Biosynthesis, isolation and properties of NAD-dependent formate dehydrogenase from the yeast Candida methylica
| Autor: | Lidia S. Ioanesyan, Aleksey M. Egorov, T.V. Avilova, Egorova Olga A |
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| Rok vydání: | 1985 |
| Předmět: |
Biology
Formate dehydrogenase Biochemistry Chromatography DEAE-Cellulose Substrate Specificity chemistry.chemical_compound Oxidoreductase Yeast extract Formate Sulfhydryl Compounds Candida chemistry.chemical_classification Chromatography Isoelectric focusing Hydrogen-Ion Concentration NAD Aldehyde Oxidoreductases Formate Dehydrogenases Yeast Molecular Weight Enzyme chemistry Sephadex Electrophoresis Polyacrylamide Gel Isoelectric Focusing |
| Zdroj: | European Journal of Biochemistry. 152:657-662 |
| ISSN: | 1432-1033 0014-2956 |
| DOI: | 10.1111/j.1432-1033.1985.tb09245.x |
| Popis: | NAD-dependent formate dehydrogenase (EC 1.2.1.2), was isolated from the methanol-utilizing yeast Candida methylica. Two purification techniques for the enzyme from the crude yeast extract have been developed: a two-step procedure, involving a sequential application of DEAE-cellulose ion-exchange chromatography and Sephadex G-200 gel filtration, and a single-step procedure, preparative isoelectric focusing in a granulated gel layer. The enzyme proved to be electrophoretically homogeneous. It consisted of two identical subunits with a relative molecular mass of 46 000, each containing one -SH group related to manifestation of the catalytic activity. The Michaelis constant was 1 X 10(-4) M for NAD and 1.3 X 10(-2) M for formate. Formate dehydrogenase was inhibited with p-chlormercuribenzoate, iodoacetamide, dithionitrobenzoate, cyanide and azide. |
| Databáze: | OpenAIRE |
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