Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is used as bac-to-bac eukaryotic protein expression system and biopesticide. However, AcMNPV isn’t widely used in protein expression system and biological control due to some drawbacks and it’s required to understand the function of AcMNPV genes comprehensively in the infection process. Ac132 is an improtant component of the budded virus (BV) nucleocapsid of AcMNPV and the function of Ac132 in regulating progeny virus generation is still unclear. In this study, the recombinant AcMNPV-Ac132-EGFP was constructed. Virus propagation assay indicated that overexpression of Ac132 could improve progeny virus production. Moreover, fluorescent microscope and western blot analysis showed AcMNPV-Ac132-EGFP could accelerate the aggregation of F-actin in the nucleus and delay the evacuation process. In addition, AcMNPV-Ac132-EGFP boosted the utilization of glucose in Sf9 cells at the aspect of energy metabolism. These results demonstrated that Ac132 played important roles in progeny virus production, F-actin formation, nucleocapsid transportion, and glucose metabolism in Sf9 cells, which provided an improved bac-to-bac eukaryotic protein expression system and biopesticide.