紫贻贝加工下脚料抗菌肽的分离及其稳定性研究  

Isolation and Stability Analysis of Antimicrobial Peptides from Mytilus edulis By-Products

作　　者：时光宇 张玥 潘渊博 赵莘芷 徐琼怡 杨玉霞 庄黛娜 SHI Guang-yu;ZHANG Yue;PAN Yuan-bo(School of Donghai Science and Technology of Zhejiang Ocean University,Zhoushan 316000,China)

机构地区：[1]浙江海洋大学东海科学技术学院,浙江舟山316000

出　　处：《浙江海洋大学学报（自然科学版）》2021年第1期9-15,共7页Journal of Zhejiang Ocean University：Natural Science

基　　金：国家级大学生创新创业训练计划项目(201810340038);浙江省自然科学基金(LY19C200007)。

摘　　要：以紫贻贝加工下脚料为原料,采用超声辅助乙酸粗提抗菌肽粗提物,筛选其抑菌活性敏感指示菌。在此基础上,以固相萃取、Sephadex G-25层析和半制备RP-HPLC对抗菌肽进行分离纯化及其稳定性分析。结果显示,金黄色葡萄球菌为最敏感菌株,获得的Sp40-2-4和Sp40-2-5组分抑菌活性显著高于抗菌肽粗提物(P<0.05),凝胶过滤色谱分析表明Sp40-2-4组分单一,相对分子质量约为1970.78 Da;Sp40-2-4组分具有一定的热稳定性(100℃、20 min,抑菌活性残留率72.42%)、反复冻融性(冻融次数≤10,抑菌活性残留率91.42%以上)和抗胃蛋白酶水解能力(抑菌活性残留率75.59%)。The crude antimicrobial peptides were extracted from Mytilus edulis by-product using ultrasonic-assisted acetate extraction,and the sensitive bacteria of the crude peptides were investigated.Using the solid-phase extraction,Sephadex G-25 chromatography and semi-preparative RP-HPLC,antimicrobial peptides were isolated and purified from crude peptides,and the stability of peptides with high purity and strong antimicrobial activity were further analyzed.The results showed that Staphylococcus aureus was the most sensitive strain,and fraction Sp40-2-4 and Sp40-2-5 had significantly higher antibacterial activity than the crude antimicrobial peptide(P<0.05).Then,a single chromatographic peak with relative molecular mass of approximately 1970.78 Da was detected in the Sp40-2-4 fraction by gel filtration chromatography(GPC).Meanwhile,the Sp40-2-4 fraction has certain thermostability(its residual rate of antimicrobial activity was up to 72.42%after heating at 100℃for 20 min),freeze-thaw stability(its residual rate of antimicrobial activity was higher than 91.42%when freeze-thaw cycles≤10),and anti-pepsin hydrolysis(its residual rate of antimicrobial activity was 75.59%after hydrolyzing by pepsin).The results provide a theoretical basis for the preparation and application of antimicrobial peptides in M.edulis by-products.

关 键 词：紫贻贝加工下脚料 抗菌肽 分离纯化 稳定性 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]