Biochemical and molecular characterization of the NAD+-dependent isocitrate dehydrogenase from the chemolithotroph Acidithiobacillus thiooxidans

Hiroyuki Inoue, Takashi Tamura, Nagisa Ehara, Akira Nishito, Yumi Nakayama, Makiko Maekawa, Katsumi Imada, Hidehiko Tanaka, Kenji Inagaki

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

An isocitrate dehydrogenase (ICDH) with an unique coenzyme specificity from Acidithiobacillus thiooxidans was purified and characterized, and its gene was cloned. The native enzyme was homodimeric with a subunit of Mr 45 000 and showed a 78-fold preference for NAD+ over NADP+. The cloned ICDH gene (icd) was expressed in an icd-deficient strain of Escherichia coli EB106; the activity was found in the cell extract. The gene encodes a 429-amino acid polypeptide and is located between open reading frames encoding a putative aconitase gene (upstream of icd) and a putative succinyl-CoA synthase β-subunit gene (downstream of icd). A. thiooxidans ICDH showed high sequence similarity to bacterial NADP+-dependent ICDH rather than eukaryotic NAD+-dependent ICDH, but the NAD+-preference of the enzyme was suggested due to residues conserved in the coenzyme binding site of the NAD+-dependent decarboxylating dehydrogenase.

Original languageEnglish
Pages (from-to)127-132
Number of pages6
JournalFEMS Microbiology Letters
Volume214
Issue number1
DOIs
Publication statusPublished - Aug 27 2002

Fingerprint

Acidithiobacillus thiooxidans
Isocitrate Dehydrogenase
Genes
Coenzymes
NAD
Aconitate Hydratase
Enzymes
Cell Extracts
NADP
Open Reading Frames
Oxidoreductases
Binding Sites
Escherichia coli
Amino Acids
Peptides

Keywords

  • Acidithiobacillus thiooxidans
  • Coenzyme specificity
  • Decarboxylating dehydrogenase
  • Enzyme purification
  • Isocitrate dehydrogenase

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Biochemical and molecular characterization of the NAD+-dependent isocitrate dehydrogenase from the chemolithotroph Acidithiobacillus thiooxidans. / Inoue, Hiroyuki; Tamura, Takashi; Ehara, Nagisa; Nishito, Akira; Nakayama, Yumi; Maekawa, Makiko; Imada, Katsumi; Tanaka, Hidehiko; Inagaki, Kenji.

In: FEMS Microbiology Letters, Vol. 214, No. 1, 27.08.2002, p. 127-132.

Research output: Contribution to journalArticle

Inoue, Hiroyuki ; Tamura, Takashi ; Ehara, Nagisa ; Nishito, Akira ; Nakayama, Yumi ; Maekawa, Makiko ; Imada, Katsumi ; Tanaka, Hidehiko ; Inagaki, Kenji. / Biochemical and molecular characterization of the NAD+-dependent isocitrate dehydrogenase from the chemolithotroph Acidithiobacillus thiooxidans. In: FEMS Microbiology Letters. 2002 ; Vol. 214, No. 1. pp. 127-132.
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