Production, Partial Purification and Characterization of Lipase Enzyme Expressed by Klebsiella pnemoniae of Vegetable Oil Contaminated Soil

Abstract

Lipases are a versatile class of enzymes owing to their ability to perform a specific range of biotransformations. Bacteria for lipase production were cultured on nutrient agar (NA) plates and identified using Gram Stain and biochemical tests. Screening for lipase producers was performed on NA media supplemented with 3% olive oil at 37 °C. Seven bacteria were isolated and identified as Bacillus megaterium, Klebsiella pneumoniae, Escherichia coli, Bacillus subtilis, Bacillus licheniformis and Micrococcus luteus. Lipase production was carried using standard methods and the best lipase producer was purified and characterized. The lipase enzyme from K. pneumoniae had a yield of 18.8% and was purified 26.3 fold. The optimum pH for the partially purified lipase was determined to be 8 with maximum activity at 30 °C. The lipase enzyme had affinity for substrates in the following order, olive oil > groundnut oil > palm oil and the activity of the enzyme was enhanced by metal ions in the following order, MgCl 2 > CaCl 2 > KCl whereas inhibitory effects were observed in the following order, CoCl 2 > HgCl 2 > CuSO 4 > FeCl 3. The lipase enzyme had V max of 0.0006 U/Sec, K m of 0.4960 mM and K cat of 0.0125 S-1 .

Optimisation of Brevibacterium, Bacillus and Pseudomonas Species for Production of Lipase Using Tobacco Seed Oil and Olive Oil as Sole Carbon Sources

“…They constitute one of the most important group of biocatalyst for biotechnological applications (Javed et al, 2017). Lipase-catalyzed processes generally offer cost-effectiveness as against the traditional downstream processing having the problems of energy consumption and toxic by-products (Moses et al, 2020). Microbial lipases are of special interest because of their stability in organic solvents and their lack of requirements for co-factors, their broad substrate specification and high enantio-selectivity (Godfrey and West, 1996).…”