Improvement of Rimocidin Biosynthesis by Increasing Supply of Precursor Malonyl-CoA via Over-expression of Acetyl-CoA Carboxylase in Streptomyces rimosus M527
Current Microbiology(2022)
摘要
Precursor engineering is an effective strategy for the overproduction of secondary metabolites. The polyene macrolide rimocidin, which is produced by Streptomyces rimosus M527, exhibits a potent activity against a broad range of phytopathogenic fungi. It has been predicted that malonyl-CoA is used as extender units for rimocidin biosynthesis. Based on a systematic analysis of three sets of time-series transcriptome microarray data of S . rimosus M527 fermented in different conditions, the differentially expressed acc sr gene that encodes acetyl-CoA carboxylase (ACC) was found. To understand how the formation of rimocidin is being influenced by the expression of the acc sr gene and by the concentration of malonyl-CoA, the acc sr gene was cloned and over-expressed in the wild-type strain S . rimosus M527 in this study. The recombinant strain S . rimosus M527-ACC harboring the over-expressed acc sr gene exhibited better performances based on the enzymatic activity of ACC, intracellular malonyl-CoA concentrations, and rimocidin production compared to S . rimosus M527 throughout the fermentation process. The enzymatic activity of ACC and intracellular concentration of malonyl-CoA of S . rimosus M527-ACC were 1.0- and 1.5-fold higher than those of S . rimosus M527, respectively. Finally, the yield of rimocidin produced by S . rimosus M527-ACC reached 320.7 mg/L, which was 34.0% higher than that of S . rimosus M527. These results confirmed that malonyl-CoA is an important precursor for rimocidin biosynthesis and suggested that an adequate supply of malonyl-CoA caused by acc sr gene over-expression led to the improvement in rimocidin production.
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