Purification, immobilization, and characterization of a specific lipase from Staphylococcus warneri EX17 by enzyme fractionating via adsorption on different hydrophobic supports
- 23 March 2011
- journal article
- research article
- Published by Wiley in Biotechnology Progress
- Vol. 27 (3), 717-723
- https://doi.org/10.1002/btpr.601
Abstract
Staphylococcus warneri strain EX17 produces three lipases with different molecular weights of 28, 30, and 45 kDa. The 45 kDa fraction (SWL‐45) has been purified from crude protein extracts by one chromatographic step based on the selective adsorption of this lipase by interfacial activation on different hydrophobic supports at low ionic strength. The adsorption of SWL‐45 on octyl‐Sepharose increased the enzyme activity by 60%, but the other lipases were also adsorbed on this support. Using butyl‐Toyopearl, which is a lesser hydrophobic support, the purification factor was close to 20, and the only protein band detected on the sodium dodecyl sulfate‐polyacrylamide electrophoresis analysis gel was that corresponding to the SWL‐45, which could be easily desorbed from the support by incubation with triton X‐100, producing a purified enzyme. SWL‐45 was immobilized under very mild conditions on cyanogen bromide Sepharose, showing similar activities and stability as for its soluble form but without intermolecular interaction. The effects of different detergents over the activity of the immobilized SWL‐45 were analyzed, which was hyperactivated by factors of 1.3 and 2.5 with 0.01% Tween 80 and 0.1% Triton X‐100, respectively, while ionic detergents produced detrimental effects on the enzyme activity even at very low concentrations. Optimal reaction conditions and the effect of other additives on the enzyme activity were also investigated. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011Keywords
This publication has 41 references indexed in Scilit:
- Activation of Bacterial Thermoalkalophilic Lipases Is Spurred by Dramatic Structural RearrangementsJournal of Biological Chemistry, 2009
- Cold active microbial lipases: Some hot issues and recent developmentsBiotechnology Advances, 2008
- Production of organic solvent tolerant lipase by Staphylococcus caseolyticus EX17 using raw glycerol as substrateJournal of Chemical Technology & Biotechnology, 2008
- Trends in lipase-catalyzed asymmetric access to enantiomerically pure/enriched compoundsTetrahedron, 2007
- Cloning, purification and characterisation of Staphylococcus warneri lipase 2Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 2001
- Bacterial Biocatalysts: Molecular Biology, Three-Dimensional Structures, and Biotechnological Applications of LipasesAnnual Review of Microbiology, 1999
- ‘Interfacial activation’ of lipases: facts and artifactsTrends in Biotechnology, 1997
- Purification and characterization of extracellular Staphylococcus warneri lipaseCurrent Microbiology, 1995
- Current progress, in crystallographic studies of new Upases from filamentous fungiProtein Engineering, Design and Selection, 1994
- News from the interface: the molecular structures of triacyglyceride lipasesTrends in Biochemical Sciences, 1993