Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain MG1655 | Neurospora crassa |
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Neurospora crassa |
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 (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain MG1655 by nickel affinity chromatography | Neurospora crassa |
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 | Comment | Organism |
---|---|---|
CAR | - |
Neurospora crassa |
carboxylate reductases | - |
Neurospora crassa |
NcCAR | - |
Neurospora crassa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | - |
assay at | Neurospora crassa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Neurospora crassa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Neurospora crassa | |
NADPH | - |
Neurospora crassa |
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 |