生物技术通报2023，Vol.39Issue(12)：200-208,9.DOI:10.13560/j.cnki.biotech.bull.1985.2023-0653
芍药苷转化酶G6046的异源表达及酶活鉴定
Heterologous Expression of Paeoniflorin Converting Enzyme G6046 and the Identification of Its Enzymatic Activities
摘要
Abstract
Paeoniflorin is a glycoside compound found in Paeonia lactiflora Pall that shows various protective effects on the cardio-cerebral vascular system and always used in traditional herbal medicine.However,the bioavailability of paeoniflorin by oral administration is low,which greatly limits its clinical application.It has been reported that the pharmacological effects of paeoniflorin might be generated by its metabolites and mainly metabolized in intestinal bacteria.In this paper,the induction expression and purification of paeoniflorin converting enzyme G6046 in Escherichia coli BL21 DE3 were studied.The induction reaction conditions were optimized to improve the enzyme yield and the optimal conditions for induction reaction were found to be 24 h induction reaction at 16℃.Enzyme production of bacterium reached 5.34 mg/g,representing a 51.69%increase in enzyme production.Based on the conditions of induction expression,the conditions(pH and temperature)for the reaction of G6046 with paeoniflorin were optimized,pH 9.0 and 45℃were proved to be the optimized conditions,resulting in a specific enzyme activity of 14.56 U/mg.The reaction system was enlarged under this optimized condition,and the reaction scale was from G6046-substrate(1∶1;mg/mg)to G6046-substrate(1∶20;mg/mg),the yield of the converted product significantly increased,which lays a foundation for the isolation of peonidin conversion product.关键词
芍药苷/芍药苷转化酶/高效液相色谱Key words
paeoniflorin/paeoniflorin converting enzyme/HPLC引用本文复制引用
韩潆仪,李章涵,曹学丽,裴海闰..芍药苷转化酶G6046的异源表达及酶活鉴定[J].生物技术通报,2023,39(12):200-208,9.基金项目
国家自然科学基金面上项目(21773014) （21773014）
