Effect of pII key nitrogen regulatory gene on strain growth and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona

PII signal transduction proteins are widely found in bacteria and plant chloroplast, and play a central role in nitrogen metabolism regulation, which interact with many key proteins in metabolic pathways to regulate carbon/nitrogen balance by sensing changes in concentrations of cell-mediated indicators such as alpha-ketoglutarate. In this study, the knockout strain Saccharopolyspora pogona-Delta pII and overexpression strain S. pogona-pII were constructed using CRISPR/Cas9 technology and the shuttle vector POJ260, respectively, to investigate the effects on the growth and secondary metabolite biosynthesis of S. pogona. Growth curve, electron microscopy, and spore germination experiments were performed, and it was found that the deletion of the pII gene inhibited the growth to a certain extent in the mutant. HPLC analysis showed that the yield of butenyl-spinosyn in the S. pogona-pII strain increased to 245% than that in the wild-type strain while that in S. pogona-Delta pII decreased by approximately 51%. This result showed that the pII gene can promote the growth and butenyl-spinosyn biosynthesis of S. pogona. This research first investigated PII nitrogen metabolism regulators in S. pogona, providing significant scientific evidence and a research basis for elucidating the mechanism by which these factors regulate the growth of S. pogona, optimizing the synthesis network of butenyl-spinosyn and constructing a strain with a high butenyl-spinosyn yield.

Automated summary

In this study, the effects of pII gene on the growth and secondary metabolite biosynthesis of Saccharopolyspora pogona were investigated. The results showed that the pII gene can promote the growth and biosynthesis of butenyl-spinosyn in S. pogona. This research provides significant scientific evidence and a research basis for elucidating the mechanism by which these factors regulate the growth of S. pogona and optimizing the synthesis network of butenyl-spinosyn.