Tomita Takeo, Yin Lulu, Nakamura Shugo, Kosono Saori, Kuzuyama Tomohisa, Nishiyama Makoto
Biotechnology Research Center, The University of Tokyo, Japan.
Faculty of Information Networking for Innovation and Design, Toyo University, Tokyo, Japan.
FEBS Lett. 2017 Jun;591(11):1611-1622. doi: 10.1002/1873-3468.12667. Epub 2017 May 25.
The NADP -dependent glutamate dehydrogenase from Corynebacterium glutamicum (CgGDH) is considered to be one of the key enzymes in the industrial fermentation of glutamate due to its high glutamate-producing activity. We determined the crystal structure of CgGDH complexed with NADP and 2-iminoglutarate. Among six subunits of hexameric CgGDH-binding NADP , only four subunits bind 2-iminoglutarate in a closed form, while the other two are in an open form. In the closed form, 2-iminoglutarate is bound to the substrate-binding site with the 2-imino group stacked by the nicotinamide ring of the coenzyme, suggesting a prehydride transfer state in a hypothesized reaction scheme with the imino intermediate. We also conducted MD simulations and provide insights into the extreme preference for the glutamate-producing reaction of CgGDH.
The atomic coordinate and structure factors have been deposited in the RCSB PDB database under the accession number 5GUD.
谷氨酸棒杆菌的依赖烟酰胺腺嘌呤二核苷酸磷酸(NADP)的谷氨酸脱氢酶(CgGDH)因其高谷氨酸生产活性,被认为是谷氨酸工业发酵中的关键酶之一。我们确定了与NADP和2-亚氨基戊二酸复合的CgGDH的晶体结构。在结合NADP的六聚体CgGDH的六个亚基中,只有四个亚基以封闭形式结合2-亚氨基戊二酸,而另外两个处于开放形式。在封闭形式中,2-亚氨基戊二酸与底物结合位点结合,其2-亚氨基基团与辅酶的烟酰胺环堆叠,这表明在具有亚氨基中间体的假设反应方案中处于预氢化物转移状态。我们还进行了分子动力学模拟,并深入了解了CgGDH对谷氨酸生产反应的极端偏好。
原子坐标和结构因子已存入RCSB蛋白质数据库(PDB),登录号为5GUD。