拟南芥中一种新的参与蔗糖代谢活动的酶蛋白的表达与鉴定  

Expression and identification of a new enzyme protein involving sucrose metabolizing in Arabidopsis thaliana

作　　者：沈健 岳智亮[1] 潘延云[1] 

机构地区：[1]河北农业大学生命科学学院,河北保定071000

出　　处：《河北农业大学学报》2017年第5期60-65,共6页Journal of Hebei Agricultural University

基　　金：河北省自然科学基金项目(C2017204095);河北省高等学校科学技术研究项目(ZD2017039)

摘　　要：为了检测拟南芥AtPGH基因—一个推测为糖苷水解酶的基因的生化活性,利用酵母真核表达系统对该基因进行了体外表达,经纯化后获得AtPGH-His重组蛋白。进一步利用3.5-二硝基水杨酸法和间苯二酚法分别检测了该重组蛋白的蔗糖酶与蔗糖合酶活性。结果发现:AtPGH-His重组蛋白既有蔗糖酶活性,也有蔗糖合酶活性。比对拟南芥已知的蔗糖酶基因和蔗糖合酶基因,发现AtPGH与这2个家族的基因序列同源性均很低。推断AtPGH是拟南芥中一个新的具有双重酶活性的蔗糖代谢酶。AtPGH,a kind of predicted glycoside hydrolase in Arabidopsis thaliana,was detected for the biochemical activity.Firstly,the heterologous AtPGH-His protein was express by Pichia pastoris and purified using Ni-Sephaore.Secondly,the activities of sucrase and sucrose synthase of the recombinant protein were detected by 3.5-dinitrosalicylic acid method and resorcin method,respectively.The results showed that AtPGH-His recombinant protein had both sucrase and sucrose synthase activity.Blast results showed that AtPGH has very low sequence homology with the sucrase gene and sucrose synthase gene known to Arabidopsis thaliana.The results showed that AtPGH might be a new sucrose-metabolizing enzyme with double enzyme activity in Arabidopsis thaliana.

关 键 词：拟南芥 酵母表达 蔗糖酶 蔗糖合酶 

分 类 号：Q943[生物学—植物学]