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Literature summary for 5.1.1.13 extracted from

  • Uda, K.; Abe, K.; Dehara, Y.; Mizobata, K.; Edashige, Y.; Nishimura, R.; Radkov, A.D.; Moe, L.A.
    Triple serine loop region regulates the aspartate racemase activity of the serine/aspartate racemase family (2017), Amino Acids, 49, 1743-1754 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Crassostrea gigas
recombinant expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Acropora millepora
sequence comparisons and phylogenetic tree of serine and aspartate racemases, recombinant expression of C-terminally His-tagged wild-type enzyme in Escherichia coli strain BL21(DE3) Penaeus monodon

Protein Variants

Protein Variants Comment Organism
additional information substituting the triple serine loop region in AspRs enhances serine racemization Penaeus monodon
additional information substituting the triple serine loop region in AspRs enhances serine racemization Crassostrea gigas
additional information substituting the triple serine loop region in AspRs enhances serine racemization Acropora millepora
P150S PCR-based site-directed mutagenesis, the mutant shows reduced aspartate and serine racemase activities compared to the wild-type enzyme Acropora millepora
S150P/S151P/S152Y PCR-based site-directed mutagenesis, the SSS to PPY mutation of Crassostrea AspR triple serine loop region drastically decreases the specific AspR activity to below 0.04% compared to wild-type, whereas it significantly increases the specific SerR and SDH activities by 6-13fold. The large change in the AspR activity in the mutant enzyme is caused by approximately 400fold decrease in kcat rather than by 22-32fold decrease in Km Crassostrea gigas
S151P/S152F PCR-based site-directed mutagenesis, the mutant is inactive as aspartate racemase, but shows serine racemase activity, EC 5.1.1.18 Acropora millepora

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of enzyme AspR with L-Ser and D-Ser as substrates, comparison to the serine racemase enzyme of Penaeus monodon, EC 5.1.1.18, overview Penaeus monodon
additional information
-
additional information kinetics of wild-type and mutant enzymes AspR with L-Ser and D-Ser as substrates, comparison to the serine racemase enzyme of Acropora millepora, overview Acropora millepora
additional information
-
additional information kinetics of wild-type and mutant enzymes AspR with L-Ser and D-Ser as substrates, comparison to the serine racemase enzyme of Crassostrea gigas, EC 5.1.1.18, overview Crassostrea gigas
1.37
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora
3.11
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora
3.2
-
L-aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
4.55
-
D-Aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
4.75
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
6.21
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
9.86
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
14.9
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
48.6
-
L-aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas
80.6
-
D-Aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-aspartate Penaeus monodon
-
D-aspartate
-
r
L-aspartate Crassostrea gigas
-
D-aspartate
-
r
L-aspartate Acropora millepora
-
D-aspartate
-
r
additional information Penaeus monodon the enzyme AspR also shows low serine racemase, SerR (EC 5.1.1.18), activity ?
-
?

Organism

Organism UniProt Comment Textmining
Acropora millepora JT020910
-
-
Crassostrea gigas K1QH67
-
-
Penaeus monodon A0A0U5A554
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography Crassostrea gigas
recombinant C-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography Acropora millepora
recombinant C-terminally His-tagged wild-type enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Penaeus monodon

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-aspartate
-
Penaeus monodon L-aspartate
-
r
D-aspartate
-
Crassostrea gigas L-aspartate
-
r
D-aspartate
-
Acropora millepora L-aspartate
-
r
L-aspartate
-
Penaeus monodon D-aspartate
-
r
L-aspartate
-
Crassostrea gigas D-aspartate
-
r
L-aspartate
-
Acropora millepora D-aspartate
-
r
additional information the enzyme AspR also shows low serine racemase, SerR (EC 5.1.1.18), activity Penaeus monodon ?
-
?
additional information the enzyme AspR also shows negligible serine racemase, SerR (EC 5.1.1.18), activity Crassostrea gigas ?
-
?
additional information the enzyme is also active in the racemisation of L- and D-serine, EC 5.1.1.18 Penaeus monodon ?
-
?

Synonyms

Synonyms Comment Organism
AspR
-
Penaeus monodon
AspR
-
Crassostrea gigas
AspR
-
Acropora millepora

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Penaeus monodon
30
-
assay at Crassostrea gigas
30
-
assay at Acropora millepora

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0228
-
L-aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas
0.0322
-
D-Aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas
8.65
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
13.1
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
14.5
-
L-aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
17.8
-
D-Aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
25.3
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora
27.6
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
33
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
57.7
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Penaeus monodon
8
-
assay at Crassostrea gigas
8
-
assay at Acropora millepora

General Information

General Information Comment Organism
evolution the enzyme belongs to the serine/aspartate racemase family, the triple serine loop region regulates the aspartate racemase activity of the serine/aspartate racemase family members. The ancestral gene of the serine/aspartate racemase family is SerR, the evolution of AspR from SerR occured in three steps: first, gene duplication of the original SerR gene. Second, introduction of the two serine residues at position 151 and 152, resulting in a drastic increase of the AspR activity, and third, formation of the complete triple serine loop region by additional introduction of serine residue at position 150, resulting in enhanced the enzyme specificity for aspartate Penaeus monodon
evolution the enzyme belongs to the serine/aspartate racemase family, the triple serine loop region regulates the aspartate racemase activity of the serine/aspartate racemase family members. The ancestral gene of the serine/aspartate racemase family is SerR, the evolution of AspR from SerR occured in three steps: first, gene duplication of the original SerR gene. Second, introduction of the two serine residues at position 151 and 152, resulting in a drastic increase of the AspR activity, and third, formation of the complete triple serine loop region by additional introduction of serine residue at position 150, resulting in enhanced the enzyme specificity for aspartate Crassostrea gigas
evolution the enzyme belongs to the serine/aspartate racemase family, the triple serine loop region regulates the aspartate racemase activity of the serine/aspartate racemase family members. The ancestral gene of the serine/aspartate racemase family is SerR, the evolution of AspR from SerR occured in three steps: first, gene duplication of the original SerR gene. Second, introduction of the two serine residues at position 151 and 152, resulting in a drastic increase of the AspR activity, and third, formation of the complete triple serine loop region by additional introduction of serine residue at position 150, resulting in enhanced the enzyme specificity for aspartate Acropora millepora
malfunction substituting the triple serine loop region in AspRs enhances serine racemization Penaeus monodon
malfunction substituting the triple serine loop region in AspRs enhances serine racemization. Single amino acid substitution mutants, H150S, P151S and N152S, reveal several effects on activity. The Ser150 substitution decreases all enzyme activities, especially the serine dehydrogenase (SDH) activity, while Ser151 dramatically increases the SerR and AspR activities with no change on SDH activity. An increase in the AspR activity is observed after introducing Ser152, while there is little or no change in the other activities Crassostrea gigas
malfunction substituting the triple serine loop region in AspRs enhances serine racemization. Single amino acid substitution mutants, H150S, P151S and N152S, reveal several effects on activity. The Ser150 substitution decreases all enzyme activities, especially the serine dehydrogenase (SDH) activity, while Ser151 dramatically increases the SerR and AspR activities with no change on SDH activity. An increase in the AspR activity is observed after introducing Ser152, while there is little or no change in the other activities Acropora millepora
additional information important role of triple serine loop region for the aspratate racemase activity in several SerRs and AspRs, structure-function analysis, overview Penaeus monodon
additional information important role of triple serine loop region for the aspratate racemase activity in several SerRs and AspRs, structure-function analysis, overview Crassostrea gigas
additional information important role of triple serine loop region for the aspratate racemase activity in several SerRs and AspRs, structure-function analysis, overview Acropora millepora

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0004
-
D-Aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas
0.0005
-
L-aspartate recombinant His-tagged mutant S150P/S151P/S152Y, pH 8.0, 30°C Crassostrea gigas
1.82
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
2.12
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Crassostrea gigas
2.22
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
2.8
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Penaeus monodon
3.9
-
D-Aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
4.7
-
L-aspartate recombinant His-tagged mutant P150S, pH 8.0, 30°C Acropora millepora
18.4
-
D-Aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora
18.7
-
L-aspartate recombinant His-tagged wild-type enzyme, pH 8.0, 30°C Acropora millepora