ABSTRACT The aim of this work was to produce enzymatic hemp seed protein hydrolysates (HPH) followed by bioassay guided fractionation to identify antioxidant and antihypertensive peptides. Therefore, simulated gastrointestinal digestion of isolated hemp seed proteins was conducted using consecutive actions of pepsin and pancreatin to produce HPH, which was then separated by membrane ultrafiltration to obtain peptide sizes of <1, 1-3, 3-5, and 5-10 kDa. Evaluation of HPH and its membrane fractions for antioxidant and antihypertensive properties showed that they significantly (P<0.05) scavenged radicals, reduced and strongly chelated metal ions as well as inhibited lipid oxidation. During a 24-hr test, the HPH reduced systolic blood pressure (SBP) of spontaneously hypertensive rats (SHR) after oral administration by a maximum of -30 mmHg when compared to -15 mmHg for the membrane fractions. To reduce production cost, hemp seed protein meal (HPM) was directly hydrolyzed to a protein hydrolysate (HMH) and was shown to also reduce SBP during 4-8 weeks of dietary feeding. The attenuation of SBP correlated to suppressed plasma levels (0.047-0.059 U/mL and 0.040-0.054 µg/mL) of angiotensin converting enzyme (ACE) and renin, respectively, when compared to the control rats (0.123 U/mL and 0.151 µg/mL). A total of 23 peptides were identified to be present in the HPH. WVYY and PSLPA showed superior in vitro antioxidant properties, while ACE activity was inhibited by WYT (89%), WVYY (91%) and PSLPA (90%). Renin activity was inhibited by WYT (77%), SVYT (87%) and IPAGV (75%). However, oral administration to SHR showed that the pentapeptides (PSLPA and IPAGV) were more effective SBP-reducing agents (-40 mm Hg) when compared to the tri- (-13 mmHg) and tetrapeptides (-36 mmHg). These results show for the first time in literature, the bioactive properties of hemp seed peptides and indicate their potential use as ingredients to formulate antioxidant and antihypertensive functional foods and nutraceuticals.

ABSTRACT The aim of this work was to produce enzymatic hemp seed protein hydrolysates (HPH) followed by bioassay guided fractionation to identify antioxidant and antihypertensive peptides. Therefore, simulated gastrointestinal digestion of isolated hemp seed proteins was conducted using consecutive actions of pepsin and pancreatin to produce HPH, which was then separated by membrane ultrafiltration to obtain peptide sizes of <1, 1-3, 3-5, and 5-10 kDa. Evaluation of HPH and its membrane fractions for antioxidant and antihypertensive properties showed that they significantly (P<0.05) scavenged radicals, reduced and strongly chelated metal ions as well as inhibited lipid oxidation. During a 24-hr test, the HPH reduced systolic blood pressure (SBP) of spontaneously hypertensive rats (SHR) after oral administration by a maximum of -30 mmHg when compared to -15 mmHg for the membrane fractions. To reduce production cost, hemp seed protein meal (HPM) was directly hydrolyzed to a protein hydrolysate (HMH) and was shown to also reduce SBP during 4-8 weeks of dietary feeding. The attenuation of SBP correlated to suppressed plasma levels (0.047-0.059 U/mL and 0.040-0.054 µg/mL) of angiotensin converting enzyme (ACE) and renin, respectively, when compared to the control rats (0.123 U/mL and 0.151 µg/mL). A total of 23 peptides were identified to be present in the HPH. WVYY and PSLPA showed superior in vitro antioxidant properties, while ACE activity was inhibited by WYT (89%), WVYY (91%) and PSLPA (90%). Renin activity was inhibited by WYT (77%), SVYT (87%) and IPAGV (75%). However, oral administration to SHR showed that the pentapeptides (PSLPA and IPAGV) were more effective SBP-reducing agents (-40 mm Hg) when compared to the tri- (-13 mmHg) and tetrapeptides (-36 mmHg). These results show for the first time in literature, the bioactive properties of hemp seed peptides and indicate their potential use as ingredients to formulate antioxidant and antihypertensive functional foods and nutraceuticals.

Il rinnovato interesse nella coltivazione della canapa industriale non è stato supportato da studi approfonditi e generalmente si basa su esperienze dirette di coltivazione. Questo studio si prefigge come obbiettivo quello di sfruttare tutte la parti della pianta di canapa per migliorare l’economicità complessiva della sua coltivazione tramite l’uso di tecniche agronomiche e di pratiche post-raccolta. La Cannabis sativa ha un vasto campo di applicazioni, tra cui le più promettenti in termini di ritorno economico sono il cosmetico e il farmaceutico. I composti naturali più interessanti della Cannabis sono i cannabinoidi non-psicoattivi cannabidiolo (CBD) e cannabigerolo (CBG) presenti in elevate concentrazioni nelle foglie e nelle brattee di Cannabis. Altri metaboliti secondari prodotti dalla canapa e di grande interesse sono le cannflavine, una classe di prenil-flavonoidi caratteristico di questa specie. Nel primo e nel secondo capitolo la dinamica temporale di accumulo dei metaboliti secondari ad elevato valore aggiunto è stata monitorata per alcune delle varietà più comuni di canapa industriale ed è stata determinata la produzione potenziale di questi composti. Nel terzo capitolo una collezione di semi di canapa è stata valutata per le caratteristiche morfologiche e per il contenuto e la qualità dell’olio. Concludendo questo studio contribuisce allo sviluppo di una coltivazione della canapa per le molteplici destinazioni d’uso dei semi e del fitocomplesso. / Rapidly growing interest into industrial hemp cultivation is not supported by strong data and generally is referred to a limited experience on field cultivation. This study aims to exploit all the parts of hemp plant, in order to ameliorate the overall economy of its cultivation and by providing information on agronomic management and post-harvest operations. Cannabis sativa has a wide range of industrial applications and the most promising, in terms of economic revenues, are the cosmetic and pharmaceutics. The most interesting phytochemicals in Cannabis are the cannabinoids, in particular the non-intoxicating compounds cannabidiol (CBD) and cannabigerol (CBG), present at high concentration in the leaves and bracts of Cannabis. Some other promising secondary metabolites of hemp are cannflavins, a class of prenyl-flavonoids characteristic of this species. In the first and second chapter the time course accumulation of high added-value compounds was monitored for some of the most common industrial hemp cultivars, and the potential production was assessed. In the third chapter a collection of hemp seeds was evaluated for its morphological characteristics and for its oil content and quality. In conclusion this thesis contributes to a further development of multipurpose hemp cultivation for seed and phytocomplex production.