| http://purl.uniprot.org/citations/11358519 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/11358519 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/11358519 | http://www.w3.org/2000/01/rdf-schema#comment | "Alpha-D-glucuronidases cleave the alpha-1,2-glycosidic bond of the 4-O-methyl-D-glucuronic acid side chain of xylan, as a part of an array of xylan hydrolyzing enzymes. The alpha-D-glucuronidase from Bacillus stearothermophilus T-6 was overexpressed in Escherichia coli using the T7 polymerase expression system. The purification procedure included two steps, heat treatment and gel filtration chromatography, and provided over 0.3 g of pure enzyme from 1 L of overnight culture. Based on gel filtration, the native protein is comprised of two identical subunits. Kinetic constants with aldotetraouronic acid as a substrate, at 55 degrees C, were a Km of 0.2 mM, and a specific activity of 42 U x mg(-1) (kcat = 54.9 s(-1)). The enzyme was most active at 65 degrees C, pH 5.5-6.0, in a 10-min assay, and retained 100% of its activity following incubation at 70 degrees C for 20 min. Based on differential scanning calorimetry, the protein denatured at 73.4 degrees C. Truncated forms of the enzyme, lacking either 126 amino acids from its N-terminus or 81 amino acids from its C-terminus, exhibited low residual activity, indicating that the catalytic site is located in the central region of the protein. To identify the potential catalytic residues, site-directed mutagenesis was applied on highly conserved acidic amino acids in the central region. The replacements Glu392-->Cys and Asp364-->Ala resulted in a decrease in activity of about five orders of magnitude, suggesting that these residues are the catalytic pair."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.org/dc/terms/identifier | "doi:10.1046/j.1432-1327.2001.02193.x"xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.org/dc/terms/identifier | "doi:10.1046/j.1432-1327.2001.02193.x"xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Baasov T."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Baasov T."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shoham Y."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shoham Y."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shulami S."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shulami S."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Golan G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Golan G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shallom D."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shallom D."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shoham G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Shoham G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Zaide G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Zaide G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Zolotnitsky G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/author | "Zolotnitsky G."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/date | "2001"xsd:gYear |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/date | "2001"xsd:gYear |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/name | "Eur. J. Biochem."xsd:string |
| http://purl.uniprot.org/citations/11358519 | http://purl.uniprot.org/core/name | "Eur. J. Biochem."xsd:string |