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