NADP nicotinamide-adenine-dinucleotide phosphate → Ethyl (3S)-4-chloro-3-hydroxybutanoate

Temperature

30 °C

To 45 ml of the cell-free extract prepared from E. coli HB101(pNTS1) by the method described above were added 5 ml of the cell-free extract of E. coli HB101(pNGLP) as prepared by the method described in Example 6, 5.5 g of glucose and 1.6 mg of NADP and, while the mixture was adjusted to pH 6.5 with a 5 M aqueous solution of sodium hydroxide and stirred at 30° C., ethyl 4-chloroacetoacetate was added in 250-mg portions at 15-minute intervals. The reaction was allowed to proceed for a total of 5 hours while adding a total amount of 5 g of ethyl 4-chloroacetoacetate in that manner. Thereafter, the reaction mixture was extracted with ethyl acetate, the solvent was then removed, and the remaining extract was analyzed by gas chromatography under the conditions given below, whereupon the formation of ethyl 4-chloro-3-hydroxybutyrate in 95% yield was revealed. The product ethyl 4-chloro-3-hydroxybutyrate was analyzed for optical purity by high-performance liquid chromatography under the conditions given below; it was found to be in the S form with 100% e.e.

1. extractionTo 45 ml of the cell-free extract
2. customprepared from E
3. additioncoli HB101(pNTS1) by the method described above were added 5 ml of the
4. extractioncell-free extract of E
5. customas prepared by the method
6. customto proceed for a total of 5 hours
7. extractionThereafter, the reaction mixture was extracted with ethyl acetate
8. customthe solvent was then removed
9. extractionthe remaining extract

ORD record

ord-2c32e9a0a8134289b0c40baa656baff4

Source dataset

uspto-grants-2016_04

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US09303280B2