This paper proposes the preparation and formula analysis of anti-biofouling Titania–polyurea (TiO2–SPUA) spray coating, which uses nano-scale antibacterial and photocatalytic agents, titanium dioxide, to construct regularly hydrophobic surface texture on the polyurea coating system. Through formulating analysis of anti-biofouling performance, it is found the causal factors include antibacterial TiO2, surface wettability and morphology in order of their importance. The most optimized formula group is able to obtain uniform surface textures, high contact angle (91.5°), low surface energy (32.5 mJ/m2), and strong hardness (74 A). Moreover, this newly fabricated coating can effectively prevent Pseudomonas aeruginosa and biofilm from enriching on the surface, and there is no toxins release from the coating itself, which makes it eco-friendly, even after long-time exposure. These studies provide insights to the relative importance of physiochemical properties of Titania–polyurea spray coatings for further use in marine, as well as bio medical engineering.