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Literature summary for 5.1.1.15 extracted from

  • Matsui, D.; Fuhshuku, K.I.; Nagamori, S.; Takata, M.; Asano, Y.
    Isolation and characterization of racemase from Ensifer sp. 23-3 that acts on alpha-aminolactams and alpha-amino acid amides (2017), J. Ind. Microbiol. Biotechnol., 44, 1503-1510 .
    View publication on PubMed
Search for more literature for 5.1.1.15

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Ensifer sp. 23-3

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis(2-nitrobenzoic acid) complete inhibition at 1 mM Ensifer sp. 23-3
8-hydroxyquinoline 59% inhibition at 1 mM Ensifer sp. 23-3
Ag+ 88% inhibition at 1 mM Ensifer sp. 23-3
Al3+ 57% inhibition at 1 mM Ensifer sp. 23-3
Cd2+ 57% inhibition at 1 mM Ensifer sp. 23-3
Co2+ 32% inhibition at 1 mM Ensifer sp. 23-3
Cr3+ 23% inhibition at 1 mM Ensifer sp. 23-3
Cu2+ complete inhibition at 1 mM Ensifer sp. 23-3
D-cycloserine 21% inhibition at 1 mM Ensifer sp. 23-3
D-penicillamine 88% inhibition at 1 mM Ensifer sp. 23-3
Hg2+ complete inhibition at 1 mM Ensifer sp. 23-3
hydroxylamine complete inhibition at 1 mM Ensifer sp. 23-3
iodoacetic acid 12% inhibition at 1 mM Ensifer sp. 23-3
Mn2+ 62% inhibition at 1 mM Ensifer sp. 23-3
additional information racemization activity can partially be recovered by the subsequent addition of 1.0 mM pyridoxal 5'-phosphate Ensifer sp. 23-3
N-ethylmaleimide 50% inhibition at 1 mM Ensifer sp. 23-3
Ni2+ 58% inhibition at 1 mM Ensifer sp. 23-3
phenylmethane sulfonyl fluoride 59% inhibition at 1 mM Ensifer sp. 23-3
Zn2+ 43% inhibition at 1 mM Ensifer sp. 23-3

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3
-
 L-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
2.1
-
 L-2-aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
3.5
-
 D-2-Aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
3.8
-
 D-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
11.2
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
 Ensifer sp. 23-3 5737
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
49800
-
 native enzyme, gel filtration Ensifer sp. 23-3

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-2-aminohexano-6-lactam Ensifer sp. 23-3
-
 L-2-aminohexano-6-lactam
-
 r

Organism

Organism UniProt Comment Textmining
Ensifer sp. 23-3
-
 isolated from soil
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme by several steps of ion exchange and hydrophobic interaction chromatography, dialysis, ultrafiltration, and gel filtration, recombinant His-tagged enzyme 31.9fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity Ensifer sp. 23-3

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
365
-
 purified recombinant His-tagged enzyme, substrate D-2-aminohexano-6-lactam, pH 7.0, 30°C Ensifer sp. 23-3

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-2-amino-delta-valerolactam best substrate Ensifer sp. 23-3 L-2-amino-delta-valerolactam
-
 r
D-2-amino-omega-octalactam 1.78% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 L-2-amino-omega-octalactam
-
 r
D-2-aminobutyric acid amide 0.19% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 L-2-aminobutyric acid amide
-
 r
D-2-aminohexano-6-lactam
-
 Ensifer sp. 23-3 L-2-aminohexano-6-lactam
-
 r
D-2-aminohexano-6-lactam high activity Ensifer sp. 23-3 L-2-aminohexano-6-lactam
-
 r
D-alanine amide 2.12% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 L-alanine amide
-
 r
L-2-amino-delta-valerolactam 17.2% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 D-2-amino-delta-valerolactam
-
 r
L-2-amino-omega-octalactam 1.41% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 D-2-amino-omega-octalactam
-
 r
L-2-aminobutyric acid amide 0.08% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 D-2-aminobutyric acid amide
-
 r
L-2-aminohexano-6-lactam high activity Ensifer sp. 23-3 D-2-aminohexano-6-lactam
-
 r
L-alanine amide 0.77% activity compared to D-2-aminohexano-6-lactam Ensifer sp. 23-3 D-alanine amide
-
 r
additional information the enzyme acts on 2-aminohexano-6-lactam and 2-amino acid amides, the enzyme prefers the D-enantiomers as substrates Ensifer sp. 23-3 ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 45000, native enzyme, SDS-PAGE Ensifer sp. 23-3

Synonyms

Synonyms Comment Organism
ACL racemase
-
 Ensifer sp. 23-3
ACLR
-
 Ensifer sp. 23-3
alpha-Amino-epsilon-caprolactam racemase
-
 Ensifer sp. 23-3

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65 70
-
 Ensifer sp. 23-3

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 95 50% of maximal activity at 30°C, about 40% at 90°C, and 20% at 95°C Ensifer sp. 23-3

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 60 purified recombinant enzyme, pH 7.0, 30 min, 90% and more activity remaining Ensifer sp. 23-3
65
-
 purified recombinant enzyme, pH 7.0, 30 min, 70% activity remaining Ensifer sp. 23-3
70
-
 purified recombinant enzyme, pH 7.0, 30 min, 30% activity remaining Ensifer sp. 23-3
75
-
 purified recombinant enzyme, pH 7.0, 30 min, 10% activity remaining Ensifer sp. 23-3
80
-
 purified recombinant enzyme, pH 7.0, 30 min, inactivation Ensifer sp. 23-3

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.7
-
 L-2-aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
1.1
-
 D-2-Aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
3.3
-
 L-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
10.2
-
 D-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
371
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 9
-
 Ensifer sp. 23-3

pH Range

pH Minimum pH Maximum Comment Organism
5.5 10.5 over 50% of maximal activity Ensifer sp. 23-3

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5
-
 purified recombinant enzyme, 30°C, 30 min, inactivation Ensifer sp. 23-3
5
-
 purified recombinant enzyme, 30°C, 30 min, about 60% activity remaining Ensifer sp. 23-3
5.5 9 purified recombinant enzyme, 30°C, 30 min, over 80% activity remaining Ensifer sp. 23-3
9
-
 purified recombinant enzyme, 30°C, 30 min, about 75% activity remaining Ensifer sp. 23-3
10
-
 purified recombinant enzyme, 30°C, 30 min, about 60% activity remaining Ensifer sp. 23-3
11
-
 purified recombinant enzyme, 30°C, 30 min, inactivation Ensifer sp. 23-3

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on, may be tightly bound to the enzyme Ensifer sp. 23-3

General Information

General Information Comment Organism
evolution the deduced amino acid sequence of ACLR from Ensifer sp. 23-3 exhibits high similarity (99%) to that of aspartate aminotransferase family protein (UniProt ID W8HW01) from Ensifer adhaerens strain OV14 Ensifer sp. 23-3

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.31
-
 D-2-Aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
0.33
-
 L-2-aminobutyric acid amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
2.54
-
 L-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
2.68
-
 D-alanine amide recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3
33.1
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.0, 30°C Ensifer sp. 23-3