波吉卵囊藻蛋白质提取方法比较及提取条件优化  

Comparison of Protein Extraction Methods and Optimization of Extraction Conditions for Oocystis borgei

作　　者：洪廷 任佳佳 张宁 李长玲[1,2,3] 黄翔鹄 HONG Ting;REN Jia-jia;ZHANG Ning;LI Chang-ling;HUANG Xiang-hu(College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,China;Shenzhen Research Institute,Guangdong Ocean University,Shenzhen 518000,China;Guangdong Research Center for Algae Culture and Applied Engineering,Zhanjiang 524025,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东海洋大学深圳研究院,广东深圳518000 [3]广东省藻类养殖及应用工程技术研究中心,广东湛江524025

出　　处：《广东海洋大学学报》2020年第3期56-64,共9页Journal of Guangdong Ocean University

基　　金：深圳市大鹏新区产业发展专项资金“活性微藻制品产业化关键技术的研究与示范”。

摘　　要：【目的】探究基于考马斯亮蓝染色法的波吉卵囊藻(Oocystis borgei)蛋白质提取方法,以快速测定波吉卵囊藻的蛋白质含量。【方法】以凯氏定氮法为基准,以考马斯亮蓝法测定经反复冻融法、超声波破碎法和碱提法提取的波吉卵囊藻(鲜品)蛋白含量,比较各提取方法的提取效果;通过单因素和响应面实验,考察碱提法中提取温度、NaOH浓度和提取时间对波吉卵囊藻蛋白提取率的影响。【结果】碱提法显著优于其他提取方法(P<0.01);碱提法中,各因素对波吉卵囊藻蛋白提取率的影响显著性由大到小依次为提取温度、NaOH浓度、提取时间;最佳提取条件:提取温度74.5℃、NaOH浓度0.35mol/L、提取时间69min,此时蛋白得率为1.726%,与模型预测值1.731%接近,以凯氏定氮法测定的藻体蛋白质含量为基准,蛋白提取率高达99.20%。【结论】碱提法是波吉卵囊藻(鲜品)蛋白的最佳提取方法,采用响应面法建立的模型可较好地预测提取温度、NaOH浓度和提取时间对波吉卵囊藻蛋白提取率的影响。【Objective】To explore the use of Coomassie blue rapid protein determination for different proteins extraction methods from Oocystis borgei to quickly determine the protein content of O.borgei.【Method】Using Kjeldahl method as standard,the Coomassie blue staining method was used to determine the protein content of O.borgei freshly extracted by repeated freeze-thaw method,ultrasonic disruption method or alkaline extraction method.The extraction effects of each method were compared.Through single-factor tests and response surface methodology,the effects of extraction temperature,NaOH concentration and extraction time on the extraction rate of O.borgei protein in alkaline extraction were investigated.【Results】The alkaline extraction method was significantly better than other extraction methods(P<0.01).In the alkaline extraction method,the effects of various factors on the extraction rate of O.borgei were in the following order:extraction temperature>NaOH concentration>extraction time.The optimal conditions were as follows:extraction temperature was 74.5℃;NaOH concentration was 0.35 mol/L;extraction time was 69 min.The yield rate of protein was 1.726%,which was closed to the model predicted value of 1.731%with Kjeldahl method as the standard and the protein extraction rate reached 99.2%.【Conclusions】Alkali extraction method is the best method for extracting O.borgei protein.The model established by response surface method can better predict the effects of extraction temperature,NaOH concentration and extraction time on the protein extraction rate of O.borgei.

关 键 词：波吉卵囊藻 蛋白提取 快速提取 考马斯亮蓝法 响应面法 

分 类 号：Q946.1[生物学—植物学] Q949.217