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.%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6.69×105cells/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.
展开▼
