Purification and characterization of buffalo liver L-arginase and its kinetic properties with dihydropyrimidine and metal ions. 2016

Shivraj Hariram Nile, and Se Won Park

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.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D011743 Pyrimidines A family of 6-membered heterocyclic compounds occurring in nature in a wide variety of forms. They include several nucleic acid constituents (CYTOSINE; THYMINE; and URACIL) and form the basic structure of the barbiturates.
D002020 Buffaloes Ruminants of the family Bovidae consisting of Bubalus arnee and Syncerus caffer. This concept is differentiated from BISON, which refers to Bison bison and Bison bonasus. Bubalus,Syncerus,Water Buffaloes,Buffalo,Water Buffalo,Buffalo, Water
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001119 Arginase A ureahydrolase that catalyzes the hydrolysis of arginine or canavanine to yield L-ornithine (ORNITHINE) and urea. Deficiency of this enzyme causes HYPERARGININEMIA. EC 3.5.3.1. Arginase A1,Arginase A4,Hepatic Proliferation Inhibitor,Liver Immunoregulatory Protein,Liver-Derived Inhibitory Protein,Liver-Derived Lymphocyte Proliferation Inhibiting Protein,Immunoregulatory Protein, Liver,Inhibitor, Hepatic Proliferation,Inhibitory Protein, Liver-Derived,Liver Derived Inhibitory Protein,Liver Derived Lymphocyte Proliferation Inhibiting Protein,Proliferation Inhibitor, Hepatic,Protein, Liver Immunoregulatory,Protein, Liver-Derived Inhibitory
D019216 Metals, Heavy Metals with high specific gravity, typically larger than 5. They have complex spectra, form colored salts and double salts, have a low electrode potential, are mainly amphoteric, yield weak bases and weak acids, and are oxidizing or reducing agents (From Grant & Hackh's Chemical Dictionary, 5th ed) Heavy Metal,Heavy Metals,Metal, Heavy

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