title: Purification and characterization of arginine amidinohydrolase from Bacillus brevis TT02-8.
abstract: A bacterial arginase was purified to homogeneity from a strain of Bacillus brevis. The native enzyme, with an estimated MW of 143,000, migrated on SDS-PAGE as a single polypeptide of estimated MW of 33,000. The enzyme, highly specific to L-arginine, showed the maximum activity at pH 11.0 in the presence of Mn2+ ions and the pI was 4.8 by isoelectric focusing. The enzyme activity was increased significantly by the addition of Mn2+, Ni2+, or Co2+ ions, and inhibited potently by chemicals such as HgCl2, N-bromosuccinimide, or glutathione. The Kms for L-arginine and L-canavanine were 0.69 and 22.2 mM, respectively. The enzyme was inhibited competitively by gamma-guanidinobutyric acid, and non-competitively by L-lysine, L-ornithine, creatine, blasticidin S, and edeine B1. Analysis of the N-terminal amino acid sequence of the purified bacterial enzyme found 33-36% homologies with the Agrobacterium, yeast, rat, and human enzymes.
authors.affiliation: Kyoritsu College of Pharmacy, Tokyo, Japan.
authors.forename: K W
authors.initials: KW
authors.lastname: Shimotohno
authors.forename: J
authors.initials: J
authors.lastname: Iida
authors.forename: N
authors.initials: N
authors.lastname: Takizawa
authors.forename: T
authors.initials: T
authors.lastname: Endo
mesh_terms: D000595:Amino Acid Sequence; D001119:Arginase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D001407:Bacillus / Q000201:enzymology*; D002412:Cations; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D002412:Cations; D010446:Peptide Fragments; D001119:Arginase