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Literature summary extracted from

  • Oda, K.; Matoba, Y.; Kumagai, T.; Noda, M.; Sugiyama, M.
    Crystallographic study to determine the substrate specificity of an L-serine-acetylating enzyme found in the D-cycloserine biosynthetic pathway (2013), J. Bacteriol., 195, 1741-1749.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.30 gene dcsE, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS Streptomyces lavendulae
2.3.1.31 gene dcsE, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS Streptomyces lavendulae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.1.30 purified recombinant wild-type and mutant G52A-P55G enzymes, sitting drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml of protein in 20 mM Tris-HCl, pH 7.6, 0.2 M NaCl, 5 mM dithiothreitol, and 1 mM EDTA, with 0.001 ml of precipitant solution containing 0.1 M Tris-HCl, pH 7.5, 25% w/v PEG 4000, and 0.2 M ammonium acetate, 3 weeks, structure modeling, X-ray diffraction structure determination and analysis at 1.7-1.8 A resolution Streptomyces lavendulae
2.3.1.31 purified recombinant wild-type and mutant G52A-P55G enzymes, sitting drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml of protein in 20 mM Tris-HCl, pH 7.6, 0.2 M NaCl, 5 mM dithiothreitol, and 1 mM EDTA, with 0.001 ml of precipitant solution containing 0.1 M Tris-HCl, pH 7.5, 25% w/v PEG 4000, and 0.2 M ammonium acetate, 3 weeks, structure modeling, X-ray diffraction structure determination and analysis at 1.7-1.8 A resolution Streptomyces lavendulae

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.30 G52A site-directed mutagenesis, no activity with L-serine, but with L-homoserine Streptomyces lavendulae
2.3.1.30 G52A/P55G site-directed mutagenesis, no activity with L-serine, but with L-homoserine Streptomyces lavendulae
2.3.1.30 P55G site-directed mutagenesis, increased activity with L-serine and with L-homoserine compared to the wild-type enzyme Streptomyces lavendulae
2.3.1.31 G52A site-directed mutagenesis, no activity with L-serine, but with L-homoserine, although the mutant is less active than the wild-type enzyme Streptomyces lavendulae
2.3.1.31 G52A/P55G site-directed mutagenesis, no activity with L-serine, but with L-homoserine, activity with L-homoserine is increased compared to the wild-type enzyme Streptomyces lavendulae
2.3.1.31 P55G site-directed mutagenesis, increased activity with L-serine and with L-homoserine compared to the wild-type enzyme Streptomyces lavendulae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.30 0.5
-
 L-serine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.30 4.9
-
 L-serine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 1
-
 L-homoserine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 2
-
 L-homoserine recombinant His-tagged mutant G52A/P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 25
-
 L-homoserine recombinant His-tagged mutant G52A enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 98
-
 L-homoserine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.30 acetyl-CoA + L-serine Streptomyces lavendulae
-
 CoA + O-acetyl-L-serine
-
 ?
2.3.1.30 acetyl-CoA + L-serine Streptomyces lavendulae ATCC 11924
-
 CoA + O-acetyl-L-serine
-
 ?
2.3.1.31 acetyl-CoA + L-homoserine Streptomyces lavendulae
-
 CoA + O-acetyl-L-homoserine
-
 ?
2.3.1.31 acetyl-CoA + L-homoserine Streptomyces lavendulae ATCC 11924
-
 CoA + O-acetyl-L-homoserine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.30 Streptomyces lavendulae D2Z028 gene dcsE; gene dcsE
-
2.3.1.30 Streptomyces lavendulae ATCC 11924 D2Z028 gene dcsE; gene dcsE
-
2.3.1.31 Streptomyces lavendulae D2Z028 gene dcsE; gene dcsE
-
2.3.1.31 Streptomyces lavendulae ATCC 11924 D2Z028 gene dcsE; gene dcsE
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.30 recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, dialysis and ultrafiltration Streptomyces lavendulae
2.3.1.31 recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, dialysis and ultrafiltration Streptomyces lavendulae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.30 acetyl-CoA + L-serine
-
 Streptomyces lavendulae CoA + O-acetyl-L-serine
-
 ?
2.3.1.30 acetyl-CoA + L-serine
-
 Streptomyces lavendulae ATCC 11924 CoA + O-acetyl-L-serine
-
 ?
2.3.1.30 additional information the enzyme prefers L-serine as a substrate, but is also active with L-homoserine, EC 2.3.1.31, substrate specificity, overview Streptomyces lavendulae ?
-
 ?
2.3.1.30 additional information the enzyme prefers L-serine as a substrate, but is also active with L-homoserine, EC 2.3.1.31, substrate specificity, overview Streptomyces lavendulae ATCC 11924 ?
-
 ?
2.3.1.31 acetyl-CoA + L-homoserine
-
 Streptomyces lavendulae CoA + O-acetyl-L-homoserine
-
 ?
2.3.1.31 acetyl-CoA + L-homoserine
-
 Streptomyces lavendulae ATCC 11924 CoA + O-acetyl-L-homoserine
-
 ?
2.3.1.31 additional information the enzyme prefers L-serine as a substrate, cf. EC 2.3.1.30, but is also active with L-homoserine, substrate specificity, overview Streptomyces lavendulae ?
-
 ?
2.3.1.31 additional information the enzyme prefers L-serine as a substrate, cf. EC 2.3.1.30, but is also active with L-homoserine, substrate specificity, overview Streptomyces lavendulae ATCC 11924 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.30 DcsE
-
 Streptomyces lavendulae
2.3.1.31 DcsE
-
 Streptomyces lavendulae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.30 30
-
 assay at Streptomyces lavendulae
2.3.1.31 30
-
 assay at Streptomyces lavendulae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.30 0.62
-
 L-serine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.30 1.58
-
 L-serine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.22
-
 L-homoserine recombinant His-tagged mutant G52A enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.63
-
 L-homoserine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.78
-
 L-homoserine recombinant His-tagged mutant G52A/P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 1.47
-
 L-homoserine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.30 8
-
 assay at Streptomyces lavendulae
2.3.1.31 8
-
 assay at Streptomyces lavendulae

General Information

EC Number General Information Comment Organism
2.3.1.30 additional information the enzyme displays a high sequence homology to L-homoserine O-acetyltransferase, but it prefers L-serine over L-homoserine as the substrate, structure analysis and modeling, overview Streptomyces lavendulae
2.3.1.31 additional information the enzyme displays a high sequence homology to L-homoserine O-acetyltransferase, but it prefers L-serine over L-homoserine as the substrate, structure analysis and modeling, overview Streptomyces lavendulae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.1.30 0.32
-
 L-serine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.30 1.23
-
 L-serine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.009
-
 L-homoserine recombinant His-tagged mutant G52A enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.015
-
 L-homoserine recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.4
-
 L-homoserine recombinant His-tagged mutant G52A/P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae
2.3.1.31 0.63
-
 L-homoserine recombinant His-tagged mutant P55G enzyme, pH 8.0, 30°C Streptomyces lavendulae