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Literature summary for 1.1.1.184 extracted from

  • Richter, N.; Hummel, W.
    Biochemical characterisation of a NADPH-dependent carbonyl reductase from Neurospora crassa reducing alpha- and beta-keto esters (2011), Enzyme Microb. Technol., 48, 472-479.
    View publication on PubMed

Cloned(Commentary)

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 [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

Organism UniProt Comment Textmining
Neurospora crassa Q7S3U4 putative
-
Neurospora crassa Q7S3U4 predicted; gene nccr
-
Neurospora crassa DSM 894 Q7S3U4 predicted; gene nccr
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme 3.1fold from Escherichia coli strain BL21(DE3) Neurospora crassa

Specific Activity [micromol/min/mg]

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

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 and Products (Substrate)

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

Subunits Comment Organism
monomer 1 * 40000, SDS-PAGE, 1 * 37533, calculated Neurospora crassa

Synonyms

Synonyms Comment Organism
NADPH-dependent carbonyl reductase
-
Neurospora crassa
NCCR
-
Neurospora crassa

Temperature Optimum [°C]

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 Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
45
-
sharp decrease in activity above Neurospora crassa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Neurospora crassa

Cofactor

Cofactor Comment Organism Structure
additional information no cofactor: NADH Neurospora crassa
NADPH
-
Neurospora crassa
NADPH dependent on Neurospora crassa

General Information

General Information Comment Organism
evolution the enzyme is a member of the aldo-keto-reductase family (AKR) Neurospora crassa