Purification and Characterization of Fumarase from Corynebacterium glutamicum
Access this Article
Search this Article
Fumarase (EC 220.127.116.11) from <I>Corynebacterium glutamicum</I> (<I>Brevibacterium flavum</I>) ATCC 14067 was purified to homogeneity. Its amino-terminal sequence (residues 1 to 30) corresponded to the sequence (residues 6 to 35) of the deduced product of the fumarase gene of <I>C. glutamicum</I> (GenBank accession no. BAB98403). The molecular mass of the native enzyme was 200 kDa. The protein was a homotetramer, with a 50-kDa subunit molecular mass. The homotetrameric and stable properties indicated that the enzyme belongs to a family of Class II fumarase. Equilibrium constants (<I>K</I><SUB>eq</SUB>) for the enzyme reaction were determined at pH 6.0, 7.0, and 8.0, resulting in <I>K</I><SUB>eq</SUB>=6.4, 6.1, and 4.6 respectively in phosphate buffer and in 16, 19, and 17 in non-phosphate buffers. Among the amino acids and nucleotides tested, ATP inhibited the enzyme competitively, or in mixed-type, depending on the buffer. Substrate analogs, <I>meso</I>-tartrate, <small>D</small>-tartrate, and pyromellitate, inhibited the enzyme competitively, and <small>D</small>-malate in mixed-type.
- Agricultural and Biological Chemistry
Agricultural and Biological Chemistry 70(5), 1102-1109, 2006-05-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry