Identification of a novel glyoxylate reductase supports phylogeny-based enzymatic substrate specificity prediction
| Autor: | Kristien Braeken, Karen Vos, Maarten Fauvart, Jean-Paul Noben, Johan Robben, Jozef Vanderleyden, Maxime Ndayizeye, Ruth Daniels, Jan Michiels |
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| Rok vydání: | 2007 |
| Předmět: |
chemistry.chemical_classification
Molecular Sequence Data Biophysics Glyoxylate cycle Dehydrogenase Sequence alignment Biology Biochemistry Rhizobium etli Analytical Chemistry Substrate Specificity Alcohol Oxidoreductases Kinetics Enzyme chemistry Phylogenetics Enzyme kinetics Amino Acid Sequence Molecular Biology Sequence Alignment Glyoxylate reductase Phylogeny |
| Zdroj: | Biochimica et biophysica acta. 1774(9) |
| ISSN: | 0006-3002 |
| Popis: | Phylogenetic analysis of the superfamily of D-2-hydroxyacid dehydrogenases identified the previously unrecognized cluster of glyoxylate/hydroxypyruvate reductases (GHPR). Based on the genome sequence of Rhizobium etli, the nodulating endosymbiont of the common bean plant, we predicted a putative 3-phosphoglycerate dehydrogenase to exhibit GHPR activity instead. The protein was overexpressed and purified. The enzyme is homodimeric under native conditions and is indeed capable of reducing both glyoxylate and hydroxypyruvate. Other substrates are phenylpyruvate and ketobutyrate. The highest activity was observed with glyoxylate and phenylpyruvate, both having approximately the same kcat/Km ratio. This kind of substrate specificity has not been reported previously for a GHPR. The optimal pH for the reduction of phenylpyruvate to phenyllactate is pH 7. These data lend support to the idea of predicting enzymatic substrate specificity based on phylogenetic clustering. |
| Databáze: | OpenAIRE |
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