天然产物研究与开发 ›› 2016, Vol. 28 ›› Issue (9): 1443-1448.doi: 10.16333/j.1001-6880.2016.9.018

• 开发研究 • 上一篇    下一篇

高压酶解卵清蛋白的工艺优化及其在BHK-21细胞培养中的初步应用

张健1,2,李明生2,3,靳冬武1,张磊1,郑荣1,马忠仁1,2,冯玉萍1,2*   

  1. 1 西北民族大学生命科学与工程学院;2 西北民族大学生物工程与技术国家民委重点实验室;3 甘肃省动物细胞工程技术研究中心,兰州 730030
  • 出版日期:2016-09-30 发布日期:2017-03-24

High Pressure Enzymatic Hydrolysis of Egg-white Protein and Its Preliminary Application in BHK-21 Cell Culture

ZHANG Jian1,2,LI Ming-sheng2,3,JIN Dong-wu1,ZHANG Lei1,ZHENG Rong1,MA Zhong-ren1,2,FENG Yu-ping1,2*   

  1. 1 Life Science and Engineering College of Northwest University for Nationalities;2 Key Laboratory of  Bioengineering & Biotechnology of State Ethnic Affair Commission,Northwest University for Nationalities; 3 Engineering Technology Research Center for Animal Cell,Lanzhou 730030,China
  • Online:2016-09-30 Published:2017-03-24

摘要: 采用高压技术水解卵清蛋白,以水解度为指标,对6种酶的水解效果进行比较,筛选出最佳水解用酶;在单因素实验的基础上,采用正交实验的方法对高压酶解卵清蛋白的工艺进行优化,并对酶解产物在BHK-21细胞培养中的应用进行了初步探讨。结果表明:6种酶中胰酶的水解效果最好,水解最佳工艺为:压力120 MPa、温度50 ℃、pH值7.0和酶/底物(E/S)=1:2.5(质量比),水解度达到45.13%;用酶解产物培养BHK-21细胞,细胞生长形态良好,在培养168 h后,试验组细胞最大密度达到6.69×105 cells/mL,是对照组的1.56倍,细胞数量增大了33.45倍,说明卵清蛋白酶解产物有明显的促细胞生长效果,为其在细胞培养中的应用提供了理论依据。

关键词: 高压, 酶解, 卵清蛋白, 细胞培养

Abstract: This research utilized high-pressure technique to enzymatic hydrolyzed egg-white protein.By comparing the hydrolysis effect of six enzymes based on the degree of hydrolysis indicator,we determined the best enzyme for the research.The process of egg-white protein proteolysis was optimized by orthogonal experiments based on single factor experiment,and hydrolyzes used for BHK-21cell culture was tested.The result showed that the hydrolysis effect of pancreatin was the best among that of the six enzymes.The optimum hydrolysis was:120 MPa,50 ℃,pH7.0 and enzyme/substrate (E/S) =1:2.5(mass ratio).The degree of hydrolysis could reach to 45.13%.The hydrolyzes was used to culture BHK-21 cells and the cell morphology was good.The maximum amount of proliferation was 6.69×105 cells/mL,which was 1.56 times of the control group and the number of cells increased 33.45 times during 168 h of culture.This study showed that enzymatic hydrolysis of egg-white protein significantly promoted cell proliferation,providing a theoretical basis for the egg-white protein peptide used as a new cell culture protein additive.

Key words: high pressure, enzymatic hydrolysis, egg white protein, cell culture

中图分类号: 

Q813.1