Cloning, expression and characterization of Ahlinactivating aiiA gene homologue in Escherichia coli
Loading...
Date
2014-08
Authors
Muniandy, Subashini
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
Cell-to-cell communication pathway of single-celled microbes is usually coordinated by quorum sensing (QS) system mediated by signal molecules. This cell density-dependent system plays a vital role in functional coordination of most pathogenic bacteria. Antagonistically, interruption of this signal pathway by inactivating the signal molecules through quorum quenching (QQ) could be an alternative strategy to control bacterial virulence. In this study, a local soil isolate identified as Bacillus cereus SM01,has been shown to possess the capability of enzymatic inactivation of acyl homoserine lactones (AHLs). This special trait of SM01 prompted the interest in testing its potential quorum quenching ability. Hence, the amplified 770 bp long AHL-lactonase gene homologue (aiiA) of SM01 was subsequently cloned into pCold expression system in order to express the desired recombinant AHL-lactonase protein. Further characterization of that the overexpressed 29 kDa recombinant protein revealed that it drastically decreased biofilm formation of Burkholderia pseudomallei UKMS-01 strain, an animal pathogen. Interestingly, the recombinant AHL-lactonase protein was observed to be stable up to 85ÂșC in temperature where most of the enzymes will be inactive. Nevertheless, it showed wide range of pH stability, from pH 5 up to 11. All these characteristics suggest that this recombinant AHL-lactonase has the potential to be used as a commercial enzyme for the control of pathogenic bacterial infection.
Description
Keywords
Ahlinactivating aiiA gene homologue , Escherichia coli