反应详情

温度

90 °C

15 mg of the N-(ferrocenyl methyl)-6-amino caproic acid (Fc-aca) prepared in (1) of Example 8, 6 mg of N-hydrosuccinimide (NHS, Fluka Corp), and 9 mg of N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC, Fluka Corp.) were dissolved in 0.3 mL of an anhydrous DMF solution and stirred for 30 minutes after the temperature was raised to 90° C. in a nitrogen atmosphere. The reaction was further performed for 30 minutes while the temperature of the solution was maintained at 80° C. 30 μL of the reaction mixture was added into a solution of 7 mg of lipase in 1 mL of a phosphate buffer solution (100 mM) at 2-minute intervals. The mixture was shaken at room temperature for a day and centrifuged to separate a precipitate. The filtrate was dialyzed in a 50-mM phosphate buffer solution (pH 7.4) for 30 hours to obtain an N-(ferrocenyl methyl)-6-amino caproic acid (Fc-aca)-bound lipase (ferrocene-bound lipase).

1. waitThe reaction was further performed for 30 minutes while the temperature of the solution
2. temperaturewas maintained at 80° C
3. stirringThe mixture was shaken at room temperature for a day
4. customto separate a precipitate
5. customThe filtrate was dialyzed in a 50-mM phosphate buffer solution (pH 7.4) for 30 hours