EC Number | Cloned (Comment) | Organism |
---|---|---|
2.5.1.47 | expressed in Escherichia coli | Acidithiobacillus ferrooxidans |
2.5.1.47 | PCR-amplification, expression of His-tagged wild type and mutant enzyme in Escherichia coli BL21(DE3) with expression vector pLM1 | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.5.1.47 | H150A | in contrast to the wild-type protein the mutant protein is colorless after purification, and UV-Vis scanning of the mutant proteins show that there are no absorptions between 300 and 500 nm, mutant protein does not show a comparable activity to the wild-type protein. This suggests that the mutated residue is crucial or pyridoxal 5'-phosphate binding and stabilization | Acidithiobacillus ferrooxidans |
2.5.1.47 | H150A | not active, crucial for cofactor binding | Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | H168A | in contrast to the wild-type protein the mutant protein is colorless after purification, and UV-Vis scanning of the mutant proteins show that there are no absorptions between 300 and 500 nm, mutant protein does not show a comparable activity to the wild-type protein. This suggests that the mutated residue is crucial or pyridoxal 5'-phosphate binding and stabilization | Acidithiobacillus ferrooxidans |
2.5.1.47 | H168A | not active, crucial for cofactor binding | Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | K30A | in contrast to the wild-type protein the mutant protein is colorless after purification, and UV-Vis scanning of the mutant proteins show that there are no absorptions between 300 and 500 nm, mutant protein does not show a comparable activity to the wild-type protein. This suggests that the mutated residue is crucial or pyridoxal 5'-phosphate binding and stabilization | Acidithiobacillus ferrooxidans |
2.5.1.47 | K30A | not active, crucial for cofactor binding | Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | K41A | mutant protein is also colourful as the wild-type protein, mutant protein does not show the same activity as the wild-type protein | Acidithiobacillus ferrooxidans |
2.5.1.47 | K41A | not active | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.5.1.47 | 32000 | - |
SDS-PAGE | Acidithiobacillus ferrooxidans |
2.5.1.47 | 32000 | - |
wild type and mutants, SDS-PAGE | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.5.1.47 | O3-acetyl-L-serine + hydrogen sulfide | Acidithiobacillus ferrooxidans ATCC 23270 | - |
L-cysteine + acetate | - |
? |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
2.5.1.47 | Glycerol | 5% glycerol and 5 mM 2-mercaptoethanol are essential for maintaining the long-term stability of the enzyme | Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | mercaptoethanol | 5% glycerol and 5 mM 2-mercaptoethanol are essential for maintaining the long-term stability of the enzyme | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.5.1.47 | Acidithiobacillus ferrooxidans | - |
- |
- |
2.5.1.47 | Acidithiobacillus ferrooxidans ATCC 23270 | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.5.1.47 | cells are harvested by centrifugation, washed with sterile water, again harvested by centrifugation, suspended in 20 mM potassium phosphate buffer, pH 7.4, containing 0.5 M NaCl, lysed with lysozyme at room temperature, one-step affinity chromatography with nickel metal-affinity resin columns, dialyzed against 20 mM potassium phosphate buffer, pH 7.5, 5% glycerol, and 5 mM 2-mercaptoethanol | Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | the soluble protein is purified by one-step affinity chromatography to apparent homogeneity | Acidithiobacillus ferrooxidans |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
2.5.1.47 | 5 | - |
wild type | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.5.1.47 | O3-acetyl-L-serine + hydrogen sulfide | - |
Acidithiobacillus ferrooxidans ATCC 23270 | L-cysteine + acetate | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.5.1.47 | ATCC 23270 protein | - |
Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | O-acetylserine sulfhydrylase | - |
Acidithiobacillus ferrooxidans |
2.5.1.47 | O-acetylserine sulfhydrylase | - |
Acidithiobacillus ferrooxidans ATCC 23270 |
2.5.1.47 | OASS | - |
Acidithiobacillus ferrooxidans |
2.5.1.47 | OASS | - |
Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
2.5.1.47 | pyridoxal 5'-phosphate | sequence alignment and site-directed mutation of the enzyme reveal that the cofactor PLP is covalently bound in Schiff base linkage with K30, as well as the two residues H150 and H168 are the crucial residues for PLP binding and stabilization | Acidithiobacillus ferrooxidans | |
2.5.1.47 | pyridoxal 5'-phosphate | enzyme sites K30, H150, and H168 are crucial for cofactor binding and stabilization | Acidithiobacillus ferrooxidans ATCC 23270 |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.5.1.47 | metabolism | key enzyme in the L-cysteine pathway | Acidithiobacillus ferrooxidans ATCC 23270 |