Work

A computational strategy to improve the activity of tyrosine phenol-lyase for the synthesis of L-DOPA

Year: 2024

Type: article

Abstract: Enzymes with high catalytic activity and stability are essential for industrial production, yet most natural enzymes do not meet these requirements. Therefore, efficient strategies for enzyme engineering are crucial. In this study, we developed a cost-effective computational design strategy to enhance the activity of tyrosine phenol-lyase (TPL) for the production of L-DOPA. By integrating structural analysis with computational design, and guided by our understanding of conformational flexibility of TPL, we identified a region where enhanced stability is most likely to facilitate enzyme activity. We screened stabilizing mutations by Cartesian_ddg in Rosetta. After identifying single stabilizing mutations, we grouped the nearby positions harboring multiple stabilizing mutations and calculated the energy of combinatorial variants. We found two promising groups where most variants exhibited lower calculated energy than the wild-type. Experimental validation showed five variants in these groups exhibit increased activity, with the two best variants showing catalytic activity enhancements of 1.8-fold and 1.6-fold compared to the wild-type enzyme.

Source: Scientific Reports

DOI: 10.1038/s41598-024-76111-8

Authors: Jiayu Xu, Sheng Ye, Fenghui Guan

Institutions: Tianjin University, Zhejiang Cancer Hospital, Hangzhou Institute of Medicine, Chinese Academy of Sciences

Language: English

Topic: Enzyme Catalysis and Immobilization

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,590

Funders: National Natural Science Foundation of China, Ministry of Science and Technology of the People's Republic of China