›› 2013, Vol. 25 ›› Issue (6): 0-1367.

• 食品科学 •    

鸡蛋清中溶菌酶的分离提取

傅冰1,季秀玲2,俞汇颖2,魏云林2
  

  1. 1丽水职业技术学院 环境工程学院, 浙江 丽水 323000;2昆明理工大学 生命科学与技术学院,云南 昆明 650500
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2013-11-25 发布日期:2013-11-25

Preliminary study on isolation and extraction of lysozyme from egg white

FU Bing;JI Xiu\|ling;YYU Hui\|ying;WEI Yun\|lin   

  1. 1Branch of Environment Engineering, Lishui Vocational & Technical College,Lishui 323000, China;2 Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-11-25 Published:2013-11-25

摘要: 采用二次硫酸铵沉淀和离子交换层析透过法分离溶菌酶,并用聚丙烯酰胺凝胶电泳(SDS\|PAGE)检测其纯度,研究鸡蛋清中溶菌酶的分离提取工艺。鸡蛋清原液用Tris\|HCl缓冲液稀释5倍,4℃下静置6 h,经50%硫酸铵沉淀后收集上清,再经80%硫酸铵收集沉淀后溶于pH 90的Tris\|HCl后经Q\|Sepharose FF阴离子交换层析直接在透过液中一步就可分离得到SDS\|PAGE电泳纯的溶菌酶,酶比活由14413 U·mg-1提高到2 235 U·mg-1,酶活提高超过15倍,回收率为1576%。通过硫酸铵两级沉淀后只需收集Q\|Sepharose FF阴离子交换层析的透过液即可分离得到溶菌酶,同时还可一次性制备其他中性和酸性蛋白,简化了工艺,降低了生产成本。

关键词: 鸡蛋清, 溶菌酶, 离子交换层析, 分离

Abstract: Lysozyme was purified by two\|step ammonium sulfates precipitation and anion\|exchange chromatography to study the separation processing of lysozyme from egg white. The purity of lysozyme was assayed by sodium dodecyl sulfate\|polyacrylamide gel electrophoresis (SDS\|PAGE). Egg white was diluted with five times volume of Tris\|HCl buffer and kept at 4℃ for 6 h. The supernatant was collected from precipitation with 50% saturation ammonium sulfate and subjected to precipitation with 80% saturation ammonium sulfate. The precipitant was dissolved into pH 90 Tris\|HCl and applied onto Q\|Sepharose FF anion\|exchange chromatography. High purity lysozyme was directly obtained from permeate. The specific activity of lysozyme was increased nearly 15 folds from 14413 U·mg-1 to 2 235 U·mg-1 with recovery ratio of 1576%. Not only lysozyme can be easily purified by two\|step ammonium sulfates precipitation and Q\|sepharose FF anion\|exchange chromatography but also other proteins such as acidic proteins can be easily purified with the same procedure. This procedure can simplify the separation process and reduce the cost in lysozyme separation.

Key words: egg white, lysozyme, anion\|exchange chromatography, separation