Semiconductor photocatalysis has recently emerged as a promising method for microbial inactivation. So far, quantum dots have generally been investigated as antibacterial suspension. Instead, here we demonstrate a InP/ZnS quantum dot nanoassembly film against both Gram-negative and Gram-positive bacteria. For effective operation in the solid phase, a thin layer of ZnS shell was grown on InP QD and the native long-chain ligand of stearic acid was replaced with sulfide that led to a high quantum yield of superoxide generation as 4.9%. QDs are assembled onto solid surfaces through sequential dip coating of positively charged poly(diallyldimethylammonium chloride) and negatively charged QDs. These QD nanoassemblies demonstrate growth inhibition against Escherichia coli and multidrug-resistant Staphylococcus aureus under illumination. Interestingly, such an approach can be directly applied to irregular surfaces, as well. This study unveils the potential of the nanoengineering of QDs for antibacterial coatings.