本科毕业论文4.docxVIP

本科毕业论文利用异丙醇甲醇进行猪血中乙酰胆碱酯酶的分离及活性测定杨阳201130700230指导教师赵赣副教授学院名称生命科学学院专业名称生物技术论文提交日期2015?年?5?月??15日论文答辩日期2015?年? ?月?? 日摘要本文以来源广泛、廉价的猪血清为材料，对乙酰胆碱酯酶进行了分离纯化以及部分酶学性质的研究，以期为该酶的来源及进一步研究提供参考。通过提取猪血清中的乙酰胆碱酯酶，然后在pH为4、5、6、7.5、8、9条件下，其他条件不变，测定乙酰胆碱酯酶的活性，以酶活力的最高值为100%。利用分光光度计检测活性和考马斯亮蓝法测定蛋白含量，同时用不同的有机溶剂浓度分别是30%，60%，80%，90%，分别用异丙醇和甲醇在这些的浓度再进行分离，最后通过SDS凝胶电泳进行检测分离的纯度，同时也要测蛋白含量和酶活性来研究有机溶剂对其的有关影响。实验结果：pH7.5为活力最适pH值比活=0.342U/mg 甲醇溶液提取的比活性逐渐升高，异丙醇溶液提取后乙酰胆碱酯酶的比活性逐渐降低，但是甲醇的总活性和总蛋白量很低，说明高浓度的甲醇和异丙醇对乙酰胆碱酯酶有明显抑制作用，用浓度为50%的甲醇提取较好。关键词：乙酰胆碱酯酶活性蛋白质含量分离The Activity and Extraction of Acetylcholinesterase from Pig BloodYang Yang(College of Science,South China Agricultural University,Guangzhou 510000 China)Abstract:The works aimed on finding a pathway to obtain AChE and provide the data for deeply investigation of AChE and application of porcine blood.It will determine the activity of acetylcholinesterase and enzyme activity of the highest value is 100%. While other conditions remain unchanged and determine pH for 4, 5, 6, 7, 8, 9, ,in the experiment.By extracting the acetylcholinesterase in pig serum.In?the?experiment?by?extracting?the?Porcine?serumacetyl?cholinesterase,?then?proceed?active?testing?under?different?pH4,?5,?6,?7.5,?8,?9?,?and?coomassie?brilliant?blue?method?for?the?determination?of?protein?content,?at?the?same?time?in?different?organic?solvents?are?respectively?30%,?60%,?80%?and?90%,?respectively,?with?the?concentration?of?isopropyl?alcohol?and?methanol?in?the?separation?again,?finally?through?sds?gel?electrophoresis?for?testing.The?result:The?dynamic?optimal?ph?=pH7.5?,Specific?activity=0.342?U?,?high?concentrations?of?methanol?and?isopropanol?on?acetylcholinesterase?have?obvious?inhibitory?effect,?better?with?a?concentration?of?50%?methanol?extract.Keywords: AcetylcholinesteraseActivity The Content Of Protein Extraction缩略语及英文对照AchE acetylcholinesterase 乙酰胆碱酯酶pH hydrogen ion concentration 溶液酸性或碱性程度的数值pbs phosphate buffered saline磷酸盐缓冲液D