Kinetic Properties of Pseudomonas sp. Producing Lipase

Abstract

Pseudomonas sp was isolated from petroleum spilled soil using culture dependent techniques. The organism was confirmed Pseudmonas aeruginosa using molecular typing (PCR and sequencing). Lipase producing capability of the organisms was screened in an optimized culture broth in the presence of p-NPP. Yellow coloration of the broth confirms the exo-lipase secretory of the bacteria. Lipase production peaked on day 5 of the fermentation; pH 7 was best for the lipase production. 60% ammonium sulphate was used in precipitation of proteins with peak lipase activity. Protein was dialyzed against gradient (0.01/0.1M) for 12 hrs for salt removal. Native sephadex G100 was used in separation of the dialysate into sizes and weight; fraction tubes of 16-25 were recorded as the elution volume of the chromatogram (VE). pH 4.5 and 60°C were characteristics of the purified protein where as Michaelis-Menten and velocity maximal (Km and Vmax) were 3.45mg/ml and 275 µmole/min. Pseudmonas aeruginosa seen from the study as a competitive bacteria for prolific lipase production to meet with the current biotechnological demand of the next generational enzyme.