天然产物研究与开发 ›› 2016, Vol. 28 ›› Issue (12): 1880-1885.doi: 10.16333/j.1001-6880.2016.12.005

• 研究论文 • 上一篇    下一篇

超高压酶解酪蛋白及其产物在动物细胞培养中的应用

靳冬武1,2,魏锁成2,冯玉萍1,2,马忠仁1,2,3,郑荣2,李明生1,2,3*   

  1. 1 西北民族大学生物工程与技术国家民委重点实验室;2西北民族大学生命科学与工程学院;3甘肃省动物细胞工程技术研究中心,兰州 730030
  • 出版日期:2016-12-31 发布日期:2017-03-31

Ultra-high Pressure Enzymatic Hydrolysis of Casein and its Application in Animal Cell Culture

JIN Dong-wu1,2,WEI Suo-cheng2,FENG Yu-ping1,2,MA Zhong-ren1,2,3,ZHENG Rong2,LI Ming-sheng1,2,3*   

  1. 1 The Key Bio-engineering and Technology Laboratory of Nationality Commission Northwest University For Nationalities;2Life Science and Engineering College of Northwest University for Nationalities; 3Engineering Technology Research Center for Animal Cell GanSu,Lanzhou 730030,China
  • Online:2016-12-31 Published:2017-03-31

摘要: 以牦牛酪蛋白为底物,游离氨基酸含量为指标,在超高压条件下分别采用胰蛋白酶、胃蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和胰酶六种不同的酶水解酪蛋白并筛选出最佳用酶,并在单因素实验的基础上采用正交实验确定最佳用酶水解酪蛋白的最佳工艺条件。最后探讨水解产物在细胞培养中的应用。结果表明,胰酶水解效果最好,用其在底物浓度15%,E/S=1:5、水解压力150 MPa,温度为45 ℃,pH 7.5条件下水解4 h后水解产物中游离氨基酸含量高达52.18%。酶解产物对BHK-21细胞没有细胞毒性,而具有促生长作用。这为酪蛋白的进一步开发利用和蛋白质水解产物在细胞培养中的应用奠定基础。

关键词: 超高压, 酶解, 酪蛋白, 氨基酸含量, 细胞培养

Abstract: Yak casein was used as the substrate,the content of free amino acids was used as evaluation index,six different enzymes,including trypsin,pepsin,papain,neutrase,alcalase and pancreatin,were used to hydrolysis casein under ultra-high pressure.The optimal process of casein hydrolysis was optimized by single factor analysis and orthogonal experiments.At the same time,the application of hydrolysates in animal cell culture was investigated.The results showed that pancreatin was more suitable for the hydrolysis of yak casein,the optimal process was as follows:substrate concentration 15%,E/S=1:5,hydrolysis pressure 150 MPa,temperature was 45 ℃,pH 7.5 and hydrolyzed for 8 h.Under these conditions,the content of free amino acids of hydrolysate was determined to be 52.18%.The hydrolysates did not appear any toxic to BHK-21 cells when supplemented in the medium,and cells growth well.The study can be served for the further exploitation and utilization of the yak casein and the application of protein hydrolysates in animal cell culture.

Key words: ultra-high pressure, enzymatic hydrolysis, casein, amino acids content, cell culture

中图分类号: 

Q814.9