Sulfuric Acid → Methyl ((2S,3S)-4-chloro-3-hydroxy-1-phenylbutan-2-yl)carbamate

A culture was obtained by the same method as in Example 1 using Candida gropengiesseri IFO 0659 as the microorganism and adjusted to pH 6 with an aqueous sodium hydroxide solution and an aqueous sulfuric acid solution. A 75-ml portion of this culture was separated into a supernatant and cells by centrifugation. To the whole amount of the cells was added a portion of the supernatant to make the total volume 25 ml. This cell suspension (25 ml), 1 g of glucose and 250 mg of (3S)-1-chloro-3-methoxycarbonylamino-4-phenyl-2-butanone were mixed together in a 500-ml Sakaguchi flask and the mixture was shaken for 1 day to allow the reaction to proceed. A further 250-mg portion of (3S)-1-chloro-3-methoxycarbonylamino-4-phenyl-2-butanone was added. The reaction was allowed to proceed by shaking for further 6 days while controlling the glucose concentration within the range of 0 to 4% by adding glucose portionwise. Water, 100 ml, was added to the reaction mixture and the whole mixture was extracted with 100 ml portion of ethyl acetate for 3 times. The organic extract layers were combined, washed with 100 ml of water and further with 100 ml of a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. This solution was concentrated to dryness to give (2S,3S)-1-chloro-3-methoxycarbonylamino-4-phenyl-2-butanol as a white solid (401 mg; yield 61%; purity 76.8 wt %; selectivity 2S,3S/2R,3S=98/2; optical purity 98.3% ee).

1. customA culture was obtained by the same method as in Example 1 using Candida gropengiesseri IFO 0659 as the microorganism
2. customA 75-ml portion of this culture was separated into a supernatant and cells by centrifugation
3. additionTo the whole amount of the cells was added a portion of the supernatant
4. additionThis cell suspension (25 ml), 1 g of glucose and 250 mg of (3S)-1-chloro-3-methoxycarbonylamino-4-phenyl-2-butanone were mixed together in a 500-ml Sakaguchi flask
5. customthe reaction
6. stirringby shaking for further 6 days
7. concentrationthe glucose concentration within the range of 0 to 4%
8. additionby adding glucose portionwise
9. additionWater, 100 ml, was added to the reaction mixture
10. extractionthe whole mixture was extracted with 100 ml portion of ethyl acetate for 3 times
11. washwashed with 100 ml of water and further with 100 ml of a saturated aqueous sodium chloride solution
12. dry with materialdried over anhydrous sodium sulfate
13. concentrationThis solution was concentrated to dryness

ORD record

ord-dfb2793f61924a0a99912919753f0f33

Source dataset

uspto-grants-1998_03

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05726047