基于固定化酶法合成肉桂醇糖基化衍生物的研究  

Research on the Synthesis of Cinnamyl Alcohol Glycosylated Derivatives Based on Immobilized Enzyme Method

作　　者：程歆然 宗卫勇 窦文芳 CHENG Xinran;ZONG Weiyong;DOU Wenfang(Life Sciences and Health Engineering College,Jiangnan University,Jiangsu Wuxi 214122,China)

机构地区：[1]江南大学生命科学与健康工程学院,江苏无锡214122

出　　处：《生物技术进展》2025年第1期110-118,共9页Current Biotechnology

摘　　要：络塞是苯丙素苷类化合物,具有多种生物活性。络塞的传统生产方式主要从植物中提取,但自然资源有限。利用来自拟南芥的糖基转移酶UGT73C5和来自大豆的蔗糖合酶GmSUS偶联催化,对2种带有His标签的酶进行固定化,偶联催化肉桂醇反应生成络塞,同时形成尿苷二磷酸葡萄糖(uridine diphosphate glucose,UDPG)的再生。对2种酶分别进行固定,确定了树脂为2 g时分别可固定5 g的UGT73C5粗酶和6 g的GmSUS粗酶,并优化固定化偶联催化反应的最适条件,测定偶联固定化酶可重复稳定反应7个批次,最后采用分批补料策略,络塞的产量可达到15.82 mmol·L^(-1),转化率为97.5%。反应后进行高效液相色谱(high performance liquid chromatography,HPLC)分析,推断出反应产物为络塞。固定化酶策略将为双酶偶联合成络塞开辟新前景,并降低游离酶在大规模工业应用中的成本。Rosin is a phenylpropanoid glycoside compound with various biological activities.The traditional way of producing ros⁃in is through plant extraction,but natural resources are limited.This article utilized the coupling catalysis of glycosyltransferase from Arabidopsis thaliana and sucrose synthase from soybean to immobilize two enzymes with His tags,converting cinnamyl alco⁃hol into a complex and regenerating uridine diphosphate glucose(UDPG).Two enzymes were fixed separately,and it was deter⁃mined that 5 g of UGT73C5 crude enzyme and 6 g of GmSUS crude enzyme could be fixed with 2 g of resin.The optimal condi⁃tions for the immobilized coupling catalytic reaction were optimized,and the coupling immobilized enzyme was tested to be repro⁃ducible and stable for seven batches.Finally,a batch feeding strategy was adopted,and the yield of the complex plug reached 15.82 mmol·L^(-1) with a conversion rate of 97.5%.After the reaction,high-performance liquid chromatography(HPLC)was per⁃formed,and it was inferred that the reaction product is rosin.The strategy of immobilizing enzymes will open up new prospects for the development of double enzyme pairs to form rosin,and reduce the cost of free enzymes in large-scale industrial applications.

关 键 词：肉桂醇 络塞 糖基化反应 蔗糖合酶 固定化酶 

分 类 号：Q559[生物学—生物化学]