赤子爱胜蚓胆碱酯酶的纯化和性质

赤子爱胜蚓的TritonX－100抽提液经PEG2000-磷酸钾缓冲液处理、离子交换层析及凝胶过滤后，获得电泳纯的胆碱酯酶．纯酶经鉴定为糖蛋白，凝胶过滤法和SDS-PAGE法测得其Mr分别为59000和58000．酸性氨基酸含量约占24％，碱性氨基酸含量约占12％，最大紫外吸收入λ＝272um；酶作用最适温度θ＝39℃，热稳定性较差；最适pH为7．8，在pH7．0～8．0酶较稳定．对碘化硫代乙酰胆碱的Km值为88μmol／L，有过量底物抑制现象．Mg2＋、Ca2＋和Mn2＋对酶活性的影响较小，DFP则对该酶有强烈抑制作用．     

褐菖鲉稚鱼对柴油污染的行为和生理响应

为研究柴油水溶性组分对褐菖鲉稚鱼行为反应和胆碱酯酶活性的影响,本研究以我国东海区典型的近岸岩礁性鱼类褐菖鲉为研究对象,以近岸海水本底调查的石油烃浓度为参考,探讨了不同浓度船用燃料油(0#柴油水溶性组分,DWSF)对褐菖鲉稚鱼行为活跃性、探索性及捕食能力的影响,并结合鳃的组织学变化与神经传递相关酶活性变化探究了石油类污染对褐菖鲉稚鱼的毒性效应.结果表明,当海水中DWSF浓度增加时,褐菖鲉稚鱼的探索性与活跃性均呈现先升高后降低的趋势.当DWSF浓度超过四类海水标准时(>0.30mg/L),稚鱼的行为活跃性受到明显抑制,捕食能力显著降低;通过组织学观察发现,当DWSF浓度达到0.45mg/L时,稚鱼的鳃组织会出现明显的病变和损伤,与神经传递相关的胆碱酯酶活性受到明显抑制,死亡率升高.综上可知,海水柴油污染能够引发褐菖鲉稚鱼的行为异常,显著降低其活饵捕食成功率,推断神经和呼吸系统功能受损可能是引发其行为异常的重要原因.为确保褐菖鲉等近岸岩礁性鱼类幼体的正常生长发育,海水中DWSF浓度应控制在0.45mg/L以下.     

蔬菜中有机磷农药残留的分光光度法快速检测

以鸡脑为酶源提取乙酰胆碱酯酶,利用酶抑制分光光度法检测蔬菜中有机磷农药残留量.用此方法测定了对硫磷、辛硫磷、氧化乐果三种农药,结果满意,回收率分别达到97%,101%和99%.     

单甲脒农药对鱼脑胆碱酯酶活性的影响

金鱼，鲤鱼在实验室驯化４－７ｄ，其脑胆碱酯酶活力在显著水平为Ｐ≤０．０５时，不同组的金鱼，鲤鱼无显著差异。离体实验条件下，单甲脒农药对金鱼，鲤鱼的半抑制浓度分别为１２５．９ｍｇ．ｌ＾－１和２５．７ｍｇ．ｌ＾－１。ＤＭＡ农药对鲤鱼的作用大于对金鱼的作用。     

Acute effects of chlorpyryphos-ethyl and secondary treated e uents on acetylcholinesterase and butyrylcholinesterase activities in Carcinus maenas

The acute effects of commercial formulation of chlorpyrifos-ethyl (Dursbanr®) and the secondary treated industrial/urban effluent (STIUE) exposure on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in hepatopancreas and gills of Mediterranean crab Carcinus maenas were investigated. After 2 d of exposure to chlorpyriphos-ethyl, the AChE activity was inhibited in both organs at concentrations of 3.12 and 7.82 μ/L, whereas the BuChE was inhibited only at higher concentration 7.82 μ/L of commercial preparation Dursbanr®. The exposure of crabs to Dursbanr® (3.12 μ/L) showed a significant decrement of AChE activity at 24 and 48 h, whereas the BuChE was inhibited only after 24 h and no inhibition for both enzymes was observed after 72 h. Moreover, a significant repression of AChE activity was observed in both organs of C. maenas exposed to 5% of STIUE. Our experiments indicated that the measurement of AChE activity in gills and hepatopancreas of C. maenas would be useful biomarker of organophosphorous (OP) and of neurotoxic effects of STIUE in Tunisia.     

应用胆碱酯酶生物传感器分析测定有机磷及重金属等潜在环境污染物

采用硝化纤维素为载体的固定化丁酰胆碱酯酶膜与银基汞膜电极耦合制得安培胆碱酯酶生物传感器。以该传感器为工作电极,饱和甘汞电极为参比电极组成双电极系统,用示波极谱仪接收电流信号,分析测定了有机磷(包括农用杀虫剂)和重金属等两类酶抑制剂,在较宽的浓度范围内示波峰电流值与抑制剂浓度的对数值之间具有良好的线性关系,最低检出下限可达到2．0×10－6mg／L．上述酶传感系统可用于毒性评价和环境监测等领域。      

鲫鱼(Carassius auratus)体内胆碱酯酶的组织分布及其对氨基甲酸酯类杀虫药剂的敏感度

鲫鱼(Carassius auratus Linnaeus)是重要的淡水鱼类,也是重要的食用鱼种.鱼体内胆碱酯酶(ChE)活性的降低是水体环境抗胆碱酯酶类药剂污染的重要生物化学标记.本文研究了鲫鱼体内乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BuChE)的组织特异性分布及其对灭多威、硫双灭多威、克百威以及丁硫克百威等4种氨基甲酸酯类药剂的敏感度.AChE主要分布在鲫鱼的脑部,提取液中含有Triton X-100时,AChE的比活力要比无Triton-100时大,说明AChE是膜结合的.鲫鱼中肌肉、胰脏、肝脏、肠道中BuChE比活力较高.灭多威、硫双灭多威、克百威、丁硫克百威对鱼脑中的AChE的抑制中浓度(IC50)分别为9.47×10-7 mol/L,1.34×10-6 mol/L,2.40×10-7mol/L,2.75×10-6mol/L.上述4种氨基甲酸酯类农药对鱼脑中的AChE的抑制具有很好的剂量关系,表明鲫鱼脑AChE可以作为水体环境中氨基甲酸酯类药污染的生物化学标记.     

铜、镉、敌敌畏和甲胺磷对南美白对虾的亚急性毒性作用

在恒温和充气条件下,研究了重金属(铜、镉)和有机磷农药(敌敌畏、甲胺磷)对南美白对虾的亚急性毒性作用及对虾体胆碱酯酶的影响.结果表明:南美白对虾幼虾在各浓度重金属(铜、镉)和有机磷农药(敌敌畏、甲胺磷)试液中经过60d的亚急性中毒试验,除铜在低浓度促进生长外,其他各组体长、体重及蜕皮次数均小于对照组,且随毒物浓度的增加而逐渐降低;有机磷农药对南美白对虾体内胆碱酯酶具有显著影响,虾体内胆碱酯酶含量随水体中有机磷农药浓度的升高而显著降低;重金属对胆碱酯酶也有一定影响,胆碱酯酶随重金属离子浓度升高也有逐渐降低的趋势,但重金属对胆碱酯酶的影响效果没有有机磷农药显著.     

Hormetic response of cholinesterase from Daphnia magna in chronic exposure to triazophos and chlorpyrifos

In vivo activity of cholinesterase (ChE) in Daphnia magna was measured at di erent time points during 21-day exposure to triazophos and chlorpyrifos ranging from 0.05 to 2.50 g/L and 0.01 to 2.00 g/L, respectively. For exposure to triazophos, ChE was induced up to 176.5% at 1.5 g/L and day 10 when measured by acetylthiocholine (ATCh), whereas it was induced up to 174.2% at 0.5 g/L and day 10 when measured by butyrylthiocholine (BTCh). For exposure to chlorpyrifos, ChE was induced up to 134.0% and 160.5% when measured by ATCh and BTCh, respectively, with both maximal inductions detected at 0.1 g/L and day 8. Obvious induction in terms of ChE activity was also detected in daphnia removed from exposures 24 hr after their birth and kept in a recovery culture for 21 days. Results indicated that the enzyme displayed symptoms of hormesis, a characteristic featured by conversion from low-dose stimulation to high-dose inhibition. In spite of that, no promotion in terms of reproduction rate and body size was detected at any tested concentrations regardless of whether the daphnia were collected at end of the 21-day exposure or at end of a 21-day recovery culture. This suggested that induction of ChE caused by anticholinesterases had nothing to do with the prosperity of the daphnia population.     

Layer—By—Layer技术构建化学传感器酶电极检测水体中有机磷的研究

基于层层累积自组装法将胆碱氧化酶和胆碱酯酶逐层固定在电极表面，制备了电流型水质有机磷检测化学传感器。讨论了自组装膜层数、pH值、温度对传感器电流响应的影响。制备的化学传感器对有机磷在浓度10^-9～10^-5mol／L呈良好的线性响应，检出限为1×10^-10mol／L。传感器的稳定性好，30天时的响应值仍保持90％。