聚乙二醇沉淀联用双水相萃取法纯化蛋清溶菌酶的研究  

Purification of Lysozyme from Egg White by Combination of Polyethylene Glycol Precipitation and Aqueous Two-phase Extraction

作　　者：闻崇炜[1] 赵烨清 石莉[1] 欧阳臻[1] 

机构地区：[1]江苏大学药学院,镇江212013

出　　处：《生物技术通报》2017年第5期89-93,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(81573529);江苏大学高级人才启动基金(1281370001);江苏省普通高校研究生实践创新计划项目(SJLX15_0512)

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36.6μg PEG 4000,生物安全性较高。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 （ NH4 ） 2SO4 solution until the mass fraction of this salt reached 4.32%. Therefore, the aqueous two-phase system of PEG/（ NH4 ） 2SO4 was established and most of lysozyme could be transferred into the top phase. It was found that 96.34% of the freeze-dried top phase was lysozyme, and the rest was PEG 4000, no any other proteins existed. The recovery rate and the specifie activity of lysozyme were 70.2% and 25 000 U/mg, respeetively. In conclusion, this purification technology was simple, accurate, safe and reliable, and eould be used for large-scale production of lysozyme.

关 键 词：聚乙二醇4000 沉淀法 双水相法 溶菌酶 

分 类 号：Q55[生物学—生物化学]