Genetics of subpeptin JM4-A and subpeptin JM4-B production by Bacillus subtilis JM4

Biochemical and Biophysical Research Communications
Shimei WuLiandong Huan

Abstract

Subpeptin JM4-A and subpeptin JM4-B are two novel antimicrobial peptides produced by Bacillus subtilis JM4. To identify putative genes involved in their production, degenerate PCR primers targeted to conserved motifs of nonribosomal peptide synthetases (NRPSs) were used. A resulting 1.2 kb PCR product had high sequence similarity to genes of NRPSs, and then a 2.8 kb DNA fragment flanking it was cloned subsequently. Gene disruption of the resulting 4 kb DNA fragment produced subpeptin-deficient mutant, suggesting that subpeptin JM4-A and subpeptin JM4-B were biosynthesized by NRPSs. Based on this result, a 48 kb gene cluster was cloned, which consisted of nine coding sequences (CDSs) involved in antimicrobial peptide biosynthesis, regulation, and resistance. Disruption of two relatively large CDSs subA and subC led to subpeptin-deficient mutants, which supported the involvement of the cloned gene cluster in subpeptin biosynthesis.

References

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Citations

Apr 5, 2011·Extremophiles : Life Under Extreme Conditions·Fumitoshi ManabeTakayoshi Wakagi
Mar 29, 2014·Probiotics and Antimicrobial Proteins·Avelino Alvarez-OrdóñezColin Hill
Aug 25, 2015·International Journal of Systematic and Evolutionary Microbiology·Christopher A DunlapSoo-Jin Kim
Dec 25, 2012·Natural Product Reports·Anne-Catrin LetzelChristian Hertweck

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