Study of Catalytic Mechanism of Hydrogen Activation by Hydrogenase using Neutron Crystal Structure Analysis

2010年首次尝试将硫酸盐还原菌接种于氘代培养基中制备完全氘代的[NiFe]氢化酶，但未能获得足够量的蛋白样品用于结晶试验。在2011-2012年，尝试在氘代缓冲溶液中使用氘代沉淀剂结晶普通（非氘代）氢化酶。我们已经成功地制备了体积为4 mm^3的单晶，该晶体在100 K下用日本原子能研究所的中子束给出了分辨率为5.0 A的中子衍射图。此外，还找到了[NiFe]氢化酶大单晶低温衍射实验的一些参数。

The perfectly deuterated[NiFe] hydrogenase was tried to prepare by inoculating the sulfate-reducing bacterium in the deuterated media in 2010.However, it was not successful to obtain enough amount of the protein sample for crystallization trial. In 2011-2012, the ordinary(not deuterated) hydrogenase was tried to crystallize using deuterated precipitant in deuterated buffer solution. We have succeeded in preparation of a single crystal with a volume of 4 mm^3.This crystal gave a neutron diffraction pattern at 5.0 Aresolution at 100 K using the neutron beam at Japan Atomic Energy Research Institute. In addition, some parameters for diffraction experiment for large single crystals of[NiFe] hydrogenase at cryo-temperature have been found.