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Literature summary for 2.5.1.47 extracted from

  • Zheng, C.; Nie, L.; Qian, L.; Wang, Z.; Liu, G.; Liu, J.
    K30, H150, and H168 Are Essential Residues for Coordinating Pyridoxal 5-Phosphate of O-Acetylserine Sulfhydrylase from Acidithiobacillus ferrooxidans (2009), Curr. Microbiol., 60, 461-465.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.5.1.47

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Acidithiobacillus ferrooxidans
PCR-amplification, expression of His-tagged wild type and mutant enzyme in Escherichia coli BL21(DE3) with expression vector pLM1 Acidithiobacillus ferrooxidans ATCC 23270

Protein Variants

Protein Variants Comment Organism
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
H150A not active, crucial for cofactor binding Acidithiobacillus ferrooxidans ATCC 23270
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
H168A not active, crucial for cofactor binding Acidithiobacillus ferrooxidans ATCC 23270
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
K30A not active, crucial for cofactor binding Acidithiobacillus ferrooxidans ATCC 23270
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
K41A not active Acidithiobacillus ferrooxidans ATCC 23270

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
 SDS-PAGE Acidithiobacillus ferrooxidans
32000
-
 wild type and mutants, SDS-PAGE Acidithiobacillus ferrooxidans ATCC 23270

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
O3-acetyl-L-serine + hydrogen sulfide Acidithiobacillus ferrooxidans ATCC 23270
-
 L-cysteine + acetate
-
 ?

Organic Solvent Stability

Organic Solvent Comment Organism
Glycerol 5% glycerol and 5 mM 2-mercaptoethanol are essential for maintaining the long-term stability of the enzyme Acidithiobacillus ferrooxidans ATCC 23270
mercaptoethanol 5% glycerol and 5 mM 2-mercaptoethanol are essential for maintaining the long-term stability of the enzyme Acidithiobacillus ferrooxidans ATCC 23270

Organism

Organism UniProt Comment Textmining
Acidithiobacillus ferrooxidans
-
-
-
Acidithiobacillus ferrooxidans ATCC 23270
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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
the soluble protein is purified by one-step affinity chromatography to apparent homogeneity Acidithiobacillus ferrooxidans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5
-
 wild type Acidithiobacillus ferrooxidans ATCC 23270

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
O3-acetyl-L-serine + hydrogen sulfide
-
 Acidithiobacillus ferrooxidans ATCC 23270 L-cysteine + acetate
-
 ?

Synonyms

Synonyms Comment Organism
ATCC 23270 protein
-
 Acidithiobacillus ferrooxidans ATCC 23270
O-acetylserine sulfhydrylase
-
 Acidithiobacillus ferrooxidans
O-acetylserine sulfhydrylase
-
 Acidithiobacillus ferrooxidans ATCC 23270
OASS
-
 Acidithiobacillus ferrooxidans
OASS
-
 Acidithiobacillus ferrooxidans ATCC 23270

Cofactor

Cofactor Comment Organism Structure
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
pyridoxal 5'-phosphate enzyme sites K30, H150, and H168 are crucial for cofactor binding and stabilization Acidithiobacillus ferrooxidans ATCC 23270

General Information

General Information Comment Organism
metabolism key enzyme in the L-cysteine pathway Acidithiobacillus ferrooxidans ATCC 23270