Engineering the 182 site of cyclodextrin glucosyltransferase for glycosylated genistein synthesis

• Published in: Sheng wu gong cheng xue bao = Chinese journal of biotechnology Vol. 38; no. 2; p. 749
• Main Authors: Chai, Baocheng, Jiang, Yulin, Ni, Ye, Han, Ruizhi
• Format: Journal Article
• Language: Chinese
• Published: China 25.02.2022
• Subjects: Cyclodextrins; Genistein; Glucosyltransferases - genetics; Glucosyltransferases - metabolism; Glycosylation; Kinetics; genistein; cyclodextrin glucosyltransferase; glycosylation; site-specific saturation mutagenesis; sophoricoside; water solubility

Genistein and its monoglucoside derivatives play important roles in food and pharmaceuticals fields, whereas their applications are limited by the low water solubility. Glycosylation is regarded as one of the effective approaches to improve water solubility. In this paper, the glycosylation of sophoricoside (genistein monoglucoside) was investigated using a cyclodextrin glucosyltransferase from ( CGTase). Saturation mutagenesis of D182 from CGTase was carried out. Compared with the wild-type (WT), the variant D182C showed a 13.42% higher conversion ratio. Moreover, the main products sophoricoside monoglucoside, sophoricoside diglucoside, and sophoricoside triglucoside of the variant D182C increased by 39.35%, 56.05% and 64.81% compared with that of the WT, respectively. Enzymatic characterization showed that the enzyme activities (cyclization, hydrolysis, disproportionation) of the variant D182C were higher than that of the WT, and the optimal pH and temperature of the variant D182C were 6 and 40℃, respective