Potentialities of aqueous extract from cultivated Onopordum tauricum (Willd.) as milk clotting agent for cheesemaking

• Cultivated Onopordum tauricum showed better clotting activity than wild thistles. • Cultivated and wild thistles showed no differences in their proteolytic activities. • The purified tauricosin was identified as a heterodimeric aspartic-type protease. • O. tauricum showed good potential as herbaceous crop for vegetable rennet production. In Western and Central Mediterranean countries proteases from wild herbaceous perennial plants commonly known as “thistles” have been used as milk coagulants in cheese-making for centuries. For the first time, the technological and biochemical traits of proteases from cultivated Onopordum tauricum Willd. (Taurian thistle, bull cottonthistle) were assessed. The optimal conditions for minimizing the clotting time and the non-specific proteolytic activity were estimated at the highest (T = 43–45 °C; [Ca 2+ ] = 11–13 mM) and the lowest (T = 35–39 °C; [Ca 2+ ] = 5 mM) temperature and calcium ion levels in the explored range respectively, thus highlighting the difficulty to set the best operative compromise in the first step of cheesemaking. In the conditions adopted in common cheesemaking practice (T = 37 °C; pH = 6.5) 1 mL of reconstituted extract from cultivated thistles coagulated 10 mL of ewe’s and goat’s milk in 114–146 and 129–167 s, respectively, and 1 mL of reconstituted extract from spontaneous thistles coagulated 10 mL of ewe’s and goat’s milk in 232–294 and 428–621 s, respectively, while no significant differences in the non-specific proteolytic activity between cultivated and spontaneous O. tauricum extracts were observed. The purified enzyme (tauricosin) was identified as an aspartic protease made up of two sub-units with molecular weights of 32 and 9.6 kDa, respectively. Experimental data encouraged the exploitation of O. tauricum as a new and sustainable non-food crop in marginal and rainfed lands of Mediterranean countries, thus reducing the potential biodiversity losses due to wild collection.