摘要
为提升表达A型口蹄疫病毒(FMDV)结构蛋白大肠杆菌的稳定性,以大肠杆菌活性为评价指标,在单因素试验基础上采用Box-Behnken响应面法优化冻干保护剂组合。利用Box-Behnken结果进行回归模型拟合,得到最佳冻干保护剂配方为3.192%(质量分数,下同)山梨醇、3.094%谷氨酸钠、6.924%乳糖、3%脱脂奶粉和5%海藻糖,并对配方进行了验证。为分析真空冷冻干燥菌种的稳定性,将冻干及未冻干的菌种置于25℃和37℃放置不同时间,重溶后测定大肠杆菌的活性和表达蛋白的水平,结果显示,在25℃和37℃条件下,冻干样品相比于未冻干样品显示出更好的活性,表明冷冻干燥可以提高表达A型FMDV结构蛋白大肠杆菌的储存时长和稳定性。本研究为提升原核表达系统的稳定性和储存期提供了新方案。
To enhance the stability of Escherichia coli expressing the structural protein of foot-and-mouth disease virus(FMDv)typeA.Using Escherichia coliactivity as the evaluation index,Box-Behnken response surface methodology was used to optimize the combination of freeze-drying protectants based on single factor experiments.Using Box-Behnken results for regression model fitting,the optimal freeze-drying protectant formula was obtained as 3.192%sorbitol,3.094%monosodium glutamate,6.924%lactose,3%skim milk powder,and 5%trehalose,and the formula was validated.To analyze the stability of vacuum freeze-dried bacterial strains,freeze-dried and non-freeze-dried strains were placed at 25℃and 37℃for different periods of time.After reconstitution,the activity and protein expression levels of Escherichia coli were measured.The results showed that freeze-dried samples exhibited better activity compared to non-freeze-dried samples at 25℃and 37℃,indicating that freeze-drying can improve the storage time and stability of Escherichia coli expressing FMDV type A structural protein.This study provides a new approach to enhance the stability and storage period of prokaryotic expression systems.
作者
郭伟
刘瑞鹏
张佳甜
尉嘉锡
郭慧琛
孙世琪
马全英
滕志东
王晓强
GUO Wei;LIU Ruipeng;ZHANG Jiatian;YU Jiaxi;CUO Huichen;SUN Shiqi;MA Quanying;TENG Zhidong;WANG Xiaoqiang(School of Biological and Pharmaceutical Engineering,Lanzhou Jiaotong University,Lanzhou 730070,China;School of Chemistry and Chemical Engineering,Lanzhou Jiaotong University,Lanzhou 730070,China;State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine,Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of A gricultural Sciences,Lanzhou 730000,China)
出处
《中国兽医科学》
北大核心
2025年第7期973-981,共9页
Chinese Veterinary Science
基金
中国博士后科学基金项目(2023M733819)
国家成都农业科技中心地方财政专项(NASC2024KR06)
兰州市人才创业创新项目(2023-RC-3)。
