反应详情

温度

22 °C

The fungi is stored at −25° C. 250 mL conical flasks each containing 50 mL seed culture medium (medium 216) are inoculated with 2 mL of fungi suspension and incubated on a rotary shaker (200 rpm at 23° C. for 3 days. 250 mL conical flasks each containing 50 mL of the same medium were inoculated with mL of the seed culture and incubated on a rotary shaker (200 rpm ) at 23° C. After 24 hours, (1R,2R,4)- (+)-Bicyclo[2.2.]hept-2-yl-(6,7-dimethoxyquinoxalin-2-yl)-amine of Example 25 is dissolved in MeOH added to the flasks to a final concentration of 300 mg/L. The cultures are harvested after 24 hours of incubation. (Medium 216: Glucose 0.4%, Yeast extract 0.05%, Soya flour 0.05%, NaCI 0.05%, KH2PO4 0.05.) The extraction is performed using 2 volumes of acetonitrile, 1 volume de tert-butylmethyl ether and volume of n-heptane were added to 1 volume of broth. After magnetic stirring at 22° C, the extract separates to 3 layers. The intermediate layer is collected and evaporated to dryness, and redissolved in ethyl acetate. The ethyl acetate extract is separated on silica gel (0.04-0.063 mm) using ethyl acetate as eluent. Fractions containing the biotransformation product are separated on C1 8 silica using a H2O/MeOH gradient as eluent. This chromatography yields the pure titled compound as an amorphous yellow powder, m.p. 190-192° C.

1. customis stored at −25° C
2. custom250 mL conical flasks each containing 50 mL seed culture medium (medium 216) are inoculated with 2 mL of fungi suspension
3. customat 23° C.
4. customfor 3 days
5. custom250 mL conical flasks each containing 50 mL of the same medium were inoculated with mL of the seed culture
6. customincubated on a rotary shaker (200 rpm ) at 23° C
7. additionadded to the flasks to a final concentration of 300 mg/L
8. waitThe cultures are harvested after 24 hours of incubation
9. extraction(Medium 216: Glucose 0.4%, Yeast extract 0.05%, Soya flour 0.05%, NaCI 0.05%, KH2PO4 0.05.)
10. extractionThe extraction
11. additionwere added to 1 volume of broth
12. customThe intermediate layer is collected
13. customevaporated to dryness
14. dissolutionredissolved in ethyl acetate
15. extractionThe ethyl acetate extract
16. customis separated on silica gel (0.04-0.063 mm)
17. additionFractions containing the biotransformation product
18. customare separated on C1 8 silica using a H2O/MeOH gradient as eluent