Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/12218
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dc.contributor.authorMiñambres Rodríguez, Baltasar-
dc.contributor.authorOlivera, Elías R.-
dc.contributor.authorJensen, Roy A.-
dc.contributor.authorLuengo, José M.-
dc.date.accessioned2009-04-13T10:36:22Z-
dc.date.available2009-04-13T10:36:22Z-
dc.date.issued2000-
dc.identifier.citationJournal of Biological Chemistry 275(50): 39529-39542 (2000)en_US
dc.identifier.issn0021-9258-
dc.identifier.urihttp://hdl.handle.net/10261/12218-
dc.description14 pages, 10 figures, 6 tables.-- PMID: 10924516 [PubMed].en_US
dc.descriptionThe nucleotide sequence reported in this paper has been submitted to the DDBJ/GenBankTM/EBI Data Bank with accession number AF218569.-
dc.description.abstractA new class of glutamate dehydrogenase (GDH) is reported. The GDH of Streptomyces clavuligerus was purified to homogeneity and characterized. It has a native molecular mass of 1,100 kDa and exists as an alpha(6) oligomeric structure composed of 183-kDa subunits. GDH, which requires AMP as an essential activator, shows a maximal rate of catalysis in 100 mm phosphate buffer, pH 7.0, at 30 degrees C. Under these conditions, GDH displayed hyperbolic behavior toward ammonia (K(m), 33 mm) and sigmoidal responses to changes in alpha-ketoglutarate (S(0.5) 1.3 mm; n(H) 1.50) and NADH (S(0.5) 20 microm; n(H) 1.52) concentrations. Aspartate and asparagine were found to be allosteric activators. This enzyme is inhibited by an excess of NADH or NH(4)(+), by some tricarboxylic acid cycle intermediates and by ATP. This GDH seems to be a catabolic enzyme as indicated by the following: (i) it is NAD-specific; (ii) it shows a high value of K(m) for ammonia; and (iii) when S. clavuligerus was cultured in minimal medium containing glutamate as the sole source of carbon and nitrogen, a 5-fold increase in specific activity of GDH was detected compared with cultures provided with glycerol and ammonia. GDH has 1,651 amino acids, and it is encoded by a DNA fragment of 4,953 base pairs (gdh gene). It shows strong sequence similarity to proteins encoded by unidentified open reading frames present in the genomes of species belonging to the genera Mycobacterium, Rickettsia, Pseudomonas, Vibrio, Shewanella, and Caulobacter, suggesting that it has a broad distribution. The GDH of S. clavuligerus is the first member of a class of GDHs included in a subfamily of GDHs (large GDHs) whose catalytic requirements and evolutionary implications are described and discussed.en_US
dc.description.sponsorshipThis investigation was supported by Comisión Interministerial de Ciencia y Tecnología, Madrid, Spain, Grant AMB97-0603-C02-01, Fondo Europeo de Desarrollo Regional Grant 1FD97-0245, and Junta de Castilla y León Grant LE 42/96.en_US
dc.format.extent94510 bytes-
dc.format.mimetypeapplication/pdf-
dc.language.isoengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biologyen_US
dc.rightsclosedAccessen_US
dc.subjectBeta-lactam-
dc.subjectCephalosporin production-
dc.subjectPyrococcus-furiosus-
dc.subjectSequence alignment-
dc.subjectClavulanic acid-
dc.subjectPurification-
dc.subjectEnzyme-
dc.subjectEvolution-
dc.subjectSynthetase-
dc.subjectMechanism-
dc.titleA new class of glutamate dehydrogenases (GDH). Biochemical and genetic characterization of the first member, the AMP-requiring NAD-specific GDH of Streptomyces clavuligerusen_US
dc.typeartículoen_US
dc.identifier.doi10.1074/jbc.M005136200-
dc.description.peerreviewedPeer revieweden_US
dc.relation.publisherversionhttp://dx.doi.org/10.1074/jbc.M005136200en_US
dc.type.coarhttp://purl.org/coar/resource_type/c_6501es_ES
item.openairetypeartículo-
item.fulltextNo Fulltext-
item.grantfulltextnone-
item.languageiso639-1en-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.cerifentitytypePublications-
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