食品与发酵工业 >

2021 , Vol. 47 >Issue 18: 175 - 180

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026837

生产与科研应用

通过发酵优化提高大肠杆菌生产L-半胱氨酸产量

  • 张博 ,
  • 史永吉 ,
  • 杨辉 ,
  • 吴梓丹 ,
  • 陈开 ,
  • 蔡雪 ,
  • 柳志强 ,
  • 郑裕国
展开
  • (浙江工业大学 生物工程学院,浙江 杭州,310023)
博士,副教授(柳志强教授为通讯作者,E-mail:microliu@zjut.edu.cn)

收稿日期: 2021-01-20

  修回日期: 2021-02-27

  网络出版日期: 2021-10-18

基金资助

国家重点基础研究发展计划(973计划)(2018YFA0901400);国家自然科学基金(32070099;31971342)

收起

Enhancement of L-cysteine production in Escherichia coli through fermentation optimization

  • ZHANG Bo ,
  • SHI Yongji ,
  • YANG Hui ,
  • WU Zidan ,
  • CHEN Kai ,
  • CAI Xue ,
  • LIU Zhiqiang ,
  • ZHENG Yuguo
Expand
  • (College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310023, China)

Received date: 2021-01-20

  Revised date: 2021-02-27

  Online published: 2021-10-18

Fold

摘要

该研究为提高菌株L-半胱氨酸合成能力,首先在摇瓶发酵中利用响应面分析方法对培养基组分进行了优化;其次,结合甜菜碱、氨基酸等物质的添加,在2 L发酵罐中进行了发酵放大优化。实验确定了较优的培养基成分,为葡萄糖(42.69 g/L)、硫酸铵(7.77 g/L)、酵母粉(6.53 g/L)、硫代硫酸钠(6.22 g/L);优化后L-半胱氨酸的产量显著提升,摇瓶发酵达到3.85 g/L,较优化前提高了169％。2 L发酵罐放大试验结果表明,结合外源物质添加,发酵47 h后L-半胱氨酸产量可达10.25 g/L。以上结果为后续L-半胱氨酸的工业化发酵生产奠定了基础。

关键词： L-半胱氨酸; 大肠杆菌; 响应面; 外源添加; 发酵优化

本文引用格式

张博 , 史永吉 , 杨辉 , 吴梓丹 , 陈开 , 蔡雪 , 柳志强 , 郑裕国 . 通过发酵优化提高大肠杆菌生产L-半胱氨酸产量[J]. 食品与发酵工业, 2021 , 47(18) : 175 -180 . DOI: 10.13995/j.cnki.11-1802/ts.026837

Abstract

Cysteine is an important amino acid with active sulfhydryl group, which plays vital role in cellular physiological functions. In this study, to improve the L-cysteine production of the Escherichia coli MCYS-7, optimization of the culture medium was firstly performed by response surface methodology in flask fermentation. With a exogenous addition of betaine, amino acids and other chemicals, the fermentation process optimization and scale-up were carried out in 2 L bioreactors. The optimum culture medium was determined to be glucose (42.69 g/L), ammonium sulfate (7.77 g/L), yeast powder (6.53 g/L), sodium thiosulfate (6.22 g/L). In the flask fermentation after medium optimization, the titer of L-cysteine was significant improved to 3.85 g/L, which was increased by 169% compared with that before optimization. The scale-up results in the 2 L bioreactors revealed that, with the addition of exogenous chemical, the titer for L-cysteine production could reach 10.25 g/L after 47 h of fermentation. Our results laid a foundation for the L-cysteine production through microbial fermentation in industrial scale.

Key words： L-cysteine; Escherichia coli; response surface; exogenous chemical addition; fermentation optimization

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