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

Temperatur

30 °C

The recombinant E. coil HB101(pNTS1G) obtained in Example 10 was inoculated in 100 ml of a 2×YT medium sterilized in a 500 ml Sakaguchi flask, and cultured with agitation at 37° C. for 13 hours. Glucose, 19.2 g, and 2.5 mg of NADP were added to 40 ml of the resultant culture. The culture was stirred at 30° C. while being adjusted at pH 6.5 with a 5 M sodium hydroxide solution. While stirring, a total of 16.1 g of ethyl 4-chloroacetoacetate was continuously added to the culture at a rate of about 2 g per hour. The reaction was performed for 24 hours. After the reaction, the reaction solution was subjected to extraction using ethyl acetate, the solvent was removed under decompression, and the concentrate was purified by silica gel column chromatography, to obtain 15.6 g of ethyl (S)-4-chloro-3-hydroxybutyrate. The optical purity of the ethyl (S)-4-chloro-3-hydroxybutyrate was analyzed by an HPLC method and found to be 100% e.e. 1H-NMR(CDC13)δ(ppm): 1.33(3H,t), 2.65(2H,d), 3.31(1H,d), 3.60(2H,d), 4.2(3H,m); Column: Chiralcel OB (0.46×25 cm) manufactured by Daicel Chemical Industries, Co., Ltd.; Column temperature: 0° C.; Eluent: n-hexane/2-propanol of 9/1; Flow rate: 0.8 ml/min.; Detection: 215 nm; Elution time: 19.2 minutes for (S), 17.0 minutes for (R).

1. customcoil HB101(pNTS1G) obtained in Example 10
2. stirringWhile stirring
3. waitThe reaction was performed for 24 hours
4. customAfter the reaction
5. extractionthe reaction solution was subjected to extraction
6. customthe solvent was removed under decompression
7. customthe concentrate was purified by silica gel column chromatography

ORD-Eintrag

ord-ac8c4e31422b493cbd93ffa08206284b

Quelldatensatz

uspto-grants-2001_04

DOI

10.6084/m9.figshare.5104873.v1

Patentnummer

US06218156B1