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Literature summary for 1.2.1.30 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
Search for more literature for 1.2.1.30

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
A922G site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
E337A site-directed mutagenesis, inactive mutant Neurospora crassa
E433A site-directed mutagenesis, inactive mutant Neurospora crassa
E441A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
F787A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G457A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
G592A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G691A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G694A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G697A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G755A site-directed mutagenesis, the mutant has activity similar to wild-type Neurospora crassa
G843A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
G882A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
H237A site-directed mutagenesis, inactive mutant Neurospora crassa
K848A site-directed mutagenesis, inactive mutant Neurospora crassa
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
N885A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
P198A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
P234A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
P285A site-directed mutagenesis, the enzyme shows altered substrate specificity compared to wild-type Neurospora crassa
P904A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
S595A site-directed mutagenesis, inactive mutant Neurospora crassa
Y542A site-directed mutagenesis, the enzyme shows reduced activity compared to wild-type Neurospora crassa
Y844A site-directed mutagenesis, inactive mutant Neurospora crassa

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Neurospora crassa

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
CAR
-
 Neurospora crassa
carboxylate reductases
-
 Neurospora crassa
NcCAR
-
 Neurospora crassa

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Neurospora crassa
NADPH
-
 Neurospora crassa

General Information

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