摘要
探讨了缓冲溶液、溶剂和表面活性剂对对羟基苯丙酸-过氧化氢-辣根过氧化物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在溶液中存在酸碱平衡,用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铵(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 the media properties on the horseradish preoxidase-analyzed flourogenic reaction is investigated. There existed acid-base equilibrium in the dimer product. For p-hydroxylphenolpropionic acid, the pKa of the product is 8.3. The product fluorescence increases with higher pH, however, longer time is needed to reach the reaction equilibrium due to the pH mismatch with the enzyme activity. Cationic micelles cetyltrimethyl ammonium bromide and cetyltrimithylammonium chloride have catalytic and enhancing effects on the reaction at the neutral pH. A more sensitive method for the determination of horseradish peroxidase is thus suggested.
出处
《化学学报》
SCIE
CAS
CSCD
北大核心
1995年第3期248-253,共6页
Acta Chimica Sinica
