Ethanol → HCID 9899181, compound

HRID 1166027Notes

Reaction equation

HRID 1166027 reaction equation
[C:1]1([O:7][C:8]([N:10]2[CH2:14][CH2:13][CH2:12][CH2:11]2)=[O:9])[CH:6]=[CH:5][CH:4]=[CH:3][CH:2]=1.C([OH:17])C>>[C:1]1([O:7][C:8]([N:10]2[CH2:14][CH2:13][C@H:12]([OH:17])[CH2:11]2)=[O:9])[CH:2]=[CH:3][CH:4]=[CH:5][CH:6]=1

Participants

3

Reactant 2

HCID 702EthanolHCID 945741-Pyrrolidinecarboxylic acid, phenyl ester

Product 1

HCID 9899181HCID 9899181, compoundYield 33%

Conditions

Temperature
21 °C

Procedure

Six Fernbach flasks, each containing 0.5 L of medium 2 as described in Example 2 were inoculated with spores of C. echinulata ATCC 8688b and incubated at 29° C. on a rotary shaker (220 rpm). After 24 hours, 5 mL of a 60 mg/mL ethanol solution of pyrrolidine-1-carboxylic acid phenylester (II) were added to each flask, and incubated for 16 more days. The bioconversion was monitored in two of the six flasks (flasks A and B) by removing samples of fermentation broth, extracting with ETOAc, and analyzing the extracts by reverse phase and chiral HPLC as described in Example 1. The results of these HPLC analyses are summarized in Table 3. The contents of all six flasks were pooled after the 16 days incubation and filtered through a triple-layer of cheesecloth to remove cells. The filtrate was then stirred with 50 g of Amberlite® XAD-16 resin (Rohm and Haas, Philadelphia, Pa.) for 4.5 hours at 21° C. The resin was collected onto filter paper and washed with 0.8 L of ETOAc. The ETOAc extract was washed with water, dried with anhydrous magnesium sulfate, and concentrated under reduced pressure to give 0.93 g of crude product. The crude product was applied to a 5 g silica Seppak® cartridge (Waters Corporation, Milford, Mass.) and eluted with mixtures of ETOAc and hexane, (2:3, 1:1) and the fractions containing the desired product were concentrated under reduced pressure to give 0.64 g (33% yield, 89% ee) of (S)-3-hydroxypyrrolidine-1-carboxylic acid phenylester (II).

Workup

  1. customincubated at 29° C. on a rotary shaker (220 rpm)
  2. customby removing samples of fermentation broth
  3. extractionextracting with ETOAc
  4. customThe results of these HPLC analyses
  5. customafter the 16 days incubation
  6. filtrationfiltered through a triple-layer of cheesecloth
  7. customto remove cells
  8. customThe resin was collected
  9. filtrationonto filter paper
  10. washwashed with 0.8 L of ETOAc
  11. extractionThe ETOAc extract
  12. washwas washed with water
  13. dry with materialdried with anhydrous magnesium sulfate
  14. concentrationconcentrated under reduced pressure
  15. customto give 0.93 g of crude product
  16. washeluted with mixtures of ETOAc and hexane, (2:3, 1:1)
  17. additionthe fractions containing the desired product
  18. concentrationwere concentrated under reduced pressure

Sources & evidence

ORD record
ord-76509b30747843738feb5ad6e68fed6c
Source dataset
uspto-grants-2003_09
Patent number
US06613550B2

Reaction notes

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