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http://dx.doi.org/10.4014/jmb.1710.10034

Scarless Genomic Point Mutation to Construct a Bacillus subtilis Strain Displaying Increased Antibiotic Plipastatin Production  

Jeong, Da-Eun (Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
So, Younju (Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
Lim, Hayeon (Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
Park, Seung-Hwan (Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
Choi, Soo-Keun (Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
Publication Information
Journal of Microbiology and Biotechnology / v.28, no.6, 2018 , pp. 1030-1036 More about this Journal
Abstract
Bacillus strains produce various types of antibiotics, and random mutagenesis has traditionally been used to overproduce these natural metabolites. However, this method leads to the accumulation of unwanted mutations in the genome. Here, we rationally designed a single nucleotide substitution in the degU gene to generate a B. subtilis strain displaying increased plipastatin production in a foreign DNA-free manner. The mutant strain (BS1028u) showed improved antifungal activity against Pythium ultimum. Notably, pps operon deletion in BS1028u resulted in complete loss of antifungal activity, suggesting that the antifungal activity strongly depends on the expression of the pps operon. Quantitative real-time PCR and lacZ assays showed that the point mutation resulted in 2-fold increased pps operon expression, which caused the increase in antifungal activity. Likewise, commercial Bacillus strains can be improved to display higher antifungal activity by rationally designed simple modifications of their genome, rendering them more efficient biocontrol agents.
Keywords
Bacillus subtilis; genome engineering; degU; antibiotics; plipastatin;
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