摘要
目的:研究鹿皮蛋白酶解相关工艺,并对其酶解产物作出功效性评价。方法:以水解度作考察指标筛选酶制剂,运用响应面实验优化水解工艺参数,并研究水解液对RAW 264.7巨噬细胞增殖、NO分泌、LZM活力以及细胞因子TNF-α和IL-6分泌表达的影响。结果:表明以碱性蛋白酶酶解鹿皮蛋白的最适合条件是酶底比5%、pH=9、酶解时间5 h、酶解温度60℃,在此参数下水解度为48.64%。鹿皮蛋白酶解产物浓度为200.00μg/mL时,可以显著提高RAW 264.7细胞的NO浓度,达到最大值(10.82 mmol/mL);显著促进RAW 264.7细胞中的LZM活性;显著增加免疫因子TNF-α、IL-6的分泌(P<0.05)。结论:响应面法可较好地应用于鹿皮蛋白酶解工艺的优化,鹿皮蛋白酶解产物具有显著的免疫活性,研究结果可为鹿皮蛋白的高值化开发提供理论基础。
Objective:To study the deer skin proteolysis-related technology and evaluate the efficacy of its enzymolysis products.Methods:Enzyme preparations were screened using the degree of hydrolysis as an index of investigation,response surface experiments were used to optimize the hydrolysis process parameters,and the hydrolysate was used to stimulate RAW 264.7 macrophage proliferation,NO secretion,LZM activity,and cytokine TNF-α,IL-6 secretion study the impact of expression.Results:It showed that the most suitable conditions for digesting buckskin protein with alkaline protease is the ratio of enzyme to substrate 5%,pH=9,enzymolysis time 5 h,enzymolysis temperature 60℃,and the degree of hydrolysis under this parameter was 48.64%.When the concentration of buckskin proteolysis product is 200.00μg/mL,it can significantly increase the NO concentration of RAW 264.7 cells to the maximum value(10.82 mmol/mL),significantly promote the LZM activity in RAW 264.7 cells,significantly increase the immune factor TNF-Secretion ofαand IL-6(P<0.05).Conclusion:The response surface method can be applied to the optimization of the deer skin proteolysis process.The deer skin proteolysis products have significant immune activity.The results of this study provide an experimental basis for the high-value development of deer skin protein.
作者
汪涛
兰梦
张辉
郭焱
WANG Tao;LAN Meng;ZHANG Hui;GUO Yan(College of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China;Jilin Xinshui Technology Development Co.,Ltd.,Changchun 130000,China.)
出处
《食品科技》
CAS
北大核心
2020年第7期166-173,共8页
Food Science and Technology
关键词
鹿皮蛋白
酶解工艺
响应面法
巨噬细胞RAW
264.7
免疫活性
deer skin protein
enzymatic hydrolysis process
response surface method
macrophage RAW 264.7
immunological activity
