PMID- 11737210
OWN - NLM
STAT- MEDLINE
DCOM- 20020115
LR  - 20190620
IS  - 0014-2956 (Print)
IS  - 0014-2956 (Linking)
VI  - 268
IP  - 24
DP  - 2001 Dec
TI  - Cloning, overproduction and characterization of cytochrome c peroxidase from the 
      purple phototrophic bacterium Rhodobacter capsulatus.
PG  - 6559-68
AB  - The bacterial cytochrome c peroxidase (BCCP) from Rhodobacter capsulatus was 
      purified as a recombinant protein from an Escherichia coli clone over-expressing 
      the BCCP structural gene. BCCP from Rb. capsulatus oxidizes the Rhodobacter 
      cytochrome c2 and reduces hydrogen peroxide, probably functioning as a 
      detoxification mechanism. The enzyme binds two haem c groups covalently. The gene 
      encoding BCCP from Rb. capsulatus was cloned through the construction of a 7-kb 
      subgenomic clone. In comparison with the protein sequence, the sequence deduced 
      from the gene has a 21-amino-acid N-terminal extension with the characteristics 
      of a signal peptide. The purified recombinant enzyme showed the same 
      physico-chemical properties as the native enzyme. Spectrophotometric titration 
      established the presence of a high-potential (Em=+270 mV) and a low-potential 
      haem (between -190 mV and -310 mV) as found in other BCCPs. The enzyme was 
      isolated in the fully oxidized but inactive form. It binds calcium tightly and 
      EGTA treatment of the enzyme was necessary to show calcium activation of the 
      mixed valence enzyme. This activation is associated with the formation of a 
      high-spin state at the low-potential haem. BCCP oxidizes horse ferrocytochrome c 
      better than the native electron donor, cytochrome c2; the catalytic activities 
      ('turnover number') are 85 800 min(-1) and 63 600 min(-1), respectively. These 
      activities are the highest ever found for a BCCP.
FAU - De Smet, L
AU  - De Smet L
AD  - Department of Biochemistry, Physiology and Microbiology, Laboratory for Protein 
      Biochemistry and Protein Engineering, University of Gent, Belgium.
FAU - Pettigrew, G W
AU  - Pettigrew GW
FAU - Van Beeumen, J J
AU  - Van Beeumen JJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Eur J Biochem
JT  - European journal of biochemistry
JID - 0107600
RN  - 0 (DNA, Bacterial)
RN  - 0 (Recombinant Proteins)
RN  - EC 1.11.1.5 (Cytochrome-c Peroxidase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Chromatography, Ion Exchange
MH  - Cloning, Molecular
MH  - Cytochrome-c Peroxidase/*genetics/isolation & purification/metabolism
MH  - DNA, Bacterial
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Escherichia coli/genetics
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Recombinant Proteins/genetics/isolation & purification/metabolism
MH  - Rhodobacter capsulatus/*enzymology
MH  - Substrate Specificity
EDAT- 2001/12/12 10:00
MHDA- 2002/01/16 10:01
CRDT- 2001/12/12 10:00
PHST- 2001/12/12 10:00 [pubmed]
PHST- 2002/01/16 10:01 [medline]
PHST- 2001/12/12 10:00 [entrez]
AID - 2610 [pii]
AID - 10.1046/j.0014-2956.2001.02610.x [doi]
PST - ppublish
SO  - Eur J Biochem. 2001 Dec;268(24):6559-68. doi: 10.1046/j.0014-2956.2001.02610.x.