1. Single residues dictate the co-evolution of dual esterases: MCP hydrolases from the α/β hydrolase family.
- Author
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Alcaide M, Tornés J, Stogios PJ, Xu X, Gertler C, Di Leo R, Bargiela R, Lafraya A, Guazzaroni ME, López-Cortés N, Chernikova TN, Golyshina OV, Nechitaylo TY, Plumeier I, Pieper DH, Yakimov MM, Savchenko A, Golyshin PN, and Ferrer M
- Subjects
- Amino Acid Sequence, Bacterial Proteins genetics, Burkholderia chemistry, Crystallography, X-Ray, Esterases genetics, Hydrolases genetics, Molecular Sequence Data, Proteobacteria chemistry, Pseudomonas chemistry, Pseudomonas genetics, Sphingomonas chemistry, Sphingomonas genetics, Bacterial Proteins chemistry, Esterases chemistry, Evolution, Molecular, Hydrolases chemistry
- Abstract
Several members of the C-C MCP (meta-cleavage product) hydrolase family demonstrate an unusual ability to hydrolyse esters as well as the MCPs (including those from mono- and bi-cyclic aromatics). Although the molecular mechanisms responsible for such substrate promiscuity are starting to emerge, the full understanding of these complex enzymes is far from complete. In the present paper, we describe six distinct α/β hydrolases identified through genomic approaches, four of which demonstrate the unprecedented characteristic of activity towards a broad spectrum of substrates, including p-nitrophenyl, halogenated, fatty acyl, aryl, glycerol, cinnamoyl and carbohydrate esters, lactones, 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate and 2-hydroxy-6-oxohepta-2,4-dienoate. Using structural analysis and site-directed mutagenesis we have identified the three residues (Ser32, Val130 and Trp144) that determine the unusual substrate specificity of one of these proteins, CCSP0084. The results may open up new research avenues into comparative catalytic models, structural and mechanistic studies, and biotechnological applications of MCP hydrolases.
- Published
- 2013
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