Purification and Characterization of Glycerol 3-Phosphate Dehydrogenase from Two Types of Acidiphilium sp

Takashi Hatta, Kenji Inagaki, Tsuyoshi Sugio, Noriaki Kishimoto, Tatsuo Tano

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Acidiphilium sp. 63 and 24R, which were isolated from strongly and weakly acidic environments, grew well on glycerol. Both glycerol kinase and glycerol 3-phosphate dehydrogenase (G3PDH) were induced during growth on glycerol. After centrifugation of the crude extracts at 100, 000 × g for 60 min, the G3PDH activity in strain 63 was found in the precipitate, on the other hand, 80 % of that in strain 24R was found in the supernatant. We solubilized the G3PDH from strains 63 and 24R with 0.1 % Brij 58, and both enzymes were purified to homogeneity. When a detergent was removed from these enzymes with cold aqueous 90% acetone, the activities were reduced to 11% and 28%, and were restored to about 55% of the original activities on addition of 0.1% Brij 58. By sonication with phospholipid, the 63-G3PDH activity was restored very well. Both purified enzymes have a molecular weight of 108, 000 and their coenzymes were FAD. These enzymes from strain 63 and 24R had a Km of 0.38 and 1.3 mM for DL-glycerol 3-phosphate and the optimal pHs of these enzymes were 7.0 and 8.8, respectively.

Original languageEnglish
Pages (from-to)651-658
Number of pages8
JournalAgricultural and Biological Chemistry
Volume53
Issue number3
DOIs
Publication statusPublished - 1989

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Acidiphilium
glycerol-3-phosphate dehydrogenase
Glycerolphosphate Dehydrogenase
Purification
Cetomacrogol
Enzymes
glycerol
enzymes
Glycerol
glycerol kinase
Glycerol Kinase
Flavin-Adenine Dinucleotide
Sonication
coenzymes
Centrifugation
Coenzymes
Acetone
Complex Mixtures
detergents
Detergents

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Purification and Characterization of Glycerol 3-Phosphate Dehydrogenase from Two Types of Acidiphilium sp. / Hatta, Takashi; Inagaki, Kenji; Sugio, Tsuyoshi; Kishimoto, Noriaki; Tano, Tatsuo.

In: Agricultural and Biological Chemistry, Vol. 53, No. 3, 1989, p. 651-658.

Research output: Contribution to journalArticle

Hatta, Takashi ; Inagaki, Kenji ; Sugio, Tsuyoshi ; Kishimoto, Noriaki ; Tano, Tatsuo. / Purification and Characterization of Glycerol 3-Phosphate Dehydrogenase from Two Types of Acidiphilium sp. In: Agricultural and Biological Chemistry. 1989 ; Vol. 53, No. 3. pp. 651-658.
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