降解苯丙氨酸的工程益生菌构建及其喂养苯丙酮尿症小鼠的效果分析  

Construction of engineered probiotic for degrading phenylalanine and analysis of its effect on feeding phenylketonuria mice

作　　者：马欣 付加芳 林赓[1,2,3] 张佩佩 曹广祥 MA Xin;FU Jiafang;LIN Geng;ZHANG Peipei;CAO Guangxiang(Biomedical Sciences College&Shandong Medicinal Biotechnology Centre,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117,China;NHC Key Laboratory of biotechnology drugs(Shandong Academy of Medical Sciences),Jinan 250117,China;Key Lab for Rare&Uncommon Diseases of Shandong Province,Jinan 250117,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)生物医学科学学院(省医药生物技术研究中心),山东济南250117 [2]国家卫健委医药生物技术重点实验室(山东省医学科学院),山东济南250117 [3]山东省罕少见病重点实验室,山东济南250117

出　　处：《食品与发酵工业》2024年第17期75-81,共7页Food and Fermentation Industries

基　　金：山东第一医科大学青年培育基金项目(202201-004);山东省重点研发计划项目(2021ZDSYS27)。

摘　　要：苯丙酮尿症(phenylketonuria,PKU)是一种常染色体隐性遗传病,低蛋白饮食是目前临床治疗的主要方法,而采用工程益生菌治疗尚处于研发阶段。为了探究工程益生菌对PKU的治疗效果,该研究基于大肠埃希菌Nissle 1917(EcN)菌株,构建了一株表达苯丙氨酸裂解酶的工程益生菌EcN-PAL。通过λ-Red同源重组系统在EcN中导入经优化设计的苯丙氨酸裂解酶基因,并通过体外降解实验和小鼠喂养实验验证其功能性。实验结果表明,工程益生菌EcN-PAL成功表达苯丙氨酸裂解酶,并且可降解苯丙氨酸和有效降低PKU小鼠的血液苯丙氨酸浓度。该研究成功构建的工程益生菌EcN-PAL对苯丙氨酸有良好的降解效果,动物实验证实可有效治疗PKU小鼠,为用于PKU患者的益生菌治疗奠定了基础。Phenylketonuria(PKU)is an autosomal recessive disease.A low-protein diet is the main treatment for PKU,and engineered probiotic is still in the stage of research and development.To explore the therapeutic effect of engineered probiotic on PKU,an engineered probiotic strain EcN-PAL expressing phenylalanine lyase was constructed based on Escherichia coli Nissle 1917(EcN).The optimized codons of phenylalanine lyase gene was integrated into the genome of EcN by using theλ-Red homologous recombination system,and its function was verified by in vitro degradation experiment and mouse feeding experiment.Results showed that the phenylalanine lyase was successfully expressed in EcN-PAL,and the phenylalanine can be effectively degradated by EcN-PAL in vitro.The blood phenylalanine concentration in PKU mice showed a significant decrease when the mice was treated with EcN-PAL.The engineered probiotic E.coli EcN-PAL exhibited good ability to degrade phenylalanine,and could effectively treat PKU mice,laying a foundation for probiotic therapy in patients with PKU.

关 键 词：苯丙酮尿症 λ-Red同源重组系统 工程益生菌 苯丙氨酸降解 苯丙酮尿症小鼠 

分 类 号：Q789[生物学—分子生物学] R596[医药卫生—内科学] TS201.3[医药卫生—临床医学]