乳酸克鲁维酵母表面展示系统研究  

Study on yeast surface display system of Kluyveromyces lactis

作　　者：姜巨全[1] 安吉一 商娜 刘文婷 郑迎迎 JIANG Juquan;AN Jiyi;SHANG Na;LIU Wenting;ZHENG Yingying(School of Life Sciences,Northeast Agricultural University,Harbin 150030,China;Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]中国科学院天津工业生物技术研究所,天津300308

出　　处：《东北农业大学学报》2024年第6期39-45,57,共8页Journal of Northeast Agricultural University

基　　金：国家自然科学基金联合基金重点项目(U23A20143);国家自然科学基金面上项目(32070031,31770051);天津市合成生物技术创新能力提升行动项目(TSBICIP-KJGG-013)。

摘　　要：酵母表面展示技术通过将目标蛋白锚定在细胞表面,广泛应用于疫苗制备、药物筛选、环境治理、菌株筛选等领域。为在乳酸克鲁维酵母中进行医药蛋白的高效生产并开展蛋白质工程研究,利用酿酒酵母来源的Sed1作为锚蛋白,His Tag作为检测靶标,在乳酸克鲁维酵母中进行表面展示系统的构建,并利用间接法免疫荧光标记和全细胞流式分析相结合的方法进行高通量表征。激光扫描共聚焦显微镜成像结果进一步证实了人血清白蛋白的成功表达。优化检测条件后,最高展示效率达89.8%。研究首次在乳酸克鲁维酵母中建立并系统优化了酵母表面展示系统,为在乳酸克鲁维酵母中开展高效蛋白质生产和蛋白质工程研究奠定了基础。Yeast surface display technology,which anchors target proteins on the cell surface,has been widely applied in the fields of vaccine preparation,drug screening,environmental management,and strain selection.In pursuit of high-efficiency production of therapeutic proteins and performing protein engineering in K.lactis,the study designed and implemented a surface display system utilizing Sed1 from Saccharomyces cerevisiae as an anchor and His Tag as a detection target.The highthroughput characterization technology that combined indirect immunofluorescence labeling and wholecell flow cytometry was employed to comprehensively evaluate the performance of the system.Laser scanning confocal microscopy imaging further confirmed the successful display of Human serum albumin as a model therapeutic protein.Following optimization of detection conditions,a remarkable maximum display efficiency of 89.8%was achieved.This study was the first time to establish and systematically optimize a yeast surface display system in K.lactis and laid the foundation for conducting protein engineering in K.lactis.

关 键 词：乳酸克鲁维酵母 酵母表面展示 优化 人血清白蛋白 

分 类 号：Q939.97[生物学—微生物学] Q789