反应详情

Palmitoyl CoA (10 mg) and 1 mg of L-serine were dissolved in 5 mL of 100 mM HEPES-NaOH buffer (containing 10 μM PLP; pH 8.0) and well stirred using a magnetic stirrer. While the stirring speed was reduced to such an extent that a mild mixing was resulted, 0.5 mL of a crude SPT enzyme solution was added and the mixture was made to react at 37° C. for 24 hours. The solution was made alkaline with 1 mL of 2N ammonia solution and the product in the solution was extracted and recovered with 5 mL of chloroform/methanol (2:1, (v/v)). The extract liquid was filtered, concentrated appropriately and analyzed by the same manner as in Example 3 whereupon the produced amount of 3-ketodihydrosphingosine was about 0.02 mg.

1. additiona mild mixing
2. customwas resulted
3. addition0.5 mL of a crude SPT enzyme solution was added
4. customto react at 37° C. for 24 hours
5. extractionthe product in the solution was extracted
6. customrecovered with 5 mL of chloroform/methanol (2:1, (v/v))
7. filtrationThe extract liquid was filtered
8. concentrationconcentrated appropriately