Use of methylumbeliferyl-derivative substrates for lipase activity characterization |
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Authors: | Nú ria Prim, Marta S nchez, Cristian Ruiz, F. I. Javier Pastor,Pilar Diaz |
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Affiliation: | Department of Microbiology, Faculty of Biology, University of Barcelona, Avenue Diagonal 645, 08028, Barcelona, Spain |
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Abstract: | Lipases and esterases have been recognized as very useful biocatalysts because of their wide-ranging versatility in industrial applications, their stability, low cost, and non-requirement for added cofactors. The physical properties of lipidic substrates, typically water insoluble, have determined a great difficulty in studying lipolytic enzymes. A method for fast and simple detection of lipolytic activity, based on the use of 4-methylumbelliferone (MUF)-derivative substrates was developed. The system has been used for the detection of lipase activity either from microbial colonies, cell culture suspensions, or from proteins separated on SDS-polyacrylamide or isoelectric focusing gels. The use of MUF-derivative substrates has also been extended to the quantitative determination of lipolytic activity from a variety of assays including optimum pH and temperature determination, growth dependency, kinetics or stability studies, or residual activity quantification after treatment with potential inhibitors. The method has shown to be a useful tool for the characterization of a variety of lipases from microbial origin, including those cloned in heterologous hosts. |
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Keywords: | Lipases Esterases MUF-butyrate MUF-oleate Bacillus Saccharomyces E. coli |
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