大豆谷丙转氨酶的结构模型构建及其应用  

Construction and application of structural model of alanine aminotransferase in soybean

作　　者：林深 钟镕华 孟繁梅[1] 郑晓如 伍俭儿[1] 张尚宏[2] LIN Shen;ZHONG Rong-hua;MENG Fan-mei;ZHENG Xiao-ru;WU Jian-er;ZHANG Shang-hong(School of Life Science,Sun Yat-sen University,Guangzhou 510275;Biotechnology Research Center,Sun Yat-sen University,Guangzhou 510275,China)

机构地区：[1]中山大学生命科学学院,广州510275 [2]中山大学生物工程研究中心,广州510275

出　　处：《生物学杂志》2020年第4期26-30,共5页Journal of Biology

基　　金：国家自然科学基金项目(30270752);中山大学本科教学改革研究课题(33000-18832618)。

摘　　要：大豆是一种重要的固氮农作物,其种子萌发时具有较强的氨基酸代谢作用,谷丙转氨酶的活性较高。而谷丙转氨酶能催化谷氨酸和丙酮酸可逆反应生成α-酮戊二酸和丙氨酸,且这一反应在生物的氨基酸代谢调节中起到十分重要的作用。通过查阅数据库,获得大豆谷丙转氨酶的可能序列,并利用Discovery Studio软件对大豆谷丙转氨酶的蛋白质序列(NCBI序列号NP001237567.2)进行了同源建模,得到了大豆谷丙转氨酶的三维结构模型。也通过分子动力学探究该模型在不同浓度的缓冲液中的状态,得到了一个稳定的溶液态模型。通过对该谷丙转氨酶的三维结构模型的改造和优化也证明了大豆谷丙转氨酶添加组氨酸标记的可行性,为后续表达和分离纯化大豆谷丙转氨酶的工作进行了理论准备。Soybean is an important nitrogen-fixing crop having strong amino acid metabolism and high activity of alanine aminotransferase during seed germination.Alanine aminotransferase can catalyze the reversible reaction of glutamic acid and pyruvate to produce alpha-ketoglutarate and alanine.This reaction is essential in the metabolism of amino acid in organisms.In this study,the possible sequence of Glycine max glutamic-alanine aminotransferase was obtained through database.We built a 3-D homology model of the protein sequence of Glycine max alanine aminotransferase(NP001237567.2)with Discovery Studio,explored the conditions of the model in different concentrations of buffers with molecular dynamic,and obtained a stable model of solution state.The modification and optimization of the three-dimensional structure model of alanine aminotransferase prove the feasibility of adding histidine labelling to Glycine max alanine aminotransferase and make a theoretical preparation for the expression and separation of Glycine max alanine transaminase later.

关 键 词：谷丙转氨酶 大豆 同源建模 组氨酸标签 

分 类 号：Q55[生物学—生物化学]