To verify the anti-fungus properties of the crop-harmful pseudocercospora, the essential oil (EO) of pomelo peel (PP) was extracted by a single factor combined with response surface optimization. Meanwhile, the composition and activity of EO were studied. The PP was squeezed by a screw extruder and pretreatmented by pectinase, then extracted by microwave-assisted steam distillation. The optimal conditions were as follows: pectinase dosage was 69.17 μmol/g, microwave power was 651.42 W and extraction time was 43.84 min. The dry weight (DW) yield of PPEO reached 14.63 mL/kg DW after BBD optimization. There were 23 compounds in the PPEO identified by GC-MS. Limonene, α-phrenbutene, and laurene in PPEO accounted for 79.31%, 4.72%, and 3.46%, respectively. In addition, the antifungal was effective when the concentration of PPEO was 3.5 mg/mL. Therefore, this study has guiding significance for the development of natural resources.