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Literature summary for 5.1.1.1 extracted from

  • Kobayashi, J.; Yukimoto, J.; Shimizu, Y.; Ohmori, T.; Suzuki, H.; Doi, K.; Ohshima, T.
    Characterization of Lactobacillus salivarius alanine racemase short-chain carboxylate-activation and the role of A131 (2015), SpringerPlus, 4, 639 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information wild-type enzyme ALR is strongly activated by low concentrations (e.g. 1 mM) of short-chain carboxylates, and is inhibited at higher concentrations (e.g. 10 mM), enzyme residue A131 mediates the activation and stabilization of enzyme ALR by short chain carboxylates Ligilactobacillus salivarius

Cloned(Commentary)

Cloned (Comment) Organism
gene alr, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-CodonPlusTM(DE3)-RIPL Ligilactobacillus salivarius

Protein Variants

Protein Variants Comment Organism
A131K site-directed mutagenesis, the mutant shows a 3fold increased activity compared to wild-type Ligilactobacillus salivarius

General Stability

General Stability Organism
acetate increases the stability of enzyme ALR, enzyme residue A131 mediates the activation and stabilization of enzyme ALR by short chain carboxylates Ligilactobacillus salivarius

Inhibitors

Inhibitors Comment Organism Structure
2-oxoglutarate
-
Ligilactobacillus salivarius
acetate
-
Ligilactobacillus salivarius
aspartate
-
Ligilactobacillus salivarius
Butyrate
-
Ligilactobacillus salivarius
DL-lactate
-
Ligilactobacillus salivarius
additional information wild-type enzyme ALR is strongly activated by low concentrations (e.g. 1 mM) of short-chain carboxylates, and is inhibited at higher concentrations (e.g. 10 mM). The enzyme mutant ALRA131K is inhibited at all carboxylate concentrations tested (1-40 mM). Both propionate and butyrate strongly inhibit mutant ALRA131K. ALR and ALRA131K are both inhibited by DL-lactate, though wild-type ALR is inhibited to a lesser degree than ALRA131K. In addition, succinate, pyruvate, 2-oxoglutarate, oxaloacetate, and aspartate also more strongly inhibit mutant ALRA131K than wild-type ALR Ligilactobacillus salivarius
oxaloacetate
-
Ligilactobacillus salivarius
propionate
-
Ligilactobacillus salivarius
pyruvate
-
Ligilactobacillus salivarius
succinate
-
Ligilactobacillus salivarius

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-alanine Ligilactobacillus salivarius
-
D-alanine
-
r
L-alanine Ligilactobacillus salivarius UCC 118
-
D-alanine
-
r

Organism

Organism UniProt Comment Textmining
Ligilactobacillus salivarius Q1WV14
-
-
Ligilactobacillus salivarius UCC 118 Q1WV14
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-CodonPlusTM(DE3)-RIPL by nickel affinity chromatography, dialysis, and ultrafiltration Ligilactobacillus salivarius

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
161
-
purified recombinant His-tagged wild-type enzyme, pH 7.0, 30°C Ligilactobacillus salivarius
438
-
purified recombinant His-tagged mutant A131K enzyme, pH 7.0, 30°C Ligilactobacillus salivarius

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-alanine
-
Ligilactobacillus salivarius D-alanine
-
r
L-alanine
-
Ligilactobacillus salivarius UCC 118 D-alanine
-
r

Subunits

Subunits Comment Organism
? x * 43000, recombinant His-tagged enzyme, SDS-PAGE Ligilactobacillus salivarius

Synonyms

Synonyms Comment Organism
ALR
-
Ligilactobacillus salivarius

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
recombinant wild-type enzyme, without acetate Ligilactobacillus salivarius
45
-
recombinant enzyme mutant A131K, with or without acetate Ligilactobacillus salivarius
50
-
recombinant wild-type enzyme, in presence of 1 mM acetate Ligilactobacillus salivarius

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
recombinant wild-type enzyme, with or without acetate Ligilactobacillus salivarius
9
-
recombinant enzyme mutant A131K, with or without acetate Ligilactobacillus salivarius

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Ligilactobacillus salivarius

General Information

General Information Comment Organism
physiological function alanine racemase (ALR) is responsible for the high D-alanine production in Lactobacillus salivarius Ligilactobacillus salivarius