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Literature summary for 1.1.1.B3 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

Application Comment Organism
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)

Cloned (Comment) Organism
expression in Escherichia coli Scheffersomyces stipitis

Inhibitors

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

KM Value [mM]

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

Organic Solvent Stability

Organic Solvent Comment Organism
dimethyl formamide 1%, 56% residual activity Scheffersomyces stipitis
dimethyl sulfoxide 1%, 67% residual activity Scheffersomyces stipitis
tetrahydrofuran 1%, 77% residual activity Scheffersomyces stipitis

Organism

Organism UniProt Comment Textmining
Scheffersomyces stipitis A3GF05
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Scheffersomyces stipitis

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
CRII
-
Scheffersomyces stipitis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Scheffersomyces stipitis

Temperature Range [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
Scheffersomyces stipitis

pH Range

pH Minimum pH Maximum Comment Organism
5.5 7 more than 80% of maximum activity within Scheffersomyces stipitis

pH Stability

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

Cofactor

Cofactor Comment Organism Structure
NADH preferred over NADPH Scheffersomyces stipitis
NADPH about 10% of the activity with NADH Scheffersomyces stipitis