This study aimed to investigate effects of heat treatment temperatures (60℃, 70℃, 80℃, 90℃, 100℃) and times (10 min, 30 min, 60 min) on the aggregation behavior of Moringa Oleiferaseed salt-soluble protein (MOSP). With the increase of heating temperature and time, the MOSP solutions were changed from a more transparent state to a cloudy and opaque state, the absorbance value of the MOSP solution increased significantly, and the particle size gradually increased to a maximum of 1220.33 nm (100℃, 30 min). The polymer dispersion index (PDI) also increased and the maximum PDI of MOSP (100℃, 10 min) was 0.53, but the absolute value of the zeta-potential was gradually decreased. With the increase of heating temperature and heating time, the β-sheet content of MOSP was increased to 30.8%, while the α-helix content of MOSP decreased. There was no obvious change in the subunit distribution of the heat-induced MOSP. The spectrum analysis displayed that heat treatment led to the exposure of the hydrophobic residues in MOSP, but as the heating temperature was further increased to 90℃, re-embedding of the hydrophobic groups was observed due to the formation of aggregates. The total sulfhydryl group (-SHT) increased with the increase of temperature, while the free sulfhydryl group (-SHF)first decreased and then increased during short-term heating (10 and 30 min). The maximum content of -SHFreached 4.40% and the content of disulfide bonds was 11.97% (90℃, 30 min). These changes could have a significant impact on protein products and might spur innovation in new protein applications.