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Literature summary extracted from
- Characterization and preliminary mutation analysis of a thermostable alanine racemase from Thermoanaerobacter tengcongensis MB4 (2013), Extremophiles, 17, 611-621.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.1.1.1 | gene MBalr2, DNA and amino acid sequence determination and analysis, expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Caldanaerobacter subterraneus subsp. tengcongensis |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.1.1.1 | H359Y | site-directed mutagenesis | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | S171A | site-directed mutagenesis | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | S171A/H359Y | site-directed mutagenesis | Caldanaerobacter subterraneus subsp. tengcongensis |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.1.1.1 | 20.16 | - |
D-alanine | pH 11.0, 70°C, recombinant enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |  |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 46000 | - |
2 * 46000, recombinant His-tagged wild-type enzyme, SDS-PAGE, 2 * 46621, recombinant His-tagged wild-type enzyme, sequence calculation, 2 * 46605, recombinant His-tagged mutant S171A, sequence calculation, 2 * 46647, recombinant double-His-tagged mutant H359Y, sequence calculation | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 46605 | - |
2 * 46000, recombinant His-tagged wild-type enzyme, SDS-PAGE, 2 * 46621, recombinant His-tagged wild-type enzyme, sequence calculation, 2 * 46605, recombinant His-tagged mutant S171A, sequence calculation, 2 * 46647, recombinant double-His-tagged mutant H359Y, sequence calculation | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 46621 | - |
2 * 46000, recombinant His-tagged wild-type enzyme, SDS-PAGE, 2 * 46621, recombinant His-tagged wild-type enzyme, sequence calculation, 2 * 46605, recombinant His-tagged mutant S171A, sequence calculation, 2 * 46647, recombinant double-His-tagged mutant H359Y, sequence calculation | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 46647 | - |
2 * 46000, recombinant His-tagged wild-type enzyme, SDS-PAGE, 2 * 46621, recombinant His-tagged wild-type enzyme, sequence calculation, 2 * 46605, recombinant His-tagged mutant S171A, sequence calculation, 2 * 46647, recombinant double-His-tagged mutant H359Y, sequence calculation | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 76000 | - |
recombinant His-tagged wild-type enzyme, gel filtration | Caldanaerobacter subterraneus subsp. tengcongensis |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.1 | L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | catalyzes the interconversion of D-alanine and L-alanine | D-alanine | - |
r | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.1.1.1 | Caldanaerobacter subterraneus subsp. tengcongensis | Q8R860 | gene MBalr2; gene MBalr2 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.1.1.1 | recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinit chromatography and gel filtration | Caldanaerobacter subterraneus subsp. tengcongensis |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 1414 | - |
purified recombinant wild-type enzyme, pH 11.0, 70°C | Caldanaerobacter subterraneus subsp. tengcongensis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.1 | L-alanine | catalyzes the interconversion of D-alanine and L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-alanine | - |
r | |
| 5.1.1.1 | L-alanine | catalyzes the interconversion of D-alanine and L-alanine, highly preferred substrate | Caldanaerobacter subterraneus subsp. tengcongensis | D-alanine | - |
r | |
| 5.1.1.1 | L-arginine | less than 10% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-arginine | - |
r | |
| 5.1.1.1 | L-leucine | 0.9% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-leucine | - |
r | |
| 5.1.1.1 | L-lysine | less than 10% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-lysine | - |
r | |
| 5.1.1.1 | L-Methionine | less than 10% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-Methionine | - |
r | |
| 5.1.1.1 | L-phenylalanine | 1.1% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-phenylalanine | - |
r | |
| 5.1.1.1 | L-proline | 1.1% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-proline | - |
r | |
| 5.1.1.1 | L-serine | less than 10% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-serine | - |
r | |
| 5.1.1.1 | L-valine | less than 10% activity compared to L-alanine | Caldanaerobacter subterraneus subsp. tengcongensis | D-valine | - |
r | |
| 5.1.1.1 | additional information | high specificity to L-alanine, low activity with L-arginine, L-methionine, L-lysine, L-serine, L-valine, L-proline, L-phenylalanine, L-leucine, respectively, no activity with L-histidine | Caldanaerobacter subterraneus subsp. tengcongensis | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.1.1.1 | dimer | 2 * 46000, recombinant His-tagged wild-type enzyme, SDS-PAGE, 2 * 46621, recombinant His-tagged wild-type enzyme, sequence calculation, 2 * 46605, recombinant His-tagged mutant S171A, sequence calculation, 2 * 46647, recombinant double-His-tagged mutant H359Y, sequence calculation | Caldanaerobacter subterraneus subsp. tengcongensis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.1.1.1 | MBAlr2 | - |
Caldanaerobacter subterraneus subsp. tengcongensis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 70 | 75 | - |
Caldanaerobacter subterraneus subsp. tengcongensis |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 30 | 90 | activity range | Caldanaerobacter subterraneus subsp. tengcongensis |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 50 | - |
purified recombinant wild-type enzyme, 12 h, completely stable, the first several hours of incubation at 50-70°C help in thermoactivation of the enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 60 | - |
purified recombinant wild-type enzyme, 3 h, completely stable, the first several hours of incubation at 50-70°C help in thermoactivation of the enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 70 | - |
purified recombinant wild-type enzyme, 3 h, completely stable, half-life is 12 h, the first several hours of incubation at 50-70°C help in thermoactivation of the enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| 5.1.1.1 | 80 | - |
purified recombinant wild-type enzyme, sharp decline during incubation at 80°C, half-life is 1.5 h | Caldanaerobacter subterraneus subsp. tengcongensis |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 11 | - |
- |
Caldanaerobacter subterraneus subsp. tengcongensis |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 6.5 | 12.5 | activity range | Caldanaerobacter subterraneus subsp. tengcongensis |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 7 | 11.2 | purified recombinant wild-type enzyme, most thermostable at, enzyme activity is lost abruptly at below pH 7.0 and above pH 11.2 | Caldanaerobacter subterraneus subsp. tengcongensis |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.1.1.1 | pyridoxal 5'-phosphate | dependent on, alanine racemase requires pyridoxal-5'-phosphate as a cofactor to form a Schiff base between pyridoxal 5'-phosphate and epsilon-amino group of the lysine residue in the active site | Caldanaerobacter subterraneus subsp. tengcongensis | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.1.1.1 | evolution | alanine racemase belongs to the fold-type III group of pyridoxal 5'-phosphate-dependent enzymes | Caldanaerobacter subterraneus subsp. tengcongensis |
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