Molecular Identification, Production and Optimization of Lipase from Oil Contaminated Soil using Submerged Fermentation

Abstract

The most important products obtained for human needs are enzymes, through microbial sources. Hydrolytic enzymes occupied major industrial share such as esterases, lipases, amylases and proteases. Applications of these enzymes are varied in medical, cosmetics, food, dairy, pharmaceutical, leather, detergents, bioremediations and paper industries. So, lipases are gaining more attention now a day. Lipases hydrolyse fats into monoglycerides, diglycerides, free fatty acids and glycerol. Bacillus and Stenotrophomonas, are found to be the efficient lipase producers. A total of 48 different lipolytic organisms were isolated from soil samples. Organisms producing lipases were selected on the basis of clear zone formation in tributyrin agar medium. Maximum enzyme yield was obtained at 40° C with 16 U/ml in Bacillus and 14 U/ml in Stenotrophomonas sp., at a pH of 7 in 3 days. The optimum parameters were studied. The yield of lipase enzyme was high while using peptone as a nitrogen source in Stenotrophomonas sp., where as in presence of CaCl2, Bacillus sp., produced more lipase enzyme. However the presence of glucose as carbon source also yielded a much closer value to be highest yield. The lipase producing strains were subjected to gram staining and biochemical characterization following Bergey’s manual of systematic bacteriology. 16SrRNA studies and physiological characterization reveals that Bacillus sp., Stenotrophomonas sp., Halomonas sp., Serratia sp., and Enterobacter sp., are the efficient producers. The sequences were deposited in Genbank.

Keywords

• enzymes
• lipase
• bacillus
• stenotrophomonas
• optimization
• applications