The efficacy of entomopathogenic fungi as pest control agents is constrained by both their physiological state and external environmental factors. This study identified synergists capable of enhancing the insecticidal activity of Beauveria bassiana (Bb) and investigated the underlying synergistic mechanisms. Our results found that among 6 potential synergists, polyethylene glycol 400 (PEG) and trehalose significantly improved Bb's lethality against Hyphantria cunea larvae, with PEG demonstrating the most pronounced effect. PEG treatment markedly increased Bb spore adhesion and germination rates, while spore hydrophobicity and growth rates remained unaffected. Moreover, PEG-treated spores exhibited higher thermal tolerance compared to untreated ones. In the Bb + PEG treatment group, the hemocyte count, encapsulation and melanization activities, and the expression of related regulatory genes were significantly lower than those in the Bb treatment group. Additionally, pathogen recognition, signal transduction, and humoral immunity effector genes expression were markedly suppressed in the Bb + PEG group. A significant reduction in the content of total amino acids, free fatty acids, glucose, and trehalose, alongside decreased expression of key regulatory genes in the tricarboxylic acid cycle and glycolysis pathways, was observed in the Bb + PEG treatment group. Furthermore, PEG enhanced Bb-induced apoptosis in H. cunea larvae, as evidenced by the upregulation of apoptosis-related genes. Notably, PEG alone did not significantly impact the innate immunity, energy metabolism, or apoptosis in H. cunea larvae. Overall, PEG exhibits considerable potential in amplifying Bb's insecticidal activity by directly optimizing spore performance and indirectly modulating the larvae's innate immunity, energy metabolism, and apoptosis.