Pomacea canaliculata, more commonly known as the golden apple snail (GAS), is a freshwater mollusc native from the Plata region comprising Argentina, Uruguay, southern Paraguay, and Brazil. Currently GAS is in the list of the top “100 world’s worst invasive alien species” since it has invaded Eastern Asia, Hawaii and Spain, where it has become a plague1. The identification of digestive enzymes in adult snails would increase the understanding of their digestive physiology and potentially generate new opportunities to eradicate and/or control this invasive species2. In this study, liquid chromatography coupled to tandem mass spectrometry was applied to define the occurrence, diversity, and origin of digestive enzymes along the digestive tract of P. canaliculata. Using a custom-built transcript database, a total of 3,541 proteins (≥ 95% confidence) were identified in the digestive gland and 2,518 were found in the intestine. Additionally, 920 and 801 proteins were identified in the digestive contents of crop and style sac, respectively. Later, 250 peptides derived from 81 glycosidases and 144 peptides of 55 proteases were selected and quantified using multiple reaction monitoring mass spectrometry. Through mass spectrometry, it was possible to obtain the proteome of a range of tissues specific to the survival of this important molluscan pest. This comprehensive study showed digestive enzymes involved in the digestive physiology of P. canaliculata which will provide the foundation for the development of strategies aiming to eradicate and/or control this invasive species