| http://purl.uniprot.org/citations/10543446 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/10543446 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/10543446 | http://www.w3.org/2000/01/rdf-schema#comment | "The Synechocystis PCC 6803 katG gene encodes a dual-functional catalase-peroxidase (EC 1.11.1.7). We have established a system for the high level expression of a fully active recombinant form of this enzyme. Its entire coding DNA was extended using a synthetic oligonucleotide encoding a hexa-histidine tag at the C-terminus and expressed in Escherichia coli [BL21-(DE3)pLysS] using the pET-3a vector. Hemin was added to the culture medium to ensure its proper association with KatG upon induction. The expressed protein was purified to homogeneity by two chromatography steps including a metal chelate affinity and hydrophobic interaction chromatography. The homodimeric acidic protein (pl = 5.4) had a molecular mass of 170 kDa and a Reinheitszahl (A406/A280) of 0.64. The recombinant protein contained high catalase activity (apparent Km = 4.9 +/-0.25 mM and apparent kcat = 3500 s(-1)) and an appreciable peroxidase activity with o-dianisidine, guaiacol and pyrogallol, but not with NAD(P)H, ferrocytochrome c, ascorbate or glutathione as electron donors. By using both conventional and sequential stopped-flow spectroscopy, formation of compound I with peroxoacetic acid was calculated to be (8.74 +/-0.26) x 10(3) M(-1) s(-1), whereas compound I reduction by o-dianisidine, pyrogallol and ascorbate was determined to be (2.71 +/-0.03) x 10(6) M(-1) S(-1), (8.62 +/- 0.21) x 10(4) M(-1) S(-1), and (5.43 +/-0.19) x 10(3) M(-1) S(-1), respectively. Cyanide binding studies on native and recombinant enzyme indicated that both have the same heme environment. An apparent second-order rate constant for cyanide binding of (4.8 +/-0.1) x 10(5) M(-1) S(-1) was obtained."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.org/dc/terms/identifier | "doi:10.1515/bc.1999.135"xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.org/dc/terms/identifier | "doi:10.1515/bc.1999.135"xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Obinger C."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Obinger C."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Regelsberger G."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Regelsberger G."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Jakopitsch C."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Jakopitsch C."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Peschek G.A."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Peschek G.A."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Dockal M."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Dockal M."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Ruker F."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/author | "Ruker F."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/date | "1999"xsd:gYear |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/date | "1999"xsd:gYear |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/name | "Biol. Chem."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/name | "Biol. Chem."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/pages | "1087-1096"xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/pages | "1087-1096"xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/title | "Catalase-peroxidase from the cyanobacterium Synechocystis PCC 6803: cloning, overexpression in Escherichia coli, and kinetic characterization."xsd:string |
| http://purl.uniprot.org/citations/10543446 | http://purl.uniprot.org/core/title | "Catalase-peroxidase from the cyanobacterium Synechocystis PCC 6803: cloning, overexpression in Escherichia coli, and kinetic characterization."xsd:string |