Position: Home > Articles > Purification of Lysozyme from Egg White by Combination of Polyethylene Glycol Precipitation and Aqueous Two-phase Extraction
Biotechnology Bulletin
2017
(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
作 者：
闻崇炜;赵烨清;石莉;欧阳臻
单 位：
江苏大学药学院
关键词：
聚乙二醇4000;沉淀法;双水相法;溶菌酶
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。
译 名：
Purification of Lysozyme from Egg White by Combination of Polyethylene Glycol Precipitation and Aqueous Two-phase Extraction
作 者：
WEN Chong-wei;ZHAO Ye-qing;SHI Li;OUYANG Zhen;School of Pharmacy,Jiangsu University;
关键词：
polyethylene glycol 4000;;precipitation;;aqueous two-phase extraction;;lysozyme
摘 要：
This work aims to establish a new technology of purifying lysozyme from egg white based on the combination of polyethyleneglycol(PEG)precipitation and aqueous two-phase extraction. The results showed that 98.1% of non-lysozyme proteins were removed bythe precipitation when PEG 4000 was added into pretreated egg white until its mass fraction reached 16%. Then,the supernatant of PEGprecipitation was collected and mixed with saturated(NH4)2SO4 solution until the mass fraction of this salt reached 4.32%. Therefore,theaqueous two-phase system of PEG/(NH4)2SO4 was established and most of lysozyme could be transferred into the top phase. It was foundthat 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 thespecific activity of lysozyme were 70.2% and 25 000 U/mg,respectively. In conclusion,this purification technology was simple,accurate,safe and reliable,and could be used for large-scale production of lysozyme.
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