鸿雁雁血多肽的制备及免疫调节作用的初步研究  

Preliminary Study on Preparation and Immunomodulatory Effect of Anser cygnoides Blood Protein Peptide

作　　者：王铮 赵宇 高晓晨 孙尧[1] 崔本海 高冷[1] WANG Zheng;ZHAO Yu;GAO Xiao-chen;SUN Yao;CUI Ben-hai;GAO Leng(School of Chemistry and Life Sciences,Changchun University of Technology,Changchun 130012,China;Jilin Institute of Ginseng Science,Changchun University of Traditional Chinese Medicine,Changchun 130012,China;Tongyu County Xianghai Cuicheng Dayan Breeding Co.,Ltd.,Baicheng 137200,China)

机构地区：[1]长春工业大学化学与生命科学学院,吉林长春130012 [2]长春中医药大学吉林省人参科学研究院,吉林长春130012 [3]通榆县向海崔成大雁养殖有限公司,吉林白城137200

出　　处：《食品工业科技》2020年第13期65-71,共7页Science and Technology of Food Industry

基　　金：吉林省科技发展计划项目(20190304086YY)。

摘　　要：本研究以酶解法提取雁血多肽,探究提取工艺对水解度的影响,以新鲜鸿雁血为原料,通过单因素实验和响应面试验优化提取工艺,并通过SDS-PAGE电泳测定分子量分布,同时采用小鼠脾淋巴细胞体外增殖检测实验,并以超滤法和凝胶过滤层析法对雁血多肽进行分离纯化。采用ELISA法检测细胞因子IL-2、IFN-γ、PGE2的含量。结果表明,最佳提取工艺为酶解温度47℃,pH7,加酶量4500 U/g,此时水解度为29.29%;复合蛋白酶酶解效果最优;雁血蛋白酶解前后在不同浓度下均可促进淋巴细胞的增殖活性;从雁血多肽中分离纯化出3种组分多肽;ACP1、ACP2、ACP3均可促进小鼠淋巴细胞分泌IL-2、IFN-γ,抑制分泌PGE2,且ACP2在100μg/mL浓度下效果最佳,由此可以判定雁血多肽具有一定的免疫调节作用,这为雁血多肽免疫机理的深度开发提供了理论依据。In this study,the polypeptide of Anser cygnoides blood was extracted by enzyme hydrolysis and the effect of extraction process on the degree of hydrolysis was investigated.Using fresh Anser cygnoides blood as raw material,the extraction process was optimized by single factor tests and response surface experiment.And the molecular weight distribution was determine by SDS-PAGE electrophoresis.At the same time,mouse spleen lymphocyte proliferation assay was used in this work.The polypeptide from Anser cygnoides blood was separated and purified by ultrafiltration and gel filtration chromatography.The contents of cytokines IL-2,IFN-γ and PGE2 were detected by ELISA.The results showed that the optimum extraction conditions were as follows:The hydrolysis temperature was 47℃,the pH was 7,the amount of enzyme was 4500 U/g and the hydrolysis degree was 29.29%.The enzyme hydrolysis effect of complex protease was the best.The proliferation of lymphocytes could be promoted at different concentrations before and after the hydrolysis of Anser cygnoides blood protein.Three component peptides were isolated and purified from the peptides of Anser cygnoides blood.ACP1,ACP2 and ACP3 could promote the secretion of IL-2 and IFN-γ by mouse lymphocytes and inhibit the secretion of PGE2.The effect of ACP2 was the best at the concentration of 100μg/mL.Thus,it could be determined that the Anser cygnoides blood protein polypeptide had a certain immunomodulatory effect.It would provide a theoretical basis for the deep development of immune mechanism of Anser cygnoides blood protein peptide.

关 键 词：鸿雁血 响应面法 小鼠脾淋巴细胞增殖 凝胶过滤层析 免疫调节 

分 类 号：TS201.1[轻工技术与工程—食品科学]