Fabrication of a rational design method for enzyme mutants based on an understanding of substrate specificity

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K15371
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: MORI YUTARO 国立研究開発法人理化学研究所, 環境資源科学研究センター, 研究員 (50758539)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 酵素変異体 / バイオプロダクション

Outline of Final Research Achievements

In this study, we aimed to modifiy and extend the substrate specificity of enzymes by deepening our intrinsic understanding. In addition, this study's objective is to establish rational rules for designing enzyme mutants to catalyze enzymatic reactions with high activity. Several types of Information were accumulated by comparing the results of enzyme screening with the constructed enzyme-substrate binding model. The results suggest that the affinity between the enzyme and the substrate is an important factor for the enzyme to be able to recognize the substrate. In fact, it is suggested that the HOT SPOT, which is likely to contribute to increased activity, can be predicted by in silico calculation. Finally, a 2.8-fold increase in activity was achieved with the enzyme mutant compared to the wild type.

Free Research Field

タンパク質工学

Academic Significance and Societal Importance of the Research Achievements

これまで生体内の反応を触媒する酵素に着目し、アミノ酸変異を導入することで、既存の酵素反応の活性向上や新規の非天然化合物の生成を達成した研究は多数報告されている。しかしながら、これらの設計は使用する酵素次第のケースバイケースであり、数百数千の変異体を作製することで高性能な変異体の取得に成功したとしても、他の鋳型酵素を使用する場合にはまた一から変異体を構築しなければならなかった。そこで本研究がもたらす合理的な酵素変異体設計ルールにより、目的の性能値を持つ酵素変異体の迅速な獲得や、非天然有用な化合物おｗ生産する酵素の短期間での開発が可能となれば、持続可能社会の実現に向けた大きな一助となると考える。