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Literature summary extracted from

  • Cao, H.; Mi, L.; Ye, Q.; Zang, G.; Yan, M.; Wang, Y.; Zhang, Y.; Li, X.; Xu, L.; Xiong, J.; Ouyang, P.; Ying, H.
    Purification and characterization of a novel NADH-dependent carbonyl reductase from Pichia stipitis involved in biosynthesis of optically pure ethyl (S)-4-chloro-3-hydroxybutanoate (2011), Biores. Technol., 102, 1733-1739.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.B3 synthesis synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate in Escherichia coli. Coexpression of carbonyl reductase CRII and a glucose dehydrogenase gives an activity of 15 U/mg protein using ethyl 4-chloro-3-oxobutanoate as a substrate in a water/butyl acetate system. The transformants give a molar yield of 91%, and an optical purity of the (S)-isomer of more than 99% enantiomeric excess Scheffersomyces stipitis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.B3 expression in Escherichia coli Scheffersomyces stipitis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.B3 EDTA 5 mM, 50% residual activity Scheffersomyces stipitis
1.1.1.B3 phenyl methyl sulfonylfluoride 5 mM, 49% residual activity Scheffersomyces stipitis
1.1.1.B3 sodium dodecylsulfate 1%, 51% residual activity Scheffersomyces stipitis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.B3 3.3
-
 ethyl 4-chloro-3-oxobutanoate pH 6.0, 30°C Scheffersomyces stipitis

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
1.1.1.B3 dimethyl formamide 1%, 56% residual activity Scheffersomyces stipitis
1.1.1.B3 dimethyl sulfoxide 1%, 67% residual activity Scheffersomyces stipitis
1.1.1.B3 tetrahydrofuran 1%, 77% residual activity Scheffersomyces stipitis

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.B3 Scheffersomyces stipitis A3GF05
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.B3 recombinant enzyme Scheffersomyces stipitis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.B3 224
-
 pH 6.0, 30°C Scheffersomyces stipitis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.B3 2,3-butanedione + NADH + H+ 9% of the activity with ethyl 4-chloro-3-oxobutanoate Scheffersomyces stipitis ? + NAD+
-
 ?
1.1.1.B3 2,3-pentanedione + NADH + H+ 82% of the activity with ethyl 4-chloro-3-oxobutanoate Scheffersomyces stipitis ? + NAD+
-
 ?
1.1.1.B3 ethyl 2-chloro-3-oxobutanoate + NADH + H+ 49% of the activity with ethyl 4-chloro-3-oxobutanoate Scheffersomyces stipitis ethyl 2-chloro-3-hydroxybutanoate + NAD+
-
 ?
1.1.1.B3 ethyl 3-oxobutanoate + NADH + H+ 28% of the activity with ethyl 4-chloro-3-oxobutanoate Scheffersomyces stipitis ethyl 3-hydroxybutanoate + NAD+
-
 ?
1.1.1.B3 ethyl 4-chloro-3-oxobutanoate + NADH + H+
-
 Scheffersomyces stipitis ethyl (S)-4-chloro-3-hydroxybutanoate + NAD+ product in more than 99% enantiomeric excess ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.B3 CRII
-
 Scheffersomyces stipitis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.B3 30
-
-
 Scheffersomyces stipitis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.B3 25 35 more than 80% of maximum activity within Scheffersomyces stipitis
1.1.1.B3 50
-
 no residual activity Scheffersomyces stipitis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.B3 6
-
-
 Scheffersomyces stipitis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.B3 5.5 7 more than 80% of maximum activity within Scheffersomyces stipitis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.B3 5.6
-
 between pH 5.6-7.0, 4°C, stable for 48 h Scheffersomyces stipitis
1.1.1.B3 7
-
 between pH 5.6-7.0, 4°C, stable for 48 h Scheffersomyces stipitis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.B3 NADH preferred over NADPH Scheffersomyces stipitis
1.1.1.B3 NADPH about 10% of the activity with NADH Scheffersomyces stipitis