Atomically precise gold nanoclusters are ideal model catalysts with well-defined compositions and tunable structures. Determination of the ligand effect on catalysis requires the use of gold nanoclusters with protecting ligands as the only variable. Two isostructural Au38 nanoclusters, [Au38(L)20(Ph3P)4]2+ (L = alkynyl or thiolate), have been synthesized by a direct reduction method, and they have an unprecedented face-centered cubic (fcc)-type Au34 kernel surrounded by 4 AuL2 staple motifs, 4 Ph3P, and 12 bridging L ligands. The Au34 kernel can be derived from the fusion of two fcc-type Au20 via sharing a Au6 face. Catalytic performance was studied with these two nanoclusters supported on TiO2 (1/TiO2 and 2/TiO2) as catalysts. The alkynyl-protected Au38 are very active (>97%) in the semihydrogenation of alkynes (including terminal and internal ones) to alkenes, whereas the thiolated Au38 showed a very low conversion (<2%). This fact suggests that the protecting ligands play an important role in H2 activation. This work presents a clear demonstration that catalytic performance of gold nanoclusters can be modulated by the controlled construction of ligand spheres.