摘要
以仿刺参肠道组织为原料,优化酶解工艺条件,研究梯级肽对小鼠原代巨噬细胞过氧化氢损伤的保护作用。采用单因素试验及响应面试验分析法对酶解工艺条件进行优化,利用超滤法获取不同截留分子质量的肽段,采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)比色法检测细胞活力。结果表明:通过响应面优化得最佳条件为:酶解温度50.10℃、酶解时间2.84 h、加酶量2.639万U时水解度可达41.82%。通过超滤制备,分别得到分子质量小于650、650~1 000 u与1 000~10 000 u肽段,各肽段均能显著提高过氧化氢损伤细胞的细胞活力值,所获肽段对过氧化氢损伤巨噬细胞具有显著保护作用,其中小于650 u肽段更佳。
This work reports the application of single factor experimental design and response surface methodology to optimize conditions for enzymatic hydrolysis of the intestinal tissue of Apostichopus japonicas and the protective effect of the resulting peptides on primary macrophages of mice from injury caused by hydrogen peroxide. Hydrolysates were fractionated by ultrafiltration membranes with different cut-off molecular weights(MWCO). The cell viability was detected by MTT(3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide) assay. Results showed that the highest degree of hydrolysis of 41.82% was attained after 2.84 h of hydrolysis at 50.10 ℃ with 26 390 U of papain. Peptides with molecular mass less than 650, 650–1 000 and 1 000–10 000 u were separated by ultrafiltration, and they all could significantly increase the viability of primary cells and show a protective effect against hydrogen peroxide-induced injury. Furthermore, the peptide with molecular mass less than 650 u showed better protective capacity.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2014年第23期182-187,共6页
Food Science
基金
国家海洋局海洋公益性行业科研专项(201205027
201105029)
山东省现代农业产业技术体系刺参产业创新团队建设项目(SDAIT-08)
海洋经济创新发展区域示范项目(20130125)
水生动物营养与饲料泰山学者岗位经费资助项目(TS 200651036)
关键词
仿刺参肠道组织
酶解
响应面
超滤
过氧化氢损伤
Apostichopus japonicus intestinal tissue
enzymolysis
response surface analysis
ultrafiltration
hydrogen peroxide-induced injury
