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<p>Itaconic acid (ITA) is an unsaturated organic acid used in industrial production due to its versatility as a polymer building block. Engineering microbial cell factories for ITA biosynthesis from cost-effective and renewable raw materials has gained significant attention. Here, we performed combinatorial engineering using <i>Saccharomyces cerevisiae</i> to improve ITA production. First, exogenous <i>cis</i>-aconitic acid decarboxylase (CAD) was integrated into <i>S. cerevisiae</i> to construct the ITA-producing chassis. Then, the rate-limiting step was eliminated by changing the promoter that drives CAD expression to optimize ITA synthesis. A mitochondrial cis-aconitate transporter MTTA was also expressed to facilitate the transport of precursor, which resulted in an ITA titer of 244&#xa0;mg/L. Furthermore, with overexpression of truncated citrate synthase tCIT2, an increased titer of 409&#xa0;mg/L was obtained. Finally, the transport protein Qdr3 was overexpressed to enhance the export of ITA, resulting in a production of 578&#xa0;mg/L in shake flask. In a 5-L bioreactor, the ITA titer reached 1.2&#xa0;g/L, representing the highest reported level in <i>S. cerevisiae</i>. Overall, an advanced recombinant yeast strain was constructed for the efficient production of ITA via combinatorial metabolic engineering.</p>

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Engineering Saccharomyces cerevisiae for improving itaconic acid production

  • 期刊论文

  • 发表日期：

    2025年2月18日

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  • Hao Xu,
  • Wenwen Yu,
  • Xuan Zhou,
  • Jiaheng Liu,
  • Xianhao Xu,
  • Yanfeng Liu,
  • Jianghua Li,
  • Guocheng Du,
  • Long Liu,
  • Xueqin Lv

摘要

Itaconic acid (ITA) is an unsaturated organic acid used in industrial production due to its versatility as a polymer building block. Engineering microbial cell factories for ITA biosynthesis from cost-effective and renewable raw materials has gained significant attention. Here, we performed combinatorial engineering using Saccharomyces cerevisiae to improve ITA production. First, exogenous cis-aconitic acid decarboxylase (CAD) was integrated into S. cerevisiae to construct the ITA-producing chassis. Then, the rate-limiting step was eliminated by changing the promoter that drives CAD expression to optimize ITA synthesis. A mitochondrial cis-aconitate transporter MTTA was also expressed to facilitate the transport of precursor, which resulted in an ITA titer of 244 mg/L. Furthermore, with overexpression of truncated citrate synthase tCIT2, an increased titer of 409 mg/L was obtained. Finally, the transport protein Qdr3 was overexpressed to enhance the export of ITA, resulting in a production of 578 mg/L in shake flask. In a 5-L bioreactor, the ITA titer reached 1.2 g/L, representing the highest reported level in S. cerevisiae. Overall, an advanced recombinant yeast strain was constructed for the efficient production of ITA via combinatorial metabolic engineering.