Purification and characterization of hydroxypyruvate reductase from a serine‐producing methylotroph, Hyphomicrobium methylovorum GM2

Yoshikazu IZUMI, Toyokazu YOSHIDA, Hiroshi Kanzaki, Shin‐ichiro ‐i TOKI, Silvia Susana MIYAZAKI, Hideaki YAMADA

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Hydroxypyruvate reductase of a serine‐producing methylotroph, Hyphomicrobium methylovorum GM2, was purified to complete homogeneity, crystallized and characterized, the first time for an enzyme from a methylotroph. The enzyme was found to be a dimer composed of identical subunits (38 kDa), the molecular mass of the enzyme being about 70 kDa. The enzyme was stable against heating at 25°C for 10 min at pH values between 5 and 9. Optimal activity was observed at pH 6.8 and around 45°C. The enzyme catalyzed the reduction of hydroxypyruvate with the oxidation of only NADH. Other than hydroxypyruvate, only glyoxylate served as a substrate. The Km values were found to be 0.175 mM for hydroxypyruvate and 10.8 mM for glyoxylate. Taking advantage of the high substrate specificity of this enzyme, a means of enzymatic determination of hydroxypyruvate was established.

Original languageEnglish
Pages (from-to)279-284
Number of pages6
JournalEuropean Journal of Biochemistry
Volume190
Issue number2
DOIs
Publication statusPublished - Jan 1 1990
Externally publishedYes

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Hydroxypyruvate Reductase
Hyphomicrobium
Purification
Enzymes
Molecular mass
Substrates
Substrate Specificity
Dimers
NAD
Heating
Oxidation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and characterization of hydroxypyruvate reductase from a serine‐producing methylotroph, Hyphomicrobium methylovorum GM2. / IZUMI, Yoshikazu; YOSHIDA, Toyokazu; Kanzaki, Hiroshi; TOKI, Shin‐ichiro ‐i; MIYAZAKI, Silvia Susana; YAMADA, Hideaki.

In: European Journal of Biochemistry, Vol. 190, No. 2, 01.01.1990, p. 279-284.

Research output: Contribution to journalArticle

IZUMI, Yoshikazu ; YOSHIDA, Toyokazu ; Kanzaki, Hiroshi ; TOKI, Shin‐ichiro ‐i ; MIYAZAKI, Silvia Susana ; YAMADA, Hideaki. / Purification and characterization of hydroxypyruvate reductase from a serine‐producing methylotroph, Hyphomicrobium methylovorum GM2. In: European Journal of Biochemistry. 1990 ; Vol. 190, No. 2. pp. 279-284.
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