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Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I
- Source :
- Frontiers in Bioengineering and Biotechnology, Vol 10 (2022)
- Publication Year :
- 2022
- Publisher :
- Frontiers Media S.A., 2022.
-
Abstract
- 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 α-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 (kcat/KM: 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.
Details
- Language :
- English
- ISSN :
- 22964185
- Volume :
- 10
- Database :
- Directory of Open Access Journals
- Journal :
- Frontiers in Bioengineering and Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.5fdc5aca55c3488988313ca2e8337f47
- Document Type :
- article
- Full Text :
- https://doi.org/10.3389/fbioe.2022.926829