Abstract:Sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE），two-dimensional electrophoresis and matrix assisted laser desorption ionization-time of fight-mass spectrometry（MALDI-TOF/TOF/MS）were used to reveal the properties and composition of donkey bone extract，which provided a basis for the deep development of donkey bone bioactive peptides with characteristic physiological activities.The results showed that the protein content of donkey milk powder was 47.82 g/100 g，the molecular weight was distributed in the range of 10 kDa-250 kDa，and the isoelectric point was uniformly distributed between 4.5-6.5.By comparing with the equine protein library，89 proteins were retrieved，and these proteins were classified by gene ontology（GO）classification and kyoto encyclopedia of genes and genomes（KEGG）signaling pathway enrichment.It was found that these proteins were involved in biological processes such as mesenchyme migration，cellular response to amino acid stimulus，nucleosome assembly，retina homeostasis，cartilage development involved in endochondral bone morphogenesis，and also participate in signal pathways such as platelet activation，focal adhesion，protein digestion and absorption.