化学酶法转化羟乙基芦丁合成槲皮素  

A combined chemical and enzymatic approach to convert hydroxyethylrutin to quercetin

作　　者：李佳瑞 顾洁 徐晓宇 聂尧[1] 徐岩 LI Jiarui;GU Jie;XU Xiaoyu;NIE Yao;XU Yan(School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学生物工程学院,江苏无锡214122

出　　处：《生物加工过程》2023年第5期574-581,共8页Chinese Journal of Bioprocess Engineering

基　　金：国家重点研发计划(2021YFC2102000);国家自然科学基金(22178147)。

摘　　要：槲皮素是一种多羟基黄酮类化合物,其药理作用包括抗过敏、抗炎、抗癌等。工业上生产曲克芦丁产生的母液中含有未被利用的羟乙基芦丁,为实现母液羟乙基芦丁的回收利用,本文从羟乙基芦丁的结构出发,提出了一种化学法与酶法结合的方法将羟乙基芦丁转化为槲皮素。首先采用化学法实现羟乙基芦丁到芦丁的转化,反应收率为89.26%。其次在毕赤酵母中异源表达芦丁降解酶,并优化其芦丁转化槲皮素的反应体系,结果表明在底物浓度为2 mmol/L、酶添加量为15%(体积分数)、pH=5.0、温度为55℃时产率最高,最终酶法产率为53.96%。最后尝试以化学法得到的芦丁为酶法的底物,最终整体的转化率为47.17%。Quercetin,a polyhydroxy flavonoid,demonstrated pharmacological effects including anti-allergy,anti-inflammatory,anti-cancer and so on.The industrial production of troxerutin always left a mixture of mono/di/tri/tetra-hydroxyethyl substituted rutinin the mother liquor.In order to turn such waste mother liquor into useful chemicals,herein we developed a chemical and enzymatic method to convert hydroxyethyl rutin into quercetin.At first,the multiple hydroxyethyl substitutions were readily cleaved through a chemical conversion to afford rutin in 89.26%yield.Then an enzymatic process was utilized to hydrolyze rutin to quercetin.Such arutin degrading enzyme(RDE)was expressed in Pichia pastoris,and the corresponding enzymatic transformation was thoroughly optimized.As a result,the highest conversion was achieved at a substrate concentration of 2 mmol/L,enzyme addition of 15%(volume fraction),pH=5.0,and temperature of 55℃,with 53.96%yield for this step.Overall,the synthesis of quercetin was accomplished through a combined chemical and enzymatic sequence in 47.17%yield.

关 键 词：槲皮素 羟乙基芦丁 芦丁降解酶 曲克芦丁 

分 类 号：Q814.9[生物学—生物工程]