Literature summary for 2.3.1.31 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
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
P55G	site-directed mutagenesis, increased activity with L-serine and with L-homoserine compared to the wild-type enzyme	Streptomyces lavendulae

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Streptomyces lavendulae	D2Z028	gene dcsE; gene dcsE	-
Streptomyces lavendulae ATCC 11924	D2Z028	gene dcsE; gene dcsE	-

Purification (Commentary)

Purification (Comment)	Organism
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)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + L-homoserine	-	Streptomyces lavendulae	CoA + O-acetyl-L-homoserine	-	?
acetyl-CoA + L-homoserine	-	Streptomyces lavendulae ATCC 11924	CoA + O-acetyl-L-homoserine	-	?
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	?	-	?
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

Synonyms	Comment	Organism
DcsE	-	Streptomyces lavendulae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Streptomyces lavendulae

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Streptomyces lavendulae

General Information

General Information	Comment	Organism
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]

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