Characterization of the hemB gene encoding δ-aminolevulinic acid dehydratase from Propionibacterium freudenreichii

Yoshiteru Hashimoto, Mitsuo Yamashita, Hisayo Ono, Yoshikatsu Murooka

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The Propionibacterium freudenreichii hemB gene that encodes δ- aminolevulinic acid (ALA) dehydratase (EC 4.2.1.24), which catalyzes the synthesis of monopyrrole porphobilinogen, a precursor of tetrapyrrole biosynthesis, was cloned and characterized. A DNA fragment that could complement the hemB mutation of Escherichia coli was sequenced. An open reading frame (ORF) located downstream from a potential ribosome-binding site encoded a polypeptide of 332 amino acid residues, with a deduced molecular mass of 35,830 Da. The predicted amino acid sequence encoded by the ORF is similar to those of other ALA dehydratases. Two regions, the putative zinc- binding site and the predicted catalytic site including the invariant lysine residue of the active site, were highly conserved. The gene was confirmed to encode ALA dehydratase by determining the enzymatic activity of the transformant. After expression of the hemB gene, the N-terminal amino acid sequence of the gene product was identified with the deduced amino acid sequence, although the gene product moved more slowly than the expected molecular size of the monomer in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The M(r) was determined to be 290 k by gel filtration. The ALA dehydratase from P. freudenreichii is thus an octamer with a subunit of 36 kDa.

Original languageEnglish
Pages (from-to)93-100
Number of pages8
JournalJournal of Fermentation and Bioengineering
Volume82
Issue number2
DOIs
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

Porphobilinogen Synthase
Gene encoding
Genes
Amino acids
Acids
Amino Acids
Binding sites
Gels
Porphobilinogen
Binding Sites
Tetrapyrroles
Polypeptides
Biosynthesis
Sodium dodecyl sulfate
Molecular mass
Polyacrylates
Electrophoresis
Sodium Dodecyl Sulfate
Escherichia coli
Lysine

Keywords

  • δ-aminolevulinic acid
  • hemB
  • Propionibacterium freudenreichii
  • tetrapyrrole

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Characterization of the hemB gene encoding δ-aminolevulinic acid dehydratase from Propionibacterium freudenreichii. / Hashimoto, Yoshiteru; Yamashita, Mitsuo; Ono, Hisayo; Murooka, Yoshikatsu.

In: Journal of Fermentation and Bioengineering, Vol. 82, No. 2, 1996, p. 93-100.

Research output: Contribution to journalArticle

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