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Impact on strain growth and butenyl-spinosyn biosynthesis by overexpression of polynucleotide phosphorylase gene in Saccharopolyspora pogona

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Abstract

Polynucleotide phosphorylase is a highly conserved protein found in bacteria and fungi that can regulate the transcription of related enzymes involved in amino acid metabolism, organic acid metabolism, and cell biosynthesis. We studied the effect of polynucleotide phosphorylase on Saccharopolyspora pogona (S. pogona) growth and the synthesis of secondary metabolites. First, we generated the overexpression vector pOJ260-PermE-pnp via overlap extension PCR. The vector pOJ260-PermE-pnp was then introduced into S. pogona by conjugal transfer, thereby generating the recombination strain S. pogona-Pnp. Results showed that engineering strains possessed higher biomass than those of the wild-type strains. Moreover, the ability of these strains to produce spores on solid medium was stronger than that of the wild-type strains. HPLC results revealed that the butenyl-spinosyn yield in S. pogona-Pnp increased by 1.92-fold compared with that of S. pogona alone. These findings revealed that overexpression of polynucleotide phosphorylase effectively promoted butenyl-spinosyn biosynthesis in S. pogona. This result may be extended to other Streptomyces for strain improvement.

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Funding

This work was supported by funding from the National Natural Science Foundation of China (31770106, 3107006), the National Basic Research Program (973) of China (2012CB722301), the National High Technology Research and Development program (863) of China (2011AA10A203), and the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486).

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Author notes

  1. Li Li and Jie Rang contributed equally to this work.

  2. Liqiu Xia and Shengbiao Hu are the co-corresponding authors.

Authors and Affiliations

  1. Hunan Provincial Key Laboratory for Microbial Molecular Biology, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, 410081, China

    Li Li, Jie Rang, Haochen He, Siying He, Zhudong Liu, Jianli Tang, Jie Xiao, Lian He, Shengbiao Hu, Ziquan Yu, Xuezhi Ding & Liqiu Xia

Authors
  1. Li Li
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  2. Jie Rang
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  3. Haochen He
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  4. Siying He
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  5. Zhudong Liu
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  8. Lian He
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  9. Shengbiao Hu
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  10. Ziquan Yu
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Contributions

L.L. and J.X. performed bacterial strain isolation. L.L., J.T., and X.L. performed HPLC. J.R., H.S., and Z.L. performed LC-MS/MS and data analysis. L.L., L.X., and X.D. designed the study and wrote the manuscript. All authors discussed the results and approved the final manuscript.

Corresponding authors

Correspondence to Shengbiao Hu or Liqiu Xia.

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The authors declare that they have no conflict of interest.

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Not applicable. This article does not contain any studies with human participants or animals performed by any of the authors.

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Li, L., Rang, J., He, H. et al. Impact on strain growth and butenyl-spinosyn biosynthesis by overexpression of polynucleotide phosphorylase gene in Saccharopolyspora pogona. Appl Microbiol Biotechnol 102, 8011–8021 (2018). https://doi.org/10.1007/s00253-018-9178-z

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  • DOI: https://doi.org/10.1007/s00253-018-9178-z

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