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Literature summary extracted from

  • Li, J.J.; Li, C.; Blindauer, C.A.; Bugg, T.D.
    Evidence for a gem-diol reaction intermediate in bacterial C-C hydrolase enzymes BphD and MhpC from 13C NMR spectroscopy (2006), Biochemistry, 45, 12461-12469.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.7.1.8
-
Escherichia coli
3.7.1.8 into the pET-14b vector, overexpression in Escherichia coli BL21(DE3) cells Paraburkholderia xenovorans

Protein Variants

EC Number Protein Variants Comment Organism
3.7.1.8 D237N reduced activity Paraburkholderia xenovorans
3.7.1.8 H114A reduced activity, is able to accept the 6-phenyl-containing substrate, on a shorter time scale Escherichia coli
3.7.1.8 H265A reduced activity Paraburkholderia xenovorans
3.7.1.8 S112A reduced activity Paraburkholderia xenovorans
3.7.1.14 H114A MhpC mutant is able to accept the 2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid, on a shorter time scale Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7.1.8 0.0017
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant D237N Paraburkholderia xenovorans
3.7.1.8 0.002
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate His6-BphD Paraburkholderia xenovorans
3.7.1.8 0.0238
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant H114A Escherichia coli
3.7.1.8 0.037
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant H265A Paraburkholderia xenovorans
3.7.1.8 0.0373
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant S112A Paraburkholderia xenovorans
3.7.1.14 0.0238
-
2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid pH 8.0, 25°C Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.7.1.8 Escherichia coli
-
-
-
3.7.1.8 Paraburkholderia xenovorans
-
LB400
-
3.7.1.14 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.7.1.8
-
Escherichia coli
3.7.1.8 by metal affinity chromatography, to more than 90% homogeneity Paraburkholderia xenovorans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.7.1.8 ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate + H2O
-
Escherichia coli benzoate + ethyl 2-(acetyloxy)penta-2,4-dienoate
-
?
3.7.1.8 ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate + H2O
-
Paraburkholderia xenovorans benzoate + ethyl-2-(acetyloxy)penta-2,4-dienoate
-
?
3.7.1.14 2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid + H2O MhpC mutant is able to accept the 2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid, on a shorter time scale Escherichia coli ?
-
?

Synonyms

EC Number Synonyms Comment Organism
3.7.1.8 BphD
-
Paraburkholderia xenovorans
3.7.1.8 MhpC
-
Escherichia coli
3.7.1.14 MhpC
-
Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.7.1.14 25
-
assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.7.1.8 0.0009
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant S112A Paraburkholderia xenovorans
3.7.1.8 0.0016
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant H265A Paraburkholderia xenovorans
3.7.1.8 0.028
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant D237N Paraburkholderia xenovorans
3.7.1.8 0.036
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate mutant H114A Escherichia coli
3.7.1.8 6.5
-
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate His6-BphD Paraburkholderia xenovorans
3.7.1.14 0.036
-
2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid pH 8.0, 25°C Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.7.1.14 8
-
assay at Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.7.1.14 1.51
-
2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid pH 8.0, 25°C Escherichia coli