The effect of the surface morphology of gold-deposited quartz substrate on the enantioselective crystallization of leucine was investigated by using quartz crystal microbalance (QCM) and X-ray diffraction methods. The gold substrate was modified with a self-assembled monolayer with covalently attached D-leucine molecules. For a rough surface with root-mean-square (RMS) roughness of 17 nm, the QCM measurement showed a continuous decrease in frequency, while for a smooth surface with RMS roughness of 2 nm, essentially no change in frequency was observed. Nevertheless, leucine crystals with high crystallinity were clearly formed on the smooth surface.