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Rational Design of Cyclodextrin Glycosyltransferase with Improved Hesperidin Glycosylation Activity.
- Source :
-
Catalysts (2073-4344) . May2023, Vol. 13 Issue 5, p885. 11p. - Publication Year :
- 2023
-
Abstract
- Cyclodextrin glycosyltransferase (CGTase) can catalyze the glycosylation of hesperidin, resulting in α-glycosyl hesperidin with significantly improved water solubility. In this study, a rational design of CGTase to improve its hesperidin glycosylation activity was investigated. The strategy we employed involved docking hesperidin in its near-attack conformation and virtually mutating the surrounding residues, followed by calculating the changes in binding energy using Rosetta flex-ddG. The mutations with a stabilization effect were then subjected to an activity assay. Starting from CGTase-Y217F, we obtained three double-point mutants, Y217F/M351F, Y217F/M351L, and Y217F/D393H, with improved hesperidin glycosylation activities after screening twenty variants. The best variant, Y217F/D393H, exhibited a catalytic activity of 1305 U/g, and its kcat/KmA is 2.36 times higher compared to CGTase-Y217F and 15.14 times higher compared to the wild-type CGTase. Molecular dynamic simulations indicated that hesperidin was repulsed by CGTase-Y217F when bound in a near-attack conformation. However, by introducing a second-point mutation with a stabilization effect, the repulsion effect is weakened, resulting in a reduction in the distances between the bond-forming atoms and, thus, favoring the reaction. [ABSTRACT FROM AUTHOR]
- Subjects :
- *CYCLODEXTRINS
*HESPERIDIN
*GLYCOSYLATION
*BINDING energy
*CATALYTIC activity
Subjects
Details
- Language :
- English
- ISSN :
- 20734344
- Volume :
- 13
- Issue :
- 5
- Database :
- Academic Search Index
- Journal :
- Catalysts (2073-4344)
- Publication Type :
- Academic Journal
- Accession number :
- 163941636
- Full Text :
- https://doi.org/10.3390/catal13050885