Reaction equation
[C:1]([O-:6])(=[O:5])[C:2]([CH3:4])=O.C([O-])(=O)C(C)O.C1[N:14]=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3O)C=2N=1.C([O-])=O.[F:60][C:61]([F:70])([F:69])[CH2:62]CC(=O)C(O)=O.NC(C(O)=O)C.CC1N=CC(COP(O)(O)=O)=C(C=O)C=1O.C([O-])=O.[Na+]>P([O-])([O-])([O-])=O.[K+].[K+].[K+]>[F:60][C:61]([F:70])([F:69])[CH2:62][CH2:4][C@@H:2]([NH2:14])[C:1]([OH:6])=[O:5]Participants
Reactant 8
Product 1
Reagents, catalysts & solvents 1
Procedure
The yields were increased by adding auxiliary enzymes to reduce pyruvate to lactate. Lactate dehydrogenase requires NADH as a cofactor. The NADH was regenerated using formate dehydrogenase. A solution containing 5,5,5-trifluoro-2-oxopentanoic acid (100 mg, 0.588 mmoles), D,L-alanine (200 mg, 2.244 mmoles), 0.02 mM pyridoxal phosphate, sodium formate (68 mg, 1 mmole), NAD (3.31 mg, 5 μmoles) L-lactate dehydrogenase cloned from rabbit muscle (Biocatalytics LDH-103, 0.107 mg, 15 units), and formate dehydrogenase (0.5 mL, 15 units cloned from Pichia pastoris and expressed in Escherichia coli) in 0.1 M potassium phosphate buffer, pH 7.5, was incubated with R-transaminase AT-103 from Biocatalytics (5 mg, 44units) or cloned R-transaminase from Bacillus thuringiensis SC16569 (0.5 mL, 10 units) at 30° C. in a total volume of 5 mL in 15 mL tubes. Reaction yields of (R)-5,5,5-trifluoro-2-aminopentanoic acid of 94% and 91% were obtained with AT-103 and BMS transaminases, respectively. Ee was 100% in both cases.
Workup
- temperatureThe yields were increased
- additionby adding auxiliary enzymes
- customReaction
Sources & evidence
- ORD record
- ord-4eea0d8855d842899a27b9e64841d610
- Source dataset
- uspto-grants-2011_12
- Patent number
- US08084477B2
Linked notes
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