Novel evolutionary design platform for engineering of diagnostic enzymes

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K20485
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0401:Materials engineering, chemical engineering, and related fields
• Research Institution: Waseda University
• Principal Investigator: Seki Takahiro 早稲田大学, 理工学術院総合研究所(理工学研究所), 次席研究員 (20836530)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: 進化分子工学 / 合成生物学 / 酵素工学 / 膜タンパク質 / ペリプラズム空間

Outline of Final Research Achievements

Evolutionary engineering for protein is a technology to create novel protein functions through repeated gene-level diversification and functional selection. However, there are many protein motifs that are difficult to express in the intracellular environment. Recently, I have developed a technique to display of arbitrary proteins in the periplasmic space of E. coli. This technology can not only evaluate the activity of the displayed protein in living cells, but also readout the stabilization of the protein by binding to its substrate through the function of a scaffold membrane protein. In this study, we attempted to use this technology to evolve the function of diagnostic enzymes, and challenged the establishment of a novel evolutionary engineering platform.

Free Research Field

進化分子工学

Academic Significance and Societal Importance of the Research Achievements

診断に利用される酵素の多くは，細胞から分泌されるタンパク質であり，そのほとんどが生細胞内での発現が困難あるいは機能選抜不可である．本研究により，診断酵素の進化工学プラットフォームが確立されたことで，これまで機能改良が困難だった多くの診断酵素の改良がハイスループットに行えるようになる．また，ペリプラズムという化学的環境を自在操作可能な空間での酵素改良が行えるようになったことで，診断酵素以外についてもさまざまな新規機能を付与可能となり，タンパク質機能の再デザインの歩留りが大きく高まるものと期待される．