聚乙二醇沉淀联用双水相萃取法纯化蛋清溶菌酶的研究 - 生物技术通报.pdf

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN ( ) 2017, 33 5 :89-93 聚乙二醇沉淀联用双水相萃取法纯化蛋清溶菌酶的研究 闻崇炜  赵烨清  石莉  欧阳臻 （江苏大学药学院，镇江 212013） 摘 要 ： 研究以聚乙二醇 （Polyethylene Glycol，PEG）沉淀法联用PEG- 硫酸铵双水相萃取体系纯化蛋清溶菌酶的工艺。结 果表明，向预处理的鸡蛋清液中加PEG 4000 至质量分数为16% 时，可选择性沉淀除去蛋清中98.1% 的杂蛋白，随后向上清液中 加硫酸铵溶液至其质量分数为4.32%， 可以构建PEG- 硫酸铵双水相体系，分离上相即得高纯度溶菌酶。该法所得产物中96.34% 为溶菌酶，其余为PEG 4000，不含有其它杂蛋白，溶菌酶总回收率达70.2%， 比活为25 000 U/mg。该法简便易行，易于放大，每 毫克精制溶菌酶中仅残留36.6 μg PEG 4000，生物安全性较高。 关键词 ： 聚乙二醇4000 ；沉淀法 ；双水相法 ；溶菌酶 DOI ：10.13560/j.cnki.biotech.bull.1985.2017.05.013 Purification of Lysozyme from Egg White by Combination of - Polyethylene Glycol Precipitation and Aqueous Two phase Extraction WEN Chong-wei ZHAO Ye-qing SHI Li OUYANG Zhen （School of Pharmacy ，Jiangsu University ，Zhenjiang 212013 ） Abstract: This work aims to establish a new technology of purifying lysozyme from egg white based on the combination of polyethylene glycol（PEG）precipitation and aqueous two-phase extraction. The results showed that 98.1% of non-lysozyme proteins were removed by the precipitation when PEG 4000 was added into pretreated egg white until its mass fraction reached 16%. Then，the supernatant of PEG precipitation was collected and mixed with saturated（NH ）SO solution until the mass fraction of this salt reached 4.32%. Therefore，the 4 2 4 aqueous two-phase system of PEG/（NH ）SO was established and most of lysozyme could be transferred into the top phase. It was found 4 2 4 that 96.34%