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