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Literature summary for 5.1.1.10 extracted from

  • Kato, S.; Oikawa, T.
    A novel bifunctional amino acid racemase with multiple substrate specificity, malY from Lactobacillus sakei LT-13 Genome-based identification and enzymological characterization (2018), Front. Microbiol., 9, 403 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene malY, DNA and amino acid sequence determination and analysis, genetic organization, sequence comparisons, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Latilactobacillus sakei

Protein Variants

Protein Variants Comment Organism
K233A site-directed mutagenesis Latilactobacillus sakei
Y123A site-directed mutagenesis Latilactobacillus sakei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
150
-
D-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei
169
-
L-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
281000
-
recombinant His6-tagged enzyme, gel filtration Latilactobacillus sakei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Latilactobacillus sakei Ls-MalY is a bifunctional amino acid racemase with multiple substrate specificity, both the amino acid racemase and beta-lyase reactions of Ls-MalY are catalyzed at the same active site ?
-
?
additional information Latilactobacillus sakei LT-13 Ls-MalY is a bifunctional amino acid racemase with multiple substrate specificity, both the amino acid racemase and beta-lyase reactions of Ls-MalY are catalyzed at the same active site ?
-
?

Organism

Organism UniProt Comment Textmining
Latilactobacillus sakei A0A223K4L6
-
-
Latilactobacillus sakei LT-13 A0A223K4L6
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis, to homogeneity Latilactobacillus sakei

Source Tissue

Source Tissue Comment Organism Textmining
additional information strain LK-145 is isolated from a Japanese sake seller as a high D-amino acid producer, and strain LT-13 is isolated from Moto, a starter of sake, as a low D-amino acid producer, using a medium of amylase digested rice as a carbon source Latilactobacillus sakei
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-alanine
-
Latilactobacillus sakei L-alanine
-
r
L-alanine
-
Latilactobacillus sakei D-alanine
-
r
L-alanine
-
Latilactobacillus sakei LT-13 D-alanine
-
r
L-arginine
-
Latilactobacillus sakei D-arginine
-
r
L-asparagine
-
Latilactobacillus sakei D-asparagine
-
r
L-glutamate
-
Latilactobacillus sakei D-glutamate
-
r
L-glutamate
-
Latilactobacillus sakei LT-13 D-glutamate
-
r
L-glutamine
-
Latilactobacillus sakei D-glutamine
-
r
L-histidine
-
Latilactobacillus sakei D-histidine
-
r
L-leucine
-
Latilactobacillus sakei D-leucine
-
r
L-lysine
-
Latilactobacillus sakei D-lysine
-
r
L-Methionine
-
Latilactobacillus sakei D-Methionine
-
r
L-Methionine
-
Latilactobacillus sakei LT-13 D-Methionine
-
r
L-serine
-
Latilactobacillus sakei D-serine
-
r
L-threonine
-
Latilactobacillus sakei D-threonine
-
r
L-tryptophan
-
Latilactobacillus sakei D-tryptophan
-
r
L-valine
-
Latilactobacillus sakei D-valine
-
r
additional information Ls-MalY is a bifunctional amino acid racemase with multiple substrate specificity, both the amino acid racemase and beta-lyase reactions of Ls-MalY are catalyzed at the same active site Latilactobacillus sakei ?
-
?
additional information enzyme Ls-MalY shows cystathionine beta-lyase (EC 4.4.1.8) and amino acid racemase activity with various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val, substrate specificity, overview. L-Tyr and D-Tyr are poor substrates. Thr is only converted from L- to D-Thr, but not vice versa. The epsilon-amino group of Lys233 in the primary structure of Ls-MalY likely bound to pyridoxal 5'-phosphate, and Lys233 is an essential residue for Ls-MalY to catalyze both the amino acid racemase and beta-lyase reactions. Tyr123 is a catalytic residue in the amino acid racemase reaction but strongly affects beta-lyase activity Latilactobacillus sakei ?
-
?
additional information Ls-MalY is a bifunctional amino acid racemase with multiple substrate specificity, both the amino acid racemase and beta-lyase reactions of Ls-MalY are catalyzed at the same active site Latilactobacillus sakei LT-13 ?
-
?
additional information enzyme Ls-MalY shows cystathionine beta-lyase (EC 4.4.1.8) and amino acid racemase activity with various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val, substrate specificity, overview. L-Tyr and D-Tyr are poor substrates. Thr is only converted from L- to D-Thr, but not vice versa. The epsilon-amino group of Lys233 in the primary structure of Ls-MalY likely bound to pyridoxal 5'-phosphate, and Lys233 is an essential residue for Ls-MalY to catalyze both the amino acid racemase and beta-lyase reactions. Tyr123 is a catalytic residue in the amino acid racemase reaction but strongly affects beta-lyase activity Latilactobacillus sakei LT-13 ?
-
?

Subunits

Subunits Comment Organism
homohexamer 6 * 46000, recombinant His6-tagged enzyme, SDS-PAGE Latilactobacillus sakei
More structural modeling of Ls-MalY, structure comparisons, overview Latilactobacillus sakei

Synonyms

Synonyms Comment Organism
LACBS_00576 locus name Latilactobacillus sakei
MalY
-
Latilactobacillus sakei
patB
-
Latilactobacillus sakei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
Latilactobacillus sakei

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 55 activity range, profile overview Latilactobacillus sakei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1830
-
D-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei
2310
-
L-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
-
Latilactobacillus sakei

pH Range

pH Minimum pH Maximum Comment Organism
8.5 10.5 activity range, profile overview Latilactobacillus sakei

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Latilactobacillus sakei

General Information

General Information Comment Organism
additional information structural modeling of Ls-MalY, structure comparisons, overview Latilactobacillus sakei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
12.2
-
D-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei
13.7
-
L-alanine pH 10.0, 35°C, recombinant His-tagged enzyme Latilactobacillus sakei