A new lipase as a pharmaceutical target for battling infections caused by Staphylococcus aureus: Gene cloning and biochemical characterization

Abstract

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.

Comprehensive Virulence Gene Profiling of Bovine Non- aureus Staphylococci Based on Whole-Genome Sequencing Data

“…The exact role of lipase in NAS pathogenicity is not understood; however, lipases are involved in release of free fatty acids (octadecanoic acid) in blood plasma (58–60). These free fatty acids affect several immune system functions and thus indirectly enhance pathogenic potential (58, 61, 62). Interestingly, SAG, SHY, and SCH contain vWbp , which along with coa and clfA were considered sufficient to convert commensal SSI into an invasive pathogen (63).…”

Staphylococcus aureuslipase: purification, kinetic characterization, crystallization and crystallographic study

“…It has been well established that colonization and infection by S. aureus are associated with atopic dermatitis (Findley & Grice, 2014). Several SAL variants have been purified and enzymatically characterized (Ü nlü et al, 2015;Rosenstein & Gö tz, 2000). However, the three-dimensional structure of SAL has not been determined.…”