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Isolated Bacillus subtilis HemY has coproporphyrinogen III to coproporphyrin III oxidase activity

Hansson, Mats LU ; Gustafsson, Mattias LU ; Kannangara, C Gamini and Hederstedt, Lars LU (1997) In BBA - Protein Structure and Molecular Enzymology 1340(1). p.97-104
Abstract
Oxidation of coproporphyrinogen III to coproporphyrin III is found in extracts of Escherichia coli cells containing the Bacillus subtilis HemY protein (M. Hansson and L. Hederstedt, J. Bacteriol. 176, 5962-5970). We have analysed whether this activity is due to the heterologous expression system, since it in vivo would lead to disruption of the heme biosynthetic pathway. B. subtilis hemY was fused in its 3'-end to a polynucleotide encoding six histidine residues and expressed from plasmids in both E. coli and B. subtilis. The His6-tagged HemY protein extracted from membranes using non-ionic detergent was purified by Ni2+ affinity chromatography. Isolated HemY fusion protein synthesised in E. coli and B. subtilis oxidised coproporphyrinogen... (More)
Oxidation of coproporphyrinogen III to coproporphyrin III is found in extracts of Escherichia coli cells containing the Bacillus subtilis HemY protein (M. Hansson and L. Hederstedt, J. Bacteriol. 176, 5962-5970). We have analysed whether this activity is due to the heterologous expression system, since it in vivo would lead to disruption of the heme biosynthetic pathway. B. subtilis hemY was fused in its 3'-end to a polynucleotide encoding six histidine residues and expressed from plasmids in both E. coli and B. subtilis. The His6-tagged HemY protein extracted from membranes using non-ionic detergent was purified by Ni2+ affinity chromatography. Isolated HemY fusion protein synthesised in E. coli and B. subtilis oxidised coproporphyrinogen III to coproporphyrin III. No direct formation of protoporphyrin IX from coproporphyrinogen III could be detected. Our results suggest that the coproporphyrinogen III to coproporphyrin III activity of HemY is either avoided in B. subtilis in vivo or that coproporphyrin III is a heme biosynthetic intermediate in this bacterium. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Protoporphyrinogen IX oxidase, Heme biosynthesis, hemY, Bacillus subtilis, Coproporphyrinogen
in
BBA - Protein Structure and Molecular Enzymology
volume
1340
issue
1
pages
97 - 104
publisher
Elsevier
external identifiers
  • pmid:9217019
  • scopus:0030918517
ISSN
0167-4838
DOI
10.1016/S0167-4838(97)00030-7
language
English
LU publication?
yes
id
bc80bdf3-f1ac-4552-a828-5c4b3aa393fa (old id 1111904)
date added to LUP
2008-07-18 16:10:04
date last changed
2017-07-31 14:59:58
@article{bc80bdf3-f1ac-4552-a828-5c4b3aa393fa,
  abstract     = {Oxidation of coproporphyrinogen III to coproporphyrin III is found in extracts of Escherichia coli cells containing the Bacillus subtilis HemY protein (M. Hansson and L. Hederstedt, J. Bacteriol. 176, 5962-5970). We have analysed whether this activity is due to the heterologous expression system, since it in vivo would lead to disruption of the heme biosynthetic pathway. B. subtilis hemY was fused in its 3'-end to a polynucleotide encoding six histidine residues and expressed from plasmids in both E. coli and B. subtilis. The His6-tagged HemY protein extracted from membranes using non-ionic detergent was purified by Ni2+ affinity chromatography. Isolated HemY fusion protein synthesised in E. coli and B. subtilis oxidised coproporphyrinogen III to coproporphyrin III. No direct formation of protoporphyrin IX from coproporphyrinogen III could be detected. Our results suggest that the coproporphyrinogen III to coproporphyrin III activity of HemY is either avoided in B. subtilis in vivo or that coproporphyrin III is a heme biosynthetic intermediate in this bacterium.},
  author       = {Hansson, Mats and Gustafsson, Mattias and Kannangara, C Gamini and Hederstedt, Lars},
  issn         = {0167-4838},
  keyword      = {Protoporphyrinogen IX oxidase,Heme biosynthesis,hemY,Bacillus subtilis,Coproporphyrinogen},
  language     = {eng},
  number       = {1},
  pages        = {97--104},
  publisher    = {Elsevier},
  series       = {BBA - Protein Structure and Molecular Enzymology},
  title        = {Isolated <em>Bacillus subtilis</em> HemY has coproporphyrinogen III to coproporphyrin III oxidase activity},
  url          = {http://dx.doi.org/10.1016/S0167-4838(97)00030-7},
  volume       = {1340},
  year         = {1997},
}