*Article* **Exploring the DPP-IV Inhibitory, Antioxidant and Antibacterial Potential of Ovine "Scotta" Hydrolysates**

**Roberto Cabizza 1, Francesco Fancello 1, Giacomo Luigi Petretto 2, Roberta Addis 2, Salvatore Pisanu 3, Daniela Pagnozzi 3, Antonio Piga 1 and Pietro Paolo Urgeghe 1,\***


**Abstract:** The aim of this work was to valorize the by-product derived from the ricotta cheese process (scotta). In this study, ovine scotta was concentrated by ultrafiltration and then subjected to enzymatic hydrolyses using proteases of both vegetable (4% E:S, 4 h, 50 ◦C) and animal origin (4% E:S, 4 h, 40 ◦C). The DPP-IV inhibitory, antioxidant, and antibacterial activities of hydrolysates from bromelain (BSPH) and pancreatin (PSPH) were measured in vitro. Both the obtained hydrolysates showed a significantly higher DPP-IV inhibitory activity compared to the control. In particular, BSPH proved to be more effective than PSPH (IC50 8.5 ± 0.2 vs. 13 ± 1 mg mL−1). Moreover, BSPH showed the best antioxidant power, while PSPH was more able to produce low-MW peptides. BSPH and PSPH hydrolysates showed a variable but slightly inhibitory effect depending on the species or strain of bacteria tested. BSPH and PSPH samples were separated by gel permeation chromatography (GPC). LC-MS/MS analysis of selected GPC fractions allowed identification of differential peptides. Among the peptides 388 were more abundant in BSPH than in the CTRL groups, 667 were more abundant in the PSPH group compared to CTRL, and 97 and 75 of them contained sequences with a reported biological activity, respectively.

**Keywords:** ovine scotta; bioactive peptides; bromelain; pancreatin; dipeptidyl peptidase IV inhibition; ovine second whey cheese; enzymatic hydrolysis
