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Purification and characterisation of a water-soluble ferrochelatase from Bacillus subtilis

Författare

Summary, in English

Bacillus subtilis ferrochelatase is encoded by the hemH gene of the hemEHY gene cluster and catalyses the incorporation of Fe2+ into protoporphyrin IX. B. subtilis ferrochelatase produced in Escherichia coli was purified. It was found to be a monomeric, water-soluble enzyme of molecular mass 35 kDa which in addition to Fe2+ can incorporate Zn2+ and Cu2+ into protoporphyrin IX. Chemical modification experiments indicated that the single cysteine residue in the ferrochelatase is required for enzyme activity although it is not a conserved residue compared to other ferrochelatases. In growing B. subtilis, the ferrochelatase constitutes approximately 0.05% (by mass) of the total cell protein, which corresponds to some 600 ferrochelatase molecules/cell. The turnover number of isolated ferrochelatase, 18-29 min-1, was found to be consistent with the rate of haem synthesis in exponentially growing cells (0.2 mol haem formed/min/mol enzyme). It is concluded that the B. subtilis ferrochelatase has enzymic properties which are similar to those of other characterised ferrochelatases of known primary structure, i.e. ferrochelatases of the mitochondrial inner membrane of yeast and mammalian cells. However, in contrast to these enzymes the B. subtilis enzyme is a water-soluble protein and should be more amenable to structural analysis.

Publiceringsår

1994

Språk

Engelska

Sidor

201-208

Publikation/Tidskrift/Serie

European Journal of Biochemistry

Volym

220

Issue

1

Dokumenttyp

Artikel i tidskrift

Förlag

Wiley-Blackwell

Ämne

  • Biochemistry and Molecular Biology

Nyckelord

  • Gene Deletion
  • Ferrochelatase/genetics/*isolation & purification/metabolism
  • Molecular
  • Escherichia coli/enzymology/genetics
  • Cloning
  • Catalysis
  • Amino Acid Sequence
  • Bacillus subtilis/*enzymology/genetics
  • Genes
  • Bacterial
  • Kinetics
  • Molecular Sequence Data
  • Molecular Weight
  • Solubility
  • Water

Status

Published

Forskningsgrupp

  • Plant Biology

ISBN/ISSN/Övrigt

  • ISSN: 0014-2956