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Literature summary extracted from

  • Buss, O.; Muller, D.; Jager, S.; Rudat, J.; Rabe, K.S.
    Improvement in the thermostability of a beta-amino acid converting omega-transaminase by using FoldX (2018), ChemBioChem, 19, 379-387 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.6.1.18 recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Variovorax paradoxus

Protein Variants

EC Number Protein Variants Comment Organism
2.6.1.18 D392K site-directed mutagenesis, slightly reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 E345F site-directed mutagenesis, reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 E391K site-directed mutagenesis, reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 G165M best DELTADELTAG prediction, the mutant shows a 3.3fold reduction in enzymatic activity and an only a slight improvement in the Tm value, which stresses the importance of experimental evaluation of the theoretical data Variovorax paradoxus
2.6.1.18 G165M site-directed mutagenesis, reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 G98M site-directed mutagenesis, slightly reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 G98M/D392K site-directed mutagenesis, reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 G98M/E345F site-directed mutagenesis, reduced activity compared to wild-type enzyme Variovorax paradoxus
2.6.1.18 additional information synthesis and optical resolution of beta-phenylalanine and other important aromatic beta-amino acids by biotransformation utilizing an omega-transaminase (omega-TA) from Variovorax paradoxus. Design of mutant variants of the omega-TA to gain higher process stability on the basis of predictions calculated by using the FoldX software. Thermostabilization of a nonthermostable S-selective omega-TA by FoldX-guided site-directed mutagenesis. The melting point (Tm) of the best-performing mutant is increased to 59.3°C, an increase of 4.0°C relative to the Tm value of the wild-type enzyme, whereby the mutant fully retains its specific activity Variovorax paradoxus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.6.1.18 additional information
-
 additional information thermodynamics of wild-type and mutant enzymes Variovorax paradoxus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.6.1.18 pyruvate + beta-alanine Variovorax paradoxus
-
 L-alanine + 3-oxopropanoate
-
 r
2.6.1.18 pyruvate + beta-phenylalanine Variovorax paradoxus
-
 L-alanine + 3-oxo-phenylpropanoate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.6.1.18 Variovorax paradoxus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.6.1.18 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration Variovorax paradoxus

Reaction

EC Number Reaction Comment Organism Reaction ID
2.6.1.18 L-alanine + 3-oxopropanoate = pyruvate + beta-alanine reaction mechanism with beta-phenylalanine as an amino donor and 2-oxoglutarate as an acceptor, detailed overview Variovorax paradoxus
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.6.1.18 24
-
 purified recombinant mutant G98M, pH 7.2, 40°C, substrate rac-beta-phenylalanine with 2-oxoglutarate Variovorax paradoxus
2.6.1.18 26
-
 purified recombinant wild-type enzyme, pH 7.2, 40°C, substrate rac-beta-phenylalanine with 2-oxoglutarate Variovorax paradoxus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.6.1.18 2-oxoglutarate + (S)-beta-phenylalanine
-
 Variovorax paradoxus L-glutamate + 3-oxo-phenylpropanoate
-
 r
2.6.1.18 2-oxoglutarate + rac-beta-phenylalanine
-
 Variovorax paradoxus L-glutamate + 3-oxo-phenylpropanoate
-
 r
2.6.1.18 pyruvate + beta-alanine
-
 Variovorax paradoxus L-alanine + 3-oxopropanoate
-
 r
2.6.1.18 pyruvate + beta-phenylalanine
-
 Variovorax paradoxus L-alanine + 3-oxo-phenylpropanoate
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.6.1.18 omega-transaminase
-
 Variovorax paradoxus
2.6.1.18 omega-VpTA
-
 Variovorax paradoxus
2.6.1.18 S-selective omega-TA
-
 Variovorax paradoxus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.6.1.18 40
-
 assay at Variovorax paradoxus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.6.1.18 54.3
-
 Tm of recombinant mutant enzyme E391K Variovorax paradoxus
2.6.1.18 55.3
-
 Tm of recombinant wild-type Variovorax paradoxus
2.6.1.18 56.2
-
 Tm of recombinant mutant enzyme E345F Variovorax paradoxus
2.6.1.18 56.7
-
 Tm of recombinant mutant enzyme D392K Variovorax paradoxus
2.6.1.18 58.8
-
 Tm of recombinant mutant enzyme G98M/D345F Variovorax paradoxus
2.6.1.18 59.3
-
 Tm of recombinant mutant enzyme G98M Variovorax paradoxus
2.6.1.18 59.4
-
 Tm of recombinant mutant enzyme G98M/E392K Variovorax paradoxus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.6.1.18 7.2
-
 assay at Variovorax paradoxus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.6.1.18 5 9 activity range of wild-type enzyme, inactive above Variovorax paradoxus

Cofactor

EC Number Cofactor Comment Organism Structure
2.6.1.18 pyridoxal 5'-phosphate PLP Variovorax paradoxus

General Information

EC Number General Information Comment Organism
2.6.1.18 additional information structure-function analysis, overview Variovorax paradoxus