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Literature summary for 1.1.1.184 extracted from

  • Yamamoto, H.; Kimoto, N.; Matsuyama, A.; Kobayashi, Y.
    Purification and properties of a carbonyl reductase useful for production of ethyl (S)-4-chloro-3-hydroxybutanoate from Kluyveromyces lactis (2002), Biosci. Biotechnol. Biochem., 66, 1775-1778.
    View publication on PubMed

Application

Application Comment Organism
synthesis enzyme is useful for production of ethyl (S)-4-chloro-3-hydroxybutanoate, which is used in the synthesis of pharmacologically and biologically important compouds Kluyveromyces lactis

Inhibitors

Inhibitors Comment Organism Structure
Barbital 18% inhibition at 1 mM Kluyveromyces lactis
Cu2+ complete inhibition at 1 mM Kluyveromyces lactis
Hg2+ complete inhibition at 0.01 mM Kluyveromyces lactis
iodoacetamide 46% inhibition at 1 mM Kluyveromyces lactis
Methyl vinyl ketone 74% inhibition at 10 mM Kluyveromyces lactis
additional information no inhibition by EDTA and crotonic acid Kluyveromyces lactis
N-ethylmaleimide 80% inhibition at 1 mM Kluyveromyces lactis
o-phenanthroline about 11% inhibition at 1 mM Kluyveromyces lactis
p-chloromercuribenzoate complete inhibition at 0.05 mM Kluyveromyces lactis
PMSF about 10% inhibition at 1 mM Kluyveromyces lactis
quercetin 31.5% inhibition at 0.1 mM Kluyveromyces lactis
Zn2+ 88% inhibition at 1 mM Kluyveromyces lactis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
the enzyme forms aggregates of high molecular weight above the size exclusion limit of native PAGE of 1300 kDa Kluyveromyces lactis
190000
-
x * 190000, SDS-PAGE Kluyveromyces lactis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ethyl 4-chloroacetoacetate + NADPH + H+ Kluyveromyces lactis enantioselective reduction ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
ir

Organism

Organism UniProt Comment Textmining
Kluyveromyces lactis
-
enzyme KLCR1 from strain NRIC 1329
-

Purification (Commentary)

Purification (Comment) Organism
380fold to homogeneity by protamine precipitation, solubilization, ammonium sulfate fractionation, solubilization with 0.5 M NaCl, ultrafiltration, butyl-resin chromatography, and gel filtration Kluyveromyces lactis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
50.8
-
purified enzyme Kluyveromyces lactis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,3-butanedione + NADPH low activity Kluyveromyces lactis ? + NADP+
-
ir
2,3-pentanedione + NADPH + H+
-
Kluyveromyces lactis ? + NADP+
-
ir
ethyl 4-chloroacetoacetate + NADPH + H+ enantioselective reduction Kluyveromyces lactis ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
ir
ethyl 4-chloroacetoacetate + NADPH + H+ highly enantioselective reduction Kluyveromyces lactis ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
ir
methyl 4-chloroacetoacetate + NADPH highly enantioselective reduction Kluyveromyces lactis methyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
ir
methylglyoxal + NADPH very low activity Kluyveromyces lactis ? + NADP+
-
ir
additional information enzyme is a reductase no dehydrogenase, no activity with phenylglyoxal, 2,4-pentanedione, ethyl 3-oxobutanoate, or methyl 3-oxobutanoate, no activity with nitrobenzaldehydes, xylose, isatin, acetone, acetophenone, propionaldehyde, and pyridine 3-aldehyde, enzyme might be an alpha-subunit of the fatty acid synthase due to amino acid sequence identity, but does not show fatty acid synthase activity Kluyveromyces lactis ?
-
?

Subunits

Subunits Comment Organism
oligomer x * 190000, SDS-PAGE Kluyveromyces lactis

Synonyms

Synonyms Comment Organism
carbonyl reductase
-
Kluyveromyces lactis
KLCR1
-
Kluyveromyces lactis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
Kluyveromyces lactis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25
-
30 min, over 80% remaining activity Kluyveromyces lactis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Kluyveromyces lactis

pH Range

pH Minimum pH Maximum Comment Organism
6 8 more than 80% activity at pH 6.0 and pH 8.0 Kluyveromyces lactis

Cofactor

Cofactor Comment Organism Structure
additional information no activity with NADH Kluyveromyces lactis
NADPH dependent on, no reverse reaction with NADP+ as cofactor Kluyveromyces lactis