Expression and characterization of aiiA gene from Bacillus subtilis BS-1

Microbiol Res. 2008;163(6):711-6. doi: 10.1016/j.micres.2007.12.002.

Abstract

AHL-lactonase (AiiA), a metallo-beta-lactamase produced by Bacillus thuringiensis, Bacillus cereus and Bacillus anthracis, specifically hydrolyzes N-acyl-homoserine lactones (AHLs) secreted by Gram-negative bacteria and thereby attenuates the symptoms caused by plant pathogens. In this study, an aiiA gene was cloned from Bacillus subtilis BS-1 by PCR with a pair of degenerate primers. The deduced 250 amino acid sequence contained two small conserved regions, 103SHLHFDH109 and 166TPGHTPGH173, which are characteristic of the metallo-beta-lactamase family. Homology comparison revealed that the deduced amino acid sequence had a high degree of similarity with those of the known AiiA proteins in the B. cereus group. Additionally, the aiiA gene was expressed in Escherichia coli BL21 (DE3) pLysS and the expressed AiiA protein could attenuate the soft rot symptoms caused by Erwinia carotovora var. carotovora.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / genetics
  • Bacillus subtilis / growth & development
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial*
  • Metalloendopeptidases* / chemistry
  • Metalloendopeptidases* / genetics
  • Metalloendopeptidases* / metabolism
  • Molecular Sequence Data
  • Pectobacterium carotovorum / growth & development*
  • Pectobacterium carotovorum / metabolism
  • Pest Control, Biological*
  • Plant Diseases / microbiology
  • Solanum tuberosum / microbiology*

Substances

  • Bacterial Proteins
  • AiiA protein, Bacillus
  • Metalloendopeptidases