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Literature summary for 1.1.1.184 extracted from

  • Morikawa, S.; Nakai, T.; Yasohara, Y.; Nanba, H.; Kizaki, N.; Hasegawa, J.
    Highly active mutants of carbonyl reductase S1 with inverted coenzyme specificity and production of optically active alcohols (2005), Biosci. Biotechnol. Biochem., 69, 544-552.
    View publication on PubMed
Search for more literature for 1.1.1.184

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain HB101 Starmerella magnoliae

Protein Variants

Protein Variants Comment Organism
additional information design and construction of mutant enzymes, e.g. S1M1 and S1M4, with inverted cofactor specificity using computer-aided methods, in silico screening, three-dimensional structure modeling, and site-sirected mutagenesis, mutant screening, overview, the mutant enzyme utilizes NADH, but not NADPH as cofactor Starmerella magnoliae
S41A/S42A/S43Q/W63I/Y64D/N65I/S66N site-sirected mutagenesis, mutant S1M1 shows reduced activity and inverted cofactor specificity, using exclusively NADH, compared to the wild-type enzyme Starmerella magnoliae
S41A/S42A/S43R/W63I/Y64D/N65I/S66N site-sirected mutagenesis, mutant S1M4 shows reduced activity and inverted cofactor specificity, using exclusively NADH, compared to the wild-type enzyme Starmerella magnoliae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information reaction and cofactor binding kinetics of wild-type and mutant enzymes, thermodynamics, overview Starmerella magnoliae
0.0026
-
 ethyl 4-chloro-3-oxobutanoate pH 6.5, 30°C, recombinant mutant S1M4 Starmerella magnoliae
0.003
-
 ethyl 4-chloro-3-oxobutanoate pH 6.5, 30°C, recombinant wild-type enzyme Starmerella magnoliae
0.0039
-
 ethyl 4-chloro-3-oxobutanoate pH 6.5, 30°C, recombinant mutant S1M1 Starmerella magnoliae
0.0167
-
 NADPH pH 6.5, 30°C, recombinant wild-type enzyme Starmerella magnoliae
0.062
-
 NADH pH 6.5, 30°C, recombinant mutant S1M4 Starmerella magnoliae
0.16
-
 NADH pH 6.5, 30°C, recombinant mutant S1M1 Starmerella magnoliae

Organism

Organism UniProt Comment Textmining
Starmerella magnoliae
-
-
-
Starmerella magnoliae AKU4643
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain HB101 by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, to homogeneity Starmerella magnoliae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.8
-
 purified recombinant mutant S1M1 Starmerella magnoliae
9.9
-
 purified recombinant mutant S1M4 Starmerella magnoliae
27.8
-
 purified recombinant wild-type enzyme Starmerella magnoliae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxyacetophenone + NADH + H+
-
 Starmerella magnoliae (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
2-hydroxyacetophenone + NADH + H+
-
 Starmerella magnoliae AKU4643 (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
ethyl 4-chloro-3-oxobutanoate + NAD(P)H + H+ stereospecific reaction Starmerella magnoliae ethyl (S)-4-chloro-3-hydroxybutanoate + NAD(P)+ optically pure (S)-enantiomer ?
ethyl 4-chloro-3-oxobutanoate + NAD(P)H + H+ stereospecific reaction Starmerella magnoliae AKU4643 ethyl (S)-4-chloro-3-hydroxybutanoate + NAD(P)+ optically pure (S)-enantiomer ?

Synonyms

Synonyms Comment Organism
More the enzyme is a member of the short-chain dehydrogenase/reductase family Starmerella magnoliae
NADPH-dependent carbonyl reductase
-
 Starmerella magnoliae
reductase S1
-
 Starmerella magnoliae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Starmerella magnoliae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
64
-
 NADH pH 6.5, 30°C, recombinant mutant S1M1 Starmerella magnoliae
120
-
 NADH pH 6.5, 30°C, recombinant mutant S1M4 Starmerella magnoliae
870
-
 NADPH pH 6.5, 30°C, recombinant wild-type enzyme Starmerella magnoliae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Starmerella magnoliae

Cofactor

Cofactor Comment Organism Structure
NADH mutant enzyme with inverted cofactor specificity, strictly specific for Starmerella magnoliae
NADPH wild-type enzyme, strictly specific for Starmerella magnoliae