Purification and characterisation of a water-soluble ferrochelatase from Bacillus subtilis
(1994) In European Journal of Biochemistry 220(1). p.201-208- Abstract
- 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... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/8001509
- author
- Hansson, Mats LU and Hederstedt, Lars LU
- organization
- publishing date
- 1994
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- 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
- in
- European Journal of Biochemistry
- volume
- 220
- issue
- 1
- pages
- 201 - 208
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:0028147501
- ISSN
- 0014-2956
- DOI
- 10.1111/j.1432-1033.1994.tb18615.x
- language
- English
- LU publication?
- yes
- additional info
- 1
- id
- 7ea39097-fe61-4f4f-9d10-479c1e1c40e5 (old id 8001509)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/8119288
- date added to LUP
- 2016-04-01 15:50:39
- date last changed
- 2021-01-06 08:21:25
@article{7ea39097-fe61-4f4f-9d10-479c1e1c40e5,
abstract = {{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.}},
author = {{Hansson, Mats and Hederstedt, Lars}},
issn = {{0014-2956}},
keywords = {{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}},
language = {{eng}},
number = {{1}},
pages = {{201--208}},
publisher = {{Wiley-Blackwell}},
series = {{European Journal of Biochemistry}},
title = {{Purification and characterisation of a water-soluble ferrochelatase from <em>Bacillus subtilis</em>}},
url = {{http://dx.doi.org/10.1111/j.1432-1033.1994.tb18615.x}},
doi = {{10.1111/j.1432-1033.1994.tb18615.x}},
volume = {{220}},
year = {{1994}},
}