Any feedback?
No. of entries
Information on EC 4.99.1.9 - coproporphyrin ferrochelatase
for references in articles please use BRENDA:EC4.99.1.9
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree 4 Lyases
4.99 Other lyases
4.99.1 Sole sub-subclass for lyases that do not belong in the other subclasses
4.99.1.9 coproporphyrin ferrochelatase
IUBMB Comments
The enzyme, present in Gram-positive bacteria, participates in heme biosynthesis. It can also catalyse the reaction of EC 4.98.1.1, protoporphyrin ferrochelatase, at a much lower level.
The enzyme appears in viruses and cellular organisms
showSynonyms
coproporphyrin ferrochelatase, more
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
hemH
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Fe-coproporphyrin III + 2 H+ = coproporphyrin III + Fe2+
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PATHWAY SOURCE
PATHWAYS
MetaCyc
heme b biosynthesis IV (Gram-positive bacteria)
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYSTEMATIC NAME
IUBMB Comments
protoheme ferro-lyase (protoporphyrin-forming)
The enzyme, present in Gram-positive bacteria, participates in heme biosynthesis. It can also catalyse the reaction of EC 4.98.1.1, protoporphyrin ferrochelatase, at a much lower level.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
protoporphyrin IX + Cu2+
Cu-protoporphyrin IX + 2 H+
-
Substrates: -
Products: -
Products: -
?
protoporphyrin IX + Fe2+
Fe-protoporphyrin IX + 2 H+
-
Substrates: the enzyme shows no detectable activity with Ca2+, Co2+, Fe3+, Mg2+, or Rb+
Products: -
Products: -
?
protoporphyrin IX + Zn2+
Zn-protoporphyrin IX + 2 H+
-
Substrates: -
Products: -
Products: -
?
additional information
?
-
Substrates: when LmCpfC is incubated with both substrates, Fe2+ and coproporphyrin III, Fe-coproporphyrin III (coproheme) is the only detected species
Products: -
Products: -
-
coproporphyrin III + Co2+
Co-coproporphyrin III + 2 H+
-
Substrates: specific activity with Co2+ is 6fold higher than with Fe2+
Products: -
Products: -
?
coproporphyrin III + Co2+
Co-coproporphyrin III + 2 H+
-
Substrates: specific activity with Co2+ is 2.8fold higher than with Fe2+
Products: -
Products: -
?
coproporphyrin III + Cu2+
Cu-coproporphyrin III + 2 H+
-
Substrates: specific activity with Cu2+ is 4.15old higher than with Fe2+
Products: -
Products: -
?
coproporphyrin III + Cu2+
Cu-coproporphyrin III + 2 H+
-
Substrates: specific activity with Cu2+ is 5.35old higher than with Fe2+
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
Substrates: the reaction is carried out anaerobically under argon flow and in the dark
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Ni2+
Ni-coproporphyrin III + 2 H+
-
Substrates: specific activity with Ni2+ is 4fold lower than with Fe2+
Products: -
Products: -
?
coproporphyrin III + Ni2+
Ni-coproporphyrin III + 2 H+
-
Substrates: specific activity with Ni2+ is 80% compared to activity with Fe2+
Products: -
Products: -
?
protoporphyrin + Fe2+
protoheme + 2 H+
Substrates: i.e. reaction of EC 4.99.1.1
Products: -
Products: -
?
protoporphyrin + Fe2+
protoheme + 2 H+
Substrates: i.e. reaction of EC 4.99.1.1
Products: -
Products: -
?
protoporphyrin IX + Zn2+
Zn protoporphyrin IX
Substrates: -
Products: -
Products: -
?
protoporphyrin IX + Zn2+
Zn protoporphyrin IX
Substrates: -
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
Substrates: -
Products: -
Products: -
?
coproporphyrin III + Fe2+
Fe-coproporphyrin III + 2 H+
-
Substrates: -
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
[2Fe-2S]-center
the enzyme has a C-terminal [2Fe-2S] cluster
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
N-ethylmaleimide
-
2 mM, 99% inactivation after incubation for 2 h. Protoporphyrin IX or Zn2+ at concentrations of 0.8 mM and 20 mM, respectively, does not protect
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.008
protoporphyrin IX
-
pH 7.2, 30°C. The Km-value for protoporphyrin IX is the same independent of whether Zn2+ or Fe2+ is used in the reaction
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
steady-state kinetics of wild-type and mutant enzymes, overview
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.4
protoporphyrin IX
-
pH 7.2, 30°C. The Km-value for protoporphyrin IX is the same independent of whether Zn2+ or Fe2+ is used in the reaction
additional information
additional information
ligand kon rates, overview
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 8
-
pH 6.5: about 50% of maximal activity, pH 8.0: about 50% of maximal activity
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
additional information
metabolism
coproporphyrin ferrochelatases (CpfCs) insert ferrous iron into coproporphyrin III yielding coproheme. CpfCs are utilized by prokaryotic, mainly monoderm (Gram-positive) bacteria within the coproporphyrin-dependent (CPD) heme biosynthesis pathway
metabolism
coproporphyrin ferrochelatase (CpfC) catalyses the insertion of ferrous iron into the porphyrin ring as the penultimate step within the coproporphyrin-dependent heme biosynthesis pathway. In the protoporphyrin-dependent (PPD) heme biosynthesis pathway, the protoporphyrin ferrochelatase (PpfC) is active, whereas in the coproporphyrin-dependent (CPD) heme biosynthesis pathway, coproporphyrin ferrochelatase (CpfC) is present, enzyme structure comparisons. CPD heme biosynthesis pathway starting from coproporphyrinogen III, overview
physiological function
-
hemH mutants accumulate all three tetrapyrroles: uroporphyrin, coproporphyrin, and protoporphyrin. Porphyrin accumulation does not occur in these mutants (in a hemA4 background) in the absence of 5-aminolevulinic acid
physiological function
a hemH mutant primarily accumulates protoporphyrin IX
physiological function
coproporphyrin ferrochelatases (CpfCs) insert ferrous iron into coproporphyrin III yielding coproheme. CpfCs are utilized by prokaryotic, mainly monoderm (Gram-positive) bacteria within the coproporphyrin-dependent (CPD) heme biosynthesis pathway
physiological function
coproporpyhrin III is the substrate of coproporphyrin ferrochelatases (CpfCs). These enzymes catalyse the insertion of ferrous iron into the porphyrin ring. This is the penultimate step within the coproporphyrin-dependent heme biosynthesis pathway. In the CPD pathway, CpfC produces iron coproporphyrin III (coproheme), which is the substrate for coproheme decarboxylase (ChdC)
physiological function
-
a hemH mutant primarily accumulates protoporphyrin IX
-
additional information
active site architecture of wild-type and R45L mutant coproporphyrin ferrochelatases with bound physiological substrate coproporphyrin III, crystal structure analysis, propionate interactions and porphyrin core deformation, detailed overview. A cleft, build by structural elements of both ferredoxin-like domains, contains several conserved amino acid residues (distal H182 and E263, proximal Y12) and is the porphyrin binding site, where catalysis happens. The monomeric enzyme exhibits two ferredoxin-like domains, each with a four-stranded parallel beta-sheet flanked by alpha-helices. In cpIII-LmCpfC, as well as in the coproheme-bound structure, the porphyrin is oriented in the active site with the propionates 2 and 4 (p2 and p4) pointing to the inner core of the protein, p6 and p7 face towards the protein surface and are much more solvent exposed. In both cpIII and coproheme complexes with LmCpfC the H-bond interactions between the four propionates and six amino acids, Thr14, Arg29, Arg45, Tyr46, Ser53, and Tyr124, are fundamental to the porphyrin stabilization
additional information
analysis of the catalytically relevant binding mode of coproporphyrin and coproheme in coproporphyrin ferrochelatase, crystal structures analysis and calorimetric methods, detailed overview. UV-vis absorption spectroscopy of LmCpfC with the substrate coproporphyrin III and the product coproheme, spectral transitions upon binding of coproporphyrin III to apo-LmCpfC shows a biphasic behavior, structure comparisons. Binding of coproporphyrin III or coproheme to apo-LmCpfC has a strong stabilizing effect on the protein, as reflected in the increased thermal stability of the resulting complexes
additional information
analysis of the interactions of the four-propionate substrate, coproporphyrin III, and the fourpropionate product, iron coproporphyrin III (coproheme), with the CpfC from Listeria monocytogenes, differences with respect to the protoporphyrin IX and heme b complexes in the different CpfC enzymes, ligand binding structures, overview. Tyrosine Y124 in LmCpfC coordinates the propionate at position 2, which is conserved in monoderm CpfCs and is highly important for binding and stabilisation. A tyrosine-serine-threonine triad coordinates the propionate at position 4
Show AA Sequence and Transmembrane Helices (5371 entries)
Longer loading times are possible. Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Longer loading times are possible. Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PDB
SCOP
CATH
UNIPROT
ORGANISM
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
35348
40500
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure in presence of iron. Only a single iron ion is found in the active site, coordinated in a square pyramidal fashion by two amino acid residues, His183 and Glu264, and three water molecules. This iron ion is not present in the structure of a His183Ala modified ferrochelatase. Insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264
the three-dimensional structure is determined at 1.9 A resolution by the method of multiple isomorphous replacement. The structural model contains 308 of the 310 amino acid residues of the protein and 198 solvent molecules
-
purified recombinant His-tagged enzyme in apoform and with bound product coproheme, mixing of 9.5 mg/ml protein with or without bound ligand in solution with 16% w/v PEG 8000, 20% v/v glycerol, and 0.04 M KH2PO4 for the apo-enzyme crystals and with 0.1 M Bis-Tris, pH 5.5, 25% PEG 3350, 0.2 M MgCl2 for the complexed enzyme, 2 days, X-ray diffraction structure determination and analysis, molecular replacement using the PDB structure 2HK6 of BsCpfC enzyme as template
purified wild-type and mutant R45L enzymes in apo and in substrate coproporphyrin III-bound forms, apoprotein-LmCpfC is dissolved in 50 mM HEPES, pH 7.4, and mixed with 0.0.05-0.010 mM cpIII in 50 mM HEPES, pH 7.4, the crystallization solution contains 15-25% PEG MME 2000, 0.1 M Bis-Tris, pH 5.8-6.3, and 0.1-0.4 M calcium acetate, mixing of 150 nl each. The LmCpfC R45L mutant in complex with cpIII is crystallized in 18% w/v PEG 8000 and 20% glycerol with a 1:1000 dilution of apo wild-type germination solution containing crushed apo wild-type crystals and 5 mM phosphate buffer, one drop contains 150 nl of 0.35 mM cpIII-R45L protein, 200 nl of mother liquor, and 30 nl of 1:1000 seed solution, X-ray diffraction structure determination and analysis at 1.51 A and 2.64 A resolution, respectively, molecular replacement using the apo-LmCpfC structure (PDB ID 6RWV) and refined to Rfree values of 0.1816 (cpIII-LmCpfC wild-type) and 0.2112 (cpIII-LmCpfC R45L) as templates
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
R29L
site-directed mutagenesis, the mutant shows altered ligand binding, kinetics, and thermostability compared to wild-type enzyme
R45L
S53A
site-directed mutagenesis, the mutant shows altered ligand binding and thermostability compared to wild-type enzyme
T14V
site-directed mutagenesis, the mutant shows altered ligand binding, kinetics, and thermostability compared to wild-type enzyme
T14V/Y46F/S53A
site-directed mutagenesis, the mutant shows altered ligand binding, kinetics, and thermostability compared to wild-type enzyme
Y124F
Y46F
site-directed mutagenesis, the mutant shows altered ligand binding and thermostability compared to wild-type enzyme
additional information
all the non-wild-type LmCpfC variants prove to have a significantly higher koff and consequently a lower affinity. This weaker binding is also reflected in the kon-rate, which is the highest for both tested porphyrin ligands for the wild-type protein
R45L
site-directed mutagenesis, the mutant lacks the H-bonding with the propionate at position 6 (p6), determination and analysis of the mutant enzyme structure as apo-enzyme and with bound substrate coproporphyrin III, structure comparison with the wild-type enzyme, overview
R45L
site-directed mutagenesis, the mutant shows altered ligand binding, kinetics, and thermostability compared to wild-type enzyme
Y124F
site-directed mutagenesis, the mutant variant shows the presence of a minor six coordinated low spin (6cLS) species in addition to the 5cHS form, the mutant shows altered ligand binding and thermostability compared to wild-type enzyme
Y124F
site-directed mutagenesis, the mutant shows altered ligand binding, kinetics, and thermostability compared to wild-type enzyme
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
binding of coproporphyrin III or coproheme to apo-LmCpfC has a strong stabilizing effect on the protein, as reflected in the increased thermal stability of the resulting complexes
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and desalting gel filtration
the recombinant reconstituted cpIII-LmCpfC complex is purified to monodispersity by gel filtration
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene cpfC, sequence comparisons and phylogenetic analysis, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
reconstitution of LmCpfC for crystallization with coproheme is performed by the addition of a 1.2 fold molar excess, and the complex is purified by gel filtration
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hansson, M.; Hederstedt, L.
Purification and characterisation of a water-soluble ferrochelatase from Bacillus subtilis
Eur. J. Biochem.
220
201-208
1994
Bacillus subtilis
brenda
Hansson, M.D.; Karlberg, T.; Rahardja, M.A.; Al-Karadaghi, S.; Hansson, M.
Amino acid residues His183 and Glu264 in Bacillus subtilis ferrochelatase direct and facilitate the insertion of metal ion into protoporphyrin IX
Biochemistry
46
87-94
2007
Bacillus subtilis (P32396), Bacillus subtilis 168 (P32396)
brenda
Xu, K.; Delling, J.; Elliott, T.
The genes required for heme synthesis in Salmonella typhimurium include those encoding alternative functions for aerobic and anaerobic coproporphyrinogen oxidation
J. Bacteriol.
174
3953-3963
1992
Salmonella enterica subsp. enterica serovar Typhimurium
brenda
Hansson, M.; Hederstedt, L.
Cloning and characterization of the Bacillus subtilis hemEHY gene cluster, which encodes protoheme IX biosynthetic enzymes
J. Bacteriol.
174
8081-8093
1992
Bacillus subtilis (P32396), Bacillus subtilis 168 (P32396)
brenda
Dailey, H.A.; Gerdes, S.; Dailey, T.A.; Burch, J.S.; Phillips, J.D.
Noncanonical coproporphyrin-dependent bacterial heme biosynthesis pathway that does not use protoporphyrin
Proc. Natl. Acad. Sci. USA
112
2210-2215
2015
Bacillus subtilis, Cutibacterium acnes, Mycobacterium tuberculosis
brenda
Al-Karadaghi, S.; Hansson, M.; Nikonov, S.; Jonsson, B.; Hederstedt, L.
Crystal structure of ferrochelatase: the terminal enzyme in heme biosynthesis
Structure
5
1501-1510
1997
Bacillus subtilis, Bacillus subtilis 168
brenda
Hofbauer, S.; Helm, J.; Obinger, C.; Djinovic-Carugo, K.; Furtmueller, P.G.
Crystal structures and calorimetry reveal catalytically relevant binding mode of coproporphyrin and coproheme in coproporphyrin ferrochelatase
FEBS J.
287
2779-2796
2020
Listeria monocytogenes (A0A5D5VUB4)
brenda
Gabler, T.; Sebastiani, F.; Helm, J.; Dali, A.; Obinger, C.; Furtmueller, P.G.; Smulevich, G.; Hofbauer, S.
Substrate specificity and complex stability of coproporphyrin ferrochelatase is governed by hydrogen-bonding interactions of the four propionate groups
FEBS J.
289
1680-1699
2022
Listeria monocytogenes (A0A5D5VUB4)
brenda
Dali, A.; Gabler, T.; Sebastiani, F.; Destinger, A.; Furtmueller, P.G.; Pfanzagl, V.; Becucci, M.; Smulevich, G.; Hofbauer, S.
Active site architecture of coproporphyrin ferrochelatase with its physiological substrate coproporphyrin III propionate interactions and porphyrin core deformation
Protein Sci.
32
e4534
2023
Listeria monocytogenes (A0A5D5VUB4)
brenda
EXTERNAL LINKS (specific for EC-Number 4.99.1.9)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2026.1 (March 2026)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
![DSMZ Digital Diversity]()
![Global Core Biodata Resource]()
![ELIXIR Core Data Resource]()
![German Network for Bioinformatics Infrastructure]()
