Purification and properties of 5,10-methenyltetrahydrofolate cyclohydrolase from Clostridium formicoaceticum
Methenyltetrahydrofolate cyclohydrolase (EC 3.5.4.9) from Clostridium formicoaceticum has been purified to a specific activity of 469 ..mu..mol min/sup -1/ mg/sup -1/ at 35/sup 0/C, pH 7.2. The purified enzyme is homogeneous as judged by polyacrylamide disc gel electrophoresis, sedimentation velocity, and gel filtration profiles. The molecular weight is 41,000 +- 200 as determined by sedimentation equilibrium centrifugation. A subunit molecular weight of approximately 25,500 was obtained using sodium dodecyl sulfate-gel electrophoresis. The enzyme apparently is a dimer. The Stokes radius determined by gel filtration is 29.6 A. The apparent K/sub m/ at pH 7.2 and 35/sup 0/C for 5,10-methenyltetrahydrofolate is 0.19 mm. The pure enzyme does not contain any 10-formyltetrahydrofolate synthetase or 5,10-methylenetetrahydrofolate dehydrogenase activities.
- Research Organization:
- Univ. of Georgia, Athens
- OSTI ID:
- 5212487
- Journal Information:
- J. Biol. Chem.; (United States), Vol. 257:7
- Country of Publication:
- United States
- Language:
- English
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