SenX3-RegX3, an Important Two-Component System, Regulates Strain Growth and Butenyl-spinosyn Biosynthesis in Saccharopolyspora pogona

Abstract

Summary Butenyl-spinosyn produced by Saccharopolyspora pogona exhibits strong insecticidal activity and a broad pesticidal spectrum. Currently, important functional genes involved in butenyl-spinosyn biosynthesis remain unknown, which leads to difficulty in efficient understanding of its regulatory mechanism and improving its production by metabolic engineering. Here, we present data supporting a role of the SenX3-RegX3 system in regulating the butenyl-spinosyn biosynthesis. EMSAs and qRT-PCR demonstrated that RegX3 positively controls butenyl-spinosyn production in an indirect way. Integrated proteomic and metabolomic analysis, regX3 deletion not only strengthens the basal metabolic ability of S. pogona in the mid-growth phase but also promotes the flow of the acetyl-CoA produced via key metabolic pathways into the TCA cycle rather than the butenyl-spinosyn biosynthetic pathway, which ultimately leads to continued growth but reduced butenyl-spinosyn production. The strategy demonstrated here may be valuable for revealing the regulatory role of the SenX3-RegX3 system in the biosynthesis of other natural products.

A TetR family transcriptional regulator, SP_2854 can affect the butenyl-spinosyn biosynthesis by regulating glucose metabolism in Saccharopolyspora pogona

“…Although much progress has been made in understanding the butenyl-spinosyn biosynthetic pathway, little is known about the regulation of butenyl-spinosyn biosynthesis. To date, only a few regulators have been identified as being involved in the regulation of butenyl-spinosyn production in S. pogona , including the SenX3/RegX3 system, Sp1418, Sp13016, SP_1288 and an AfsR regulatory factor [ 7 – 11 ]. A better understanding of the regulatory mechanisms of butenyl-spinosyn biosynthesis will provide important insights for strain improvement by metabolic engineering.…”

The Global Regulator PhoU Positively Controls Growth and Butenyl-Spinosyn Biosynthesis in Saccharopolyspora pogona

“…Conversely, low phosphate levels has been shown to promote the transition phase to aerial mycelium generation ( Tenconi et al, 2012 )and the biosynthesis of secondary metabolites ( Rodriguez-Garcia et al, 2007 ). Recently, Rang et al (2020) found that the growth of S. pogona and the biosynthesis of butenyl-spinosyn were inhibited to a certain extent with the an increase in phosphate levels in the culture medium. There results suggested that phosphorus, as an essential element for organisms, plays a key role in regulating the growth and the biosynthesis of secondary metabolites.…”

Bacterioferritin: a key iron storage modulator that affects strain growth and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona

“…Song, et al used RedEx technology to replace different modules based on the gene cluster of spinosyn and performed heterologous expression in Streptomyces albicans , successfully synthesizing butenyl-spinosyn [ 20 ]. In addition, the concentrations of inorganic salts, such as phosphate and iron salt necessary for microorganisms in culture medium affected the growth and secondary metabolism in Streptomyces [ 21 – 23 ].…”