Isolation, Optimization and Purification of Lipase from Myroides Odoratimimus

Lipases (Triacylglycerol acylhydrolase) are industrially important enzymes that are mainly used in the food and pharmaceutical industry. Lipase enzymes are also used in food materials, detergent, medicine, cosmetics, leather, perfumery and other items. The enzyme was isolated from Myroides odoratimimus and optimized to different physiochemical characteristics, giving its optimum temperature 37°C, active over a wide range pH values (pH 8.2) and increase in substrate (Olive Oil) concentration that increases the activity of lipase. The enzyme lipase activity was stimulated by Ca and Cu, inhibited by BaCl2. All these characteristics are considered favorable for industrial processing. Lipase Activity has been measured by titrimetric, and colorimetric method, and was purified by CM-Cellulose, DEAE –Cellulose, Sephadex G-75 Chromatography, respectively; the purified enzyme was increased by gel filtration. For the protein determination (molecular weight) of our enzyme, we under went for SDS –PAGE, which showed approximately 40KDa. Zymography has been performed showing the maximum activity of the purified lipase, resulting in a clear zone of layer. Native Polyacrylamide Gel Electrophoresis (PAGE) was also performed, in which the gels were overlaid with the molten Chromogenic substrate(40°C), which was then allowed to solidify and incubated, depending on the amount of lipase, the activity was observed as yellow band over a pink background. Keywords— Lipase, Chromatography (CM – cellulose, DEAE – cellulose, Sephadex G-75), SDSPAGE, NATIVE PAGE, Zymogram.