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Information on EC 4.99.1.3 - sirohydrochlorin cobaltochelatase and Organism(s) Methanosarcina barkeri and UniProt Accession P61816

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IUBMB Comments
This enzyme, which forms part of the anaerobic (early cobalt insertion) cobalamin biosynthesis pathway, is an ATP-independent type II chelatase. Two distinct forms are known - a primordial form named CbiX, which is most common in archaea, and a strictly bacterial form named CbiK. See EC 6.6.1.2, cobaltochelatase, for the cobaltochelatase that participates in the aerobic cobalamin biosynthesis pathway.
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This record set is specific for:
Methanosarcina barkeri
UNIPROT: P61816
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Word Map
The taxonomic range for the selected organisms is: Methanosarcina barkeri
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
cobalt chelatase, cbixl, cbixs, sirohydrochlorin-ferrochelatase, sirohydrochlorin cobaltochelatase, anaerobic cobalt chelatase, archaeal cobaltochelatase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CbiXS
small, single-domain chelatase
sirohydrochlorin cobalt-lyase
-
sirohydrochlorin cobaltochelatase
-
anaerobic cobalt chelatase
-
-
-
-
archaeal cobaltochelatase
-
-
CbiK
-
-
-
-
CbiXS
cobaltochelatase
sirohydrochlorin cobaltochelatase
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
cobalt-sirohydrochlorin cobalt-lyase (sirohydrochlorin-forming)
This enzyme, which forms part of the anaerobic (early cobalt insertion) cobalamin biosynthesis pathway, is an ATP-independent type II chelatase. Two distinct forms are known - a primordial form named CbiX, which is most common in archaea, and a strictly bacterial form named CbiK. See EC 6.6.1.2, cobaltochelatase, for the cobaltochelatase that participates in the aerobic cobalamin biosynthesis pathway.
CAS REGISTRY NUMBER
COMMENTARY hide
81295-49-0
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
sirohydrochlorin + Co2+
cobalt-sirohydrochlorin + 2 H+
show the reaction diagram
-
-
-
?
sirohydrochlorin + Co2+
cobalt-sirohydrochlorin + H+
show the reaction diagram
sirohydrochlorin + Fe2+
siroheme + H+
show the reaction diagram
-
CbiXS can act as a ferrochelatase in the biosynthesis of siroheme in vivo
-
?
additional information
?
-
-
precorrin-2 is no substrate
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
sirohydrochlorin + Co2+
cobalt-sirohydrochlorin + H+
show the reaction diagram
-
cobalamin biosynthetic pathway
-
?
sirohydrochlorin + Fe2+
siroheme + H+
show the reaction diagram
-
CbiXS can act as a ferrochelatase in the biosynthesis of siroheme in vivo
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
insertion of cobalt into sirohydrochlorin
Fe2+
-
able to chelate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00043 - 0.00062
sirohydrochlorin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0056
His-tagged protein of mutant M508
0.007
His-tagged protein of mutant M518
0.0075
His-tagged protein of mutant M54
0.01098
His-tagged protein of mutant M136
0.0135
His-tagged protein of mutant M507
0.0155
His-tagged protein of mutant M519
0.0164
His-tagged protein of mutant M10
0.0255
His-tagged protein of mutant M510
0.0272
His-tagged protein of mutant M51
0.0938
His-tagged protein of mutant M200
0.1197
His-tagged wild type protein
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
14300
SDS-PAGE, His-tagged protein of mutant M76
14900
SDS-PAGE, His-tagged wild type protein
17200
SDS-PAGE, His-tagged protein of mutant M200
17500
SDS-PAGE, His-tagged protein of mutant M54
17800
SDS-PAGE, His-tagged protein of mutant M136
18400
SDS-PAGE, His-tagged protein of mutant M150
18600
SDS-PAGE, His-tagged protein of mutant M10
18700
SDS-PAGE, His-tagged protein of mutant M508
20100
SDS-PAGE, His-tagged protein of mutant M507
25300
SDS-PAGE, His-tagged protein of mutant M518
27000
SDS-PAGE, His-tagged protein of mutant M51
14000
-
gene-predicted molecular mass
15000
-
SDS-PAGE
40000
-
gel filtration
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
using the sitting drop vapor diffusion technique in 96-well plates, in two different crystal forms consisting of a central mixed b-sheet flanked by four alpha helices
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H12A
wild type enzyme
H192A
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by using metal chelate affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
overexpression in siroheme deficient Escherichia coli after random in vitro gene sequence rearrangements in order to screen the generated library for Escherichia coli cells that are able to insert both cobalt and to a lesser extend iron into its tetrapyrrole substrate sirohydrochlorin
overproduced in Escherichia coli
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Brindley, A.A.; Raux E.; Leech, H.K.; Schubert, H.L.; Warren, M.J.
A story of chelatase evolution: Identification and characterisation of a small 13-15 kDa 'ancestral' cobaltochelatase (CbiXS) in the Archaea
J. Biol. Chem.
278
22388-22395
2003
Methanobacter thermoautotrophicum, Methanosarcina barkeri, Methanosarcina barkeri Fusaro / DSM 804
Manually annotated by BRENDA team
Pisarchik, A.; Petri, R.; Schmidt-Dannert, C.
Probing the structural plasticity of an archaeal primordial cobaltochelatase CbiXS
Protein Eng. Des. Sel.
20
257-265
2007
Methanosarcina barkeri (P61816)
Manually annotated by BRENDA team