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Literature summary 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

Application

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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.1.15 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
5.1.1.15 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.1.15 2.17
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 5.3
-
 D-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.1.1.15 D-2-aminohexano-6-lactam Achromobacter obae
-
 L-2-aminohexano-6-lactam
-
 r
5.1.1.15 L-2-aminohexano-6-lactam Achromobacter obae
-
 D-2-aminohexano-6-lactam
-
 r
5.1.1.15 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

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

Purification (Commentary)

EC Number Purification (Comment) Organism
5.1.1.15 recombinant His-tagged enzyme 129fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Citreicella sp. SE45
5.1.1.15 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.1.1.15 0.04
-
 purified recombinant enzyme, substrate L-valine amide, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 0.06
-
 purified recombinant enzyme, substrate L-valine amide, pH 7.5, 30°C Achromobacter obae
5.1.1.15 0.1
-
 purified recombinant enzyme, substrate L-phenylglycine, pH 7.5, 30°C Achromobacter obae
5.1.1.15 0.1
-
 purified recombinant enzyme, substrate L-phenylglycine, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 0.2
-
 purified recombinant enzyme, substrate L-phenylalanine, pH 7.5, 30°C Achromobacter obae
5.1.1.15 0.3
-
 purified recombinant enzyme, substrate L-phenylalanine, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 0.8
-
 purified recombinant enzyme, substrate L-methionine amide, pH 7.5, 30°C Achromobacter obae
5.1.1.15 0.9
-
 purified recombinant enzyme, substrate L-methionine amide, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 1.5
-
 purified recombinant enzyme, substrate L-leucine amide, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 1.8
-
 purified recombinant enzyme, substrate L-leucine amide, pH 7.5, 30°C Achromobacter obae
5.1.1.15 5.3
-
 crude recombinant enzyme, substrate L-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 16
-
 purified recombinant enzyme, substrate L-serine amide, pH 7.5, 30°C Achromobacter obae
5.1.1.15 16.7
-
 purified recombinant enzyme, substrate L-alanine amide, pH 7.5, 30°C Achromobacter obae
5.1.1.15 17.5
-
 purified recombinant enzyme, substrate L-alanine amide, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 21.6
-
 purified recombinant enzyme, substrate L-serine amide, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 443
-
 purified recombinant enzyme, substrate L-aspartate, pH 7.5, 30°C Achromobacter obae
5.1.1.15 627
-
 purified recombinant enzyme, substrate D-aspartate, pH 7.5, 30°C Achromobacter obae
5.1.1.15 681
-
 purified recombinant enzyme, substrate L-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45
5.1.1.15 772
-
 purified recombinant enzyme, substrate D-2-aminohexano-6-lactam, pH 7.5, 30°C Citreicella sp. SE45

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.1.15 D-2-aminohexano-6-lactam
-
 Achromobacter obae L-2-aminohexano-6-lactam
-
 r
5.1.1.15 D-2-aminohexano-6-lactam preferred substrate Achromobacter obae L-2-aminohexano-6-lactam
-
 r
5.1.1.15 D-2-aminohexano-6-lactam preferred substrate Citreicella sp. SE45 L-2-aminohexano-6-lactam
-
 r
5.1.1.15 L-2-aminohexano-6-lactam
-
 Achromobacter obae D-2-aminohexano-6-lactam
-
 r
5.1.1.15 L-2-aminohexano-6-lactam high activity Achromobacter obae D-2-aminohexano-6-lactam
-
 r
5.1.1.15 L-2-aminohexano-6-lactam high activity Citreicella sp. SE45 D-2-aminohexano-6-lactam
-
 r
5.1.1.15 L-alanine amide
-
 Achromobacter obae D-alanine amide
-
 r
5.1.1.15 L-alanine amide
-
 Citreicella sp. SE45 D-alanine amide
-
 r
5.1.1.15 L-leucine amide
-
 Achromobacter obae D-leucine amide
-
 r
5.1.1.15 L-leucine amide
-
 Citreicella sp. SE45 D-leucine amide
-
 r
5.1.1.15 L-methionine amide
-
 Achromobacter obae D-methionine amide
-
 r
5.1.1.15 L-methionine amide
-
 Citreicella sp. SE45 D-methionine amide
-
 r
5.1.1.15 L-phenylalanine amide
-
 Achromobacter obae D-phenylalanine amide
-
 r
5.1.1.15 L-phenylalanine amide
-
 Citreicella sp. SE45 D-phenylalanine amide
-
 r
5.1.1.15 L-phenylglycine amide
-
 Achromobacter obae D-phenylglycine amide
-
 r
5.1.1.15 L-phenylglycine amide
-
 Citreicella sp. SE45 D-phenylglycine amide
-
 r
5.1.1.15 L-serine amide
-
 Achromobacter obae D-serine amide
-
 r
5.1.1.15 L-serine amide
-
 Citreicella sp. SE45 D-serine amide
-
 r
5.1.1.15 L-valine amide
-
 Achromobacter obae D-valine amide
-
 r
5.1.1.15 L-valine amide
-
 Citreicella sp. SE45 D-valine amide
-
 r
5.1.1.15 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 ?
-
 ?
5.1.1.15 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 ?
-
 ?
5.1.1.15 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

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.1.15 30
-
 assay at Achromobacter obae
5.1.1.15 70
-
-
 Citreicella sp. SE45

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5.1.1.15 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5.1.1.15 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.1.1.15 465
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.1.15 7.5
-
 assay at Achromobacter obae
5.1.1.15 10
-
-
 Citreicella sp. SE45

pH Range

EC Number pH Minimum pH Maximum Comment Organism
5.1.1.15 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

EC Number pH Stability pH Stability Maximum Comment Organism
5.1.1.15 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

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1.1.15 214
-
 L-2-aminohexano-6-lactam recombinant enzyme, pH 7.5, 30°C Citreicella sp. SE45