Abstract

Whole-genome sequencing of a soil isolate Bacillus pumilus, strain 7P, and its streptomycin-resistant derivative, B. pumilus 3-19, showed genome sizes of 3,609,117 bp and 3,609,444 bp, respectively. Annotation of the genome showed 3794 CDS (3204 with predicted function) and 3746 CDS (3173 with predicted function) in the genome of strains 7P and 3-19, respectively. In the genomes of both strains, the prophage regions Bp1 and Bp2 were identified. These include 52 ORF of prophage proteins in the Bp1 region and 38 prophages ORF in the Bp2 region. Interestingly, more than 50% of Bp1 prophage proteins are similar to the proteins of the phi105 in B. subtilis. The DNA region of Bp2 has 15% similarity to the DNA of the Brevibacillus Jimmer phage. Degradome analysis of the genome of both strains revealed 148 proteases of various classes. These include 60 serine proteases, 48 metalloproteases, 26 cysteine proteases, 4 aspartate proteases, 2 asparagine proteases, 3 threonine proteases, and 2 unclassified proteases. Likewise, three inhibitors of proteolytic enzymes were found. Comparative analysis of variants in the genomes of strains 7P and 3-19 showed the presence of 81 nucleotide variants in the genome 3-19. Among them, the missense mutations in the rpsL, comA, spo0F genes and in the upstream region of the srlR gene were revealed. These nucleotide polymorphisms may have affected the streptomycin resistance and overproduction of extracellular hydrolases of the 3-19 strain. Finally, a plasmid DNA was found in strain 7P, which is lost in its derivative, strain 3-19. This plasmid contains five coding DNA sequencing (CDS), two regulatory proteins and three hypothetical proteins.

Highlights

  • The genus Bacillus is defined as Gram positive, aerobic or facultative anaerobic, motile and endospore-forming rod-shaped microorganisms [1]

  • Whole-genome sequencing of the B. pumilus 7P soil isolate and its streptomycinresistant mutant B. pumilus 3-19 allowed us to detect plasmid DNA in the 7P strain of 6019 bp, which is absent in the 3-19 strain

  • Proteins of cell defense mechanisms are widely represented, as well as toxins with nuclease activity to limit the growth of competitors

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Summary

Introduction

The genus Bacillus is defined as Gram positive, aerobic or facultative anaerobic, motile (peritrichous flagella) and endospore-forming rod-shaped microorganisms [1]. Members of the Bacillus genus have been isolated from diverse habitats including soil, plant tissues [2], marine sediments [3] and extreme environmental conditions [4]. They could provoke food poisoning [5], animal and human diseases [1,6]. Many Bacillus species synthesize a wide variety of metabolites with antimicrobial activity (antibiotics, antimicrobial peptides), extracellular proteins (mainly proteases, lipase, amylase, cellulose, etc.) [7]. Food and Drug Administration (FDA) gave the GRAS-status (the acronym for “generally recognized as safe”) for B. subtilis, B. licheniformis and B. pumilus species [GRAS Notice

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