title: [Soluble expression of AiiA protein and analysis of its antimicrobial activity].
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.
authors.affiliation: College of Life Sciences, Fujian Normal University, Fuzhou 350108, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Yang
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Bin Hui
authors.initials: BH
authors.lastname: Lin
authors.forename: Xin Hua
authors.initials: XH
authors.lastname: Su
authors.forename: Li Qing
authors.initials: LQ
authors.lastname: Guo
authors.forename: Zhi Peng
authors.initials: ZP
authors.lastname: Huang
authors.forename: Xiong
authors.initials: X
authors.lastname: Guan
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism / Q000494:pharmacology*; D001413:Bacillus thuringiensis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D016973:Pectobacterium carotovorum / Q000187:drug effects; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D011994:Recombinant Proteins