Characterization of crystalline formate dehydrogenase from Candida methanolica

Y. Izumi, Hiroshi Kanzaki, S. Morita, H. Futazuka, H. Yamada

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The crystalline formate dehydrogenase from Candida methanolica, which showed the highest specific activity (7.52 U/mg) so far reported, was characterized in detail. The enzyme is a dimer composed of identical subunits, each containing one SH group related to the catalytic activity. The molecular mass of the enzyme is about 82-86 kDa. The K(m) values were found to be 3.0 mM for formate and 0.11 mM for NAD+. Even if the enzyme was incubated at pH 6.5-9.5 or at 55°C, the activity remained at 100%. Hg2+, Ni2+, NaCN, NaN3 and p-chloromercuribenzoate strongly inhibited the enzyme activity, while the enzyme showed relatively high resistance to various chelating agents. The amino acid composition and some other physicochemical properties of the enzyme were studied. Immunological studies revealed that formate dehydrogenases of methanol-utilizing yeasts immunologically more or less resemble each other, but differ from those of methanol-utilizing bacteria. Furthermore, yeast formate dehydrogenases can be immunologically classified into three types: (a) the Candida type, (b) the Torulopsis/Hansenula/Pichia type and (c) the formaldehyde-resistant yeast type. For simple and large-scale preparation of the enzyme for practical use, treatment of cells of C. methanolica with the commercial cationic detergent, 'Benzalkonium' cation, is useful: the total and specific activities of the enzyme are 1.17-fold and 3.10-fold higher than those of the crude cell-free extract, respectively.

Original languageEnglish
Pages (from-to)333-341
Number of pages9
JournalEuropean Journal of Biochemistry
Volume182
Issue number2
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

Formate Dehydrogenases
Candida
Crystalline materials
Enzymes
Yeast
formic acid
Pichia
Yeasts
Methanol
Chloromercuribenzoates
Benzalkonium Compounds
Sodium Azide
Enzyme activity
Molecular mass
Chelating Agents
Detergents
Dimers
NAD
Formaldehyde
Cell Extracts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Izumi, Y., Kanzaki, H., Morita, S., Futazuka, H., & Yamada, H. (1989). Characterization of crystalline formate dehydrogenase from Candida methanolica. European Journal of Biochemistry, 182(2), 333-341.

Characterization of crystalline formate dehydrogenase from Candida methanolica. / Izumi, Y.; Kanzaki, Hiroshi; Morita, S.; Futazuka, H.; Yamada, H.

In: European Journal of Biochemistry, Vol. 182, No. 2, 1989, p. 333-341.

Research output: Contribution to journalArticle

Izumi, Y, Kanzaki, H, Morita, S, Futazuka, H & Yamada, H 1989, 'Characterization of crystalline formate dehydrogenase from Candida methanolica', European Journal of Biochemistry, vol. 182, no. 2, pp. 333-341.
Izumi Y, Kanzaki H, Morita S, Futazuka H, Yamada H. Characterization of crystalline formate dehydrogenase from Candida methanolica. European Journal of Biochemistry. 1989;182(2):333-341.
Izumi, Y. ; Kanzaki, Hiroshi ; Morita, S. ; Futazuka, H. ; Yamada, H. / Characterization of crystalline formate dehydrogenase from Candida methanolica. In: European Journal of Biochemistry. 1989 ; Vol. 182, No. 2. pp. 333-341.
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