New Insights into the Alternative d-Glucarate Degradation Pathway
Autor: | Marielle Besnard-Gonnet, Véronique de Berardinis, Annett Kreimeyer, Peggy Sirven, Gabor Gyapay, Delphine Muselet, Marcel Salanoubat, Asadollah Aghaie, Alain Perret, Sabine Tricot, Christophe Lechaplais |
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Přispěvatelé: | Syndrome de Usher et autres atteintes rétino-cochléaires, Institut de la Vision, Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre Hospitalier de Valenciennes, Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Génomique métabolique (UMR 8030), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS) |
Jazyk: | angličtina |
Rok vydání: | 2008 |
Předmět: |
Transcription
Genetic [SDV]Life Sciences [q-bio] Mutant Bacillus subtilis Biology medicine.disease_cause Biochemistry Genome Gene Expression Regulation Enzymologic Glutarates 03 medical and health sciences Bacterial Proteins Protein purification medicine [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Molecular Biology Gene Escherichia coli Hydro-Lyases ComputingMilieux_MISCELLANEOUS 030304 developmental biology Genetics 0303 health sciences Acinetobacter 030302 biochemistry & molecular biology Gene Expression Regulation Bacterial Cell Biology biology.organism_classification Recombinant Proteins Metabolism and Bioenergetics Dehydratase Alternative complement pathway Gene Deletion Genome Bacterial |
Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2008, 283 (23), pp.15638-15646. ⟨10.1074/jbc.M800487200⟩ Journal of Biological Chemistry, 2008, 283 (23), pp.15638-15646. ⟨10.1074/jbc.M800487200⟩ |
ISSN: | 0021-9258 1083-351X |
Popis: | Although the d-glucarate degradation pathway is well characterized in Escherichia coli, genetic and biochemical information concerning the alternative pathway proposed in Pseudomonas species and Bacillus subtilis remains incomplete. Acinetobacter baylyi ADP1 is a Gram-negative soil bacterium possessing the alternative pathway and able to grow using d-glucarate as the only carbon source. Based on the annotation of its sequenced genome (1), we have constructed a complete collection of singlegene deletion mutants (2). High throughput profiling for growth on a minimal medium containing d-glucarate as the only carbon source for ∼2450 mutants led to the identification of the genes involved in d-glucarate degradation. Protein purification after recombinant production in E. coli allowed us to reconstitute the enzymatic pathway in vitro. We describe here the kinetic characterization of d-glucarate dehydratase, d-5-keto-4-deoxyglucarate dehydratase, and of cooperative α-ketoglutarate semialdehyde dehydrogenase. Transcription and expression analyses of the genes involved in d-glucarate metabolism within a single organism made it possible to access information regarding the regulation of this pathway for the first time. |
Databáze: | OpenAIRE |
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