摘要
目的:优化胰蛋白酶(PTN 6.0S)酶解酪蛋白酸钠制备高免疫活性肽的酶解条件。方法:以四甲基偶氮唑盐微量酶反应比色法(MTT法)测得的酶解产物对小鼠脾细胞增殖程度(刺激指数SI)及氨基氮(AN)含量为指标,通过单因素试验及响应曲面法优化酶解条件。结果:在pH7.0、温度55.0℃、时间15h、底物质量浓度3.5g/100mL、酶与底物比3.8:1000的酶解条件下,酶解产物的SI值达到0.184±0.002,AN含量为(0.795±0.012)g/L。结论:以酶解产物的活性为指标,可直接获得免疫活性最高的乳源多肽的酶法制备条件。
Objective： To optimize the tryptic（PTN 6.0S） hydrolysis conditions of sodium caseinate for preparing peptides with high-immune activity.Methods： One-factor-at-a-time experimental design combined with response surface methodology was employed systematically to develop optimal hydrolysis conditions of sodium caseinate.The effect of hydrolyzed sodium caseinate on proliferation degree（stimulation index,SI） and ammonia nitrogen（AN） of mouse spleen cells was measured by tetrazolium salt-based colorimetric（MTT） assay.Results： The optimal hydrolysis conditions were substrate concentration of 3.5 g/100 mL,enzyme/substrate ratio of 3.8：1000,hydrolysis time of 15 h,hydrolysis temperature of 55.0 ℃ and hydrolysis pH of 7.0.Under the optimal hydrolysis conditions,the maximum SI was achieved to be 0.184±0.002 and the ammonia nitrogen content was up to（0.795±0.012） g/L.Conclusion： Enzymatic hydrolysis is a better way to prepare high-immune activity peptides from sodium caseinate.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2011年第19期174-179,共6页
Food Science
基金
江苏省高校自然科学基础研究项目(08KJD550004)
扬州市-扬州大学合作基金项目(YZ2008089)
关键词
酪蛋白酸钠
免疫肽
MTT法
响应曲面法
sodium caseinate
immune peptides
tetrazolium salt-based colorimetric（MTT） assay
response surface methodology
