© 2020 John Wiley & Sons, Ltd.A novel Ni(II) complex containing 2-fluorobenzoate and 3-hydroxypyridine ligands was synthesized and characterized using elemental analysis, Fourier transform infrared (FT-IR) spectroscopy and single-crystal X-ray diffraction. FT-IR peaks show that the carboxylate group from 2-fluorobenzoate has a monodentate coordination mode. The coordination environment around the Ni2+ ion is of distorted octahedral geometry. The octahedral geometry consists of a total of four oxygen atoms from two carboxylate groups of two fluorobenzoate anions, two water molecules and two nitrogen atoms from two 3-hydroxypyridine ligands. The complex shows excellent inhibitory effects against some metabolic enzymes. Ki values for the complex were found as 108.17 ± 25.63, 124.88 ± 36.20, 28.11 ± 2.87, 20.95 ± 5.65 and 32.63 ± 9.67 μM against human carbonic anhydrase I, human carbonic anhydrase II, α-glycosidase, acetylcholinesterase and butyrylcholinesterase, respectively. In addition, geometry optimization and vibration frequencies were calculated, and single point energy was studied based on optimization. Experimental and theoretical data were compared. The B3LYP/6-31G(d,p) basis set was used for all calculations.