Abstract
BackgroundAcetoin utilization protein (acuC) is a type I histone deacetylase which is highly conserved in bacteria. The acuC gene is related to the acetylation/deacetylation posttranslational modification (PTM) system in S. spinosa. Spinosyns, the secondary metabolites produced by Saccharopolyspora spinosa, are the active ingredients in a family of insect control agents. However, the specific functions and influences of acuC protein in S. spinosa are yet to be characterized.ResultsThe knockout strain and overexpression strain were constructed separately with the shuttle vector pOJ260. The production of spinosyns A and D from S. spinosa-acuC were 105.02 mg/L and 20.63 mg/L, which were 1.82-fold and 1.63-fold higher than those of the wild-type strain (57.76 mg/L and 12.64 mg/L), respectively. The production of spinosyns A and D from S. spinosa-ΔacuC were 32.78 mg/L and 10.89 mg/L, respectively. The qRT-PCR results of three selected genes (bldD, ssgA and whiA) confirmed that the overexpression of acuC affected the capacities of mycelial differentiation and sporulation. Comparative proteomics analysis was performed on these strains to investigate the underlying mechanism leading to the enhancement of spinosad yield.ConclusionsThis study first systematically analysed the effects of overexpression acuC on the growth of S. spinosa and the production of spinosad. The results identify the differentially expressed proteins and provide evidences to understand the acetylation metabolic mechanisms which can lead to the increase of secondary metabolites.
Highlights
Acetoin utilization protein is a type I histone deacetylase which is highly conserved in bacteria
Liu et al Microb Cell Fact (2021) 20:141 encoding type I polyketide synthase (PKS); four genes involved in the intramolecular C–C bond formation; four genes responsible for rhamnose attachment and methylation; six genes involved in forosamine biosynthesis; four genes involved in spinosyn branched chain biosynthesis and glycosylation modification
The results showed that the logarithmic growth phases of S. spinosa and S. spinosa-acuC were basically synchronous, and the wild-type and S. spinosa-acuC strains entered the stationary growth phase after 60 h
Summary
Acetoin utilization protein (acuC) is a type I histone deacetylase which is highly conserved in bacteria. The acuC gene is related to the acetylation/deacetylation posttranslational modification (PTM) system in S. spinosa. The secondary metabolites produced by Saccharopolyspora spinosa, are the active ingredients in a family of insect control agents. The specific functions and influences of acuC protein in S. spinosa are yet to be characterized. Saccharopolyspora spinosa is a gram-positive bacterium that is non-acid-resistant and aerobic [1]. Spinosyns are synthesized by fermentation of S. spinosa under aerobic conditions [2], and they are novel macrolide compounds. S. spinosa CCTCC M206084 isolated by our lab in southern China has the ability to produce spinosad. The fermentation period is relatively long and restricts the production of spinosad [7]. The overexpression of acuC affects the gene expressions related with the mycelial differentiation and sporulation in S. spinosa as well as the production of spinosad
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