反应详情

Cells were prepared as described in Example 4. 1 mg 7-xylosyl-10-deacetyltaxol, 200 mg (108 milliunits) C-13 deacylase (described in Example 4), 0.2 ml methanol, 0.2 ml 1 M potassium phosphate buffer pH 7, and 3.6 ml water were mixed end-over-end at 48 rpm for 17 hours at 24° C. After 15 hours, HPLC analysis by Method 1 showed complete removal of C-13 side chain from the substrate. 2 ml of the solution now containing 7-xylosyl-10-deacetylbaccatin-III was added to 1 ml of the above Moraxella sp. cells (0.24 g wet cell weight) in 50 mM potassium phosphate buffer pH 7 and mixed end-over-end for 24 hours at 48 rpm. Reaction mixtures were extracted with 6 ml CH2Cl2. Extracts were dried and redissolved in methanol for HPLC analysis by Method 1. 0.099 mg/ml 10-deacetylbaccatin-III was found (103% yield based on initial 7-xylosyl-10-deacetyltaxol concentration).

1. customCells were prepared
2. customHPLC analysis by Method 1 showed complete removal of C-13 side chain from the substrate
3. addition2 ml of the solution now containing 7-xylosyl-10-deacetylbaccatin-III was added to 1 ml of the above Moraxella sp
4. additioncells (0.24 g wet cell weight) in 50 mM potassium phosphate buffer pH 7 and mixed end-over-end for 24 hours at 48 rpm
5. customReaction mixtures
6. extractionwere extracted with 6 ml CH2Cl2
7. customExtracts were dried
8. dissolutionredissolved in methanol for HPLC analysis by Method 1
9. concentration(103% yield based on initial 7-xylosyl-10-deacetyltaxol concentration)