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Literature summary 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

EC Number Application Comment Organism
1.1.1.184 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

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.184 additional information
-
 the enzyme forms aggregates of high molecular weight above the size exclusion limit of native PAGE of 1300 kDa Kluyveromyces lactis
1.1.1.184 190000
-
 x * 190000, SDS-PAGE Kluyveromyces lactis

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.184 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.184 50.8
-
 purified enzyme Kluyveromyces lactis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.184 2,3-butanedione + NADPH low activity Kluyveromyces lactis ? + NADP+
-
 ir
1.1.1.184 2,3-pentanedione + NADPH + H+
-
 Kluyveromyces lactis ? + NADP+
-
 ir
1.1.1.184 ethyl 4-chloroacetoacetate + NADPH + H+ enantioselective reduction Kluyveromyces lactis ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
 ir
1.1.1.184 ethyl 4-chloroacetoacetate + NADPH + H+ highly enantioselective reduction Kluyveromyces lactis ethyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
 ir
1.1.1.184 methyl 4-chloroacetoacetate + NADPH highly enantioselective reduction Kluyveromyces lactis methyl (S)-4-chloro-3-hydroxybutanoate + NADP+
-
 ir
1.1.1.184 methylglyoxal + NADPH very low activity Kluyveromyces lactis ? + NADP+
-
 ir
1.1.1.184 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

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

Synonyms

EC Number Synonyms Comment Organism
1.1.1.184 carbonyl reductase
-
 Kluyveromyces lactis
1.1.1.184 KLCR1
-
 Kluyveromyces lactis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.184 45
-
-
 Kluyveromyces lactis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.184 25
-
 30 min, over 80% remaining activity Kluyveromyces lactis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.184 6.5
-
-
 Kluyveromyces lactis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.184 6 8 more than 80% activity at pH 6.0 and pH 8.0 Kluyveromyces lactis

Cofactor

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