Abstract

PURIFICATION, CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY OF ENZYMATIC WEAPON - LIPASE FROM VENOMOUS CONE SNAIL AGAINST BLOOD CANCER PATHOGEN PSEUDOMONAS AERUGINOSA

*Vijayalakshmi S., Farheena M. I., Chithira A. and Mohankumar A.

Abstract

Nowadays, microbial lipases vary widely in enzymatic properties and substrate specificity. A relatively smaller number of bacterial lipases have been well studied if compared to plant and fungal lipases. Consequently, they are currently receiving much attention because of their potential applications in various industrial and pharmaceutical processes. So, the present study was aimed to isolate the lipase producing bacterial strain were carried out from pharmaceutically active venom duct of cone snail collected from coral reef island of Gulf of Mannar against blood stream pathogen Pseudomonas aeruginosa. The isolates were positive on differential medium of lipolytic agar plates and thus selected as lipase produced strain. The organisms were tested for various biochemical tests, which leads to identification of marine Bacillus megaterium producing lipase enzyme. After the enzyme production was carried in 1 liter of optimized in fermentation medium at 37◦C, for 48hours at pH 8.0. Harvested lipase product was purified by ammonium sulphate precipitation method. Finally, the enzyme purification was carried out with Sephacryl G-200 using Gel Filtration Chromatography to obtain pure and homogenous enzyme. Further the antimicrobial activity of halophilic lipase was done by well diffusion assay. The diameter of zone of inhibition was observed. The result proved the lipase enzyme is able to degrade the cell of pathogenic microbe. Further the molecular weight of the enzyme was determined by SDS PAGE. The purified halophilic lipase activity was observed in 0.70 at 600nm. Total protein content of the lipase was done by Folin Ciocalteau method using Bovine Albumin (BSA) as a standard. This result showed that Bacillus megaterium under study is a good producer of extra cellular lipase, which can be beneficial for biomedical industry for making cancer therapeutic drugs.

Keywords: Cone snail, Bacillus megaterium, Halophilic lipase, Blood stream pathogen – Pseudomonas aeruginosa, Ammonium Sulphate Precipitation, SDS PAGE, FTIR.