Isolation, identification and characterization of novel Bacillus subtilis

J Vet Med Sci. 2018 Mar 24;80(3):427-433. doi: 10.1292/jvms.16-0572. Epub 2018 Jan 23.

Abstract

In this study, we have identified a bacterium that can inhibit the growth of Staphylococcus aureus, and further analyzed its antibacterial activity and other biological characteristics and laid the foundation for its future application. Through isolation and culture of the unknown bacteria, the culture characteristics, morphology observation, biochemical test, preliminary antibacterial test, 16S rRNA PCR amplification, sequence analysis, and homology analysis were performed. It was found that the bacteria are Gram positive spore chain Bacillus. The bacteria could only ferment glucose for acid production, but could not utilize lactose and maltose. The VP test for this bacteria was positive, while indole and methyl red tests were negative. Further analysis showed that these bacteria shared a homology up to 99.4% with Bacillus subtilis DQ198162.1. Thus, this newly identified bacterium was classified as Bacillus subtilis. Importantly, the crude bacteriocin of this Bacillus subtilis could inhibit the growth of Staphylococcus aureus, Escherichia coli, Enterococcus and Salmonella, which implies its potential usage in the future.

Keywords: Bacillus subtilis; bacteriocins; identification.

MeSH terms

  • Bacillus subtilis / genetics
  • Bacillus subtilis / isolation & purification*
  • Bacillus subtilis / physiology
  • Bacteriocins / metabolism
  • Escherichia coli / growth & development
  • Glucose / metabolism
  • Microbial Sensitivity Tests
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Staphylococcus aureus / growth & development

Substances

  • Bacteriocins
  • RNA, Ribosomal, 16S
  • Glucose