Hou Feng, Miyakawa Takuya, Takeshita Daijiro, Kataoka Michihiko, Uzura Atsuko, Nagata Koji, Shimizu Sakayu, Tanokura Masaru
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Oct 1;68(Pt 10):1237-9. doi: 10.1107/S1744309112034951. Epub 2012 Sep 29.
(R)-3-Quinuclidinol is a useful chiral building block for the synthesis of various pharmaceuticals and can be produced from 3-quinuclidinone by asymmetric reduction. A novel 3-quinuclidinone reductase from Agrobacterium tumefaciens (AtQR) catalyzes the stereospecific reduction of 3-quinuclidinone to (R)-3-quinuclidinol with NADH as a cofactor. Recombinant AtQR was overexpressed in Escherichia coli, purified and crystallized with NADH using the sitting-drop vapour-diffusion method at 293 K. Crystals were obtained using a reservoir solution containing PEG 3350 as a precipitant. X-ray diffraction data were collected to 1.72 Å resolution on beamline BL-5A at the Photon Factory. The crystal belonged to space group P2(1), with unit-cell parameters a = 62.0, b = 126.4, c = 62.0 Å, β = 110.5°, and was suggested to contain four molecules in the asymmetric unit (V(M) = 2.08 Å(3) Da(-1)).
（R）-3-奎宁环醇是用于合成各种药物的有用手性构建单元，可通过不对称还原由3-奎宁环酮制得。一种来自根癌农杆菌的新型3-奎宁环酮还原酶（AtQR）以NADH作为辅因子催化3-奎宁环酮立体定向还原为（R）-3-奎宁环醇。重组AtQR在大肠杆菌中过表达，纯化后于293K使用坐滴气相扩散法与NADH一起结晶。使用含有PEG 3350作为沉淀剂的储液获得晶体。在光子工厂的BL-5A光束线上收集了分辨率为1.72Å的X射线衍射数据。该晶体属于空间群P2(1)，晶胞参数a = 62.0、b = 126.4、c = 62.0Å，β = 110.5°，且不对称单元中建议含有四个分子（V(M)= 2.08Å(3)Da(-1)）。
