Objectives The cell wall of microalgae is a necessary barrier for its survival in aquatic environments. This barrier affects the recovery of lipids, representing one of the main difficulties in the development of microalgal biofuel production technology. In this context, this work aimed to test the effect of a new method based on melittin peptide as a pretreatment to cause cellular disruption in N. oleoabundans cultivated under different temperatures and nutrient concentrations. Results Neochloris oleoabundans cells were grown at 32°C in the presence of high concentration of nitrate-phosphate, cell disruption degree being recorded at 83.6%. Also, two-fold increase in lipid recovery following melittin treatment and solvent extraction was observed. Furthermore, it was possible to verify the effect of melittin, both before and after treatment on the morphology of the cells. Scanning electron microscopy (SEM) and confocal images of the melittin-treated microalgae revealed extensive cell damage with degradation of the cell wall and release of intracellular material. Conclusions Melittin peptide produced a selective cell wall rupture effect with the highest percentage of cell disruption degree when N. oleoabundans under culture condition with nitrate-phosphate repletion at 32 °C favors cell wall susceptibility to cell disruption by melittin peptide, representing the first report on lipid recovery from microalgae. Melittin is a promising option for the enhancement of the recovery of lipids from microalgae.