不同蛋白酶海参酶解液的制备及其体外免疫调节活性  

Preparation of Hydrolysates of Sea Cucumber with Different Proteases and the in vitro Immunomodulatory Activities

作　　者：杨军楠 栾俊家 崔方超 王金厢 励建荣 李英美 李学鹏 YANG Jun-nan;LUAN Jun-jia;CUI Fang-chao;WANG Jin-xiang;LI Jian-rong;LI Ying-mei;LI Xue-peng(School of Food Science and Engineering,Bohai University,Jinzhou 121013,Liaoning,China;Dalian Food(Jinzhou)Co.,Ltd.,Jinzhou 121200,Liaoning,China;Provincial and Ministerial Collaborative Innovation Center for Key Technologies of Marine Food Intensive Processing,Dalian Un)

机构地区：[1]渤海大学食品科学与工程学院,辽宁锦州121013 [2]达莲食品(锦州)有限公司,辽宁锦州121200 [3]大连工业大学海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034

出　　处：《食品研究与开发》2023年第11期43-52,共10页Food Research and Development

基　　金：辽宁省海洋经济发展专项“海参精深加工关键技术研发及产业化”(辽财指环[2022]33号)。

摘　　要：以海参体壁为原料,采用5种商业用酶(中性蛋白酶、碱性蛋白酶、胰蛋白酶、木瓜蛋白酶、胃蛋白酶)对其进行酶解,测定酶解液的水解度和分子量分布,并重点分析分子量小于3 kDa超滤组分的氨基酸组成、结构特性及其对巨噬细胞的免疫调节活性。结果表明:中性蛋白酶对海参体壁的水解能力最高,反应300 min水解度达17.18%;中性蛋白酶酶解液中小于3 k Da超滤组分(SCH-N-I)占96.33%,必需氨基酸高达89.52 mg/g,高于其余4种蛋白酶酶解液。紫外光谱和荧光光谱结果显示,中性蛋白酶酶解液超滤组分SCH-N-I较其它酶解液超滤组分具有较高的表面疏水性、紫外吸光度和内源荧光强度,且最大紫外吸收波长和最大荧光峰向高波长方向偏移,表明其可能具有更高的免疫生物活性。RAW264.7巨噬细胞免疫活性实验结果显示,当SCH-N-I浓度为200μg/mL时,巨噬细胞表现出良好的存活率[(121.79±4.98)%]及较高的吞噬力(0.1077±0.0010)和NO释放量[(15.39±0.33)μmol/L];SCH-N-I的免疫活性高于其他4种蛋白酶酶解液超滤组分。The body wall of sea cucumber was hydrolyzed by five commercially available enzymes(neutral protease,alkaline protease,trypsin,papain,pepsin),respectively.The degree of hydrolysis(DH)and molecular weight distribution of the hydrolysates were determined,and the amino acid composition and structural properties of the fraction in the ultrafiltrate with molecular weight less than 3 kDa and its regulation of macrophages were investigated.The results showed that neutral protease had the strongest hydrolysis capacity(DH=17.18%in 300 min).The fraction less than 3 kDa(SCH-N-I)accounted for 96.33%and the essential amino acids hit 89.52 mg/g in the hydrolysate prepared with neutral protease,which were higher than those in the hydrolysates yielded by the rest four proteases.According to the ultraviolet(UV)spectra and fluorescence spectra,SCH-N-I had higher surface hydrophobicity,UV absorption,and endogenous fluorescence intensity than fractions in ultrafiltrates yielded by other enzymes,and exhibited a red shift of the maximum absorption wavelength and the maximum intrinsic fluorescence,indicating that it might have higher immunobiological activity.The results of the macrophage immunoactivity assay showed that when the concentration of SCH-N-I was 200μg/mL,macrophages demonstrated high survival rate[(121.79±4.98)%],strong phagocytosis(0.1077±0.0010),and high nitric oxide release[(15.39±0.33)μmol/L]in the LPS-induced immunization model.SCH-N-I had higher immunoactivity than fractions in ultrafiltrates prepared with the rest four proteases.

关 键 词：海参 酶解液 氨基酸组成 结构特性 免疫调节活性 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]