Conversion of a Carboxylesterase into a Triacylglycerol Lipase by a Random Mutation

A true convert: The carboxylesterase enzyme R.34 (see picture) can be converted into a triacylglycerol lipase without modification of the shape, size, or hydrophobicity of the substrate binding site. The substitution of Asn33 by Asp results in a salt bridge between the Asp33 and Arg49, which causes...

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Published inAngewandte Chemie International Edition Vol. 44; no. 46; pp. 7553 - 7557
Main Authors Reyes-Duarte, Dolores, Polaina, Julio, López-Cortés, Nieves, Alcalde, Miguel, Plou, Francisco J., Elborough, Kieran, Ballesteros, Antonio, Timmis, Kenneth N., Golyshin, Peter N., Ferrer, Manuel
Format Journal Article
LanguageEnglish
Published Weinheim WILEY-VCH Verlag 25.11.2005
WILEY‐VCH Verlag
Subjects
Carboxylesterase - antagonists & inhibitors
Carboxylesterase - chemistry
Carboxylesterase - genetics
Enzyme Activation - drug effects
Enzyme Inhibitors - pharmacology
enzymes
esterases
Lipase - antagonists & inhibitors
Lipase - chemistry
Lipase - genetics
lipases
Mutagenesis
Mutation
Protein Conformation
Protein Structure, Tertiary
regiospecificity
Structure-Activity Relationship
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Summary:A true convert: The carboxylesterase enzyme R.34 (see picture) can be converted into a triacylglycerol lipase without modification of the shape, size, or hydrophobicity of the substrate binding site. The substitution of Asn33 by Asp results in a salt bridge between the Asp33 and Arg49, which causes a distortion of the enzyme structure that makes the catalytic site more accessible to larger substrates but also more labile.
Bibliography:istex:1F14D76CB04E4C39C4EF14D786A7C62922BA378E
ark:/67375/WNG-H047H1RK-T
This research was supported by EC Project MERG-CT-2004-505242 "BIOMELI", Spanish CICYT Projects BIO2002-00337 and BIO2004-03773-C04-02, and the BMBF "GenoMik" initiative. M.F. thanks the European Commission for a Marie Curie postdoctoral fellowship and the Spanish Ministerio de Ciencia y Tecnología. The authors also thank ViaLactia Biosciences Ltd. (New Zealand) for financial support. K.N.T. gratefully acknowledges the generous support by the Fonds der Chemischen Industrie.
ArticleID:ANIE200502461
This research was supported by EC Project MERG‐CT‐2004‐505242 “BIOMELI”, Spanish CICYT Projects BIO2002‐00337 and BIO2004‐03773‐C04‐02, and the BMBF “GenoMik” initiative. M.F. thanks the European Commission for a Marie Curie postdoctoral fellowship and the Spanish Ministerio de Ciencia y Tecnología. The authors also thank ViaLactia Biosciences Ltd. (New Zealand) for financial support. K.N.T. gratefully acknowledges the generous support by the Fonds der Chemischen Industrie.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.200502461