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Literature summary for 1.1.1.14 extracted from

  • Cui, Z.; Zhang, J.; Fan, X.; Zheng, G.; Chang, H.; Wei, W.
    Highly efficient bioreduction of 2-hydroxyacetophenone to (S)- and (R)-1-phenyl-1,2-ethanediol by two substrate tolerance carbonyl reductases with cofactor regeneration (2017), J. Biotechnol., 243, 1-9 .
    View publication on PubMed
Search for more literature for 1.1.1.14

Application

Application Comment Organism
synthesis two co-expressed enantiocomplementary carbonyl reductases, BDHA (2,3-butanediol dehydrogenase from Bacillus subtilis) and GoSCR (polyol dehydrogenase from Gluconobacter oxydans) are used for asymmetric reduction of 2-hydroxyacetophenone (2-HAP) to (R)-1-phenyl-1,2-ethanediol ((R)-PED) or (S)-1-phenyl-1,2-ethanediol ((S)-PED) with excellent stereochemical selectivity and coupled with cofactor regeneration by GDH. Enantiomerically pure (R)-1-phenyl-1,2-ethanediol ((R)-PED) can be used as a building block for the preparation of (R)-norfluoxetine, (R)-fluoxetine, and beta-lactam antibiotics Gluconobacter oxydans

Cloned(Commentary)

Cloned (Comment) Organism
gene GoSCR, recombinant expression of His-tagged enzyme GoSCR in Escherichia coli strain BL21 (DE3), coexpression with 2,3-butanediol dehydrogenase (BDHA) from Bacillus subtilis for asymmetric reduction of 2-hydroxyacetophenone (2-HAP) to (R)-1-phenyl-1,2-ethanediol ((R)-PED) or (S)-1-phenyl-1,2-ethanediol ((S)-PED) Gluconobacter oxydans

Protein Variants

Protein Variants Comment Organism
additional information in vitro bioreduction of 2-hydroxyacetophenone (2-HAP) is catalyzed by GoSCR coupled with glucose dehydrogenase (GDH) from Bacillus subtilis for cofactor regeneration. The two coexpressed enantiocomplementary carbonyl reductases, BDHA (2, 3-butanediol dehydrogenase from Bacillus subtilis) and GoSCR are used for asymmetric reduction of 2-hydroxyacetophenone (2-HAP) to (R)-1-phenyl-1,2-ethanediol ((R)-PED) or (S)-1-phenyl-1,2-ethanediol ((S)-PED) with excellent stereochemical selectivity, method optimization, overview. Products (R)-PED and (S)-PED are obtained with 99% yield, over 99% enantiomeric excess and 18.0 g/l/h volumetric productivity. The reaction is carried out in 5 ml sodium phosphate buffer (pH 7.0, 100 mM) at 30°C, containing 10 U/ml BDHA (cell free extract of Escherichia coli (BDHA)), 15 U/ml GoSCR (cell free extract of Escherichia coli (GoSCR)), 10 U/ml GDH (cell free extract of Escherichia coli (GDH)), 50-200 mM 2-HAP (with 10% DMSO as cosolvent), 60-250 mM D-glucose. Strong tolerance of BDHA and GoSCR against high substrate concentration Gluconobacter oxydans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Gluconobacter oxydans
0.8
-
 2-hydroxyacetophenone pH 7.0, 25°C, recombinant His-tagged enzyme Gluconobacter oxydans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-hydroxyacetophenone + NADH + H+ Gluconobacter oxydans
-
 (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
2-hydroxyacetophenone + NADH + H+ Gluconobacter oxydans 621H
-
 (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?

Organic Solvent Stability

Organic Solvent Comment Organism
additional information DMSO is selected as the co-solvent, it does not affect the activity at up 30% v/v, but is inhibitory above Gluconobacter oxydans

Organism

Organism UniProt Comment Textmining
Gluconobacter oxydans Q5FNX9
-
-
Gluconobacter oxydans 621H Q5FNX9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged GoSCR from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and dialysis Gluconobacter oxydans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxyacetophenone + NADH + H+
-
 Gluconobacter oxydans (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
2-hydroxyacetophenone + NADH + H+ 99% enantiomeric excess Gluconobacter oxydans (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
2-hydroxyacetophenone + NADH + H+
-
 Gluconobacter oxydans 621H (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
2-hydroxyacetophenone + NADH + H+ 99% enantiomeric excess Gluconobacter oxydans 621H (S)-1-phenyl-1,2-ethanediol + NAD+
-
 ?
additional information an enantiocomplementary carbonyl reductase, polyol dehydrogenase (GoSCR) from Gluconobacter oxydans is discovered to convert 2-hydroxyacetophenone (2-HAP) to (S)-1-phenyl-1,2-ethanediol ((S)-PED) with excellent stereochemical selectivity. No activity with NADPH Gluconobacter oxydans ?
-
-
additional information an enantiocomplementary carbonyl reductase, polyol dehydrogenase (GoSCR) from Gluconobacter oxydans is discovered to convert 2-hydroxyacetophenone (2-HAP) to (S)-1-phenyl-1,2-ethanediol ((S)-PED) with excellent stereochemical selectivity. No activity with NADPH Gluconobacter oxydans 621H ?
-
-

Synonyms

Synonyms Comment Organism
GoSCR
-
 Gluconobacter oxydans
More cf. EC 1.1.1.76 Gluconobacter oxydans
polyol dehydrogenase
-
 Gluconobacter oxydans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 reduction of 2-HAP Gluconobacter oxydans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 purified recombinant His-tagged enzyme, inactivation within 2 h Gluconobacter oxydans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 reduction of 2-HAP Gluconobacter oxydans

pH Stability

pH Stability pH Stability Maximum Comment Organism
7
-
 purified recombinant His-tagged enzyme, more than 80% residual activity after 18 h at pH 7.0. The activity decreases significantly in acidic circumstances Gluconobacter oxydans

Cofactor

Cofactor Comment Organism Structure
additional information no activity with NADPH Gluconobacter oxydans
NADH dependent on Gluconobacter oxydans