Tromethamine hydrochloride → 7-Amino-4-methylcoumarin

Participants

Reaction Steps

50 μl of the solution of the trypsin-like protease obtained in Example 1 was added to 1.5 ml of 0.1M Tris-HCl buffer (pH 8.6) containing 100 μM of a synthetic substrate for trypsin Boc-Phe-Ser-Arg-MCA (MCA=methylcoumarinamide), and the mixture was subjected to incubation at 37° C. for 1 hour. 1 ml of 30% acetic acid was then added, the amount of 7-amino-4-methylcoumarin (AMC) formed was determined by fluorescent assay (fluorescence 440 nm, excitating light 380 nm), and the activity of the enzyme was calculated based thereon. An activity of forming 1 pM of AMC in 1 minute is defined as 1 unit (1 unit=1 pM AMC/min).

Provenance & Evidence