天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (9): 1627-1633.doi: 10.16333/j.1001-6880.2018.9.026

• 开发研究 • 上一篇    下一篇

酶解大豆蛋白的制备工艺研究及其在细胞培养中的应用研究

郑荣1,2,3,何云富3,张邵博3,李倬3,冶眩青3,程浩3,丁功涛3,冯玉萍1,2,3,马忠仁1,2,3,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西北民族大学甘肃省动物细胞工程技术研究中心; 3西北民族大学生命科学与工程学院,兰州,730030
  • 出版日期:2018-10-08 发布日期:2018-10-10
  • 基金资助:

    中央高校项目(31920150231);甘肃省科技计划(17YFI WA166);甘肃省科技计划(1504WK CA094);兰州市科技计划(2014-1-158)

Preparation Process of Enzymatic Hydrolysis of Soy Protein and Its Application in Cell Culture  

ZHENG Rong1,2,3,HE Yun-fu3,ZHANG Shao-bo3,LI Zhuo3,YE Xuan-qing3,CHENG Hao3,DING Gong-tao3,FENG Yu-ping1,2,3,MA Zhong-ren1,2,3,LI Ming-sheng1,2,3*   

  1. 1Key Laboratory of Bioengineering & Biotechnology of State Ethnic Affair Commission,Northwest University for Nationalities; 2Engineering Technology  Research Center for Animal Cell; 3Life Science and Engineering College of Northwest University for Nationalities,Lanzhou 730030,China
  • Online:2018-10-08 Published:2018-10-10

摘要: 采用两种非动物源性蛋白酶水解大豆蛋白,以水解度为指标,对不同比例4种酶两两组合测试其水解度,选出水解最佳用酶;在单因素试验基础上,采用正交试验对酶解大豆蛋白的工艺条件进行优化,并将产物进行动物细胞的生长进行检测。结果表明:复合蛋白酶与碱性蛋白酶以等比例进行混合,其水解效果最佳;水解最佳工艺为:温度60  ℃、pH 值7.0 和酶/底物( E/S) = 1∶4(质量比),水解时间为4 h,其水解度为43.7%。将酶解产物培养BHK-21细胞,并与Hypep1510进行对比,细胞生长状态良好,其细胞密度在120 h时最大,为3.96×105 cells/mL,与Hypep1510相比,最大细胞密度并无明显差异,其倍增时间为26.2 h,高于阴性对照组与阳性对照组,且细胞形态基本未发生改变,表明酶解大豆蛋白所的产物能够缩短细胞倍增的时间,且具有一定的促细胞生长作用,为其在动物细胞大规模培养中的应用提供了理论依据。

关键词: 酶解, 非动物源性, 大豆蛋白, 细胞培养

Abstract: Two kinds of non-animal-derived protease were used to hydrolyze soy protein,the degree of hydrolysis was taken as an index,the degree of hydrolysis of four kinds of enzymes in different proportions was tested in combination with each other,and the optimal enzymes for hydrolysis were selected.On the basis of single factor test,The orthogonal test was used to optimize the enzymatic hydrolysis of soybean protein,and the enzymolysis product was added to the medium to detect the growth of animal cells.The results showed that the optimum hydrolysis conditions were as follows:temperature 60  ℃,pH value 7.0 and enzyme/substrate (E/S) = 1∶4 (Mass ratio),hydrolysis time of 4 h,the degree of hydrolysis of 43.7%.BHK-21 cells were cultured and cultured in the medium,and the cell growth was good compared with that of Hypep1510.The cell density reached the maximum of 3.96×105 cells/mL at 120 h with a doubling time of 26.2 h.Which were higher than that of the negative control group and the positive control group,indicating that enzymolysised soy protein products can shorten the cell doubling time and promote cell growth,which provides a theoretical basis for its application in animal cell culture.

Key words: enzymatic hydrolysis, animal origin free, soy protein, cell culture  

中图分类号: 

R965 Q813.1