Purification and properties of 5,10-methenyltetrahydrofolate synthetase from Lactobacillus casei.
J Biol Chem
; 259(5): 2728-33, 1984 Mar 10.
Article
em En
| MEDLINE
| ID: mdl-6421815
ABSTRACT
5,10-Methenyltetrahydrofolate synthetase (EC 6.3.3.2), which catalyzes the ATP- and Mg2+ -dependent isomerization of 5-formyl- to 5,10-methenyltetrahydrofolate, has been purified 10,000-fold from Lactobacillus casei using sequential affinity chromatography on immobilized 5-formyltetrahydrofolate and ATP. The enzyme is homogeneous when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is monomeric with a molecular mass of 23,000 Da, and contains a high proportion of hydrophobic amino acids and a single cysteine residue. At 30 degrees C, the turnover number is 88 min-1, and the Km values at pH 6 for 5-formyltetrahydrofolate and Mg-ATP are 0.6 and 1.0 microM, respectively. The enzyme is specific for (6S)-5-formyltetrahydrofolate, but ATP can be replaced by other nucleoside 5'-triphosphates with varying efficiency. The purified enzyme is markedly stabilized by the non-ionic detergent, Tween 20.
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01-internacional
Base de dados:
MEDLINE
Assunto principal:
Carbono-Nitrogênio Ligases
/
Lacticaseibacillus casei
/
Ligases
Idioma:
En
Revista:
J Biol Chem
Ano de publicação:
1984
Tipo de documento:
Article