Elucidation of the bottlenecks in the yeast protein secretion process using the novel gene-overexpression library

2023 Fiscal Year Final Research Report

• Project/Area Number: 20K05229
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Kobe University
• Principal Investigator: ITO YOICHIRO 神戸大学, 先端バイオ工学研究センター, 特命准教授 (50379153)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: タンパク質分泌発現 / 酵母 / ピキアパストリス / 過剰発現ライブラリ / 遺伝子欠損ライブラリ

Outline of Final Research Achievements

The process of the yeast protein secretion has been studied so far, however the detailed picture does not become clear.　In this study, a general study was performed by exploration and comparison of secretion factors which were found from the screening of originally developed gene-overexpression library for antibody-producing yeast strains (Komagataella phaffi). In the secretion factors obtained from the screening, although there were some factors which increased protein productivity around 2 times, the bottlenecked factor to increase dramatically could not find. The accumulations of the secretion factors in one yeast strain were effective strategy for improving antibody productivity. Thus, it was suggested that some rate-limiting steps would exist in various process of the yeast protein secretion.

Free Research Field

進化工学

Academic Significance and Societal Importance of the Research Achievements

酵母での遺伝子欠損ライブラリは古くより作製方法も含めていくつか報告例があるが、過剰発現ライブラリはツールとして利用できるものは少なかった。そのため、酵母における過剰発現ライブラリ作製法の開発に成功したことは学術的にも重要な成果である。また本研究の成果は、学術研究としてのタンパク質の分泌発現機構の理解だけでなく、産業上重要な難分泌タンパク質の生産性の高度化に繋げられる。