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Literature summary for 5.1.1.21 extracted from

  • Mutaguchi, Y.; Ohmori, T.; Wakamatsu, T.; Doi, K.; Ohshima, T.
    Identification, purification, and characterization of a novel amino acid racemase, isoleucine 2-epimerase, from Lactobacillus species (2013), J. Bacteriol., 195, 5207-5215.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Lentilactobacillus buchneri

Inhibitors

Inhibitors Comment Organism Structure
hydroxylamine almost fully inactivated by the first dialysis against buffer containing 50 mM hydroxylamine, after which about 75% of the activity is recovered by a subsequent dialysis against buffer containing 1 mM pyridoxal 5'-phosphate Lentilactobacillus buchneri
hydroxylamine
-
Lentilactobacillus otakiensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5
-
L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
13.2
-
D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
49000
-
4 * 49000, SDS-PAGE Lentilactobacillus buchneri
200000
-
gel filtration Lentilactobacillus buchneri

Organism

Organism UniProt Comment Textmining
Lentilactobacillus buchneri M1GRN3
-
-
Lentilactobacillus buchneri JCM 1115 M1GRN3
-
-
Lentilactobacillus otakiensis
-
-
-
Lentilactobacillus otakiensis JCM 15040
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Lentilactobacillus buchneri
-
Lentilactobacillus otakiensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
32
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-methionine Lentilactobacillus buchneri
34.3
-
pH 5.5, 37°C Lentilactobacillus otakiensis
35.7
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-phenylalanine Lentilactobacillus buchneri
44.2
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-leucine Lentilactobacillus buchneri
44.5
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-isoleucine Lentilactobacillus buchneri
47.7
-
pH 5.5, 37°C, recombinant enzyme, substrate: (S)-2-aminobutyric acid Lentilactobacillus buchneri
54.8
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-leucine Lentilactobacillus buchneri
66.3
-
pH 5.5, 37°C, recombinant enzyme, substrate: (R)-2-aminobutyric acid Lentilactobacillus buchneri
71.7
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-valine Lentilactobacillus buchneri
74.1
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-norleucine Lentilactobacillus buchneri
83.4
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-norvaline Lentilactobacillus buchneri
93.7
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-methionine Lentilactobacillus buchneri
116
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-norleucine Lentilactobacillus buchneri
131
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-valine Lentilactobacillus buchneri
149
-
pH 5.5, 37°C, recombinant enzyme, substrate: L-isoleucine Lentilactobacillus buchneri
183
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-norvaline Lentilactobacillus buchneri
221
-
pH 5.5, 37°C, recombinant enzyme, substrate: D-allo-isoleucine Lentilactobacillus buchneri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-2-aminobutyric acid activity is 42% compared to D-allo-isoleucine Lentilactobacillus buchneri (S)-2-aminobutyric acid
-
r
(S)-2-aminobutyric acid activity is 32% compared to L-isoleucine Lentilactobacillus buchneri (R)-2-aminobutyric acid
-
r
D-allo-isoleucine
-
Lentilactobacillus buchneri L-isoleucine
-
r
D-allo-isoleucine
-
Lentilactobacillus otakiensis L-isoleucine
-
?
D-allo-isoleucine
-
Lentilactobacillus otakiensis JCM 15040 L-isoleucine
-
?
D-leucine activity is 30% compared to D-allo-isoleucine Lentilactobacillus buchneri L-leucine
-
?
D-methionine activity is 20% compared to D-allo-isoleucine Lentilactobacillus buchneri L-methionine
-
?
D-norleucine activity is 59% compared to D-allo-isoleucine Lentilactobacillus buchneri L-norleucine
-
r
D-norvaline activity is 83% compared to D-allo-isoleucine Lentilactobacillus buchneri L-norvaline
-
r
D-norvaline activity is 83% compared to D-allo-isoleucine Lentilactobacillus buchneri JCM 1115 L-norvaline
-
r
D-phenylalanine activity is 25% compared to D-allo-isoleucine Lentilactobacillus buchneri L-phenylalanine
-
?
D-valine activity is 52% compared to D-allo-isoleucine Lentilactobacillus buchneri L-Val
-
r
L-isoleucine
-
Lentilactobacillus buchneri D-allo-isoleucine
-
r
L-isoleucine
-
Lentilactobacillus otakiensis D-allo-isoleucine
-
?
L-isoleucine
-
Lentilactobacillus otakiensis JCM 15040 D-allo-isoleucine
-
?
L-leucine activity is 30% compared to L-isoleucine Lentilactobacillus buchneri D-leucine
-
?
L-Methionine activity is 19% compared to L-isoleucine Lentilactobacillus buchneri D-Methionine
-
?
L-Methionine activity is 19% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Methionine
-
?
L-Norleucine activity is 50% compared to L-isoleucine Lentilactobacillus buchneri D-Norleucine
-
r
L-Norleucine activity is 50% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Norleucine
-
r
L-Norvaline activity is 56% compared to L-isoleucine Lentilactobacillus buchneri D-Norvaline
-
r
L-Norvaline activity is 56% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Norvaline
-
r
L-phenylalanine activity is 24% compared to L-isoleucine Lentilactobacillus buchneri D-phenylalanine
-
?
L-valine activity is 48% compared to L-isoleucine Lentilactobacillus buchneri D-Val
-
r
additional information the enzyme specifically catalyzes racemization of nonpolar amino acids at the C-2 position. No activity toward the L- and D-forms of Asp, Glu, Lys, Arg, His, Orn, Thr, Asn, Gln, Trp, and tert-Leu Lentilactobacillus buchneri ?
-
?
additional information the enzyme specifically catalyzes racemization of nonpolar amino acids at the C-2 position. No activity toward the L- and D-forms of Asp, Glu, Lys, Arg, His, Orn, Thr, Asn, Gln, Trp, and tert-Leu Lentilactobacillus buchneri JCM 1115 ?
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 49000, SDS-PAGE Lentilactobacillus buchneri

Synonyms

Synonyms Comment Organism
BCAA racemase
-
Lentilactobacillus buchneri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Lentilactobacillus buchneri
37
-
assay at Lentilactobacillus otakiensis
45
-
highest activity for the reaction with D-allo-L-isoleucine Lentilactobacillus buchneri
50
-
highest activity for the reaction with L-isoleucine Lentilactobacillus buchneri

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
30 min, the enzyme retains more than 80% of its activity Lentilactobacillus buchneri

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
502
-
L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
939
-
D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
assay at Lentilactobacillus buchneri
5.5
-
assay at Lentilactobacillus otakiensis

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 11 the enzyme retains about 80% of its activity after incubation for 30 min Lentilactobacillus buchneri

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Lentilactobacillus buchneri
pyridoxal 5'-phosphate
-
Lentilactobacillus otakiensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
71.4
-
D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
101
-
L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri