水解乳蛋白的制备及在细胞培养中的初步应用

Preparation of Lacto-protein Hydrolysate and Study on Its Preliminary Application in Cell Culture

采用正交法优化复合酶(胰酶和中性蛋白酶)水解凝乳酶干酪素制备水解乳蛋白(lacto-protein hydrolysate,LPH)的制备工艺,通过BHK和Vero细胞的培养效果检测水解乳蛋白的促细胞生长作用。结果显示水解乳蛋白的最佳制备工艺为:胰酶∶中性蛋白酶=2∶1(质量比),40℃,pH 7.0,酶/底物(E/S)=1∶5(质量比)水解6 h。所得产物氨基氮含量3.21 g/L,多肽含量35.63 g/L,游离氨基酸含量14.17 mg/mL,收率达40.20%。BHK和Vero细胞培养24 h,细胞密度均为各自空白的2.0倍,分别为Hyclone产品的1.0倍和0.86倍;培养48h,分别为各自空白的5.0倍和4.0倍,均为Hyclone产品的0.88倍。因此,该工艺简单,耗时短,制备得水解乳蛋白对BHK和Vero细胞生长有明显的促进作用。

In this study, complex enzyme （panereatin and neutral protease） hydrolyzed rennet casein was used to prepare laeto-protein hydrolysate. The process was optimized by orthogonal experiments. The effects of promoting eell growth of the laeto-protein hydrolysate were assayed by cultivating BHK and Vero eells. The results showed that the optimal processes of preparing laeto-protein hydrolysate was ：panereatin：neutral protease = 2：1 （mass ratio） ,40 ℃, pH 7.0, en- zyme/substrate （E/S） = 1： 5 （ mass ratio）, hydrolysis time of 6 h. In the hydrolyzed product, the eontent of amino nitro- gen was determined to be 3.21 g/L, peptide was 35.63 g/L, free amino acids was 14. 17 mg/mL, the yield was 40. 20%. In the 24 h lacto-protein hydrolysate cultured BHK and Vero cells ,the cell densities were 2.0 times to respec- tive blank control, 1.0 and 0.86 times to Hyclone products ;In the 48 h laeto-protein hydrolysate cultured BHK and Vero cells,the cell densities were 5.0 times and 4.0 times to respective blank control;and both 0.88 times to Hyelone prod- ucts. Based ：these experimental results, it was concluded that the optimal preparation process was simple and short time- consuming;the laeto-protein hydrolysate promoted cell growth for BHK and Vero cells.