红毛藻R-藻红蛋白的提取纯化及在Cu^(2+)检测中的应用  

Extraction and Purification of R-phycoerythrin in Bangia fusco-purpurea and Its Application in Cu^(2+) Detection

作　　者：蔡思学 李月 王芳 姜泽东 王力 Cai Sixue;Li Yue;Wang Fang;Jiang Zedong;Wang Li(College of Oceanology and Food Science,Quanzhou Normal University,Quanzhou 362000,Fujian;College of Food and Biological Engineering,Jimei University,Xiamen 361021,Fujian)

机构地区：[1]泉州师范学院海洋与食品学院,福建泉州362000 [2]集美大学食品与生物工程学院,福建厦门361021

出　　处：《中国食品学报》2022年第9期276-284,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：福建省自然科学基金项目(2022J01132845)。

摘　　要：采用纤维素酶法提取红毛藻藻红蛋白,并对其进行纯化。利用Cu^(2+)能使R-藻红蛋白发生荧光猝灭的特性,建立一种荧光检测Cu^(2+)含量的方法。研究结果表明:当提取温度40℃、提取时间5 h、加酶量2.5%时,提取的R-藻红蛋白质量浓度为43.64 mg/mL。使用饱和度为60%的硫酸铵沉淀和DEAE Sepharose FF柱纯化,得到纯度为2.1(A_(620)/A_(280))的药品级R-藻红蛋白,其对Cu^(2+)有良好的选择性,最佳检测条件为:R-藻红蛋白质量浓度1.2 mg/mL、水浴温度30℃、反应时间15 min、缓冲液pH 7.0,在Cu^(2+)0.5~50μmol/L范围内,相对荧光强度与Cu^(2+)浓度之间存在良好的线性关系,最低检出限(S/N=3)0.06308μmol/L。将此方法用于实际样品中Cu^(2+)含量的检测,样品回收率为94.50%~106.03%。方法具有绿色安全、检测时间短、灵敏度高、选择性好等优势,对水产品中重金属Cu^(2+)含量的检测有广泛的应用前景。This paper used cellulase method to extract R-phycoerythrin and purified it.A fluorescence method for detecting the content of Cu^(2+)was established based on the fluorescence quenching of R-phythrocytin by Cu^(2+).The results of the study showed that the concentration of R-phycoerythrin extracted by the cellulase method was 43.64 mg/mL when the extraction temperature was 40℃,the extraction time was 5 hours,and the amount of enzyme was 2.5%.Purification of phycoerythrin by using 60%ammonium sulfate precipitation and DEAE Sepharose FF column to obtain pharmaceutical grade R-phycoerythrin with a purity of 2.1,and this R-phycoerythrin had good selectivity to Cu^(2+).The best detection conditions were R-phycoerythrin concentration 1.2 mg/mL,water bath temperature 30℃,reaction time 15 min,buffer pH 7.0.There was a good linear relationship between relative fluorescence intensity and Cu^(2+)concentration in the range of 0.5~50μmol/L Cu^(2+),and the lowest detection limit(S/N=3)can reach 0.06308μmol/L.This method was applied to the detection of Cu^(2+)content in actual samples,and the sample recovery rate was 94.50%~106.03%.This method has the advantages of green safety,short detection time,high sensitivity,good selectivity,Etc,and the detection of heavy metal Cu^(2+)content in aquatic products has broad application prospects.

关 键 词：R-藻红蛋白 红毛藻 提取纯化 荧光探针 Cu^(2+) 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程] O657.3[轻工技术与工程—食品科学与工程]