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Literature summary 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)

EC Number Cloned (Comment) Organism
5.1.1.21 expressed in Escherichia coli Lentilactobacillus buchneri

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.1.1.21 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
5.1.1.21 hydroxylamine
-
 Lentilactobacillus otakiensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.1.21 5
-
 L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
5.1.1.21 13.2
-
 D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.1.1.21 49000
-
 4 * 49000, SDS-PAGE Lentilactobacillus buchneri
5.1.1.21 200000
-
 gel filtration Lentilactobacillus buchneri

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
5.1.1.21
-
 Lentilactobacillus buchneri
5.1.1.21
-
 Lentilactobacillus otakiensis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.1.1.21 32
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-methionine Lentilactobacillus buchneri
5.1.1.21 34.3
-
 pH 5.5, 37°C Lentilactobacillus otakiensis
5.1.1.21 35.7
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-phenylalanine Lentilactobacillus buchneri
5.1.1.21 44.2
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-leucine Lentilactobacillus buchneri
5.1.1.21 44.5
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-isoleucine Lentilactobacillus buchneri
5.1.1.21 47.7
-
 pH 5.5, 37°C, recombinant enzyme, substrate: (S)-2-aminobutyric acid Lentilactobacillus buchneri
5.1.1.21 54.8
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-leucine Lentilactobacillus buchneri
5.1.1.21 66.3
-
 pH 5.5, 37°C, recombinant enzyme, substrate: (R)-2-aminobutyric acid Lentilactobacillus buchneri
5.1.1.21 71.7
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-valine Lentilactobacillus buchneri
5.1.1.21 74.1
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-norleucine Lentilactobacillus buchneri
5.1.1.21 83.4
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-norvaline Lentilactobacillus buchneri
5.1.1.21 93.7
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-methionine Lentilactobacillus buchneri
5.1.1.21 116
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-norleucine Lentilactobacillus buchneri
5.1.1.21 131
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-valine Lentilactobacillus buchneri
5.1.1.21 149
-
 pH 5.5, 37°C, recombinant enzyme, substrate: L-isoleucine Lentilactobacillus buchneri
5.1.1.21 183
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-norvaline Lentilactobacillus buchneri
5.1.1.21 221
-
 pH 5.5, 37°C, recombinant enzyme, substrate: D-allo-isoleucine Lentilactobacillus buchneri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.1.21 (R)-2-aminobutyric acid activity is 42% compared to D-allo-isoleucine Lentilactobacillus buchneri (S)-2-aminobutyric acid
-
 r
5.1.1.21 (S)-2-aminobutyric acid activity is 32% compared to L-isoleucine Lentilactobacillus buchneri (R)-2-aminobutyric acid
-
 r
5.1.1.21 D-allo-isoleucine
-
 Lentilactobacillus buchneri L-isoleucine
-
 r
5.1.1.21 D-allo-isoleucine
-
 Lentilactobacillus otakiensis L-isoleucine
-
 ?
5.1.1.21 D-allo-isoleucine
-
 Lentilactobacillus otakiensis JCM 15040 L-isoleucine
-
 ?
5.1.1.21 D-leucine activity is 30% compared to D-allo-isoleucine Lentilactobacillus buchneri L-leucine
-
 ?
5.1.1.21 D-methionine activity is 20% compared to D-allo-isoleucine Lentilactobacillus buchneri L-methionine
-
 ?
5.1.1.21 D-norleucine activity is 59% compared to D-allo-isoleucine Lentilactobacillus buchneri L-norleucine
-
 r
5.1.1.21 D-norvaline activity is 83% compared to D-allo-isoleucine Lentilactobacillus buchneri L-norvaline
-
 r
5.1.1.21 D-norvaline activity is 83% compared to D-allo-isoleucine Lentilactobacillus buchneri JCM 1115 L-norvaline
-
 r
5.1.1.21 D-phenylalanine activity is 25% compared to D-allo-isoleucine Lentilactobacillus buchneri L-phenylalanine
-
 ?
5.1.1.21 D-valine activity is 52% compared to D-allo-isoleucine Lentilactobacillus buchneri L-Val
-
 r
5.1.1.21 L-isoleucine
-
 Lentilactobacillus buchneri D-allo-isoleucine
-
 r
5.1.1.21 L-isoleucine
-
 Lentilactobacillus otakiensis D-allo-isoleucine
-
 ?
5.1.1.21 L-isoleucine
-
 Lentilactobacillus otakiensis JCM 15040 D-allo-isoleucine
-
 ?
5.1.1.21 L-leucine activity is 30% compared to L-isoleucine Lentilactobacillus buchneri D-leucine
-
 ?
5.1.1.21 L-Methionine activity is 19% compared to L-isoleucine Lentilactobacillus buchneri D-Methionine
-
 ?
5.1.1.21 L-Methionine activity is 19% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Methionine
-
 ?
5.1.1.21 L-Norleucine activity is 50% compared to L-isoleucine Lentilactobacillus buchneri D-Norleucine
-
 r
5.1.1.21 L-Norleucine activity is 50% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Norleucine
-
 r
5.1.1.21 L-Norvaline activity is 56% compared to L-isoleucine Lentilactobacillus buchneri D-Norvaline
-
 r
5.1.1.21 L-Norvaline activity is 56% compared to L-isoleucine Lentilactobacillus buchneri JCM 1115 D-Norvaline
-
 r
5.1.1.21 L-phenylalanine activity is 24% compared to L-isoleucine Lentilactobacillus buchneri D-phenylalanine
-
 ?
5.1.1.21 L-valine activity is 48% compared to L-isoleucine Lentilactobacillus buchneri D-Val
-
 r
5.1.1.21 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 ?
-
 ?
5.1.1.21 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

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

Synonyms

EC Number Synonyms Comment Organism
5.1.1.21 BCAA racemase
-
 Lentilactobacillus buchneri

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.1.21 37
-
 assay at Lentilactobacillus buchneri
5.1.1.21 37
-
 assay at Lentilactobacillus otakiensis
5.1.1.21 45
-
 highest activity for the reaction with D-allo-L-isoleucine Lentilactobacillus buchneri
5.1.1.21 50
-
 highest activity for the reaction with L-isoleucine Lentilactobacillus buchneri

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5.1.1.21 60
-
 30 min, the enzyme retains more than 80% of its activity Lentilactobacillus buchneri

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.1.1.21 502
-
 L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
5.1.1.21 939
-
 D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.1.21 5.5
-
 assay at Lentilactobacillus buchneri
5.1.1.21 5.5
-
 assay at Lentilactobacillus otakiensis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
5.1.1.21 5 11 the enzyme retains about 80% of its activity after incubation for 30 min Lentilactobacillus buchneri

Cofactor

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1.1.21 71.4
-
 D-allo-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri
5.1.1.21 101
-
 L-isoleucine pH 5.5, 37°C Lentilactobacillus buchneri