Details

Autor(en) / Beteiligte

• Genda, T.(Yamaguchi Univ. (Japan). Faculty of Education)
• Watabe, S
• Ozaki, H

Titel

Purification and characterization of fumarase from Corynebacterium glutamicum

Ist Teil von

• Bioscience, biotechnology, and biochemistry, 2006-05, Vol.70 (5), p.1102-1109

Ort / Verlag

Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry

Erscheinungsjahr

2006

Quelle

MEDLINE

Beschreibungen/Notizen

• Fumarase (EC 4.2.1.2) from Corynebacterium glutamicum (Brevibacterium flavum) 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 C. glutamicum (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 (Ksub(eq)) for the enzyme reaction were determined at pH 6.0, 7.0, and 8.0, resulting in Ksub(eq)=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, meso-tartrate, D-tartrate, and pyromellitate, inhibited the enzyme competitively, and D-malate in mixed-type.