A novel bifunctional amino acid racemase with multiple substrate specificity, malY from Lactobacillus sakei LT-13 Genome-based identification and enzymological characterization
Kato, S.; Oikawa, T.; Front. Microbiol. 9, 403 (2018) 
Data extracted from this reference:
Cloned(Commentary)
Cloned (Commentary)
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
Engineering
Protein Variants
Commentary
Organism
K233A
site-directed mutagenesis, the mutant is only active with D-Ser, but inactive with L- or D-Cys, and L-Ser, in contrast to the wild-type enzyme
Latilactobacillus sakei
Y123A
site-directed mutagenesis, the mutant shows highly reduced activity with L-Cys compared to the wild-type enzyme, and is active with L-Ser in contrast to wild-type
Latilactobacillus sakei
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
11.5
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
281000
-
recombinant His6-tagged enzyme, gel filtration
Latilactobacillus sakei
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-cystathionine + H2O
Latilactobacillus sakei
overall reaction
L-homocysteine + pyruvate + NH3
-
-
?
L-cysteine + H2O
Latilactobacillus sakei
-
? + NH3
-
-
?
L-cystine + H2O
Latilactobacillus sakei
-
? + NH3
-
-
?
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
?
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Latilactobacillus sakei
A0A223K4L6
-
-
Latilactobacillus sakei LT-13
A0A223K4L6
-
-
Purification (Commentary)
Purification (Commentary)
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
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
L-cystathionine + H2O
overall reaction
747857
Latilactobacillus sakei
L-homocysteine + pyruvate + NH3
-
-
-
?
L-cysteine + H2O
-
747857
Latilactobacillus sakei
? + NH3
-
-
-
?
L-cystine + H2O
-
747857
Latilactobacillus sakei
? + NH3
-
-
-
?
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
747857
Latilactobacillus sakei
?
-
-
-
?
additional information
enzyme Ls-MalY shows cystathionine beta-lyase and amino acid racemase activity (EC 5.1.1.10) with various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val. The beta-lyase shows activity toward L-Cys, but not toward D-Cys, L-Ser, and D-Ser, substrate specificity, overview. Ls-MalY also shows beta-lyase activity with L-cystine and L-cystathionine. 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
747857
Latilactobacillus sakei
?
-
-
-
?
Subunits
Subunits
Commentary
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
Commentary
Organism
beta-lyase
-
Latilactobacillus sakei
LACBS_00576
locus name
Latilactobacillus sakei
MalY
-
Latilactobacillus sakei
patB
-
Latilactobacillus sakei
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
40
-
-
Latilactobacillus sakei
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
20
45
activity range, profile overview
Latilactobacillus sakei
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
930
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
-
Latilactobacillus sakei
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7.5
11.5
activity range, profile overview
Latilactobacillus sakei
Cofactor
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
-
Latilactobacillus sakei
Cloned(Commentary) (protein specific)
Commentary
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
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
-
Latilactobacillus sakei
Engineering (protein specific)
Protein Variants
Commentary
Organism
K233A
site-directed mutagenesis, the mutant is only active with D-Ser, but inactive with L- or D-Cys, and L-Ser, in contrast to the wild-type enzyme
Latilactobacillus sakei
Y123A
site-directed mutagenesis, the mutant shows highly reduced activity with L-Cys compared to the wild-type enzyme, and is active with L-Ser in contrast to wild-type
Latilactobacillus sakei
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
11.5
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
281000
-
recombinant His6-tagged enzyme, gel filtration
Latilactobacillus sakei
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-cystathionine + H2O
Latilactobacillus sakei
overall reaction
L-homocysteine + pyruvate + NH3
-
-
?
L-cysteine + H2O
Latilactobacillus sakei
-
? + NH3
-
-
?
L-cystine + H2O
Latilactobacillus sakei
-
? + NH3
-
-
?
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
?
-
-
?
Purification (Commentary) (protein specific)
Commentary
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 (protein specific)
Source Tissue
Commentary
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) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
L-cystathionine + H2O
overall reaction
747857
Latilactobacillus sakei
L-homocysteine + pyruvate + NH3
-
-
-
?
L-cysteine + H2O
-
747857
Latilactobacillus sakei
? + NH3
-
-
-
?
L-cystine + H2O
-
747857
Latilactobacillus sakei
? + NH3
-
-
-
?
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
747857
Latilactobacillus sakei
?
-
-
-
?
additional information
enzyme Ls-MalY shows cystathionine beta-lyase and amino acid racemase activity (EC 5.1.1.10) with various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val. The beta-lyase shows activity toward L-Cys, but not toward D-Cys, L-Ser, and D-Ser, substrate specificity, overview. Ls-MalY also shows beta-lyase activity with L-cystine and L-cystathionine. 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
747857
Latilactobacillus sakei
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homohexamer
6 * 46000, recombinant His6-tagged enzyme, SDS-PAGE
Latilactobacillus sakei
More
structural modeling of Ls-MalY, structure comparisons, overview
Latilactobacillus sakei
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
40
-
-
Latilactobacillus sakei
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
20
45
activity range, profile overview
Latilactobacillus sakei
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
930
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
-
Latilactobacillus sakei
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7.5
11.5
activity range, profile overview
Latilactobacillus sakei
General Information
General Information
Commentary
Organism
additional information
structural modeling of Ls-MalY, structure comparisons, overview
Latilactobacillus sakei
General Information (protein specific)
General Information
Commentary
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
Commentary
Organism
Structure
80.87
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
80.87
-
L-cysteine
pH 10.0, 35°C, recombinant His-tagged enzyme
Latilactobacillus sakei
Other publictions for EC 4.4.1.8
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
747857
Kato
A novel bifunctional amino ac ...
Latilactobacillus sakei, Latilactobacillus sakei LT-13
Front. Microbiol.
9
403
2018
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