Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I

作者：Huang, Jia-Jun; Hu, Hao-Xuan; Lu, Yu-Jing; Bao, Ya-Dan; Zhou, Jin-Lin^*; Huang, Mingtao^*

来源：Frontiers in Bioengineering and Biotechnology, 2022, 10: 926829.

DOI：10.3389/fbioe.2022.926829

• icariside I
• icaritin
• homology modeling
• molecular docking
• a-L-rhamnosidase

Icariside I, the glycosylation product of icaritin, is a novel effective anti-cancer agent with immunological anti-tumor activity. However, very limited natural icariside I content hinders its direct extraction from plants. Therefore, we employed a computer-aided protein design strategy to improve the catalytic efficiency and substrate specificity of the alpha-L-rhamnosidase from Thermotoga petrophila DSM 13995, to provide a highly-efficient preparation method. Several beneficial mutants were obtained by expanding the active cavity. The catalytic efficiencies of all mutants were improved 16-200-fold compared with the wild-type TpeRha. The double-point mutant DH was the best mutant and showed the highest catalytic efficiency (k(cat )/K-M : 193.52 s(-1) M-1) against icariin, which was a 209.76-fold increase compared with the wild-type TpeRha. Besides, the single-point mutant H570A showed higher substrate specificity than that of the wild-type TpeRha in hydrolysis of different substrates. This study provides enzyme design strategies and principles for the hydrolysis of rhamnosyl natural products.

• 单位
  广东工业大学