High-efficient and low-cost catalyst on hydrogen isotope separation between hydrogen and water is an essential factor in industrial application for water detritiation in the future fusion power plants. In the past studies, impregnated nickel catalysts exist the problems of agglomeration and low Ni utilization for vapor phase catalytic exchange (VPCE). Therefore, to solve this problem, in-situ grown Ni-based catalysts derived from layered double hydroxide (LDH) were fabricated and first applied in VPCE. Its uniquely layered structure and steady homogenous highly dispersed metallic phase in LDH-derived catalysts enhanced the dynamic contact between gaseous hydrogen and vaporous water in catalyst surface, leading to the higher activity and quicker dissociative chemisorption in throughout H-D exchange process. Additionally, we find the dissociative chemisorption of HDO and H2 happened on the Ni (111) or NiO surface forming chemisorbed H(ads), D(ads) and H(ads). The results highlight that both of Ni2+ and Ni0 possess catalytic activities for VPCE.