A soy protein isolate was hydrolyzed with Alcalase^®, Flavourzyme^® and their combination, and the resulting hydrolysates (A, F and A + F) were ultrafiltered and analyzed through SDS-PAGE. Fractions with MW < 1 kDa were investigated for their ACE-inhibitory activity, and the most active one (A < 1 kDa) was purified by semi-preparative RP-HPLC, affording three further subfractions. NMR analysis and Edman degradation of the most active subfraction (A1) enabled the identification of four putative sequences (ALKPDNR, VVPD, NDRP and NDTP), which were prepared by solid-phase synthesis. The comparison of their ACE-inhibitory activities suggested that the novel peptide NDRP might be the main agent responsible for A1 fraction ACE inhibition (ACE inhibition = 87.75 ± 0.61%; IC₅₀ = 148.28 ± 9.83 μg mL⁻¹). NDRP acts as a non-competitive inhibitor and is stable towards gastrointestinal simulated digestion. The Multiple Reaction Monitoring (MRM) analysis confirmed the presence of NDRP in A < 1 kDa.

In this study, we investigated the influence of pedological parameters and variation of altitude on the mineral nutrients, phenolic compounds, and antioxidant activities of olive leaves. Samples of the Chetoui cultivar were collected from eight geographical locations with different altitudes. Levels of phenolic compounds varied according to the altitude. Classification of the locations revealed that altitude 1 (>500 m) was characterized by high levels of secoiridoids and simple phenols, while altitude 2 (500–300 m) and altitude 3 (<300 m) were higher in flavonoids. Levels of Mn, Ca and B in the leaves and level of Zn in the soil were significantly correlated with the abundance of oleuropein and luteolin-7-O glucoside, the most important phenols in Chetoui olive leaves. The results suggest that, in addition to pedological criteria, environmental conditions also influence the formation of phenolic compounds.

For the first time the effects of different sampling periods and their interaction with five major autochthonous Croatian Istrian olive cultivars and the Italian cultivar ‘Leccino’ on the quantity and composition of olive leaf phenolic compounds and mineral nutrients were investigated. For that purpose, olive leaves were sampled in two collecting periods, in October and March, coinciding with the harvesting and pruning periods, respectively. All selected cultivars had a higher oleuropein leaf content in the pruning collecting period, with the highest levels noted for the ‘Leccino’ and ‘Buža’ cultivars. Cultivar significantly affected almost all the investigated phenols, with higher concentrations of these valuable compounds in the pruning than in the harvesting period. Differences observed in leaf mineral composition were closely related to the differences in phenolic profiles and were significantly affected by genotype. Some of the studied mineral nutrients, such as P, Cu and B, were found to be significantly correlated with the most abundant olive leaf phenolic compounds, oleuropein and verbascoside.

Manganese (Mn) is an essential element that intervenes in several plant metabolic processes. The olive tree, and its fruits and leaves, are known as a source of nutraceuticals since they are rich in biophenols. However, there is still a serious lack of data about biophenolic distribution in olive stems and roots under Mn fertilisation. In this context, our study aimed to examine the effects of Mn fertilisation on the biophenolic profile in the leaves, stems, and roots of the ‘Istarska bjelica’ olive cultivar. The experiment was set up in a greenhouse, during a period of five months, as a random block design consisting of three treatments with varying Mn concentrations in full-strength Hoagland’s nutrient solution (0.2 µM Mn, 12 µM Mn, and 24 µM Mn). The obtained results indicate that the amount of Mn in the examined olive plant tissues was significantly higher under 12 µM Mn and 24 µM Mn treatments compared to that of the 0.2 µM Mn treatment. While the concentration of biophenols varied in roots depending on the compound in question, a strong positive impact of the increased Mn concentration in nutrient solution (12 µM Mn and 24 µM Mn) on the concentrations of the main biophenolic compounds was observed in stems. The concentration of oleuropein in leaves almost doubled at 24 µM Mn, with the highest Mn concentration, as compared to the 0.2 µM Mn treatment. The obtained results led to the conclusion that the supply of Mn could enhance the concentration of some biologically active compounds in olives grown hydroponically, implying a critical need for further investigation of Mn fertilisation practices in the conventional olive farming system.

This study investigates the effect of combined sulfur (S) and nitrogen (N) foliar fertilization on leaf S and N concentration, as well as on the growth of olive fruit and on the quantity and quality of olive oil, obtained from two olive cultivars ‘Istarska bjelica’ and ‘Leccino’ in two consecutive years. S and N are some of the most important nutrients, and both play a crucial role in plant oil production. The here-reported fertilization program significantly increased S concentration in leaves without affecting N concentration, which led to an increase in fruit yield and improvement of all fruit morphological parameters. The best oil yield per tree was obtained under the treatment with the highest S/N dose. Oil quality was not affected by S and N supply, and this allowed us to classify all our oil samples as extra virgin (EVOO). Regarding the content of total phenols (TPC) and composition of fatty acid methyl esters (FAME), they remained unaltered under the applied treatments. All investigated fruit morphological parameters, as well as fruit and oil yield, were highly cultivar-dependent. ‘Istarska bjelica’ was characterized as a cultivar with higher fruit mass and pulp percentage, while its stone parameters were lower than those of ‘Leccino’. Consequently, the extraction oil yield obtained from ‘Istarska bjelica’ fruits was much higher. Moreover, environmental conditions had a great impact on fruit and oil quantity. The here-obtained results led us to the conclusion that supply of S and N can enhance oil production without affecting its nutritional quality, a finding that could generate large long-term effects on economic growth in the olive oil sector.