Cloning and expression of luxS and pfs and in vitro biosynthesis autoinducer 2 of avian pathogenic Escherichia coli from Anhui Province

Abstract:

Abstract:［Objective］The LuxS /AI-2 quorum sensing (QS) system shared by Gram-positive and Gram-negative bacteria involves the production of autoinducer-2 (AI-2)． In this study，the method of biosynthesis of AI-2 was established using recombinant LuxS and Pfs of avian pathogenic Escherichia coli (APEC)，which will be benefit for future study of the role of AI-2 in APEC． ［Methods］ We investigated AI-2 production in APEC by vibrio harveyi BB170 (BB170)．Furthermore，APEC LuxS and Pfs were expressed，purified and used to investigate the production of AI-2 in vitro． LuxS and Pfs were incubated with S-ribosylhomocysteine (SAH)，the reaction was detected for the production of luminescence of BB170． ［Results］APEC can produce AI-2 by BB170 bioassay． Purified LuxS and Pfs enzymes incubated with SAH and produced 300μmol/L AI-2 in the reaction products． ［Conclusion］The results demonstrated that recombinant Pfs and LuxS synthesize AI-2 in vitro from SAH． These findings will be of benefit to future studies of the role of AI-2 in APEC．