食品科学 ›› 2017, Vol. 38 ›› Issue (22): 95-101.doi: 10.7506/spkx1002-6630-201722015

• 生物工程 • 上一篇    下一篇

鸡蛋黄蛋白质水解产物制备及其对MC3T3-E1细胞增殖分化的影响

陈宏杰,金永国,马美湖   

  1. (华中农业大学食品科技学院,湖北?武汉 430070)
  • 出版日期:2017-11-25 发布日期:2017-11-03
  • 基金资助:
    公益性行业(农业)科研专项(201303084)

Preparation of Egg Yolk Protein Hydrolysate and Its Effect on Proliferation and Differentiation of MC3T3-E1 Cells

CHEN Hongjie, JIN Yongguo, MA Meihu   

  1. (College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China)
  • Online:2017-11-25 Published:2017-11-03

摘要: 鸡蛋黄经脱脂得到蛋黄蛋白质,直接以MC3T3-E1细胞增殖活力为指标,对水解条件进行优化,从而制备蛋黄蛋白质水解产物(egg yolk proteins hydrolysate,EYPH)并对其促MC3T3-E1细胞增殖分化的活性进行研究。结果表明:在对比胰蛋白酶与风味蛋白酶水解过程中,发现单一胰蛋白酶反应得到产物活性最好,胰蛋白酶反应的最佳水解条件为水解时间6?h、水解温度37?℃、酶与底物质量比1∶50、pH?8.0。EYPH使MC3T3-E1细胞增殖活力得到提高,为129.89%,细胞周期分析得到S期细胞增加了4.69%,这2?个方面证实EYPH对MC3T3-E1细胞的增殖活力有显著的提高。EYPH对MC3T3-E1细胞分化矿化有显著影响,碱性磷酸酶活力提高了9.1%;刺激骨钙素分泌,其含量上升2?mg/mL;形成更多的矿化结节。此外,EYPH还能刺激RUNX2特异性转录因子的基因表达,表达量提高了6.12倍。结果说明EYPH对成骨细胞MC3T3-E1增殖分化有显著的促进作用,因此,EYPH对于抗骨质疏松方面有较好的应用潜力,为开发其功能性食品提供一定的数据参考。

关键词: 蛋黄蛋白质水解产物, 水解条件, MC3T3-E1细胞, 增殖, 分化

Abstract: In this study, the hydrolysis conditions for preparing egg yolk protein hydrolysate (EYPH) from defatted egg yolk were optimized based on the viability of MC3T3-E1 cells when cultured in the presence of EYPH. Additionally, we also evaluated the promoting effect of EYPH on proliferation and differentiation of MC3T3-E1 cells. The results showed that the hydrolysis with trypsin alone yielded a product with stronger cell proliferation promoting activity than when it was used in combination with flavourzyme. The optimum hydrolysis conditions for trypsin were as follows: hydrolysis time, 6 h; temperature, 37 ℃; enzyme-to-substrate ratio, 1:50; and pH, 8.0. EYPH prepared under the optimized conditions increased the viability of MC3T3-E1 cells by 9.1% and the proportion of S-phase cells by 4.69% indicating that EYPH significantly increased the proliferation of MC3T3-E1 cells. EYPH significantly affected differentiation and mineralization of MC3T3-E1 cells as indicated by a 9.1% increase in alkaline phosphatase activity, an increase in osteocalcin secretion of 2 mg/mL, and the formation of more mineralized nodules of MC3T3-E1. Furthermore, EYPH also could stimulate the expression level of RUNX2 gene by 6.12 times. These finding showed that EYPH could significantly promote proliferation and differentiation in osteoblastic MC3T3-E1 cells. Therefore, EYPH has the potential to be used as a functional food ingredient for its anti-osteoporosis activity.

Key words: egg yolk protein hydrolysate, hydrolysis conditions, MC3T3-E1 cells, proliferation, differentiation

中图分类号: