动物肝酯酶对西维因的抑制响应
Inhibitory response to carbaryl of animal liver esterase
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摘要: 采用硫酸铵分级盐析后透析的方法分别从鹅肝、鸡肝和猪肝中提取和纯化酯酶,对其进行蛋白组分的定性和定量分析及结构表征,并以西维因作为农药模型进行抑制响应研究。结果表明:鹅肝酯酶得率最高(14.58%),鸡肝酯酶得率最低(8.82%)但热稳定性最好,猪肝酯酶的蛋白质量浓度最高(12.47 mg/mL);3种动物肝酯酶的蛋白分子质量均为30~110 kDa,主要蛋白组分的分子质量均为55 kDa,且该蛋白组分在鸡肝酯酶中的相对含量最高(16.74%),在猪肝酯酶中的相对含量最低(14.31%);3种动物肝酯酶的蛋白颗粒大小不一,均呈片状结构,表面较光滑;在西维因质量浓度为0.5~10.0 mg/L时,鸡肝酯酶的线性范围(0.5~5.0 mg/L)、检测下限(0.29 mg/L)和IC50(7.74 mg/L)均最低,可作为西维因快速检测的潜在敏感酶源。Abstract: Through graded salting out with ammonium sulfate and dialysis, esterases were extracted and purified from goose liver, chicken liver and pig liver respectively, then their structures were characterized, and their inhibition responses to carbaryl as a pesticide model were further investigated. Results showed that the yield of goose liver esterase was the highest (14.58%), and that of chicken liver esterase was the lowest (8.82%) with the best thermal stability, while the protein mass concentration of pig liver esterase was the highest (12.47 mg/mL); the protein molecular weights of all three kinds of animal liver esterases were 30~110 kDa with the leading one of 55 kDa, and that in chicken liver esterase was the highest (16.74%) while the lowest was in that of pig liver esterase (14.31%); these three kinds of animal liver esterases showed different sizes of protein particles with flake structures and smooth surfaces; in the carbaryl concentration range of 0.5~10.0 mg/L, the linear range (0.5~5.0 mg/L), the detection limit (0.29 mg/L) and IC50 (7.74 mg/L) of chicken liver esterase were the lowest, which could be used as a potential sensitive enzyme source for rapid detection of carbaryl.
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