title: Purification, properties and alternate substrate specificities of arginase from two different sources: Vigna catjang cotyledon and buffalo liver.
abstract: Arginase was purified from Vigna catjang cotyledons and buffalo liver by chromatographic separations using Bio-Gel P-150, DEAE-cellulose and arginine AH Sepharose 4B affinity columns. The native molecular weight of an enzyme estimated on Bio-Gel P-300 column for Vigna catjang was 210 kDa and 120 kDa of buffalo liver, while SDS-PAGE showed a single band of molecular weight 52 kDa for cotyledon and 43 kDa for buffalo liver arginase. The kinetic properties determined for the purified cotyledon and liver arginase showed an optimum pH of 10.0 and pH 9.2 respectively. Optimal cofactor Mn(++) ion concentration was found to be 0.6 mM for cotyledon and 2 mM for liver arginase. The Michaelis-Menten constant for cotyledon arginase and hepatic arginase were found to be 42 mM and 2 mM respectively. The activity of guanidino compounds as alternate substrates for Vigna catjang cotyledon and buffalo liver arginase is critically dependent on the length of the amino acid side chain and the number of carbon atoms. In addition to L-arginine cotyledon arginase showed substrate specificity towards agmatine and L-canavanine, whereas the liver arginase showed substrate specificity towards only L-canavanine.
authors.affiliation: Department of Biochemistry, Dr BA Marathwada University, Aurangabad-400001 (MS), India. snehal_214@rediffmail.com
authors.forename: Snehal
authors.initials: S
authors.lastname: Dabir
authors.forename: Pankaj
authors.initials: P
authors.lastname: Dabir
authors.forename: Baburao
authors.initials: B
authors.lastname: Somvanshi
mesh_terms: D000818:Animals; D001119:Arginase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002020:Buffaloes; D002413:Cations, Divalent; D002853:Chromatography, Liquid; D018548:Cotyledon / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000201:enzymology*; D049670:History, 17th Century; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D013379:Substrate Specificity
chemical_list: D002413:Cations, Divalent; D001119:Arginase