Whole Genome Sequencing of Bacillus subtilis SNBS-3 and Prediction of Its Antimicrobial Substances

Abstract

As an extension of the previous research, this study aimed to comprehensively characterize the genome of Bacillus subtilis SNBS-3. Illumina second-generation sequencing technology and the third-generation high-throughput Pacbio sequencing platform were used for whole-genome sequencing of B. subtilis SNBS-3 isolated from traditional bean paste to obtain the key information of genome characteristics, gene function annotation and classification, phylogenetic evolution, and secondary metabolites. The results showed that the genome of SNBS-3 was a closed circular DNA of 4 076 387 bp in length containing 4 000 protein-coding genes. A total of 3 209, 2 824, 2 560, 147, 12 and 4 functional genes were annotated in the Clusters of Orthologous Groups (COG), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Carbohydrate-Active Enzymes (CAZyme), Comprehensive Antibiotic Resistance Database (CARD) and Virulence Factor Database (VFDB), respectively. Using the online software AntiSMASH and Bagel4, we found that it contained genes related to the synthesis of surfactin, mycosubtilin, plipastatin, bacilysin and bacillaene, as well as a complete gene cluster for the synthesis of the bacteriocin subtilosin A. Based on the results of antimicrobial test and proteinase K test, it was hypothesized that B. subtilis SNBS-3 had the ability to synthesize subtilosin A. In conclusion, the whole genome sequencing results of B. subtilis SNBS-3 show that it can produce a variety of bacteriostatic substances and thus have biocontrol potential. The results from this study provide a theoretical basis for further development and application of various bacteriostatic substances including the bacteriocin subtilosin A.

Keywords

• bacillus subtilis; whole genome; bacteriocin; synthetic gene cluster