The fungal strain BS5 was isolated from a soil sample collected in the Tibetan Plateau, which displayed good insecticidal activity and was identified as Talaromyces purpureogenus based on morphological and molecular analysis. This study aimed to evaluate the insecticidal activity and identify the active compound of the strain BS5 against the locust Locusta migratoria manilensis. The insecticidal activity of the fermented broth of BS5 was at 100% after 7 days against locusts. We extracted the fermented broth of BS5 and then evaluated the insecticidal activity of the extracts against locusts. The ethyl acetate extract exhibited promising activity levels with an LC50 value of 1077.94 μg/mL and was separated through silica gel column chromatography. The UPLC-Q-Exactive Orbitrap/MS system was employed to analyze the active fraction Fr2.2.2 (with an LC50 value of 674.87 μg/mL), and two compounds were identified: phellamurin and rubratoxin B.