虎杖苷十一碳烯酸酯的酶法合成与动力学研究  

Enzymatic Synthesis and Kinetic of Undecenoyl-polydatin

作　　者：刁怡涵 李明依 毕艳红 杜文迎 杨荣玲 王朝宇 DIAO Yihan;LI Mingyi;BI Yanhong;DU Wenying;YANG Rongling;WANG Zhaoyu(School of Life Science and Food Engineering,Huaiyin Institute of Technology,Huai'an,Jiangsu 223003)

机构地区：[1]淮阴工学院生命科学与食品工程学院,江苏淮安223003

出　　处：《核农学报》2022年第2期369-375,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金项目(21676114);江苏省六大人才高峰项目(SWYY-011、YY-061);江苏省青蓝工程青年骨干教师项目。

摘　　要：为建立绿色、高效、高选择性合成虎杖苷十一碳烯酸酯双前药的生物催化体系,本研究以棉状嗜热丝孢菌(Thermomyces lanuginosus)脂肪酶(TLL)为催化剂,以反应介质、底物摩尔比、温度及反应时间等因素为变量,探究了TLL催化虎杖苷十一碳烯酰化反应的特性。结果表明,TLL可在丙酮中实现对虎杖苷的高效转化,产物均为6″-单酯(100%);在最优条件下,该酶促反应的初速度、底物转化率和6″-区域选择性分别达20.8 mmol·L^(-1)·h^(-1)、100%和100%。TLL在不同有机介质中催化虎杖苷酰化反应的动力学特征差别较大,但在丙酮中的K_(m)值最小、V_(max)和V_(max)/K_(m)最大,分别为24.7 mmol·L^(-1)、68.9 mmol·L^(-1)·h^(-1)和2.79 h^(-1),表明在丙酮中酶与底物的亲和力更强,催化效率更高,因此丙酮是该酶促反应的适宜溶剂。本研究不仅丰富了非水相酶学基础理论知识,更为绿色高效的酶催化技术改造传统化学合成过程提供了借鉴。To establish a green and efficient synthesis of the undecenoyl-polydatin double prodrug Thermomyces lanuginosus lipase(TLL)was used as the catalyst for the first time.Some crucial factors such as the optimal reaction medium,molar ratio of polydatin to vinyl undecenoate,reaction temperature and time were investigated to characterize the behavior of the catalytic procedure.The results revealed that the immobilized TLL showed the highest regioselectivity towards the 6″-OH(100%)in acetone and the dominant product was proved to be 6″-monoester.Under the optimal conditions,the reaction rate,substrate conversion and 6″-regioselectivity were 20.8 mmol·L^(-1)·h^(-1),100%,and 100%,respectively.However,the kinetic characteristics of the TLL enzyme in different organic media were significantly different for the acylation of polydatin.The minimum K_(m) and the maximum V_(max) and V_(max)/K_(m) obtained in acetone were 24.7 mmol·L^(-1),68.9 mmol·L^(-1)·h^(-1),and 2.79 h^(-1),respectively.It indicated that the enzyme exerted an extremely higher affinity for the substrate and catalytic efficiency in acetone as compared to other organic solvents,which was the suitable solvent for the enzymatic reaction.This study not only enriched the knowledge of nonaqueous fundamental enzymology,but also provide reference for changing traditional chemical synthesis processes using more green and efficient enzyme catalytic technology.

关 键 词：虎杖苷 棉状嗜热丝孢菌脂肪酶 十一碳烯酸乙烯酯 酰化 动力学 

分 类 号：TQ460.1[化学工程—制药化工]