Purification and properties of carboxypeptidase G2 from Pseudomonas sp. strain RS-16. Use of a novel triazine dye affinity method.
Eur J Biochem
; 148(3): 447-53, 1985 May 02.
Article
em En
| MEDLINE
| ID: mdl-3838935
ABSTRACT
A folate-degrading enzyme, carboxypeptidase G2, has been purified on a large scale from Pseudomonas sp. strain RS-16. Homogeneous enzyme was obtained by a three-step procedure involving ion-exchange chromatography and a novel triazine dye (affinity) chromatography step which utilizes Zn2+ to promote adsorption of the enzyme. Enzyme was selectively eluted by the use of a chelating agent (EDTA) and a step change in pH. The enzyme is a dimeric protein (Mr 83000) with two identical subunits of 41800 and contains four atoms of zinc per enzyme molecule, which are required for full activity. The enzyme follows Michaelis-Menten kinetics with Km values of 4.0 microM for folate, 8.0 microM for methotrexate and 34.0 microM for 5-methyltetrahydrofolate, the predominant form of reduced folate found in plasma.
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01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pseudomonas
/
Triazinas
/
Carboxipeptidases
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Gama-Glutamil Hidrolase
/
Corantes
Idioma:
En
Revista:
Eur J Biochem
Ano de publicação:
1985
Tipo de documento:
Article