CHARACTERIZATION OF PARTIALLY PURIFIED LIPASE FROM SACCHAROMYCES CEREVISIAE
Keywords:
Ammonium sulphate, Nil, Gel filtration, Lipase, SDS-PAGE, Metal ionsAbstract
Objectives: This study was aimed for the purification and characterization of lipase from Saccharomyces cerevisiae and its activity.
Methods: Extracellular lipase enzyme was extracted from the culture filtrate of baker's yeast Saccharomyces cervisiae grown on lipase production medium. The enzyme was purified by ammonium sulphate fractionation attaining maximum activity at 75% w/v saturation. The dialyzed sample was then subjected to sephadex G 100 column chromatography. The enzyme was purified 4.1 fold with a specific activity of 30 U/mg.
Results: The molecular mass of this enzyme was estimated to be approximately 46 kDa by SDS-PAGE. The enzyme was stable even after exposure to a pH range of 4.0-8.0 and at 45oCtemperature for 1 hour. The enzyme exhibited optimum activity at pH 5.0-6.0 and at temperature 30oC. Its activity was greatly enhanced by the addition of 10 mM 2- mercaptoethanol and completely inhibited by Hg+2 indicating that the enzyme contains sulfhydryl group at its active site.
Conclusion: The significance of controlling parameters for lipase activity, and it's the effect on different parameters like temperature, PH and metal ions on partially purified lipase enzyme having molecular weight of approximately 46 kDa.
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References
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