Cytosolic Aspartate Aminotransferase from Grey Mullet (Mugil auratus Risso ) Red Muscle : Isolation and Properties (CROSBI ID 79686)
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Podaci o odgovornosti
Petrović, Siniša ; Ozretić, Bartolo ; Krajnović- Ozretić, Mirjana
engleski
Cytosolic Aspartate Aminotransferase from Grey Mullet (Mugil auratus Risso ) Red Muscle : Isolation and Properties
Aspartate aminotransferase isoenzymes play a central role in the metabolism of amino acids and participate in maintaining redox balance via an aspartate-malate shutle in aerobic tissues such as red muscle. The aim of this study was to purify and characterize cytosolic isoenzyme of aspartate aminotransferase from the red muscle of grey mullet , Mugil auratus Risso. The properties of the fish enzyme and its possible physiological role are discussed and compared with homotopic isoenzymes from other species. The enzyme was purified to homogeneity by using different column chromatography techniques and further characterized with gel electrophoresis. It was found to be composed of two identical subunits, M t = 41.000 ą 882 (SEM, n=3). Electrophoretic analysis revealed several active subforms of isolated cytosolic isoenzyme with pI in the pH range 5.2-5.6 .The isolated enzyme was stable in the pH range 5.0- 10.0 and between 0 and 300C. It showed optimum activity at pH 7.5-9.0. Km values of substrates L- aspartate and -ketoglutarate were 1.01 ą 0.082 and 0.031 ą 0.0035 mM, respectively. In the reverse reaction Km values of substrates L- glutamate and oxaloacetate were estimated to be 4.65ą0.90 and 1.22ą0.41mM, respectively. The isolated enzyme was responsible for the transamination of taurine intermediate, L- cysteine sulfinate, in the cytosol of red muscle with apparent Km of 3.08ą0.35 mM. This study demonstrated that cytosolic aspartate aminotransferase isoenzyme from gery mullet red muscle resembles vertebrate cytosolic isoenzymes in its general physicochemical and catalytic properties.
aspartate aminotransferase ; cytosol ; enzyme purification ; enzyme characterization ; grey mullet
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Podaci o izdanju
28 (8)
1996.
873-881
objavljeno
1357-2725
1878-5875
10.1016/1357-2725(96)00033-7