Antibacterial coating is necessary to prevent biofilm-forming bacteria from colonising medical tools causing infection and sepsis in patients. The recent coating strategies such as immobilisation of antimicrobial materials and low-pressure plasma polymerisation may require multiple processing steps involving a high-vacuum system and time-consuming process. Some of those have limited efficacy and durability. Here, we report a rapid and one-step atmospheric pressure plasma polymerisation (APPP) of D-limonene to produce nano-thin films with hydrophobic-like properties for antibacterial applications. The influence of plasma polymerisation time on the thickness, surface characteristic, and chemical composition of the plasma-polymerised films was systematically investigated. Results showed that the nano-thin films deposited at 1 min on glass substrate are optically transparent and homogenous, with a thickness of 44.3 ± 4.8 nm, a smooth surface with an average roughness of 0.23 ± 0.02 nm. For its antimicrobial activity, the biofilm assay evaluation revealed a significant 94% decrease in the number of Escherichia coli (E. coli) compared to the control sample. More importantly, the resultant nano-thin films exhibited a potent bactericidal effect that can distort and rupture the membrane of the treated bacteria. These findings provide important insights into the development of bacteria-resistant and biocompatible coatings on the arbitrary substrate in a straightforward and cost-effective route at atmospheric pressure.