介质环境对辣根国氧化物酶催化荧光反应体系的影响

探讨了缓冲溶液,溶剂和表面活性剂对对羟基苯丙酸-国氧化氢- 辣根国沿化区酶反应体系反应平衡及产物荧光性质的影响,表明二聚体荧光产物中存在酸碱平衡,应荧光滴定法求得其酸离解常数pK~a≈8.3.碱式产物的荧光强于酸式, 碱性环境使酶活性降低,反应平衡时间延长,阳离子胶束溴化十六烷基三甲胺(CTMAB) 和氯化十六烷基三甲铵(CTMAC)对该体系兼有催化和增敏作用,据此提出一种更为灵敏的测定辣根过氧化物酶的方法.

The effect of media properties on the horseradish peroxidase-catalyzed fluorogenic reaction

The effect of the media properties on the horseradish peroxidase-catalyzed fluorogenic reaction is investigated. The dimer product exhibited acid-base equilibrium For p-hydroxyphenylpropionic acid, the pKa of the product is 8.3. The fluorescence of the product increased with higher pH; however, a longer time was required to reach reaction equilibrium due to the mismatch of this pH value with that associated with enzyme activity. The cationic micelles cetyltrimethylammonium bromide and cetyltrimethylammonium chloride displayed catalytic and enhancing effects on the reaction at neutral pH. A more sensitive method for the determination of horseradish peroxidase is thus suggested.