天然产物研究与开发 ›› 2014, Vol. 26 ›› Issue (1): 100-104.

• 开发研究 • 上一篇    下一篇

水解乳蛋白的制备及在细胞培养中的初步应用

田伟1,2,冯玉萍1,2,李明生1,2,崔梦楠1,靳冬武2,龙仕和1,马忠仁1,2*   

  1. 1 西北民族大学生命科学与工程学院;2 甘肃省动物细胞工程技术研究中心,兰州 730030
  • 出版日期:2014-01-31 发布日期:2014-12-15

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

TIAN Wei1,2, FENG Yu-ping1,2, LI Ming-sheng1,2, CUI Meng-nan1, JIN Dong-wu2, LONG Shi-he1, MA Zhong-ren1,2*   

  1. 1 College of Life Science and Engineering,Northwest University for Nationalities;2 Gansu Engineering Research Center for Animal Cell,Lanzhou 730030,China
  • Online:2014-01-31 Published:2014-12-15

摘要: 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48 h,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。

关键词: 水解乳蛋白, 制备, 细胞培养, 应用

Abstract: In this study,complex enzyme (pancreatin and neutral protease) hydrolyzed rennet casein was used to prepare lacto-protein hydrolysate.The process was optimized by orthogonal experiments.The effects of promoting cell growth of the lacto-protein hydrolysate were assayed by cultivating BHK and Vero cells.The results showed that the optimal processes of preparing lacto-protein hydrolysate was:pancreatin:neutral protease=2:1(mass ratio),40 ℃,pH 7.0,enzyme/substrate (E/S) =1:5 (mass ratio),hydrolysis time of 6 h.In the hydrolyzed product,the content of amino nitrogen 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 respective blank control,1.0 and 0.86 times to Hyclone products;In the 48 h lacto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 Hyclone products.Based these experimental results,it was concluded that the optimal preparation process was simple and short timeconsuming;the lactoprotein hydrolysate promoted cell growth for BHK and Vero cells.

Key words: lacto-protein hydrolysate, preparation, cell culture, application

中图分类号: 

Q813.1