红毛藻R-藻红蛋白的提取纯化及在Cu2+检测中的应用
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(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)

作者简介:

蔡思学(1997—),女,硕士生

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基金项目:

福建省自然科学基金项目(2022J01132845)


Extraction and Purification of R-phycoerythrin in Bangia fusco-purpurea and Its Application in Cu2+ Detection
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(1.College of Oceanology and Food Science, Quanzhou Normal University, Quanzhou 362000, Fujian;2.College of Food and Biological Engineering, Jimei University, Xiamen 361021, Fujian)

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    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Cu2+能使R-藻红蛋白发生荧光猝灭的特性,建立一种荧光检测Cu2+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(A620/A280) 的药品级R-藻红蛋白,其对Cu2+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Cu2+ 0.5~50 μmol/L范围内,相对荧光强度与Cu2+浓度之间存在良好的线性关系,最低检出限(S/N=3)0.06308 μmol/L。将此方法用于实际样品中Cu2+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Cu2+含量的检测有广泛的应用前景。

    Abstract:

    This paper used cellulase method to extract R-phycoerythrin and purified it. A fluorescence method for detecting the content of Cu2+ was established based on the fluorescence quenching of R-phythrocytin by Cu2+. 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 Cu2+. 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 Cu2+ concentration in the range of 0.5~50 μmol/L Cu2+, and the lowest detection limit (S/N=3) can reach 0.06308 μmol/L. This method was applied to the detection of Cu2+ 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 Cu2+ content in aquatic products has broad application prospects.

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蔡思学,李月,王芳,姜泽东,王力.红毛藻R-藻红蛋白的提取纯化及在Cu2+检测中的应用[J].中国食品学报,2022,22(9):276

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  • 收稿日期:2021-09-05
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  • 在线发布日期: 2022-10-26
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