Glutamate:4,5-dioxovaleric acid transaminase from Euglena gracilis. Kinetic studies |
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| Authors: | M E Lombardo L S Araujo A A Juknat A M Batlle |
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| Institution: | Centro de Investigaciones sobre Porfirinas y Porfirias, Faculdad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina. |
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| Abstract: | The kinetic properties of the enzyme L-glutamate:4,5-dioxovaleric acid aminotransferase (Glu:DOVA transaminase) from Euglena gracilis have been studied. 5-Aminolevulinic acid formation was linear with time for at least 45 min at 37 degrees C and L-glutamate was the most effective amino-group donor. Lineweaver-Burk double-reciprocal plots suggested a ping-pong reaction mechanism, with Km values for L-glutamate and DOVA of 1.92 mM and 0.48 mM respectively. Competitive parabolic substrate inhibition by DOVA at concentrations greater than 3.5-4.5 mM was observed. Glyoxylate (4-10 mM) was found to be a competitive inhibitor with respect to DOVA, whereas at low concentrations (0-4 mM) noncompetitive plots were obtained. An analysis of the possible enzyme forms involved, was carried out. In more crude preparations most of the enzyme is found to be in the form of an enzyme-glutamate complex. |
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