Separation and identification of peptides in hydrolysed protein extracts from edible macroalgae by HPLC-ESI-QTOF/MS

Abstract

Macroalgae contain significant amounts of high-quality proteins which, because of their structural diversity, contain a range of yet undiscovered peptides within their primary structures. In this work, an analytical methodology was developed for the separation and identification of peptides present in protein hydrolysates from three different edible macroalgae used for human consumption (Saccharina latissima (brown macroalga), Codium spp. (green macroalga), and Mastocarpus stellatus (red macroalga)). The extraction of aqueous and alkaline soluble proteins was carried out followed by their precipitation with HCl or acetone. The protein extracts obtained were submitted to enzymatic digestion with alcalase and subsequently analyzed by reversed-phase high-performance liquid chromatography-quadrupole-time-of flight mass spectrometry (RP-HPLC-QTOF/MS) and de novo sequencing tool to separate and identify different short chain peptides. Thirty-seven peptides were identified in the hydrolysed protein extracts from the three macroalgae, five of them being common in brown and red macroalgae. After checking against BIOPEP database, several sequenced peptides were found within longer peptides with potential antibacterial activity. Any of the identified peptides had previously been identified in macroalgae.