Isolation of proteins from spent coffee grounds. Polyphenol removal and peptide identification in the protein hydrolysates by RP-HPLC-ESI-Q-TOF

Several works have been focused on the extraction of polysaccharides, polyphenols and caffeine from spent coffee grounds (SCG) and their application in food formulations, but the peptide bioactivity from SCG protein hydrolysates has never been addressed. In the present work and for the first time, two different methods to isolate proteins from SCG have been compared, demonstrating that a urea-based extraction buffer provides a higher yield. This extraction method was then applied to compare the protein content in SCG from different coffee-brewing preparations, showing a higher protein content in SCG from espresso coffee machines. In addition, a polyphenol extraction step to remove interferences has been evaluated and the hydrolysis of the extracted proteins using alcalase and thermolysin enzymes has been compared. The effect of roasting degree on the antioxidant and in vitro angiotensin-converting enzyme (ACE)-inhibitory activity has been evaluated. The results show that the ACE-inhibitory activity is higher when SCG proteins are obtained from medium and dark roasted coffees and then hydrolyzed with thermolysin. Finally, the peptides contained in these hydrolysates have been identified by reversed-phase high-performance liquid chromatography coupled via electrospray ionization to a quadrupole time-of-flight mass spectrometer (RP-HPLC-ESI-Q-TOF).