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Literature summary extracted from

  • Stolterfoht, H.; Steinkellner, G.; Schwendenwein, D.; Pavkov-Keller, T.; Gruber, K.; Winkler, M.
    Identification of key residues for enzymatic carboxylate reduction (2018), Front. Microbiol., 9, 250 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.30 recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain MG1655 Neurospora crassa

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.30 A922G site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 E337A site-directed mutagenesis, inactive mutant Neurospora crassa
1.2.1.30 E433A site-directed mutagenesis, inactive mutant Neurospora crassa
1.2.1.30 E441A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
1.2.1.30 F787A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G457A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
1.2.1.30 G592A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G691A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G694A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G697A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G755A site-directed mutagenesis, the mutant has activity similar to wild-type Neurospora crassa
1.2.1.30 G843A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 G882A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 H237A site-directed mutagenesis, inactive mutant Neurospora crassa
1.2.1.30 K848A site-directed mutagenesis, inactive mutant Neurospora crassa
1.2.1.30 additional information the replacement of Gly184, Arg870, and Trp978 by alanine does not result in soluble expression. Pro904 is located close to Trp978, and its substitution by Ala yields significantly less soluble protein. Replacement of His237, Glu433, Ser595, Tyr844, and Lys848 by Ala abolishes CAR activity. Substitutions with alanine (P189A, P234A, P285A, E441A, and G457A) enhanced the specific activity toward hexanoic acid compared to the wild-type Neurospora crassa
1.2.1.30 N885A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 P198A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
1.2.1.30 P234A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
1.2.1.30 P285A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
1.2.1.30 P904A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 S595A site-directed mutagenesis, inactive mutant Neurospora crassa
1.2.1.30 Y542A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
1.2.1.30 Y844A site-directed mutagenesis, inactive mutant Neurospora crassa

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.2.1.30 Mg2+ required Neurospora crassa

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.30 Neurospora crassa Q7RW48
-
-
1.2.1.30 Neurospora crassa 74-OR23-1A Q7RW48
-
-
1.2.1.30 Neurospora crassa ATCC 24698 Q7RW48
-
-
1.2.1.30 Neurospora crassa CBS 708.71 Q7RW48
-
-
1.2.1.30 Neurospora crassa DSM 1257 Q7RW48
-
-
1.2.1.30 Neurospora crassa FGSC 987 Q7RW48
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.30 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain MG1655 by nickel affinity chromatography Neurospora crassa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa CBS 708.71 cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa 74-OR23-1A cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa DSM 1257 cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa ATCC 24698 cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 cinnamate + NADPH + H+ + ATP
-
Neurospora crassa FGSC 987 cinnamaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa CBS 708.71 hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa 74-OR23-1A hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa DSM 1257 hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa ATCC 24698 hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 hexanoate + NADPH + H+ + ATP
-
Neurospora crassa FGSC 987 hexanal + NADP+ + AMP + diphosphate
-
?
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa ?
-
-
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa CBS 708.71 ?
-
-
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa 74-OR23-1A ?
-
-
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa DSM 1257 ?
-
-
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa ATCC 24698 ?
-
-
1.2.1.30 additional information NcCAR wild-type and mutants efficiently reduce aliphatic acids Neurospora crassa FGSC 987 ?
-
-
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa piperonylaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa CBS 708.71 piperonylaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa 74-OR23-1A piperonylaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa DSM 1257 piperonylaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa ATCC 24698 piperonylaldehyde + NADP+ + AMP + diphosphate
-
?
1.2.1.30 piperonylate + NADPH + H+ + ATP
-
Neurospora crassa FGSC 987 piperonylaldehyde + NADP+ + AMP + diphosphate
-
?

Synonyms

EC Number Synonyms Comment Organism
1.2.1.30 CAR
-
Neurospora crassa
1.2.1.30 carboxylate reductases
-
Neurospora crassa
1.2.1.30 NcCAR
-
Neurospora crassa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.30 28
-
assay at Neurospora crassa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.30 6
-
assay at Neurospora crassa

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.30 ATP
-
Neurospora crassa
1.2.1.30 NADPH
-
Neurospora crassa

General Information

EC Number General Information Comment Organism
1.2.1.30 evolution analysis of highly conserved signature sequences of CARs, overview Neurospora crassa
1.2.1.30 malfunction replacement of His237, Glu433, Ser595, Tyr844, and Lys848 by Ala abolishes CAR activity Neurospora crassa
1.2.1.30 additional information structure homology modeling Neurospora crassa