A new lipase as a pharmaceutical target for battling infections caused by Staphylococcus aureus: Gene cloning and biochemical characterization.
Staphylococcus aureus lipases along with other cell-wall-associated proteins and enzymes (i.e., catalase, coagulase, protease, hyaluronidase, and β-lactamase) play important roles in the pathogenesis of S. aureus and are important subject of drug targeting. The appearance of antibiotic-resistant types of pathogenic S. aureus (e.g., methicillin-resistant S. aureus, MRSA) is a worldwide medical problem. In the present work, a novel lipase from a newly isolated MRSA strain from a cow with subclinical mastitis was cloned and biochemically characterized. The mature part of the lipase was expressed in Escherichia coli and purified by nickel affinity chromatography. It displays a high lipase activity at pH 8.0 and 25 °C using p-nitrophenyl palmitate and has a preference for medium-long-chain substrates of p-nitrophenyl esters (pNPC10-C16). Furthermore, in search of inhibitors, the effect of farnesol on the growth of S. aureus and the lipase activity was also studied. Farnesol inhibits the growth of S. aureus and is a mixed-type inhibitor with Ki and Ki (') values of 0.2 and 1.2 mmol L(-1), respectively. A lipase with known properties could not only serve as a template for developing inhibitors for S. aureus but also a valuable addition to enzyme toolbox of biocatalysis. The discovery of this lipase can be potentially important and could provide a new target for pharmaceutical intervention against S. aureus infection.
Department of Chemistry, Gebze Institute of Technology, Gebze, Kocaeli, Turkey.
Aişe
A
Ünlü
Department of Chemistry, Gebze Institute of Technology, Gebze, Kocaeli, Turkey.
Aziz
A
Tanriseven
Department of Molecular Biology and Genetics, Gebze Institute of Technology, Gebze-Kocaeli, Turkey.
I Yavuz
IY
Sezen
Department of Chemistry, Gebze Institute of Technology, Gebze, Kocaeli, Turkey.
Ayhan
A
Çelik
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008413:Mastitis / Q000382:microbiology; D055624:Methicillin-Resistant Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics*; D058990:Molecular Targeted Therapy; D013203:Staphylococcal Infections / Q000188:drug therapy*; D013379:Substrate Specificity; D013696:Temperature
D008049:Lipase
MRSA; S. aureus; expression; farnesol; lipases; subclinical mastitis