The development of a facile and reliable assay for arsenic (As) detection is crucial for its pollution control, which is a well-known heavy-metal pollutant in the environment. In this work, an efficient colorimetric method was established for As(V) detection based on a novel cuboid-shape Mn-Urea nanozyme (UMnzyme) with arsenic-enhanced oxidase-like activity. The UMnzyme was prepared through the coordination of Mn ions and carbonyl groups in urea, which possesses outstanding oxidase-like activity that can generate the superoxide anion free radical (O₂˙⁻) by catalyzing the dissolved O₂, triggering the 3,3′,5,5′-tetramethylbenzidine (TMB) chromogenic reaction. Furthermore, the free oxygen atoms in UMnzyme provide potential sites to bind with As, in which the formation of As–O will not destroy the morphology and structure of UMnzyme. The capture of As by UMnzyme results in the improvement of its oxidase-like activity; therefore, a colorimetric method for As(V) detection was developed with a linear range of 12.5–250 μg L⁻¹. The limit of detection was determined to be 2.4 μg L⁻¹ (S/N = 3), which is lower than the threshold (10 ppb) recommended by the World Health Organization. Moreover, the developed method was applied in the detection of As(V) in environmental water, and the recoveries are 89.5–103.6%. In short, this study provides an efficient and reliable strategy for As(V) detection based on a novel Mn-based nanozyme and a new idea for the reasonable design of Mn-based nanozymes with enhanced catalytic performance.