Oxaloacetate decarboxylase from Pseudomonas stutzeri: Purification and characterization

dc.contributor.author	Labrou, NE	en
dc.contributor.author	Clonis, YD	en
dc.date.accessioned	2014-06-06T06:43:58Z
dc.date.available	2014-06-06T06:43:58Z
dc.date.issued	1999	en
dc.identifier.issn	00039861	en
dc.identifier.uri	http://dx.doi.org/10.1006/abbi.1999.1144	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/1590
dc.subject	Affinity chromatography	en
dc.subject	Biomimetic adsorbent	en
dc.subject	Biomimetic dye	en
dc.subject	Enzyme kinetics	en
dc.subject	Enzyme purification	en
dc.subject	Oxaloacetate decarboxylase	en
dc.subject	Pseudomonas stutzeri	en
dc.subject.other	acetic acid	en
dc.subject.other	adenosine diphosphate	en
dc.subject.other	avidin	en
dc.subject.other	carbon dioxide	en
dc.subject.other	carboxylyase	en
dc.subject.other	divalent cation	en
dc.subject.other	magnesium ion	en
dc.subject.other	malic acid	en
dc.subject.other	manganese	en
dc.subject.other	metal ion	en
dc.subject.other	monomer	en
dc.subject.other	monovalent cation	en
dc.subject.other	oxalic acid	en
dc.subject.other	oxaloacetic acid	en
dc.subject.other	phosphoenolpyruvate	en
dc.subject.other	protein	en
dc.subject.other	pyruvic acid	en
dc.subject.other	article	en
dc.subject.other	controlled study	en
dc.subject.other	decarboxylation	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme inhibition	en
dc.subject.other	enzyme kinetics	en
dc.subject.other	enzyme purification	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	pH	en
dc.subject.other	priority journal	en
dc.subject.other	pseudomonas stutzeri	en
dc.subject.other	Bicarbonates	en
dc.subject.other	Calcium	en
dc.subject.other	Carboxy-Lyases	en
dc.subject.other	Cations, Divalent	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Heat	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Kinetics	en
dc.subject.other	Metals, Heavy	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Pseudomonas	en
dc.subject.other	Bacteria (microorganisms)	en
dc.subject.other	Negibacteria	en
dc.subject.other	Pseudomonas	en
dc.subject.other	Pseudomonas stutzeri	en
dc.title	Oxaloacetate decarboxylase from Pseudomonas stutzeri: Purification and characterization	en
heal.type	journalArticle	en
heal.identifier.primary	10.1006/abbi.1999.1144	en
heal.publicationDate	1999	en
heal.abstract	Oxaloacetate decarboxylase (OXAD), the enzyme that catalyzes the decarboxylation of oxaloacetate to pyruvic acid and carbon dioxide, was purified 245-fold to homogeneity from Pseudomonas stutzeri. The three-step purification procedure comprised anion-exchange chromatography, metal-chelate affinity chromatography, and biomimetic-dye affinity chromatography. Estimates of molecular mass from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and native high-performance gel-filtration liquid chromatography were, respectively, 63 and 64 kDa, suggesting a monomeric protein. OXAD required for maximum activity divalent metal cations such as Mn2+ and Mg2+ but not monovalent cations. The enzyme is not inhibited by avidin, but is competitively inhibited by adenosine 5'-diphosphate, acetic acid, phosphoenolpyruvate, malic acid, and oxalic acid. Initial velocity, product inhibition, and dead-end inhibition studies suggested a rapid- equilibrium ordered kinetic mechanism with Mn2+ being added to the enzyme first followed by oxaloacetate, and carbon dioxide is released first followed by pyruvate. Inhibition data as well as pH-dependence profiles and kinetic parameters are reported and discussed in terms of the mechanism operating for oxaloacetate decarboxylation.	en
heal.journalName	Archives of Biochemistry and Biophysics	en
dc.identifier.issue	1	en
dc.identifier.volume	365	en
dc.identifier.doi	10.1006/abbi.1999.1144	en
dc.identifier.spage	17	en
dc.identifier.epage	24	en