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当前位置: 首页 > 文章 > 银杏叶片可溶性蛋白质测定非直线性原因分析 果树学报 2002,19 (1) 32-35
Position: Home > Articles > Analysis of the Reasons Caused the Protein Assay of Ginkgo L. Leaves notto be Expressed as Linearity Journal of Fruit Science 2002,19 (1) 32-35

银杏叶片可溶性蛋白质测定非直线性原因分析

作  者：
汪良驹;姜卫兵;祝海阔;范黄斌;何歧峰
单  位：
南京农业大学园艺学院
关键词：
银杏叶片;双缩脲法;考马斯亮蓝法;蛋白质测定;蛋白水解酶
摘  要：
曾报告用考马斯亮蓝法测定银杏叶片可溶性蛋白质含量缺乏直线性,猜想其可能具有较高的蛋白水解酶活性。研究表明,银杏叶片蛋白水解酶用ｐＨ７．０缓冲液提取时活性最高,酶活最适ｐＨ５．５~６．０,因而不能解释ｐＨ８．３缓冲液提取出的蛋白质含量下降的原因。比较不同的蛋白质测定方法表明,双缩脲法测得的ＯＤ５４０值相当稳定,而考马斯亮蓝法测定的ＯＤ５９５值随时间延长而升高;同样比色５ｍｉｎ,双缩脲法测得的蛋白质浓度比考马斯亮蓝法高２０~３０倍,说明银杏叶片蛋白质不能迅速溶解于酸性考马斯亮蓝显色液是其测定非直线性的主要原因。
译  名：
Analysis of the Reasons Caused the Protein Assay of Ginkgo L. Leaves notto be Expressed as Linearity
作  者：
Wang Liangju,Jiang Weibing,Zhu Haikuo,Fan Huangbin,and He Qifeng(College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095)
关键词：
Ginkgo L. leaves;Biuret;Commassie brillant blue;Protein analysis;Proteinase
摘  要：
In this paper,citric-phosphoric buffers with pH 5.0~8.0 were used to extract proteinases from Cinkgo L. leaves,and the results showed that when the buffer pH was 7.0, the enzyme activity was the highest in pH 5.5 citric-phosphoric re-action system. When the extraction buffer at pH 7.0 was used, the activities of the proteinase were the highest at pH 5.5~6.0.Thus, the former hypothesis of proteinases could not be used to explain the decrease of proteins extracted by pH 8.3 buffer.The comparison between different measurements showed that OD540 of the soluble proteins was stable in biuret method. whilethe OD595 consisted to increase with reaction time hy the Coomassie brilliant blue assay. At the same reaction time (5 min),the protein eoncentrations by biuret were 20~30 times as high as that by Coomassie brilliant blue.These facts implied that theunlinearity of protein assay by Commassie brilliant blue as reported previously was mainly caused by the dissolution of Gink-go L.proteins in acid reaction system.

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