Abstract
An extracellular lipase from Psedoxanthomonas sp. was purified 47.3 folds with an overall yield of 27% through purification procedure of acetone precipitation, ion exchange and gel filtration chromatography. Protein precipitation using acetone fractionation showed that the enzyme was precipitated at the fraction of 0 to 40%. Further purification of the enzyme by ion exchange followed by gel filtration chromatography showed that there were two types of lipases with similar molecular size at around 50 kDa. Characterization of optimum pH, temperature, and substrate specificity were carried out against the isolated lipase. Lip1 showed specific substrate preferences towards p-nitro phenyl myristate meanwhile Lip2 exhibited hydrolytic activity towards short and medium acyl chain of the substrate. Further analysis of both enzyme activities on variation of pH and temperature showed that the optimum pH and temperature for Lip1 were at pH 10.0 and 70°C, respectively while pH 8.0 and 50°C were the optimum pH and temperature for Lip2 respectively. Lyophilized lipase isolated from acetone fraction showed lipolytic activity under variation of methanol concentration up to 30%. Key words: Pseudoxanthomonas lipase, ion exchange chromatography, gel filtration chromatography.
Highlights
Lipase is one of the hydrolase enzymes used as biocatalysts in industry
Further purification of the enzyme by ion exchange followed by gel filtration chromatography showed that there were two types of lipases with similar molecular size at around 50 kDa
Lip1 showed specific substrate preferences towards p-nitro phenyl myristate Lip2 exhibited hydrolytic activity towards short and medium acyl chain of the substrate. Further analysis of both enzyme activities on variation of pH and temperature showed that the optimum pH and temperature for Lip1 were at pH 10.0 and 70°C, respectively while pH 8.0 and 50°C were the optimum pH and temperature for Lip2 respectively
Summary
Lipase is one of the hydrolase enzymes used as biocatalysts in industry. The ability of lipase in catalyzing reactions in nonpolar environments makes lipase as the preferable catalyst in organic reactions. Thermostable and solvent tolerant lipases play important roles in industrial processes, since the enzymes are applicable in the enzymatic processing of lipids, even at high temperature and alkaline condition (Lotti et al, 2015). Most of the synthetic reactions on industrial scale are carried out in organic solvents due to the easy solubility of non-polar compounds (Kumar et al, 2016). This approach requires an enzyme which is stable in the organic solvents. General enzymes show very low rate of reactions or inactive in non-aqueous media; search for solvent stable enzymes has been an extensive area of research (Yilmaz and Sayar, 2015; Bisht et al, 2013; Rahman et al, 2005)
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