Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Neurospora crassa |
gene nccr, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) | Neurospora crassa |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33700 | - |
gel filtration | Neurospora crassa |
37533 | - |
1 * 40000, SDS-PAGE, 1 * 37533, calculated | Neurospora crassa |
40000 | - |
1 * 40000, SDS-PAGE, 1 * 37533, calculated | Neurospora crassa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Neurospora crassa | Q7S3U4 | putative | - |
Neurospora crassa | Q7S3U4 | predicted; gene nccr | - |
Neurospora crassa DSM 894 | Q7S3U4 | predicted; gene nccr | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme 3.1fold from Escherichia coli strain BL21(DE3) | Neurospora crassa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3.5 | - |
purified recombinant enzyme, substrate 3,4-hexandione, pH 7.0, 30°C | Neurospora crassa |
80 | - |
purified recombinant enzyme, substrate 2,3-hexandione, pH 7.0, 30°C | Neurospora crassa |
89 | - |
purified recombinant enzyme, substrate 2,3-heptandione, pH 7.0, 30°C | Neurospora crassa |
468 | - |
purified recombinant enzyme, substrate dihydroxyacetone, pH 7.0, 30°C | Neurospora crassa |
468 | - |
substrate dihydroxyacetone, 30°C, pH 7.0 | Neurospora crassa |
Storage Stability | Organism |
---|---|
-20°C, stable for at least 21 days without notable loss of activity | Neurospora crassa |
30°C, 21 days, 25% residual activity | Neurospora crassa |
4°C, stable for at least 21 days without notable loss of activity | Neurospora crassa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,3-heptandione + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
2,3-heptandione + NADPH + H+ | - |
Neurospora crassa DSM 894 | ? + NADP+ | - |
? | |
2,3-heptanedione + NADPH + H+ | 18.8% of the activity with dihydroxyacetone | Neurospora crassa | ? + NADP+ | - |
? | |
2,3-hexandione + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
2,3-hexandione + NADPH + H+ | - |
Neurospora crassa DSM 894 | ? + NADP+ | - |
? | |
2,3-hexanedione + NADPH + H+ | 17.3% of the activity with dihydroxyacetone | Neurospora crassa | ? + NADP+ | - |
? | |
3,4-hexandione + NADPH + H+ | low activity | Neurospora crassa | ? + NADP+ | - |
? | |
dihydroxyacetone + NADPH + H+ | - |
Neurospora crassa | glycerol + NADP+ | - |
? | |
dihydroxyacetone + NADPH + H+ | - |
Neurospora crassa DSM 894 | glycerol + NADP+ | - |
? | |
dihydroxyacetone + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 2-chloro-3-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 2-oxo-4-phenylbutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 2-oxophenylacetate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 3-methyl-2-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 3-methyl-2-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ethyl (2R)-2-hydroxy-3-methylbutanoate + NADP+ | 95% conversion, 98.3% enantiomeric excess | ? | |
ethyl 3-oxo-3-phenylpropionate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 3-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 4,4,4-trifluoro-3-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | the enantioselectivity of NcCR for the reduction of ethyl 4-chloro-3-oxobutanoate shows a strong dependence on temperature. The enantiomeric excess is substantially increased by decreasing the temperature from 40°C (78.8%) to -3°C (98.0%) | Neurospora crassa | ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | the enantioselectivity of NcCR for the reduction of ethyl 4-chloro-3-oxobutanoate shows a strong dependence on temperature. The enantiomeric excess is substantially increased by decreasing the temperature from 40°C (78.8%) to -3°C (98.0%) | Neurospora crassa DSM 894 | ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | enantioselectivity shows a strong dependence on temperature. Under optimal conditions of -3°C, (S)-4-chloro-3-hydroxybutanoate in 98% enantiomeric excess is produced | Neurospora crassa | ethyl (3S)-4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
hydroxyacetone + NADPH + H+ | - |
Neurospora crassa | glycerol + NADP+ | - |
? | |
methyl 2-oxopropanoate + NADPH + H+ | - |
Neurospora crassa | methyl (2S)-2-hydroxypropanoate + NADP+ | 67% conversion, 99.9% enantiomeric excess | ? | |
methyl-2-oxobutanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
methyl-2-oxopropanoate + NADPH + H+ | - |
Neurospora crassa | ? + NADP+ | - |
? | |
additional information | the recombinant enzyme exhibits a restricted substrate spectrum towards various ketones, and the highest activity is observed with dihydroxyacetone, while hydroxyacetone is reduced with only 0.2% of the activity achieved with dihydroxyacetone, among aliphatic and aromatic ketones and aldehydes. The enzyme proves to be very selective in the reduction of different alpha- and beta-keto esters. Several compounds are converted to the corresponding hydroxy ester in high enantiomeric excess at high conversion rates. Diketones are also accepted by the enzyme. Highest activities are obtained with alpha,beta-diketones carrying the two ketogroups near the end of the aliphatic chain. No or poor activity with 2,5-hexandione, 2-cyclohexen-1-one, cyclohexanone, 2-methylcyclohexanone, 2-pentanone, 4-phenyl-2-butanone, cyclopropyl methyl ketone, trans-3-nonen-2-one, 2-butanone, 2-methyl-2-cyclopenten-1-one, and beta-ionone. The enantioselectivity of the enzyme reaction is highly increased at lower temperatures, overview | Neurospora crassa | ? | - |
? | |
additional information | the recombinant enzyme exhibits a restricted substrate spectrum towards various ketones, and the highest activity is observed with dihydroxyacetone, while hydroxyacetone is reduced with only 0.2% of the activity achieved with dihydroxyacetone, among aliphatic and aromatic ketones and aldehydes. The enzyme proves to be very selective in the reduction of different alpha- and beta-keto esters. Several compounds are converted to the corresponding hydroxy ester in high enantiomeric excess at high conversion rates. Diketones are also accepted by the enzyme. Highest activities are obtained with alpha,beta-diketones carrying the two ketogroups near the end of the aliphatic chain. No or poor activity with 2,5-hexandione, 2-cyclohexen-1-one, cyclohexanone, 2-methylcyclohexanone, 2-pentanone, 4-phenyl-2-butanone, cyclopropyl methyl ketone, trans-3-nonen-2-one, 2-butanone, 2-methyl-2-cyclopenten-1-one, and beta-ionone. The enantioselectivity of the enzyme reaction is highly increased at lower temperatures, overview | Neurospora crassa DSM 894 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 40000, SDS-PAGE, 1 * 37533, calculated | Neurospora crassa |
Synonyms | Comment | Organism |
---|---|---|
NADPH-dependent carbonyl reductase | - |
Neurospora crassa |
NCCR | - |
Neurospora crassa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
the enantioselectivity of the enzyme reaction is highly increased at lower temperatures, overview | Neurospora crassa |
25 | - |
recombinant enzyme | Neurospora crassa |
40 | - |
- |
Neurospora crassa |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
sharp decrease in activity above | Neurospora crassa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Neurospora crassa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no cofactor: NADH | Neurospora crassa | |
NADPH | - |
Neurospora crassa | |
NADPH | dependent on | Neurospora crassa |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the aldo-keto-reductase family (AKR) | Neurospora crassa |