科微学术

微生物学报

产γ-氨基丁酸基因工程重组大肠杆菌的构建及其发酵特性
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

浙江省自然科学基金(LY21C200006)


Construction and fermentation characterization of γ-aminobutyrate-producing recombinant strains of Escherichia coli
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    【目的】构建高产γ-氨基丁酸的基因工程重组大肠杆菌(E.coli),并研究其发酵特性。【方法】首先通过分子生物学方法构建重组质粒pTrc99a-gadB和pTrc99a-gadB-SNO1-SNZ1,然后分别将它们转入基因敲除菌株E.coli ΔgabPΔpuuE/pTrc99a-gadB在含20 g/L底物l-谷氨酸钠的1 L发酵液中的扩大培养结果表明,重组菌培养24 h时,其发酵液中γ-氨基丁酸的含量达到最高值9.4 g/L。【结论】经基因工程构建的重组大肠杆菌产γ-氨基丁酸的能力明显提高,该研究结果为γ-氨基丁酸的产业化生产提供了良好基础。

    Abstract:

    [Objective] To construct the γ-aminobutyrate-producing recombinant strains of Escherichia coli and investigate their fermentation characteristics. [Methods] We constructed two recombinant plasmids pTrc99a-gadB and pTrc99a-gadB-SNO1-SNZ1 and then respectively transformed them into the gene-knockout strain E. coli K12/ΔgabTΔgabPΔpuuE. We investigated and optimized the fermentation process of the recombinant strains for producing γ-aminobutyrate. [Results] The target proteins were highly expressed in the recombinant strains harboring the constructed plasmids. The highest concentration of γ-aminobutyrate was 4.6 g/L in the fermentation broth of E. coli K12ΔgabTΔgabPΔpuuE/pTrc99a-gadB cultured in the medium containing 10 g/L l-monosodium glutamate and was 21.9 times higher than that in the fermentation broth of the wild type strain. At the l-monosodium glutamate concentration of 20 g/L, the conversion rate of substrate was the highest and the concentration of γ-aminobutyrate reached 8.4 g/L. The concentration of γ-aminobutyrate was slightly lower in the fermentation broth of the recombinant strain E. coli K12ΔgabTΔgabPΔpuuE/pTrc99a-gadB-SNO1-SNZ1, probably due to the excessive consumption of energy. The highest concentration of γ-aminobutyrate was 9.4 g/L when E. coli K12ΔgabTΔgabPΔpuuE/pTrc99a-gadB was cultured in 1 L fermentation medium containing 20 g/L l-monosodium glutamate. [Conclusion] We obviously increased the yield of γ-aminobutyrate produced by the recombinant strain. This finding lays a foundation for the industrial production of γ-aminobutyrate.

    参考文献
    相似文献
    引证文献
引用本文

管付瑶,陆佳杰,朱志文,王配泽,鄢楚洋,于平. 产γ-氨基丁酸基因工程重组大肠杆菌的构建及其发酵特性. 微生物学报, 2024, 64(2): 489-501

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2023-06-26
  • 最后修改日期:2023-09-22
  • 录用日期:
  • 在线发布日期: 2024-01-31
  • 出版日期: 2024-02-04