01
  • Invited
• [Presentation] The Contribution of translocation domain in the assembly of trimeric autotransporter adhesins (2016)
  • Author(s): J. Kanie, Y. Ohara, H. Nakatani, K. Hori
  • Organizer: 1st Zing Protein Secretion in Bacteria Conference
  • Place of Presentation: Tampa, Florida, USA
  • Year and Date: 2016-11-09
  • Int'l Joint Research
• [Presentation] グラム陰性菌由来のファイバータンパク質による微生物の固定化と表層提示技術の統合 (2016)
  • Author(s): 中谷肇、小原優季、堀 克敏
  • Organizer: 第68回日本生物工学会大会
  • Place of Presentation: 富山
  • Year and Date: 2016-09-28