食品与发酵工业 >

2024 , Vol. 50 >Issue 3: 38 - 44

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035424

研究报告

己二酸生物合成的途径改造以及发酵条件优化

  • 支睿 ,
  • 李国辉 ,
  • 毛银 ,
  • 邓禹
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  • 1(江南大学 粮食发酵与食品生物制造国家工程研究中心,江苏 无锡,214122)
    2(江南大学 生物工程学院,江苏 无锡,214122)
第一作者:硕士研究生(李国辉副研究员和邓禹教授为通信作者,E-mail:guohuili@jiangnan.edu.cn;dengyu@jiangnan.edu.cn)

收稿日期: 2023-03-09

  修回日期: 2023-03-31

  网络出版日期: 2024-03-15

基金资助

国家重点研发计划项目(2021YFC2100700);国家自然科学基金项目(22008088,21877053);中国博士后科学基金资助项目(2020M681485,2021T140277);江苏省博士后科研资助计划项目(2020Z012)

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Metabolic pathway and fermentation optimization of the biosynthesis of adipic acid

  • ZHI Rui ,
  • LI Guohui ,
  • MAO Yin ,
  • DENG Yu
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  • 1(National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China)
    2(School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2023-03-09

  Revised date: 2023-03-31

  Online published: 2024-03-15

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摘要

逆己二酸降解途径因可用于己二酸合成受到了广泛的关注,但因其代谢途径较长、途径基因表达不平衡,难以对其异源的表达进行调控。因此,该研究将逆己二酸降解途径模块化,更换各模块的启动子以实现该途径在底盘细胞中表达水平的调控,从而提高己二酸产量。利用己二酸生物传感器开展高通量筛选验证,获得最优组合模块菌株己二酸产量达到120.66 mg/L,提高至对照的8.35倍。通过理性分析确定了合成途径关键酶,最终实现了异源基因的有效表达调控。在此基础上,开展发酵条件优化,己二酸产量达到240.49 mg/L,较对照提高了16.65倍。该研究通过模块化改造,协调平衡异源途径基因表达,为己二酸生物合成研究提供了新思路。

关键词： 己二酸; 组成型表达; 逆己二酸降解途径; 高通量筛选; 模块化改造

本文引用格式

支睿 , 李国辉 , 毛银 , 邓禹 . 己二酸生物合成的途径改造以及发酵条件优化[J]. 食品与发酵工业, 2024 , 50(3) : 38 -44 . DOI: 10.13995/j.cnki.11-1802/ts.035424

Abstract

Employing the reverse adipate degradation pathway to synthesize adipic acid draws significant attention, but it's difficult to modify this metabolic pathway because of the long length and the imbalance heterologous expression of the pathway genes. To further increase the production of adipic acid, this study therefore modularized the reverse adipate degradation pathway, substituting the promoters of each module in order to regulate the expression profile of the pathway genes. By utilizing an adipic acid biosensor-based high-throughput screening method, the best combination resulted in a titer of 120.66 mg/L, which achieved an 8.35-fold improvement comparing to the control group. Moreover, the key enzymes of the pathway were confirmed through rational analysis and an efficiently regulation of the heterologous-expressed genes was eventually realized. Based on this, fermentation optimization was conducted and a titer of 240.49 mg/L adipic acid was eventually obtained, which was 16.67-fold higher than that of the control group. This study provided a new thought on the biosynthesis of adipic acid by modulating the pathway and regulating the gene expression profile.

Key words： adipic acid; constitutive expression; the reverse adipate degradation pathway; high-throughput screening; module modification

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