食品科学

• 生物工程 • 上一篇    下一篇

仿刺参肠道组织酶解工艺优化及肽段分析

王 颖1,2,张 健2,3,王茂剑2,3,*,王共明4,李 倩1,2,刘 昕2,侯志刚1,2   

  1. 1.上海海洋大学食品学院,上海 200000;2.山东省海洋资源与环境研究院,山东 烟台 264006;
    3.山东省海洋生态修复重点实验室,山东 烟台 264006;4.山水海产有限公司,山东 烟台 264006
  • 发布日期:2014-12-15

Optimal Preparation and Characterization of Peptides from Enzymatic Hydrolysis of Apostichopus japonicus Intestinal Tissue

WANG Ying1,2, ZHANG Jian2,3, WANG Mao-jian2,3,*, WANG Gong-ming4, LI Qian1,2, LIU Xin2, HOU Zhi-gang1,2   

  1. 1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 200000, China;
    2. Shandong Marine Resource and Environment Research Institute, Yantai 264006, China;
    3. Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Yantai 264006, China;
    4. Shan Shui Co. Ltd. Shandong Province, Yantai 264006, China
  • Published:2014-12-15

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。

关键词: 仿刺参肠道组织, 酶解, 响应面, 超滤, 过氧化氢损伤

Abstract:

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.

Key words: Apostichopus japonicus intestinal tissue, enzymolysis, response surface analysis, ultrafiltration, hydrogen peroxide-induced injury

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