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

2010年，尝试在氘化培养基中接种硫酸盐还原菌制备完全氘化[NiFe]氢化酶。然而，没有成功地获得足够数量的蛋白质样品进行结晶试验。2011-2012年，尝试用氘沉淀剂在氘缓冲溶液中结晶普通（未氘化）氢化酶。我们成功地制备了体积为4 mm^3的单晶。利用日本原子能研究所的中子束，该晶体在100 K下给出了5.0倍分辨率的中子衍射图。此外，还获得了[NiFe]氢化酶大单晶在低温下的衍射实验参数。

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.