丰年虫水溶性蛋白的分离纯化及体外抗氧化性研究-食品科学.PDF

122 2012, Vol. 33, No. 15 食品科学 ※基础研究 聂 路，张建新 * ，李文学，刘 胐 (西北农林科技大学食品科学与工程学院，陕西 杨凌 712100) 目的：对丰年虫水溶性蛋白进行分离纯化，获得体外抗氧化活性最好的水溶性蛋白组分，并测定其分子 质量。方法：丰年虫水溶性粗蛋白经硫酸铵沉淀、透析、DEAE-Sepharose FF 离子交换层析和 Sephadex G-100 凝 胶层析分离纯化，对纯化后的蛋白质进行体外抗氧化活性的研究，利用 SDS电泳测定蛋白质分子质量。结 果：丰年虫水溶性蛋白经 DEAE-Sepharose FF 离子交换层析，梯度洗脱得到 4 个洗脱峰(峰 1～4) ，峰2 的体外抗 氧化活性最强。峰 2 经 Sephadex G-100 凝胶层析分离纯化得到 3 个洗脱峰(峰2-1～2-3) ，峰2-1 的体外抗氧化活性最 强。利用 SDS对峰 2-1 进行鉴定，得到单一蛋白条带，纯度较高，蛋白分子质量为 82.47kD 。体外抗氧化 活性比较，峰 2-1 远远强于水溶性粗蛋白。结论：蛋白组分峰 2-1 体外抗氧化活性最强，为单一蛋白组分，其分 子质量为 82.47kD 。 丰年虫；水溶性蛋白；分离纯化；抗氧化活性 Antioxidant Activity of Water-Soluble Protein Purified from Eubranchipus vernalis NIE Lu ，ZHANG Jian-xin* ，LI Wen-xue ，LIU Fei (College of Food Science and Engineering, Northwest AF University, Yangling 712100, China) Abstract ：Objective: To isolate and purify water-soluble protein with the highest antioxidant activity from Eubranchipus vernalis and determine its molecular weight. Methods: Crude water-soluble protein from Eubranchipus vernalis was extracted and purified by ammonium sulfate precipitation, salting out, ion-exchange column chromatography, DEAE-fast-flow Sepharose anion-exchange chromatography and Sephadex G-100 column chromatography. The antioxidant activities of different protein fractions were compared and their molecular weights were determined by SDS. Results: The prepared crude water- soluble protein was mainly composed of 4 fractions named as Peak-1, Peak-2, Peak-3 and Peak-4 in DEAE-Sepharose fast-flow column chromatography and Peak-2 revealed higher antioxidant activity than other 3 fractions. In addition, Peak-2 was mainly composed of 3 sub-fractions including Peak 2-1, Pea