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Literature summary extracted from

  • Venkitasubramanian, P.; Daniels, L.; Rosazza, J.P.
    Reduction of carboxylic acids by Nocardia aldehyde oxidoreductase requires a phosphopantetheinylated enzyme (2007), J. Biol. Chem., 282, 478-485.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.2.1.30 phosphopantetheine transferase required for activating the enzyme. Recombinant car is an apoenzyme that requires phosphopantetheinylation for conversion to a fully active holoenzyme Nocardia iowensis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.30 recombinant car expressed in Escherichia coli BL21-CodonPlus(DE3)-RP/pHAT305 and in Escherichia coli BL21-CodonPlus(DE3)-RP/pPV1.184 cells Nocardia iowensis

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.30 S689A completely inactive Nocardia iowensis
1.2.1.30 S691A has 39% of the activity of recombinant car Nocardia iowensis
1.2.1.30 S694A has 50% of the activity of recombinant car Nocardia iowensis
1.2.1.30 S696A has 76% of the activity of recombinant car Nocardia iowensis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.2.1.30 Mg2+
-
 Nocardia iowensis

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.30 Nocardia iowensis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.30 by gel filtration Nocardia iowensis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.2.1.30 0.3
-
 recombinant car incubated with CoA Nocardia iowensis
1.2.1.30 0.5
-
 Nocardia cell free extract incubated with recombinant car Nocardia iowensis
1.2.1.30 1.5
-
 recombinant car incubated with CoA and Escherichia coli MV1190/pUC8-sfp cell free extract Nocardia iowensis
1.2.1.30 1.6
-
 Nocardia cell free extract incubated with recombinant car and CoA. Recombinant car incubated with CoA and Npt Nocardia iowensis
1.2.1.30 2.1
-
 car purified from BL21-CodonPlus(DE3)-RP/pPV1.184 (car and sfp) Nocardia iowensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.30 benzoate + NADPH + ATP
-
 Nocardia iowensis benzaldehyde + NADP+ + AMP + phosphate
-
 ?
1.2.1.30 vanillic acid + NADPH + H+ + ATP with Escherichia coli BL21-CodonPlus(DE3)-RP/pPV2.83, in which recombinant Npt is expressed along with recombinant car, vanillic acid is reduced to vanillin and vanillyl alcohol, with vanillin (80%) as the major product. Escherichia coli BL21-CodonPlus(DE3)-RP/pHAT305 (expressing only recombinant Car) reduce only 50% of the vanillic acid starting material, with vanillyl alcohol being the major metabolite. With Escherichia coli BL21-CodonPlus(DE3)-RP/pPV2.83, in which recombinant car is presumed to be in the fully active, phosphopantetheinylated holo form, the rate of reduction of vanillic acid is much faster than that of vanillin to vanillyl alcohol by endogenous Escherichia coli aldehyde dehydrogenase Nocardia iowensis vanillin + vanillyl alcohol + NADP+ + AMP + diphosphate
-
 ?

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.30 ATP
-
 Nocardia iowensis
1.2.1.30 NADPH
-
 Nocardia iowensis