Kim_2005_Protein.Expr.Purif_39_124

Reference

Title : Purification, refolding, and characterization of recombinant Pseudomonas fluorescens lipase - Kim_2005_Protein.Expr.Purif_39_124
Author(s) : Kim KR , Kwon DY , Yoon SH , Kim WY , Kim KH
Ref : Protein Expr Purif , 39 :124 , 2005
Abstract :

Thermostable Pseudomonas fluorescens SIK W1 lipase (PFL), which is responsible for the spoilage of milk, was overexpressed as inclusion bodies in Escherichia coli. Renaturation of solubilized PFL was achieved by using size-exclusion protein refolding chromatography. The renatured enzyme was purified homogeneously using a combination of gel filtration and ion-exchange FPLC. Its specific activity was found to be enhanced in the presence of Ca2+. Secondary structural changes induced by Ca2+ were monitored by circular dichroism, which demonstrated that the activity increase of PFL in the presence of Ca2+ is strongly correlated with significant increases in alpha-helix and beta-sheet content. In the presence of Ca2+, the PFL structure was found resistant to denaturation by guanidine hydrochloride and to enzyme activity loss due to cosolvents like DMSO and trifluoroethanol, suggesting that Ca2+ plays an important role in inducing conformational changes and consequently in maintaining enzyme structural stability.

PubMedSearch : Kim_2005_Protein.Expr.Purif_39_124
PubMedID: 15596368

Related information

Citations formats

Kim KR, Kwon DY, Yoon SH, Kim WY, Kim KH (2005)
Purification, refolding, and characterization of recombinant Pseudomonas fluorescens lipase
Protein Expr Purif 39 :124

Kim KR, Kwon DY, Yoon SH, Kim WY, Kim KH (2005)
Protein Expr Purif 39 :124