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Production and Partial Characterization of Lipase from Pseudomonas putida

Huda Fatima, Natasha Khan, Asad Ur Rehman, Zahid Hussain

The production of lipase from Pseudomonas putida 922 was optimized by modifying various physical parameters such as carbon source, nitrogen source, pH, salt concentration and biochemical parameters of the production medium such as temperature and incubation time of the growth medium. Oil cakes were also used as carbon source to check for an increased production of the enzyme. The bacterium was found to have a maximal growth at pH 10 with the enzyme production being highest (24 U/ml) after 48 hours at 30°C and pH 10. The optimum composition of the medium was mustard oil cake as carbon source, yeast extract or peptone as nitrogen source and 1% sodium chloride concentration. Partial characterization of the enzyme was carried out where the optimum working pH and temperature was found to be 10 and 40ºC, respectively. Enzyme stability was found to lie in the pH and temperature ranges of 5-11 and 30-40ºC, respectively. Partial purification of the enzyme was carried out at 80% ammonium sulphate saturation. Molecular mass of lipase was determined by SDS PAGE and found to be 45 kDa.