马氏珠母贝免疫活性肽的纯化与鉴定  

Purification and Identification of Immunomodulating Peptides from Enzymatic Hydrolysates of Pinctada martensi

作　　者：邓志程 张迪[1] 吉宏武[1,2,3,4] 曹文红[1] 苏伟明[1,2] DENG Zhi-cheng;ZHANG Di;JI Hong-wu;CAO Wen-hong;SU Wei-ming(College of food Science and Technology,Guangdong Ocean University, Zhanjiang 524088, China;Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Zhanjiang 524088,China;Marine Food Engineering and Technology Research Center of Guangdong Provence,Zhanjiang 524088, China;Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution, Zhanjiang 524088, China)

机构地区：[1]广东海洋大学食品科技学院,广东湛江524088 [2]广东省水产品加工与安全重点实验室,广东湛江524088 [3]广东省海洋食品工程技术研究中心,广东湛江524088 [4]广东普通高等学校水产品深加工重点实验室,广东湛江524088

出　　处：《广东海洋大学学报》2017年第4期78-86,共9页Journal of Guangdong Ocean University

基　　金：国家海洋公益性行业科研专项(201305018);广东省高等学校学科建设项目(2013CXZDA020);广东省部产学研合作专项资金项目(2013B090600155);广东海洋大学创新强校工程项目(GDOU2014050203)

摘　　要：为制备马氏珠母贝免疫活性肽,以马氏珠母贝全脏器为原料,利用体外模拟消化的方法对其进行酶解,然后通过超滤系统收集分子量小于3 ku的酶解液。采用细胞培养方法,以体外免疫活性为指标,利用Sephadex G-25凝胶柱层析、Capto Q强阴离子交换色谱等对马氏珠母贝酶解液进行分离纯化与筛选,获得2个具有免疫活性的肽段,利用电喷雾静电场离子阱串联质谱分别对2个活性肽序列进行测定。结果表明:一个肽的相对分子质量为245.0,肽序列为Ala-Arg/Pro-Met;另一个别肽的相对分子质量为274.0,肽序列为Val-Arg。在浓度0.3 mg/m L时,2种肽能显著促进脾淋巴细胞的增殖与提高巨噬细胞的吞噬能力,表明这2种肽均具有较好的免疫活性。To prepare immunomodulating peptides from Pinctada martensi entrails,in vitro simulatedgastrointestinal tract digestive method was employed for enzymatic hydrolysis,which was followed bypassing the digest through a multistep ultrafiltration system;the permeate(<3ku peptides)wascollected as the Pinctada martensi hydrolysate(PEP).The immunomodulating activities of thehydrolysates were determined by their effects on splenic lymphocytes and peritoneal macrophages.Thepeptide fractions which exhibited the highest activity were further purified using Sephadex G-25gelfiltration chromatography,Capto Q ion exchange chromatography and so on.The peptides wereidentified using ESI LTQ-Orbitrap mass spectrometry.Finally,two immunomodulating peptides wereidentified,and the amino acid sequences were Ala-Arg/Pro-Met and Val-Arg,whose molecular weightwere245.0and274.0,respectively.Therefore,the results demonstrated that vitro immunomodulating activities were greatly enhanced in the presence of0.3mg/mL purified peptides.

关 键 词：马氏珠母贝 免疫活性肽 分离纯化 氨基酸序列 

分 类 号：S931.4[农业科学—渔业资源]