红米稻过氧化物还原酶OsPrx3增强秀丽隐杆线虫抗氧化功能  

Red Rice Peroxiredoxin OsPrx3 Enhances the Resistance to Oxidative Stress in Caenorhabditis elegans

作　　者：汪海洋 王喆皓 黄卉 沈焕然 宋小雨 王智勇 卢延克 周志 方庆 WANG Haiyang;WANG Zhehao;HUANG Hui;SHEN Huanran;SONG Xiaoyu;WANG Zhiyong;LU Yanke;ZHOU Zhi;FANG Qing(College of Biological and Food Engineering,Hubei Minzu University,Enshi 445000,China)

机构地区：[1]湖北民族大学生物与食品工程学院,湖北恩施445000

出　　处：《食品科学》2023年第6期197-204,共8页Food Science

基　　金：硒食品营养与健康智能技术湖北省工程中心开放基金项目(PT082209)。

摘　　要：目的:红米稻中OsPrx3基因预编码一个过氧化物还原酶,探究该基因原核细胞表达产物能否增强动物抗氧化的功能。方法:分子克隆OsPrx3基因,采用大肠杆菌表达目的蛋白,对摄入目的蛋白的模式动物-秀丽隐杆线虫(Caenorhabditis elegans),鉴别运动能力、体长、抗氧化应激能力、体内活性氧与脂肪含量以及与氧化相关的基因表达水平等。结果:借助大肠杆菌细胞成功表达红米稻过氧化物还原酶基因OsPrx3,分离目的蛋白,体外实验显示该蛋白具有较高酶活性,5 min内1μg/mL OsPrx3对H_(2)O_(2)相对清除率可达40%以上;与对照相比,摄入目的蛋白的秀丽隐杆线虫,运动能力和体长虽无明显变化,但抵抗外部氧化应激的能力却在氧化处理的15 min内明显增强,而且体内活性氧、细胞脂肪含量显著降低;另外,部分调节氧化的关键基因表达明显上调。结论:基因工程策略成功表达红米稻过氧化物还原酶OsPrx3基因,其产物OsPrx3体外具有较高抗氧化活性,秀丽隐杆线虫摄入该蛋白能显著增强抗氧化能力。Objective:To determine whether the prokaryotic expression product of the red rice OsPrx3 gene,which encodes a peroxiredoxin,can enhance oxidative stress resistance in animals.Methods:OsPrx3 was cloned and expressed in Escherichia coli.After ingestion of the target protein,the animal model Caenorhabditis elega was evaluated for its motility,body length,oxidative stress resistance,reactive oxygen species(ROS)and fat contents,and oxidation-related gene expression levels.Results:This gene was cloned and expressed in E.coli successfully,and the purified expressed protein had high antioxidant activity,which at 1μg/mL scavenged more than 40%of H_(2)O_(2) in five minutes.Ingestion of the target protein significantly increased the resistance to oxidative stress within 15 minutes after oxidative treatment and reduced ROS and cellular lipid content in C.elegans compared to the control group,although there were no significant changes in motility or body length.Additionally,the expression levels of several critical antioxidant genes were evidently increased in the treated C.elegans.Conclusion:The OsPrx3 protein had high in vitro antioxidant potency,and when ingested by C.elegans,significantly enhanced the resistance to oxidative stress.

关 键 词：过氧化物还原酶 红米稻 秀丽隐杆线虫 抗氧化 

分 类 号：Q81[生物学—生物工程]