The disclosure discloses a
cyclodextrin glycosyltransferase with enhanced
solvent tolerance and preparation thereof, belonging to the technical fields of
enzyme engineering and
genetic engineering. The disclosure constructs four
cyclodextrin glycosyltransferase mutants with enhanced
organic solvent tolerance. Among them, the
mutant with the optimal tolerance to DMSO and
methanol is G539I / R146F / D147N, which is 1.6 times and 1.7 times higher than that of WT, respectively; the
mutant with the optimal tolerance to
ethanol is R146F, which is 1.4 times higher than that of WT; the
mutant with the optimal tolerance to
acetone is G539I / R146F, which is 1.5 times higher than that of WT. The disclosure helps to expand the application of glycosyltransferases in
organic reaction systems, improves the enzymatic efficiency of CGTase on natural hydrophobic substrates, and has great application prospects.