苦瓜蛋白提取方法的初步研究(毕业论文11000字)
摘 要：本试验为从苦瓜果肉提取纯化降血糖的蛋白，选择新鲜苦瓜，去籽、破碎、匀浆，酸乙醇浸泡，有机溶剂沉淀.在有机溶剂沉淀这一步骤中，做了对比:一种采用乙醚﹑乙醇沉淀,另一种只采用乙醇沉淀,得粗蛋白。试验采用考马斯亮蓝G-250法测定蛋白含量,经SDS—PAGE初步鉴定提取的苦瓜蛋白,结果表明:苦瓜中采用了乙醚提取的蛋白含量比只采用了乙醇提取蛋白含量要高;提取的苦瓜蛋白经SDS-PAGE电泳，证实为一条带，分子量在15.77KD左右,且采用了乙醚提取的蛋白条带的清晰度明显较高于只用乙醇提取的蛋白条带,计算得提取的苦瓜蛋白相对分子质量与文献中的降血糖的蛋白的相对分子量24KD有差距，结果仍需进一步鉴定。
关键词：苦瓜，苦瓜蛋白 ，提取，考马斯亮蓝G-250, SDS-PAGE电泳
The extraction methods for Momordica charantia protei
Abstract: This experiment for falls the blood sugar from the balsam pear fruit pulp extraction purification the protein, the choice fresh balsam pear, goes to the seed, stave, the refining, sour ethyl alcohol immersion, organic solvent precipitation. Precipitates in this step in the organic solvent, has made the contrast: A kind of ether by ethanol precipitation, and the other using only ethanol precipitation, was crude protein. Experiment with Coomassie Brilliant Blue G-250 Determination of protein content by SDS-PAGE identification of proteins extracted from bitter melon, the results showed that: In the balsam pear, using ether extraction and protein extraction using only the protein content of ethanol were higher; Bitter melon extract proteins by SDS-PAGE electrophoresis, proved to be a molecular weight around in 15.77KD, and extracted using ether protein band resolution significantly higher in the ethanol extract only the protein bands, calculated extraction Bitter protein molecular weight and the literature of plants to lower blood glucose are weight 24KD, The result still need identification.
Keywords: Momordiea charantia; Momordica charantia Protein; Coomassie Brilliant Blue G-250; SDS-PAGEcataphoresis
目 录
摘要……………………………………………………………………………………1
关键词…………………………………………………………………………………1
1 前言…………………………………………………………………………………2
1.1 苦瓜概况………………………………………………………………2
1.2 苦瓜的营养成分…………………………………………………………2
1.3 苦瓜的化学成分…………………………………………………………3
1.4 苦瓜的药理作用…………………………………………………………3
1.5 苦瓜蛋白及降血糖作用………………………………………………5
2 材料与方法…………………………………………………………………………6
2.1 材料………………………………………………………………………6
2.1.1 材料………………………………………………………………6
2.1.2 主要仪器设备………………………………………………………6
2.1.3 主要试剂……………………………………………………………7
2.2 方法…………………………………………………………………………7
2.2.1 主要试剂配制………………………………………………………7
2.2.2 鲜苦瓜粗蛋白提取…………………………………………………8
2.2.3 透析浓缩苦瓜原液…………………………………………………8
2.2.4 考马斯亮蓝G-250法测定苦瓜蛋白含量……………………9
2.2.5 SDS-PAGE测定苦瓜蛋白质分子量………………………………10
3 结果与分析………………………………………………………………………12
3.1 提取蛋白质含量测定结果与分析…………………………………12
3.1.1苦瓜粗蛋白样品…………………………………………………12
3.1.2 标准曲线中蛋白OD测定结果……………………………………12
3.1.3 苦瓜蛋白含量的测定结果……………………………………… 13
3.1.4 各组分蛋白质得率结果…………………………………………13
3.2 苦瓜蛋白SDS聚丙烯酰胺凝胶电泳结果及分析………………14
3.2.1 提取的苦瓜蛋白的分子量粗计算………………………………14
3.2.2 数据分析…………………………………………………………15
4 讨论………………………………………………………………………………15
5结论………………………………………………………………………………16
参考文献……………………………………………………………………………16
致 谢……………………………………………………………………………18
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