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

Research output: Contribution to journalArticle

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.

Details

Authors
Organisations
External organisations
  • Carlsberg Research Center / Carlsberg Laboratory
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biological Sciences

Keywords

  • Protoporphyrinogen IX oxidase, Heme biosynthesis, hemY, Bacillus subtilis, Coproporphyrinogen
Original languageEnglish
Pages (from-to)97-104
JournalBBA - Protein Structure and Molecular Enzymology
Volume1340
Issue number1
Publication statusPublished - 1997
Publication categoryResearch
Peer-reviewedYes