小黄鱼免疫肽制备条件的响应面优化

摘要

Objective： To optimize process conditions for the hydrolysis of small yellow croaker by trypsin（PTN 6.0S） to prepare peptides with high immuno-activity.Methods： the proliferative effect（expressed as stimulation index,SI） of small yellow croaker hydrolysate on mouse spleen cells was assessed by the MTT assay.Based on the index,single-factor experiments followed by response surface analysis were carried out for the optimization of hydrolysis conditions.Results： The optimal hydrolysis conditions for preparing immune peptides were hydrolysis at 41.66 ℃ for 18.24 h at a substrate concentration of 6 g/100 mL and an enzyme-to-substrate mass ratio of 3.13‰.Under these conditions,the predicted SI of small yellow croaker hydrolysate was 0.387.When the temperature,time and substrate concentration were modified as 42 ℃,18.2 h and 3‰,the actual SI was 0.369± 0.003,which was slightly lower than the predicted value.Thus,appropriate process conditions were achieved.%优化胰蛋白酶（PTN 6.0S）酶解小黄鱼制备免疫肽的制备条件。MTT法检测酶解产物对小鼠脾细胞的增殖程度,并以脾细胞增殖指数（SI）为指标,通过单因素试验及响应曲面法优化酶解条件。结果显示：在温度41.66℃、时间18.24h、底物质量浓度6g/100mL、酶与底物的质量比（酶底比）3.13‰条件下,模拟得出多肽产物的SI值为0.387。验证实验发现,在温度42℃、时间18.2h、底物质量浓度6g/100mL、酶底比3‰条件下,SI值为0.369±0.0 0 3,略低于模拟值,所得条件较为理想。