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Literature summary for 4.99.1.3 extracted from

  • Pisarchik, A.; Petri, R.; Schmidt-Dannert, C.
    Probing the structural plasticity of an archaeal primordial cobaltochelatase CbiXS (2007), Protein Eng. Des. Sel., 20, 257-265.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
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 Methanosarcina barkeri

Crystallization (Commentary)

Crystallization (Comment) Organism
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 Methanosarcina barkeri

Protein Variants

Protein Variants Comment Organism
H12A wild type enzyme Methanosarcina barkeri
H192A double mutation in M51 mutant Methanosarcina barkeri
H192A double mutation in M518 mutant Methanosarcina barkeri
H78A double mutation in M51 mutant Methanosarcina barkeri
H78A wild type enzyme Methanosarcina barkeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00043
-
sirohydrochlorin mutant M51 Methanosarcina barkeri
0.00062
-
sirohydrochlorin wild type protein Methanosarcina barkeri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
14300
-
SDS-PAGE, His-tagged protein of mutant M76 Methanosarcina barkeri
14900
-
SDS-PAGE, His-tagged wild type protein Methanosarcina barkeri
17200
-
SDS-PAGE, His-tagged protein of mutant M200 Methanosarcina barkeri
17500
-
SDS-PAGE, His-tagged protein of mutant M54 Methanosarcina barkeri
17800
-
SDS-PAGE, His-tagged protein of mutant M136 Methanosarcina barkeri
18400
-
SDS-PAGE, His-tagged protein of mutant M150 Methanosarcina barkeri
18600
-
SDS-PAGE, His-tagged protein of mutant M10 Methanosarcina barkeri
18700
-
SDS-PAGE, His-tagged protein of mutant M508 Methanosarcina barkeri
20100
-
SDS-PAGE, His-tagged protein of mutant M507 Methanosarcina barkeri
25300
-
SDS-PAGE, His-tagged protein of mutant M518 Methanosarcina barkeri
27000
-
SDS-PAGE, His-tagged protein of mutant M51 Methanosarcina barkeri

Organism

Organism UniProt Comment Textmining
Methanosarcina barkeri P61816
-
-

Purification (Commentary)

Purification (Comment) Organism
by using metal chelate affinity chromatography Methanosarcina barkeri

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0056
-
His-tagged protein of mutant M508 Methanosarcina barkeri
0.007
-
His-tagged protein of mutant M518 Methanosarcina barkeri
0.0075
-
His-tagged protein of mutant M54 Methanosarcina barkeri
0.01098
-
His-tagged protein of mutant M136 Methanosarcina barkeri
0.0135
-
His-tagged protein of mutant M507 Methanosarcina barkeri
0.0155
-
His-tagged protein of mutant M519 Methanosarcina barkeri
0.0164
-
His-tagged protein of mutant M10 Methanosarcina barkeri
0.0255
-
His-tagged protein of mutant M510 Methanosarcina barkeri
0.0272
-
His-tagged protein of mutant M51 Methanosarcina barkeri
0.0938
-
His-tagged protein of mutant M200 Methanosarcina barkeri
0.1197
-
His-tagged wild type protein Methanosarcina barkeri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sirohydrochlorin + Co2+
-
Methanosarcina barkeri cobalt-sirohydrochlorin + 2 H+
-
?

Synonyms

Synonyms Comment Organism
CbiXS small, single-domain chelatase Methanosarcina barkeri
sirohydrochlorin cobalt-lyase
-
Methanosarcina barkeri
sirohydrochlorin cobaltochelatase
-
Methanosarcina barkeri