title: Purification and characterization of buffalo liver L-arginase and its kinetic properties with dihydropyrimidine and metal ions.
abstract: Arginase (L-arginine amidinohydrolase, EC.3.5.3.1) from animal tissues such as, liver and kidney has been partially characterized by many researchers. In this study, we purified arginase to homogeneity from buffalo liver with about ~2857 purification fold and a 20% recovery by chromatographic and spectroscopic analysis were obtained. The molecular mass determined by gel filtration and SDS-PAGE was found to be 118 kDa and 47 kDa, respectively. The optimal pH and temperature of the arginase was 9.5 and 40°C, respectively. Kinetic parameters (Km and Vmax) showed activation of arginase in the reaction medium with decrease in Km (7.14, 5.26, 4.0 and control 3.22 mM) and Vmax (0.05, 0.035, 0.027 and control 0.021 mg/mL/min), while co-factor activity of arginase was optimized using metal ions like Mn²⁺ and Mg²⁺ at 2 mM, which revealed an increase in Vmax values (0.011, 0.013, 0.015 and control 0.010 mg/mL/min) and a decrease in Km values (2.22, 2.12, 1.88 and control 1.66 mM). The kinetic data suggested that the arginase activity is enhanced in the presence of dihydropyrimidine derivative and metal ions, indicating essential mode of activation.
authors.forename: Shivraj Hariram
authors.initials: SH
authors.lastname: Nile
authors.forename: Se Won
authors.initials: SW
authors.lastname: Park
mesh_terms: D000818:Animals; D001119:Arginase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002020:Buffaloes; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D019216:Metals, Heavy / Q000737:chemistry / Q000378:metabolism; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism
chemical_list: D019216:Metals, Heavy; D011743:Pyrimidines; D001119:Arginase