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Literature summary extracted from
- Probing the structural plasticity of an archaeal primordial cobaltochelatase CbiXS (2007), Protein Eng. Des. Sel., 20, 257-265.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 4.99.1.3 | 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)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 4.99.1.3 | 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
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 4.99.1.3 | H12A | wild type enzyme | Methanosarcina barkeri |
| 4.99.1.3 | H192A | double mutation in M51 mutant | Methanosarcina barkeri |
| 4.99.1.3 | H192A | double mutation in M518 mutant | Methanosarcina barkeri |
| 4.99.1.3 | H78A | double mutation in M51 mutant | Methanosarcina barkeri |
| 4.99.1.3 | H78A | wild type enzyme | Methanosarcina barkeri |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.99.1.3 | 0.00043 | - |
sirohydrochlorin | mutant M51 | Methanosarcina barkeri |  |
| 4.99.1.3 | 0.00062 | - |
sirohydrochlorin | wild type protein | Methanosarcina barkeri | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 4.99.1.3 | 14300 | - |
SDS-PAGE, His-tagged protein of mutant M76 | Methanosarcina barkeri |
| 4.99.1.3 | 14900 | - |
SDS-PAGE, His-tagged wild type protein | Methanosarcina barkeri |
| 4.99.1.3 | 17200 | - |
SDS-PAGE, His-tagged protein of mutant M200 | Methanosarcina barkeri |
| 4.99.1.3 | 17500 | - |
SDS-PAGE, His-tagged protein of mutant M54 | Methanosarcina barkeri |
| 4.99.1.3 | 17800 | - |
SDS-PAGE, His-tagged protein of mutant M136 | Methanosarcina barkeri |
| 4.99.1.3 | 18400 | - |
SDS-PAGE, His-tagged protein of mutant M150 | Methanosarcina barkeri |
| 4.99.1.3 | 18600 | - |
SDS-PAGE, His-tagged protein of mutant M10 | Methanosarcina barkeri |
| 4.99.1.3 | 18700 | - |
SDS-PAGE, His-tagged protein of mutant M508 | Methanosarcina barkeri |
| 4.99.1.3 | 20100 | - |
SDS-PAGE, His-tagged protein of mutant M507 | Methanosarcina barkeri |
| 4.99.1.3 | 25300 | - |
SDS-PAGE, His-tagged protein of mutant M518 | Methanosarcina barkeri |
| 4.99.1.3 | 27000 | - |
SDS-PAGE, His-tagged protein of mutant M51 | Methanosarcina barkeri |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 4.99.1.3 | Methanosarcina barkeri | P61816 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 4.99.1.3 | by using metal chelate affinity chromatography | Methanosarcina barkeri |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 4.99.1.3 | 0.0056 | - |
His-tagged protein of mutant M508 | Methanosarcina barkeri |
| 4.99.1.3 | 0.007 | - |
His-tagged protein of mutant M518 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0075 | - |
His-tagged protein of mutant M54 | Methanosarcina barkeri |
| 4.99.1.3 | 0.01098 | - |
His-tagged protein of mutant M136 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0135 | - |
His-tagged protein of mutant M507 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0155 | - |
His-tagged protein of mutant M519 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0164 | - |
His-tagged protein of mutant M10 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0255 | - |
His-tagged protein of mutant M510 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0272 | - |
His-tagged protein of mutant M51 | Methanosarcina barkeri |
| 4.99.1.3 | 0.0938 | - |
His-tagged protein of mutant M200 | Methanosarcina barkeri |
| 4.99.1.3 | 0.1197 | - |
His-tagged wild type protein | Methanosarcina barkeri |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.99.1.3 | sirohydrochlorin + Co2+ | - |
Methanosarcina barkeri | cobalt-sirohydrochlorin + 2 H+ | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 4.99.1.3 | CbiXS | small, single-domain chelatase | Methanosarcina barkeri |
| 4.99.1.3 | sirohydrochlorin cobalt-lyase | - |
Methanosarcina barkeri |
| 4.99.1.3 | sirohydrochlorin cobaltochelatase | - |
Methanosarcina barkeri |
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Release 2023.1 (January 2023)
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