fulltext.study @t Gmail

Lid hinge region of Penicillium expansum lipase affects enzyme activity and interfacial activation

Paper ID Volume ID Publish Year Pages File Format Full-Text
34309 45017 2015 6 PDF Available
Title
Lid hinge region of Penicillium expansum lipase affects enzyme activity and interfacial activation
Abstract

•Sites displaying the highest B factors in the lid were selected for mutagenesis.•Mutants showed properties remarkably different from those of wild-type PEL.•Mutation shifted the chain-length specificity.•One mutant lacked interfacial activation.•We provide a new clue for selecting critical amino acid residues for the enzyme design.

Saturation mutagenesis at sites displaying the highest B factors in the lid and the hinge regions of Penicillium expansum lipase (PEL) has been employed to improve the efficiency of the lipase in biocatalysis. Replacements of amino acid on beneficial mutants were identified as T66L/D70N, T66V/D70N, E83K, E83H and E83N. In substrate specificity assays, T66L/D70N was significantly more active than wild-type PEL on substrates with medium and long chain lengths. In addition this mutant also displayed a 136.4-fold increase in activity on p-nitrophenyl palmitate. Remarkably, E83K lacked interfacial activation while it was observed in wild-type PEL and the other mutants. Insight into the relation between the mutations and enzymatic properties was gained by modeling and docking studies. All these mutants showed an enhanced catalytic activity, indicating their potential in further application. Therefore, these results indicate the amino acid composition of the lid hinge region plays an extremely important role in the interfacial activation, activity and substrate specificity of PEL. Moreover, the results in this work provide a new clue for selecting critical amino acid residues for the enzyme design.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide

Keywords
Penicillium expansum; Lipase; Lid hinge; Mutagenesis
First Page Preview
Lid hinge region of Penicillium expansum lipase affects enzyme activity and interfacial activation
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 50, Issue 8, August 2015, Pages 1218–1223
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us