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A dinuclear nickel compound that features vanillin (van) and acetate (OAc) bridging ligands, [Ni2(μ-van)2(μ-OAc)(NCS)3(H2O)]·5H2O 1, was synthesized and characterized using various techniques. Crystal structure analysis revealed a pair of Ni(II) ions bridged by an acetate anion through a μ2-η1:η1 coordination mode. At the same time, vanillin bridges both nickel(II) centers through phenolic oxygen, separating the two Ni ions by a distance of 3.001(7) Å. The temperature-dependent magnetism indicates weak antiferromagnetic behaviour in compound 1, following the Curie–Weiss law with Curie constant C = 1.0145 cm3 K mol−1 and Weiss constant = −4.1 K. Compound 1 was evaluated for its ability to mimic the catechol oxidase enzyme using 3,5-di-tert-butylcatechol as a model substrate with a Kcat value of 157.1 × 102 h−1. Furthermore, compound 1 was tested for its cytotoxicity against HepG2 cancer cells and DNA cleavage properties. [ABSTRACT FROM AUTHOR]