食品与发酵工业 >

2026 , Vol. 52 >Issue 5: 156 - 164

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042126

研究报告

阿魏酸脱羧酶的异源表达及酶学性质表征

  • 丁合霞 ,
  • 杨威 ,
  • 曾茂蝶 ,
  • 李炫辰 ,
  • 程迈 ,
  • 黄卫泉 ,
  • 邱树毅 ,
  • 周剑丽
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  • 1(贵州大学 酿酒与食品工程学院,贵州 贵阳,550000)
    2(贵州大学 贵州省发酵工程与生物制药重点实验室,贵州 贵阳,550000)
    3(贵州省酿酒工业协会,贵州 贵阳,550000)
第一作者:硕士研究生(周剑丽副教授为通信作者,E-mail:zhoujl@gzu.edu.cn)

收稿日期: 2025-01-15

  修回日期: 2025-07-03

  网络出版日期: 2026-03-25

基金资助

贵州省科技计划项目(黔科合基础-ZK[2022]一般190)

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Heterologous expression and characterization of enzymatic properties of ferulic acid decarboxylase

  • DING Hexia ,
  • YANG Wei ,
  • ZENG Maodie ,
  • LI Xuanchen ,
  • CHENG Mai ,
  • HUANG Weiquan ,
  • QIU Shuyi ,
  • ZHOU Jianli
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  • 1(School of Liquor and Food Engineering, Guizhou University, Guiyang 550000, China)
    2(Key Laboratory of fermentation Engineering and Biological Pharmacy of Guizhou Province, Guizhou University, Guiyang 550000, China)
    3(Guizhou Province Alcoholic Drinks Industry Association, Guiyang 550000, China)

Received date: 2025-01-15

  Revised date: 2025-07-03

  Online published: 2026-03-25

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摘要

阿魏酸脱羧酶(ferulic acid decarboxylase,FDC)可以生物催化阿魏酸生成广泛应用于食品、医药、化妆品等行业的4-乙烯基愈创木酚。为研究来源于非酿酒酵母中FDC的酶学性质和催化功能,从美国国家生物技术信息中心(NCBI)数据库中筛选出异常威克汉姆酵母(Wickerhamomyces anomalus)中阿魏酸脱羧酶XM,实现在毕赤酵母中的异源表达。阿魏酸脱羧酶XM与4种酸(咖啡酸、阿魏酸、对香豆酸和芥子酸)进行分子对接,预测5种氨基酸残基(酪氨酸、苯丙氨酸、组氨酸、谷氨酰胺、谷氨酸)可能改变阿魏酸脱羧酶XM的功能。结果表明,该重组蛋白在摇瓶中和在发酵罐中诱导84 h后的酶活力分别为(19.72±0.32) IU/mg和(136.39±2.94) IU/mg。重组酶在40 ℃和pH 6.0下显示出最大活力。该酶具有广泛的底物特异性,能够在底物浓度为4 mmol/L的条件下高效脱羧4种酸。咖啡酸、阿魏酸、对香豆酸和芥子酸为底物的动力学常数Km的值分别为(151.94±9.30)、(133.70±5.62)、(152.09±5.33)、(170.89±8.76) μmol/L;Vmax值分别为(156.23±2.19)、(158.16±1.52)、(156.86±1.14)、(128.64±0.96) IU/mg。该研究可为进一步深入探究阿魏酸脱羧酶XM的催化机制奠定基础。

关键词： 阿魏酸脱羧酶; 异源表达; 酶学性质; 底物特异性; 动力学常数

本文引用格式

丁合霞 , 杨威 , 曾茂蝶 , 李炫辰 , 程迈 , 黄卫泉 , 邱树毅 , 周剑丽 . 阿魏酸脱羧酶的异源表达及酶学性质表征[J]. 食品与发酵工业, 2026 , 52(5) : 156 -164 . DOI: 10.13995/j.cnki.11-1802/ts.042126

Abstract

Ferulic acid decarboxylase (FDC) can biocatalyze ferulic acid to produce 4-vinylguaiacol, which was widely used in food, medicine, cosmetics and other industries.To study the enzymatic properties and catalytic function of ferulic acid decarboxylase from non-Saccharomyces yeasts, ferulic acid decarboxylase XM from Wickerhamomyces anomalus was screened from the National Center for Biotechnology Information (NCBI) database to achieve heterologous expression in Pichia pastoris.Molecular docking of ferulic acid decarboxylase XM with four acids (caffeic, ferulic, p-coumaric, and sinapic acids) predicted five amino acid residues (tyrosine, phenylalanine, histidine, glutamine, and glutamate) that might change the function of ferulic acid decarboxylase XM.The results showed that the enzyme activity of the recombinant protein were (19.72±0.32) IU/mg and (136.39±2.94) IU/mg in shake flasks and in fermenters after 84 h of induction, respectively.The recombinant enzyme showed maximum activity at 40 ℃ and pH 6.0.The enzyme had broad substrate specificity and was able to efficiently decarboxylate four acids at a substrate concentration of 4 mmol/L.The Km values of caffeic acid, ferulic acid, p-coumaric acid, and sinapic acid were (151.94±9.30), (133.70±5.62), (152.09±5.33), and (170.89±8.76) μmol/L, respectively, and the Vmax values were (156.23±2.19), (158.16±1.52), (156.86±1.14) and (128.64±0.96) IU/mg, respectively.This study laid a foundation for further exploring the catalytic mechanism of ferulic acid decarboxylase XM.

Key words： ferulic acid decarboxylase; heterologous expression; enzymatic properties; substrate specificity; kinetic constants

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