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Improvement of Rimocidin Biosynthesis by Increasing Supply of Precursor Malonyl-CoA via Over-expression of Acetyl-CoA Carboxylase in Streptomyces rimosus M527

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Abstract

Precursor engineering is an effective strategy for the overproduction of secondary metabolites. The polyene macrolide rimocidin, which is produced by Streptomyces rimosus M527, exhibits a potent activity against a broad range of phytopathogenic fungi. It has been predicted that malonyl-CoA is used as extender units for rimocidin biosynthesis. Based on a systematic analysis of three sets of time-series transcriptome microarray data of S. rimosus M527 fermented in different conditions, the differentially expressed accsr gene that encodes acetyl-CoA carboxylase (ACC) was found. To understand how the formation of rimocidin is being influenced by the expression of the accsr gene and by the concentration of malonyl-CoA, the accsr gene was cloned and over-expressed in the wild-type strain S. rimosus M527 in this study. The recombinant strain S. rimosus M527-ACC harboring the over-expressed accsr gene exhibited better performances based on the enzymatic activity of ACC, intracellular malonyl-CoA concentrations, and rimocidin production compared to S. rimosus M527 throughout the fermentation process. The enzymatic activity of ACC and intracellular concentration of malonyl-CoA of S. rimosus M527-ACC were 1.0- and 1.5-fold higher than those of S. rimosus M527, respectively. Finally, the yield of rimocidin produced by S. rimosus M527-ACC reached 320.7 mg/L, which was 34.0% higher than that of S. rimosus M527. These results confirmed that malonyl-CoA is an important precursor for rimocidin biosynthesis and suggested that an adequate supply of malonyl-CoA caused by accsr gene over-expression led to the improvement in rimocidin production.

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Funding

This work was supported by the National Natural Science Foundation of China (31772213, 31972320), the Zhejiang Province Natural Science Foundation (LY20C140006), and the Key Program of Zhejiang Province Natural Science Foundation (LZ22C140002).

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  1. Zhijun Liao and Jinyao Zhang have contributed equally to this article.

Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, College of Life Sciences, China Jiliang University, Xueyuan Street, Xiasha Higher Education District, Hangzhou, 310018, Zhejiang Province, People’s Republic of China

    Zhijun Liao, Jinyao Zhang, Yue Shi, Yongyong Zhang, Zheng Ma & Xiaoping Yu

  2. Institute for Pharmaceutical Sciences, Pharmaceutical Biology and Biotechnology, University of Freiburg, 79104, Freiburg, Germany

    Andreas Bechthold

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  1. Zhijun Liao
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  7. Xiaoping Yu
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ZJL, JYZ, and YS conducted experiments. ZM and YYZ designed the experiments and wrote this article. AB and XPY checked the final version.

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Correspondence to Zheng Ma.

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Liao, Z., Zhang, J., Shi, Y. et al. Improvement of Rimocidin Biosynthesis by Increasing Supply of Precursor Malonyl-CoA via Over-expression of Acetyl-CoA Carboxylase in Streptomyces rimosus M527. Curr Microbiol 79, 174 (2022). https://doi.org/10.1007/s00284-022-02867-9

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