4-Methylumbelliferyl phosphate → 4-Methylumbelliferone
반응식
[CH3:1][C:2]1[C:8]2[CH:9]=[CH:10][C:11]([O:13]P(O)(O)=O)=[CH:12][C:7]=2[O:6][C:4](=[O:5])[CH:3]=1.[Mg]>>[CH3:1][C:2]1[C:8]2[CH:9]=[CH:10][C:11]([OH:13])=[CH:12][C:7]=2[O:6][C:4](=[O:5])[CH:3]=1참가 물질
3
실험 절차
The procedures used in FIG. 5 could not be applied to 4-MUP for two reasons. First, the background reading obtained from a 1 mM solution was off scale on the Turner model 111. If the concentration was reduced to 0.030 mM then the background reading was comparable to a 1 mM solution of BBTP or ABTP. Since the Km for 4-MUP in this buffer is approximately 30 micromolar then the turnover of 4-MUP is reduced thus lowering the sensitivity of the substrate for AP. It was found that with freshly prepared 30 micromolar solutions of 4-MUP that it gave approximately one half to one third the signal obtained with a 1.0 mM solution of BBTP. Secondly, it was found that basic aqueous solutions of 4-MUP containing magnesium cause the 4-MUP to hydrolyze and form 4-methyl umbelliferone. This results in a significant background increase. This limits the test to having the 4-MUP being reconstituted with buffer usually within 8 hours of use even when this solution is stored at 4° C.