以壳聚糖微球为载体的固定化乙酰胆碱酯酶的基本性质  被引量:3

CHARACTERIZATION OF ACETYLCHOLINESTERASE IMMOBILIZED ON CHITOSAN MICROSPHERES

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作  者:张爱静[1] 孟范平[1] 杨菲菲[1] 

机构地区:[1]中国海洋大学海洋环境与生态教育部重点实验室,青岛266100

出  处:《环境化学》2011年第6期1068-1074,共7页Environmental Chemistry

基  金:国家高技术研究发展计划项目(2001AA635130)资助

摘  要:为探讨以壳聚糖微球为载体制备的固定化乙酰胆碱酯酶(AChE)用于监测海水有机磷农药的可行性,研究了固定化酶在海水中的基本性质,如环境适应性、动力学性质、稳定性、对农药响应规律等.结果表明,与溶液酶相比,固定化酶对海水温度、盐度、pH值等因素的适应能力提高;固定化酶分子构象的改变增加了酶与底物之间的空间障碍,米氏常数增加到46.18 mmo·lL-1;固定化酶热稳定性和储存稳定性明显增强,4℃下保存60 d后活力仅损失17%.固定化AChE活性抑制率与农药浓度对数具有良好的线性相关性,固定化酶对甲基对硫磷响应最为敏感,检测下限为5.34μg·L-1.In order to evaluate the feasibility of using immobilized acetylcholinesterase(AChE) on chitosan microspheres to monitor organophosphorus pesticides(OPs) in seawater,basic properties of the immobilized enzyme were investigated,such as environmental adaptability,dynamic property,stability,response characteristics to OPs.The results indicated that the immobilized enzyme was more adaptive to the temperature,salinity,pH of seawater compared with the solution enzyme.The Michaelis constant of the immobilized AChE increased to 46.18 mmol·L-1,which may be caused by the steric hindrance between the immobilized enzyme and its substrate.Thermal stability and storage stability were also enhanced after immobilization.The immobilized AChE lost only 17% of its initial activity after 60 days storage at 4 ℃.Moreover,there was a good linear correlation between the activity inhibition rate of immobilized AChE and the logarithmic value of OPs concentration.The immobilized enzyme was most sensitive to methyl-parathion,with a detection limit of 5.34 μg·L-1.The results of this research suggested that the immobilized enzyme have a good prospect in monitoring OPs in seawater due to its good performance and sensitive response.

关 键 词:壳聚糖 固定化 乙酰胆碱酯酶 性质 海水 

分 类 号:Q814.2[生物学—生物工程]

 

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