银耳芽孢总蛋白提取方法的建立

doi:10.7506/spkx1002-6630-201609007

摘要/Abstract

摘要：

采用超声波与高压均质破碎细胞，通过显微观察细胞破碎效果、Bradford法测定蛋白含量并进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析，比较了三羟甲基氨基甲烷（tris(hydroxymethyl)methyl aminomethane，Tris）-饱和酚法、三氯乙酸（trichloroacetic acid，TCA）-丙酮沉淀法及Tris-饱和酚结合TCA-丙酮沉淀法提取银耳芽孢总蛋白的效果。结果表明：高压均质的细胞破碎效果优于超声波，3 种方法所得到的银耳芽孢总蛋白质量浓度分别为2.33、1.26、1.02 mg/mL，且Tris-饱和酚法经传统考马斯亮蓝染色法染色所得电泳条带亮度及清晰度均优于其他两种方法。因此，Tris-饱和酚法是进行银耳芽孢总蛋白提取的一种有效方法。

关键词: 银耳, 芽孢, 蛋白提取方法

Abstract:

In order to compare extraction efficiencies of total protein from the yeast-like conidia of Tremella fuciformis by
Tris-saturated phenol method, trichloroacetic acid (TCA)-acetone precipitation method and their combination, the cells were
disrupted by ultrasound and high-pressure homogenization, the efficiency of cell disruption was observed by microscope,
protein content was determined by the Bradford method, and SDS-PAGE gel electrophoresis analysis was performed. The
results showed that the efficiency of cell disruption by high pressure homogenization was better than that by ultrasound. The
protein contents obtained by three extraction methods were 2.33, 1.26 and 1.02 mg/mL, respectively. The brightness and
resolution of electrophoresis bands with traditional coomassie brilliant blue staining by Tris-saturated phenol method were
better than those obtained by two other methods. Hence Tris-saturated phenol method can be considered as a feasible method
for protein extraction from the yeast-like conidia of T. fuciformis.

Key words: Tremella fuciformis, yeast-like conidia, protein extraction method

中图分类号:

TS201

刘洋，朱涵予，许丹云，张丽姣，马爱民. 银耳芽孢总蛋白提取方法的建立[J]. 食品科学, doi: 10.7506/spkx1002-6630-201609007.

LIU Yang, ZHU Hanyu, XU Danyun, ZHANG Lijiao, MA Aimin. Development of an Efficient Method for Total Protein Extraction from Yeast-Like Conidia of Tremella fuciformis[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201609007.

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银耳芽孢总蛋白提取方法的建立

Development of an Efficient Method for Total Protein Extraction from Yeast-Like Conidia of Tremella fuciformis

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