Because of their biocompatibility and biodegradability in vivo, natural polysaccharides are effective nanocarriers for delivery of active ingredients or drugs. Moreover, bioactive polysaccharides, such as tea, Ganoderma lucidum, and Momordica charantia polysaccharides (TP, GLP, and MCP), have antibacterial, antioxidant, antitumor, and antiviral properties. In this study, tea, Ganoderma lucidum, and Momordica charantia polysaccharide nanoparticles (TP-NPs, GLP-NPs, and MCP-NPs) were prepared via the nanoprecipitation approach. When the ethanol to water ratio was 10:1, the diameter of the spherical polysaccharide nanoparticles was the smallest, and the mean particle size of the TP-NPs, GLP-NPs, and MCP-NPs was 99 ± 15, 95 ± 7, and 141 ± 9 nm, respectively. When exposed to heat, increased ionic strength and pH levels, the nanoparticles exhibited superior stability and higher activity than the corresponding polysaccharides. In physiological conditions (pH 7.4), the nanoparticles underwent different protein adsorption capacities in the following order: MCP-NPsTP-NPsGLP-NPs. Moreover, the 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical, and superoxide anion radical scavenging rates of the nanoparticles were increased by 9-25% as compared to the corresponding polysaccharides. Compared to the bioactive polysaccharides, the nanoparticles enhanced antimicrobial efficacy markedly and exhibited long-acting antibacterial activity.