反应详情

A solution of 7-amino-2-propylheptanoic acid hydrochloride (prepared in EXAMPLE 1, 11.2 mg) in water (0.5 ml) was neutralized with sodium hydrogen carbonate. To the solution was added a solution of m-MBS(15.5 mg) in dioxane (0.5 ml) and allowed to react for 30 minutes at ambient temperature. Then the solution (0.4 ml) was added to a mixture of β-galactosidase (Boeringer Mannheim G.m.b.H., 5 mg/ml suspension in ammonium sulfate solution, 100 μl) in 0.1 M phosphate buffer (pH 7.0, 2 ml) and allowed to react for an hour at ambient temperature. The reaction mixture was applied on a BioGel P-4 (BioRad Corp.) type I column (2.5×25 cm) equilibrated with 0.9% sodium chloride-0.02 M phosphate buffer, and eluted with the same buffer to obtain 5 ml fractions. The β-galactosidase activity of each fractions were measured. Fractions (Nos. 6-8) with high enzyme activity were combined and stored in a 0.1% sodium azide (preservative) and 0.1% BSA (stabilizer) solution.

1. customto react for 30 minutes at ambient temperature
2. customto react for an hour at ambient temperature
3. washeluted with the same buffer
4. customto obtain 5 ml fractions