食品与发酵工业 >

2022 , Vol. 48 >Issue 3: 38 - 43

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.028306

研究报告

玉米黄烷酮-3-羟化酶原核表达条件对香橙素生成量的影响

  • 黄旭 ,
  • 王振 ,
  • 刘娟 ,
  • 肖妃垚 ,
  • 田苗苗 ,
  • 郭佳婧 ,
  • 单杨
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  • 1(湖南大学 隆平分院,湖南 长沙,410082)
    2(湖南省农业科学院 农产品加工研究所/果蔬贮藏加工与质量安全湖南省重点实验室/湖南省果蔬加工与质量安全国际科技创新合作基地,湖南 长沙,410125)
硕士研究生(郭佳婧助理研究员和单杨研究员为共同通信作者,E-mail:guojiajing1986@163.com;sy6302@sohu.com)

收稿日期: 2021-06-08

  修回日期: 2021-07-13

  网络出版日期: 2022-03-04

基金资助

湖南省科技厅重点领域研发技术项目(2019NK2041);湖南省农业科技创新资金重点项目(2020CX47)

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Effect of prokaryotic expression conditions of Zea mays flavanone-3-hydroxylase on production of aromadendrin

  • HUANG Xu ,
  • WANG Zhen ,
  • LIU Juan ,
  • XIAO Feiyao ,
  • TIAN Miaomiao ,
  • GUO Jiajing ,
  • SHAN Yang
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  • 1(Longping Branch Graduate School,Hunan University,Changsha 410082,China)
    2(Institute of Agricultural Product Processing,Hunan Academy of Agricultural Sciences/Hunan Province Key Lab of Fruits & Vegetables Storage,Processing,Quality and Safety/Hunan Province International Joint Lab on Fruits & Vegetables Processing,Quality and Safety,Changsha 410125,China)

Received date: 2021-06-08

  Revised date: 2021-07-13

  Online published: 2022-03-04

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摘要

香橙素是类黄酮代谢途径中重要的中间化合物,黄烷酮-3-羟化酶(flavanone-3-hydroxylase,F3H)能以柚皮素为底物催化C3位接羟基从而生成香橙素。为实现香橙素绿色发展,该研究利用微生物法,对来自玉米的Zmf3h构建大肠杆菌原核表达系统后诱导ZmF3H蛋白表达。通过发酵实验验证ZmF3H在胞内可溶性表达并成功催化生成香橙素。为提高香橙素产量,对蛋白表达阶段的菌体密度、异丙基-β-D-硫代半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)诱导剂浓度、诱导时间和诱导温度等条件进行优化实验。单因素试验结果表明各实验组分别在菌种密度OD600=0.8~1.0,IPTG终浓度为0.6 mmol/L,诱导温度在23 ℃,诱导8 h时可明显提高香橙素生成量,综合各单因素优化条件进行实验验证后发现香橙素生成量为(32.27±1.98)mg/L,与初始条件相比提高了60.39%。

关键词： 香橙素; 黄烷酮-3-羟化酶; 原核表达; 表达条件优化

本文引用格式

黄旭 , 王振 , 刘娟 , 肖妃垚 , 田苗苗 , 郭佳婧 , 单杨 . 玉米黄烷酮-3-羟化酶原核表达条件对香橙素生成量的影响[J]. 食品与发酵工业, 2022 , 48(3) : 38 -43 . DOI: 10.13995/j.cnki.11-1802/ts.028306

Abstract

Aromadendrin is an important intermediate compound in flavonoid metabolism pathway. Flavanone-3-hydroxylase (F3H) catalyzes hydroxyl grafting of naringin at C3 site. Microbial method was used to achieve the green development of aromadendrin. ZmF3H protein was expressed by Escherichia coli containing Zmf3h from Zea mays. Fermentation experiments confirmed that ZmF3H was soluble in intracellular and successfully catalyzed the production of aromadendrin. The results showed that the production of aromadendrin was significantly increased when the final IPTG concentration was 0.6 mmol/L, the induction temperature was 23 ℃, the induction time was 8 h, and the cell density was between 0.8 to 1.0. The production of aromadendrin was (32.27±1.98) mg/L after optimization which was 60.39% higher than that of initial conditions.

Key words： aromadendrin; flavanone-3-hydroxylase; prokaryotic expression; expression optimization

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