Characterization and complete sequence of lactonase enzyme from Bacillus weihenstephanensis isolate P65 with potential activity against acyl homoserine lactone signal molecules.
Acyl homoserine lactones (AHLs) are the most common class of quorum sensing signal molecules (autoinducers) that have been reported to be essential for virulence of many relevant pathogenic bacteria such as Pseudomonas aeruginosa. New approach for controlling infections of such bacteria is through quorum quenching. In this study, the acyl homoserine lactone inhibitory activity of the crude enzyme from a Bacillus weihenstephanensis-isolate P65 was characterized. The crude enzyme was found to have relatively high thermal stability and was stable in pH range 6 to 9. The crude enzyme extract was found to have lactonase activity of 36.3 U/mg total protein. Maximum enzyme activity was achieved within a range of 28-50°C and pH 6-9. None of the metals used enhanced the activity neither did EDTA inhibit it. However, a concentration of 10 mM Fe(+2) reduced the activity to 73.8%. Catalytic activity and kinetic constants were determined using hexanoyl homoserine lactone as a substrate. Studying enzyme substrate specificity using synthetic standard signals displayed broad spectrum of activity. The enzyme was found to be constitutive. Isolation and complete nucleotide sequence of the respective lactonase gene were done and submitted to the Genbank database under accession code KC823046.
Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University, Organization of African Unity Street, Abbassia, Cairo, Egypt.
Masarra Mohammed
MM
Sakr
Khaled Mohamed Anwar
KM
Aboshanab
Mohammad Mabrouk
MM
Aboulwafa
Nadia Abdel-Haleem
NA
Hassouna
D054742:Acyl-Butyrolactones / Q000737:chemistry* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology* / Q000472:pathogenicity; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics; D002384:Catalysis; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D053038:Quorum Sensing / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
D054742:Acyl-Butyrolactones; D002265:Carboxylic Ester Hydrolases; C020579:gluconolactonase