Hamzah R Y, Tu S C
J Biol Chem. 1981 Jun 25;256(12):6392-4.
[3,5-2H]Salicylate was obtained by a hydrogen/deuterium exchange reaction carried out in 2H2O. Using this deuterated salicylate as a substrate for salicylate hydroxylase, the product catechol was isolated and characterized with respect to the positions of deuterium retention. Based on nuclear magnetic resonance and mass spectroscopy analyses, the product was identified as [3,5-2H]catechol. It is thus unequivocally demonstrated that salicylate is decarboxylated and hydroxylated at the same ring position. Also consistent with this conclusion is the lack of a kinetic isotope effect for the deuterated salicylate substrate.
[3,5-²H]水杨酸盐是通过在重水中进行氢/氘交换反应获得的。以这种氘代水杨酸盐为水杨酸盐羟化酶的底物，分离出产物儿茶酚，并对氘保留位置进行了表征。基于核磁共振和质谱分析，该产物被鉴定为[3,5-²H]儿茶酚。因此明确证明水杨酸盐在同一环位置发生脱羧和羟化反应。氘代水杨酸盐底物缺乏动力学同位素效应也与这一结论一致。
