[Soluble expression of AiiA protein and analysis of its antimicrobial activity]

Fen Zi Xi Bao Sheng Wu Xue Bao. 2008 Dec;41(6):465-72.
[Article in Chinese]

Abstract

AHLs are signaling molecules produced by Gram-negative bacterium during their proliferation. These molecules are closely related with bacteria pathogenicity. AiiA protein, functioning as a intracellular lactonase, hydrolyses AHLs at lactone loop in the molecule to reduce its biological activity and also dramatically reduces pathogenicity of bacterium at the same time. In this study, DNA fragment encoding aiiA gene was amplified from the plasmid derived from Bacillus thuringiensis by PCR. Expression vector, pET29a-aiiA, was constructed and transformed into bacteria strain of E. coli BL21 (DE3) for expression of AiiA protein. After 25 h induction with 0.8 mmol/L IPTG at 20 degrees C, the protein AiiA expressed in soluble form by recombinant strain reached about 54.4 microg/mL. The recombinant protein was purified with Ni-affinity chromatography. Biological activity analysis showed that AiiA protein had a capability to hydrolyse AHLs, and strong antimicrobial activity for Eriwinia carotovora.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents / metabolism
  • Anti-Infective Agents / pharmacology*
  • Bacillus thuringiensis / genetics
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Bacterial Proteins / pharmacology*
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Genetic Vectors / genetics
  • Pectobacterium carotovorum / drug effects
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacology

Substances

  • Anti-Infective Agents
  • Bacterial Proteins
  • Recombinant Proteins