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Literature summary for 5.1.1.15 extracted from

  • Payoungkiattikun, W.; Okazaki, S.; Ina, A.; H-Kittikun, A.; Asano, Y.
    Characterization of an alpha-amino-epsilon-caprolactam racemase with broad substrate specificity from Citreicella sp. SE45 (2017), J. Ind. Microbiol. Biotechnol., 44, 677-685 .
    View publication on PubMed
Search for more literature for 5.1.1.15

Application

Application Comment Organism
synthesis the enzyme might be useful for applications in dynamic kinetic resolution for D- or L-amino acid production Achromobacter obae
synthesis the enzyme might be useful for applications in dynamic kinetic resolution for D- or L-amino acid production Citreicella sp. SE45

Cloned(Commentary)

Cloned (Comment) Organism
gene CSE45_2055, sequence comparisons, recombinant expression of codon-optimized His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain JM109 Achromobacter obae
gene CSE45_2055, sequence comparisons, recombinant expression of codon-optimized His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain JM109 Citreicella sp. SE45

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ 44% inhibition at 5 mM Citreicella sp. SE45
Co2+ 59% inhibition at 5 mM Citreicella sp. SE45
Cu2+ 92% inhibition at 5 mM Citreicella sp. SE45
Fe2+ 97% inhibition at 5 mM Citreicella sp. SE45
Mg2+ 45% inhibition at 5 mM Citreicella sp. SE45
Mn2+ 49% inhibition at 5 mM Citreicella sp. SE45
additional information poor inhibition by EDTA at 2-10 mM Citreicella sp. SE45
Ni2+ 55% inhibition at 5 mM Citreicella sp. SE45

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.17
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45
5.3
-
 D-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-2-aminohexano-6-lactam Achromobacter obae
-
 L-2-aminohexano-6-lactam
-
 r
L-2-aminohexano-6-lactam Achromobacter obae
-
 D-2-aminohexano-6-lactam
-
 r
additional information Citreicella sp. SE45 the enzyme encoded by gene CSE45_2055 is a 2,2-dialkylglycine decarboxylase, EC 4.1.1.64 (UniProt ID D0D2T1), that shows enhanced alpha-amino-epsilon-caprolactam racemase activity on D- and L-2-aminohexano-6-lactams ?
-
 ?

Organism

Organism UniProt Comment Textmining
Achromobacter obae Q7M181
-
-
Citreicella sp. SE45
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme 129fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Citreicella sp. SE45
recombinant His-tagged enzyme 27.9fold from Escherichia coli strain BL21(DE3) by heat treatment, nickel affinity chromatography, and dialysis Achromobacter obae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.04
-
 purified recombinant enzyme, substrate L-valine amide, pH 7.5, 30°C Citreicella sp. SE45
0.06
-
 purified recombinant enzyme, substrate L-valine amide, pH 7.5, 30°C Achromobacter obae
0.1
-
 purified recombinant enzyme, substrate L-phenylglycine, pH 7.5, 30°C Achromobacter obae
0.1
-
 purified recombinant enzyme, substrate L-phenylglycine, pH 7.5, 30°C Citreicella sp. SE45
0.2
-
 purified recombinant enzyme, substrate L-phenylalanine, pH 7.5, 30°C Achromobacter obae
0.3
-
 purified recombinant enzyme, substrate L-phenylalanine, pH 7.5, 30°C Citreicella sp. SE45
0.8
-
 purified recombinant enzyme, substrate L-methionine amide, pH 7.5, 30°C Achromobacter obae
0.9
-
 purified recombinant enzyme, substrate L-methionine amide, pH 7.5, 30°C Citreicella sp. SE45
1.5
-
 purified recombinant enzyme, substrate L-leucine amide, pH 7.5, 30°C Citreicella sp. SE45
1.8
-
 purified recombinant enzyme, substrate L-leucine amide, pH 7.5, 30°C Achromobacter obae
5.3
-
 crude recombinant enzyme, substrate L-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45
16
-
 purified recombinant enzyme, substrate L-serine amide, pH 7.5, 30°C Achromobacter obae
16.7
-
 purified recombinant enzyme, substrate L-alanine amide, pH 7.5, 30°C Achromobacter obae
17.5
-
 purified recombinant enzyme, substrate L-alanine amide, pH 7.5, 30°C Citreicella sp. SE45
21.6
-
 purified recombinant enzyme, substrate L-serine amide, pH 7.5, 30°C Citreicella sp. SE45
443
-
 purified recombinant enzyme, substrate L-aspartate, pH 7.5, 30°C Achromobacter obae
627
-
 purified recombinant enzyme, substrate D-aspartate, pH 7.5, 30°C Achromobacter obae
681
-
 purified recombinant enzyme, substrate L-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45
772
-
 purified recombinant enzyme, substrate D-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-2-aminohexano-6-lactam
-
 Achromobacter obae L-2-aminohexano-6-lactam
-
 r
D-2-aminohexano-6-lactam preferred substrate Achromobacter obae L-2-aminohexano-6-lactam
-
 r
D-2-aminohexano-6-lactam preferred substrate Citreicella sp. SE45 L-2-aminohexano-6-lactam
-
 r
L-2-aminohexano-6-lactam
-
 Achromobacter obae D-2-aminohexano-6-lactam
-
 r
L-2-aminohexano-6-lactam high activity Achromobacter obae D-2-aminohexano-6-lactam
-
 r
L-2-aminohexano-6-lactam high activity Citreicella sp. SE45 D-2-aminohexano-6-lactam
-
 r
L-alanine amide
-
 Achromobacter obae D-alanine amide
-
 r
L-alanine amide
-
 Citreicella sp. SE45 D-alanine amide
-
 r
L-leucine amide
-
 Achromobacter obae D-leucine amide
-
 r
L-leucine amide
-
 Citreicella sp. SE45 D-leucine amide
-
 r
L-methionine amide
-
 Achromobacter obae D-methionine amide
-
 r
L-methionine amide
-
 Citreicella sp. SE45 D-methionine amide
-
 r
L-phenylalanine amide
-
 Achromobacter obae D-phenylalanine amide
-
 r
L-phenylalanine amide
-
 Citreicella sp. SE45 D-phenylalanine amide
-
 r
L-phenylglycine amide
-
 Achromobacter obae D-phenylglycine amide
-
 r
L-phenylglycine amide
-
 Citreicella sp. SE45 D-phenylglycine amide
-
 r
L-serine amide
-
 Achromobacter obae D-serine amide
-
 r
L-serine amide
-
 Citreicella sp. SE45 D-serine amide
-
 r
L-valine amide
-
 Achromobacter obae D-valine amide
-
 r
L-valine amide
-
 Citreicella sp. SE45 D-valine amide
-
 r
additional information the enzyme encoded by gene CSE45_2055 is a 2,2-dialkylglycine decarboxylase, EC 4.1.1.64 (UniProt ID D0D2T1), that shows enhanced alpha-amino-epsilon-caprolactam racemase activity on D- and L-2-aminohexano-6-lactams Citreicella sp. SE45 ?
-
 ?
additional information The enzyme acts on a broad range of amino acid amides, particularly unbranched amino acid amides including L-alanine amide and L-serine amide. No activity with L-tyrosine amide Achromobacter obae ?
-
 ?
additional information The enzyme acts on a broad range of amino acid amides, particularly unbranched amino acid amides including L-alanine amide and L-serine amide. No activity with L-tyrosine amide Citreicella sp. SE45 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 45000, recombinant His-tagged enzyme, SDS-PAGE Achromobacter obae
? x * 47400, recombinant His-tagged enzyme, SDS-PAGE Citreicella sp. SE45

Synonyms

Synonyms Comment Organism
ACL racemase
-
 Achromobacter obae
ACL racemase
-
 Citreicella sp. SE45
ACLR
-
 Achromobacter obae
ACLR
-
 Citreicella sp. SE45
alpha-Amino-epsilon-caprolactam racemase
-
 Achromobacter obae
alpha-Amino-epsilon-caprolactam racemase
-
 Citreicella sp. SE45
More cf. EC 5.1.1.10 Achromobacter obae
More cf. EC 5.1.1.10 Citreicella sp. SE45

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Achromobacter obae
70
-
-
 Citreicella sp. SE45

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 90 recombinant enzyme, activity range, enzyme activity rapidly increases between 40°C and 60°C and reaches its maximum at 70°C, above 70°C, the enzyme activity declines Citreicella sp. SE45

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 purified recombinant His-tagged enzyme, the enzyme retains more than 90% of its activity after being incubated at 50°C for 50 min, and enzyme activity significantly decreases at temperatures greater than 50°C Citreicella sp. SE45

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
465
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Achromobacter obae
10
-
-
 Citreicella sp. SE45

pH Range

pH Minimum pH Maximum Comment Organism
4.5 12 active in a broad pH range, with more than 80 % activity in the pH range of pH 7.5-11.0, maximal activity at pH 10.0 Citreicella sp. SE45

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 10 purified recombinant His-tagged enzyme, highly stable after 1 h incubation at 4°C, not stable under acidic conditions Citreicella sp. SE45

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
 Citreicella sp. SE45
pyridoxal 5'-phosphate PLP, the enzyme is a fold-type I PLP-dependent enzyme Achromobacter obae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
214
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45