Efficient Biosynthesis of Gastrodin by UDP-Glycosyltransferase from Rauvolfia serpentina
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Gastrodin, the primary bioactive constituent of Gastrodia elata, possesses numerous remarkable pharmacological properties. In this investigation, UDP-glycosyltransferase from Rauvolfia serpentina (RsUGT) was expressed, subsequently purified and characterized. The maximum yield of the enzyme was 17.57 mU/ml and possessed a relative molecular weight of approximately 77.7 kDa. Utilizing GST affinity resin, RsUGT was purified 20.8-fold, with an overall recovery rate of 58.6% and specific activity of 79.2 mU/mg. The optimal temperature and pH for RsUGT was identified as 40o C and 10.0, respectively. Notably, 2% DMSO could increase the RsUGT activity by 12.15%. The Michaelis-Menten constants KM and Vmax were determined to be 0.50 mM and 171.60 mU/mg. By optimizing the conditions for the enzymatic biosynthesis of gastrodin by RsUGT, the highest gastrodin production was 285.35 mg/l, accompanied by a molar conversion rate of 99.67%. In addition, the conditions of gastrodin biosynthesis by recombinant strain BL-RsUGT were also studied. The highest gastrodin production was 225.99 mg/l, and the corresponding pHBA conversion rate was 98.00%. These findings confirmed the promising potential of RsUGT in the production of gastrodin.
Abstract
Gastrodin, the primary bioactive constituent of Gastrodia elata, possesses numerous remarkable pharmacological properties. In this investigation, UDP-glycosyltransferase from Rauvolfia serpentina (RsUGT) was expressed, subsequently purified and characterized. The maximum yield of the enzyme was 17.57 mU/ml and possessed a relative molecular weight of approximately 77.7 kDa. Utilizing GST affinity resin, RsUGT was purified 20.8-fold, with an overall recovery rate of 58.6% and specific activity of 79.2 mU/mg. The optimal temperature and pH for RsUGT was identified as 40o C and 10.0, respectively. Notably, 2% DMSO could increase the RsUGT activity by 12.15%. The Michaelis-Menten constants KM and Vmax were determined to be 0.50 mM and 171.60 mU/mg. By optimizing the conditions for the enzymatic biosynthesis of gastrodin by RsUGT, the highest gastrodin production was 285.35 mg/l, accompanied by a molar conversion rate of 99.67%. In addition, the conditions of gastrodin biosynthesis by recombinant strain BL-RsUGT were also studied. The highest gastrodin production was 225.99 mg/l, and the corresponding pHBA conversion rate was 98.00%. These findings confirmed the promising potential of RsUGT in the production of gastrodin.