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Title: | Identification of a novel glyoxylate reductase supports phylogeny-based enzymatic substrate specificity prediction | Authors: | Fauvart, Maarten Braeken, Kristien DANIELS, Ruth Vos, Karen Ndayizeye, Maxime NOBEN, Jean-Paul ROBBEN, Johan Vanderleyden, Jos Michiels, Jan |
Issue Date: | 2007 | Publisher: | ELSEVIER SCIENCE BV | Source: | BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 1774(9). p. 1092-1098 | Abstract: | 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 k(cat)/K-m 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. (c) 2007 Elsevier B.V. All rights reserved. | Notes: | Katholieke Univ Leuven, Ctr Microbial & Plant Genet, B-3001 Heverlee, Belgium. Univ Hasselt, Biomed Onderzoek Inst, B-3590 Diepenbeek, Belgium.MICHIELS, J, Katholieke Univ Leuven, Ctr Microbial & Plant Genet, Kasteelpk Arenberg 20, B-3001 Heverlee, Belgium.jan.michiels@biw.kuleuven.be | Keywords: | phenylpyruvate; phenyllactate; glyoxylate; glycolate; hydroxypyruvate; D-glycerate; D-2-hydroxyacid dehydrogenase; substrate specificity; phylogeny; rhizobium etli | Document URI: | http://hdl.handle.net/1942/4100 | ISSN: | 1570-9639 | e-ISSN: | 1878-1454 | DOI: | 10.1016/j.bbapap.2007.06.009 | ISI #: | 000249840000003 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2008 |
Appears in Collections: | Research publications |
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