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Literature summary extracted from

  • Lu, M.; Xu, B.Y.; Zhou, K.; Cheng, W.; Jiang, Y.L.; Chen, Y.; Zhou, C.Z.
    Structural and biochemical analyses of Microcystis aeruginosa O-acetylserine sulfhydrylases reveal a negative feedback regulation of cysteine biosynthesis (2014), Biochim. Biophys. Acta, 1844, 308-315.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.47 gene cysK1, sequence comparisons, recombinant expression of N-terminally His6-tagged isozyme CysK1 in Escherichia coli strain BL21 (DE3) Microcystis aeruginosa
2.5.1.47 gene cysK2, sequence comparisons, recombinant expression of N-terminally His6-tagged isozyme CysK2 in Escherichia coli strain BL21 (DE3) Microcystis aeruginosa

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.47 purified isozyme CysK1, hanging drop vapor diffusion technique, mixing of 10 mg/ml protein solution with an equal volume of reservoir solution containing 0.1 M MOPS, pH 6.0, 60% 2-methyl-2,4-pentanediol, 16°C, X-ray diffraction structure determination and analysis at 2.30 A resolution, molecular replacement using the coordinates of Mycobacterium tuberculosis OASS, PDB ID 2Q3B, as the search model Microcystis aeruginosa
2.5.1.47 purified isozyme CysK2 in complex with cystine, hanging drop vapor diffusion technique, mixing of 10 mg/ml protein solution with an equal volume of reservoir solution containing 1.5 M (NH4)2SO4, 0.1M Bis-Tris-propane, pH 7.0, and 10 mM cystine, 16°C, X-ray diffraction structure determination and analysis at 1.91 A resolution, molecular replacement using the coordinates of Mycobacterium tuberculosis OASS, PDB ID 2Q3B, as the search model Microcystis aeruginosa

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.47 cystine competitive versus O-acetyl-L-serine, the cystine-binding residues are highly conserved in all OASS proteins; competitive versus O-acetyl-L-serine, the cystine-binding residues are highly conserved in all OASS proteins. Active site of CysK2–cystine binding structure, overview. Cystine occupies the substrate/product binding site of the enzyme Microcystis aeruginosa
2.5.1.47 trichloroacetic acid inactivation at 16.6% v/v; inactivation at 16.6% v/v Microcystis aeruginosa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.47 1
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa
2.5.1.47 1.1
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa
2.5.1.47 1.8
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 2.3
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 3.5
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa
2.5.1.47 4.8
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.47 O-acetyl-L-serine + hydrogen sulfide Microcystis aeruginosa
-
 L-cysteine + acetate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.47 Microcystis aeruginosa A8YBP8 gene cysK2, IPF_3084, or CAO86589
-
2.5.1.47 Microcystis aeruginosa A8YBS4 gene cysK1, IPF_2058, or cao86616
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.47 recombinant N-terminally His6-tagged isozyme CysK1 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration Microcystis aeruginosa
2.5.1.47 recombinant N-terminally His6-tagged isozyme CysK2 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration Microcystis aeruginosa

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.47 235
-
 substrate O-acetyl-L-serine, purified recombinant isozyme CysK1, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 412
-
 substrate O-acetyl-L-serine, purified recombinant isozyme CysK2, pH 7.0, 25°C Microcystis aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.47 O-acetyl-L-serine + hydrogen sulfide
-
 Microcystis aeruginosa L-cysteine + acetate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.47 O-acetylserine sulfhydrylase
-
 Microcystis aeruginosa
2.5.1.47 OASS
-
 Microcystis aeruginosa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.47 25
-
-
 Microcystis aeruginosa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.47 132
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa
2.5.1.47 136
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 140
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa
2.5.1.47 211
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 231
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa
2.5.1.47 232
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.47 7
-
 assay at Microcystis aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.47 pyridoxal 5'-phosphate dependent on, binding site structure analysis in a cleft between the N- and C-terminal domains, the phosphate group of PLP interacts with the highly conserved Gly/Thr-rich loop (G181TGGT185) and a water molecule, overview Microcystis aeruginosa

General Information

EC Number General Information Comment Organism
2.5.1.47 evolution the enzyme belongs to the family of fold-type II PLP-dependent enzymes. The cyanobacterium Microcystis aeruginosa PCC 7806 encodes three putative OASSs: CAO86616, CAO86589 and CAO86970, sequence comparisons Microcystis aeruginosa
2.5.1.47 metabolism the O-acetylserine sulfhydrylase catalyzes the final step of cysteine biosynthesis from O-acetylserine and inorganic sulfide, negative feedback regulation of the pathway. Autoinhibition by cystine might be a universal mechanism of cysteine biosynthesis pathway Microcystis aeruginosa
2.5.1.47 metabolism the O-acetylserine sulfhydrylase catalyzes the final step of cysteine biosynthesis from O-acetylserine and inorganic sulfide, negative feedback regulation of the pathway. Autoinhibition by cystine might be a universal mechanism of cysteine biosynthesis pathway, redox-dependent autoregulation Microcystis aeruginosa
2.5.1.47 additional information three-dimensional structure comparisons of isozymes CysK1 and CysK2, overview Microcystis aeruginosa

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.47 39.8
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa
2.5.1.47 48.1
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with glutathione, GSSG Microcystis aeruginosa
2.5.1.47 76.5
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 91.3
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C Microcystis aeruginosa
2.5.1.47 136
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa
2.5.1.47 219
-
 O-acetyl-L-serine recombinant enzyme, pH 7.0, 25°C, with DTT Microcystis aeruginosa