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Background <p>Heterologous expression in <i>Streptomyces</i> provides a platform for mining natural products (NPs) encoded by cryptic biosynthetic gene clusters (BGCs) of bacteria. The BGCs are first engineered in hosts with robust recombineering systems, such as <i>Escherichia coli</i>, followed by expression in optimized heterologous hosts, such as <i>Streptomyces</i>, with defined metabolic backgrounds.</p> Results <p>We developed a highly efficient heterologous expression platform, named Micro-HEP (<i>micro</i>bial <i>h</i>eterologous <i>e</i>xpression <i>p</i>latform), that uses versatile <i>E. coli</i> strains capable of both modification and conjugation transfer of foreign BGCs and optimized chassis <i>Streptomyces</i> strain for expression. The stability of repeat sequences in these <i>E. coli</i> strains was superior to that of the commonly used conjugative transfer system <i>E. coli</i> ET12567 (pUZ8002). For optimizing expression of foreign BGCs, the chassis strain <i>S. coelicolor</i> A3(2)-2023 was generated by deleting four endogenous BGCs followed by introducing multiple recombinase-mediated cassette exchange (RMCE) sites in the <i>S. coelicolor</i> A3(2) chromosome. Additionally, modular RMCE cassettes (Cre-lox, Vika-vox, Dre-rox, and phiBT1-attP) were constructed for integrating BGCs into the chassis strain. Micro-HEP was tested using BGCs for the anti-fibrotic compound xiamenmycin and griseorhodins. Two to four copies of the <i>xim</i> BGC were integrated by RMCE, with increasing copy number associated with increasing yield of xiamenmycin. The <i>grh</i> BGC was also efficiently expressed, and the new compound griseorhodin H was identified.</p> Conclusion <p>We demonstrated that our Micro-HEP system enables the efficient expression of foreign BGCs, facilitating the discovery of new NPs and increasing yields.</p> Graphical Abstract <p></p>

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A highly efficient heterologous expression platform to facilitate the production of microbial natural products in Streptomyces

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年5月14日

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  • Xiuling Wang,
  • Ping Lin,
  • Qiyao Shen,
  • Xueyan Feng,
  • Shouying Xu,
  • Qijun Zhang,
  • Yang Liu,
  • Cailing Ren,
  • Daojing Yong,
  • Qiong Duan,
  • Liujie Huo,
  • Youming Zhang,
  • Gang Li,
  • Jun Fu,
  • Ruijuan Li

摘要

Background

Heterologous expression in Streptomyces provides a platform for mining natural products (NPs) encoded by cryptic biosynthetic gene clusters (BGCs) of bacteria. The BGCs are first engineered in hosts with robust recombineering systems, such as Escherichia coli, followed by expression in optimized heterologous hosts, such as Streptomyces, with defined metabolic backgrounds.

Results

We developed a highly efficient heterologous expression platform, named Micro-HEP (microbial heterologous expression platform), that uses versatile E. coli strains capable of both modification and conjugation transfer of foreign BGCs and optimized chassis Streptomyces strain for expression. The stability of repeat sequences in these E. coli strains was superior to that of the commonly used conjugative transfer system E. coli ET12567 (pUZ8002). For optimizing expression of foreign BGCs, the chassis strain S. coelicolor A3(2)-2023 was generated by deleting four endogenous BGCs followed by introducing multiple recombinase-mediated cassette exchange (RMCE) sites in the S. coelicolor A3(2) chromosome. Additionally, modular RMCE cassettes (Cre-lox, Vika-vox, Dre-rox, and phiBT1-attP) were constructed for integrating BGCs into the chassis strain. Micro-HEP was tested using BGCs for the anti-fibrotic compound xiamenmycin and griseorhodins. Two to four copies of the xim BGC were integrated by RMCE, with increasing copy number associated with increasing yield of xiamenmycin. The grh BGC was also efficiently expressed, and the new compound griseorhodin H was identified.

Conclusion

We demonstrated that our Micro-HEP system enables the efficient expression of foreign BGCs, facilitating the discovery of new NPs and increasing yields.

Graphical Abstract