Monday, April 30, 2007
3-13

Extracellular lipase(s) from microbe(s)isolated from hot spring in Karachi Pakistan

Furqan Sami, Abid Ali Syed, and Atiya Abbasi. HEJ research institute of chemistry, International center for chemical and biological sciences, university of karachi, KArachi, Pakistan

Significant level of lipolytic enzyme(s) produced in batch cultivation of microbe(s) isolated from the hot springs in Karachi. The selected strains usually able to grow on a variety of substrates such as, oils (e.g. mustard oil, olive oil, coconut oil), fatty acid (e.g. oleic acid) and emulsifiers (Tween 20, Tween 80) grown in medium supplemented with various concentration of lipidic compounds. Maximum enzyme activity was observed in cultures supplemented with Mustard oil and Tween-20. The optimization studies have also been performed revealing the effect of aeration, optimum pH of the culture medium, and incubation time and temperature as well as enzyme production in relation to phase growth. Results of these studies for the optimization of lipase enzyme production with carbon source(s) revealed maximum value enzyme Units/ml during 24 and 72 hours, respectively. An optimum oil concentration of was selected for the bulk production of enzyme. Lipolytic enzyme(s) purified to apparent homogeneity by sequential steps of ammonium sulphate precipitation (70%), ion exchange (DEAE Sephacel) and size exclusion (Sephadex G-200) chromatography which results an increase of 48.2-folds of purification with 2.5% of the total yield. The purity of the enzyme was established polyacrylamide gel electrophoresis and gel overlay zymography on Tween incorporated agar plate.

 



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