Ganoderma lucidum: A Novel Study for Inhibiting Protease Virulence Weapon of Multiple Drug Resistant and Extended Spectrum Beta-Lactamase Pseudomonas aeruginosa Clinical Isolate

Abstract

Purpose: This study aimed to inhibit protease enzyme which considered one of the most virulence factors produced by multidrug resistant Pseudomonas aeruginosa clinical isolate using Gandoderma lucidium extract. Methodology: Production of protease enzyme from Pseudomonas aeruginosa followed by purification using ammonium sulphate precipitation and DEAE-Sephadex A-50 chromatography. Antibacterial activity of G. lucidum methanol extract on P. Aeruginosa growth and protease activity were detected. Also, the mode of inhibition on P. aeruginosa purified protease by G. lucidum extract was represented by reciprocal plot. Results: In purification, 70% ammonium sulphate fractionation gave maximum protease activity 39 u/ml. The maximum protease specific activity of DEAE-Sephadex A-50 chromatography purified sample was found to be 84.6 u/mg with purification fold of 9.4. The molecular weight of the enzyme was estimated to be approximately 40 KDa as shown by SDS electrophoretic analysis. Methanol extract of Ganoderma Lucidum fruiting bodies showed high antibacterial activity against P. aeruginosa. Then its effect on purified protease enzyme indicated that it is an irreversible competitive protease inhibitor (kis = 0.45 mg/ml) with high promising activity. Conclusions: The positive results of screening the antibacterial activity of G. lucidum extract by inhibition of protease form a primary platform for further phytochemical studies and development of new drugs for therapy of skin burn infections.