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Calcium-Phosphate Combination Enhances Spinosad Production in Saccharopolyspora spinosa via Regulation of Fatty Acid Metabolism

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Abstract

Phosphate concentration above 10 mM reduces the production of many secondary metabolites; however, the phenomenon is not mechanistically understood yet. Specifically, the problem of phosphorus limitation in antibiotic production remains unresolved. This study investigates the phosphorus inhibition effect on spinosad production and alleviates it by calcium and phosphate supplementation to fermentation media. Furthermore, we examined the mechanism of fatty acids–induced increase in polyketides production. Four phosphates that were supplemented into the fermentation media include NaH2PO4, Na2HPO4, KH2PO4, and K2HPO4 and NaH2PO4 was found to be the most effective phosphate. Under the optimal phosphate condition of supplementing 20 mM NaH2PO4 on the fourth day and 5 g/L CaCO3, the maximal spinosad production reached 520 mg/L, showing a 1.65-fold increase over the control treatment. In the NaH2PO4-CaCO3 system, the de novo fatty acid biosynthesis was significantly downregulated while spinosad biosynthesis and β-oxidation were upregulated. The coordination of de novo fatty acid biosynthesis and β-oxidation promoted intracellular acetyl-CoA concentration. The results demonstrate that NaH2PO4-CaCO3 combined addition is a simple and effective strategy to alleviate phosphorus inhibition effect through the regulation of fatty acid metabolism and accumulation of immediate precursors. This information improves our understanding of phosphates’ influence on the large-scale production of polyketides.

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All datasets generated for this study are included in the article.

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Funding

This research was financially supported by the National Natural Science Foundation of China (Nos. 21937002,), and the National Science and Technology Major Project for New Drug Research and Development of MOST (2019ZX09735002-005).

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Author notes

  1. Mi-Yang Wan and Cheng Peng contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Medicines & Shanghai Engineering Research Center of Immunotherapeutics, School of Pharmacy, Fudan University, Shanghai, 201203, China

    Mi-Yang Wan, Cheng Peng, Wen-Xin Ding, Meng-Ran Wang & Jiyang Li

  2. Department of Natural Medicine, School of Pharmacy, Fudan University, Shanghai, 201203, China

    Jinfeng Hu

Authors
  1. Mi-Yang Wan
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  2. Cheng Peng
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  3. Wen-Xin Ding
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  4. Meng-Ran Wang
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Contributions

CP performed the data analysis and wrote the first draft of the manuscript. M-YW and CP performed the experiments and data interpretation. M-RW and W-XD provided technical assistance to CP and J-FH. J-YL supervised the experimental design, data analysis, manuscript writing, and revision. All authors read and agreed on the final text.

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Correspondence to Jiyang Li.

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Wan, MY., Peng, C., Ding, WX. et al. Calcium-Phosphate Combination Enhances Spinosad Production in Saccharopolyspora spinosa via Regulation of Fatty Acid Metabolism. Appl Biochem Biotechnol 194, 2528–2541 (2022). https://doi.org/10.1007/s12010-022-03799-7

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  • DOI: https://doi.org/10.1007/s12010-022-03799-7

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