※基础研究食品科学 2016, Vol.37, No.07 33
仿刺参体壁中抗菌肽的分离及抑菌活性
孟春英1,2，王茂剑2,3，张 健2,3,*，王共明2，刘 昕2，井月欣2，王 婷1,2
（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）
摘 要：从仿刺参体壁中分离纯化出抗菌肽并研究其抑菌活性。以鲜活仿刺参体壁为原料，采用5%乙酸浸提法进行粗提，以抑菌活性为指标进行分离。经超滤系统超滤，大孔吸附树脂柱层析分离纯化，采用酶标仪比浊法检测各组分对大肠埃希氏菌（Escherichia coli）、肠炎沙门氏菌（Salmonella eternitidis）、金黄色葡萄球菌（Staphylococcusaureus）和枯草芽孢杆菌（Bacillus subtilis）的抑菌活性。采用Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis，Tricine-SDS-PAGE）检测抗菌肽分子质量。结果表明：通过超滤分离，分别得到分子质量＜1、1～5 kD和5～10 kD组分，其中分子质量＜1 kD组分的抑菌活性最高，选用该组分继续进行大孔吸附树脂柱层析分离，得抑菌活性最高的组分，其Tricine-SDS-PAGE结果表现为单一条带，分子质量约为4.35 kD。
关键词：仿刺参体壁；抗菌肽；分离纯化；抑菌活性；酶标仪比浊法
Isolation and Activity of Antimicrobial Peptide from Body Wall of Apostichopus japonicas
MENG Chunying1,2, WANG Maojian2,3, ZHANG Jian2,3,*, WANG Gongming2, LIU Xin2, JING Yuexin2, WANG Ting1,2
(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)
Abstract: The antimicrobial peptide was obtained from the body wall of Apostichopus japonicas and its activity was evaluated. The crude peptide was extracted from the body wall with 5% acetic acid and subjected to bioassay-guided fractionation for antimicrobial activity. The purification was completed by ultra-filtration and chromatographic separation with macroporous resin. A turbidimetric method was applied to evaluate the inhibitory activity of each sample against bacteria including Escherichia coli, Salmonella eternitidis, Staphylococcus aureus and Bacillus subtilis. By using Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE), the molecular weight of the purified antimicrobial peptide was measured. The results showed that three components with different molecular weights < 1 kD, 1–5 kD and 5–10 kD were obtained after ultra-filtration. The component with molecular weight of < 1 kD had the strongest antibacterial activity, and then it was chosen for further chromatographic separation with macroporous resin column. The subcomponent F3 with the highest antimicrobial activity was obtained and its molecular weight was identified as 4.35 kD.
Key words: Apostichopus japonicas body wall; antimicrobial peptides; separation and purification; antimicrobial activity; turbidimetric method
DOI:10.7506/spkx1002-6630-201607007
中图分类号：TS254.1 文献标志码：A 文章编号：1002-6630（2016）07-0033-06引文格式：
孟春英, 王茂剑, 张健, 等. 仿刺参体壁中抗菌肽的分离及抑菌活性[J]. 食品科学, 2016, 37(7): 33-38. DOI:10.7506/spkx1002-6630-201607007. //m.kkreddy.com *通信作者：张健（1980—），男，助理研究员，博士，研究方向为海洋食品科学。E-mail：zjsd408@163.com
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