| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | All reagents were of analytical grade. Water was daily obtained with a Milli-Q system from Millipore (Bedford, MA, USA). Tris(hydroxymethyl) aminomethane (Tris), hydrochloric acid, sodium phosphate, and sodium dodecyl sulphate (SDS) were from Merck (Darmstadt, German). Acetic acid (AA), N-(2-hydroxyethyl)-1-piperazinyl-N’-(2-ethanesulfonic) acid (HEPES), tripeptide hippuryl-histidyl-leucine (HHL), trifluorocetic acid (TFA), angiotensin converting enzyme (ACE) from rabbitlung, Thermolysin, pancreatic and pepsin enzymes, dimethylsulphoxide (DMSO) concentrated type-I collagen solution, propidium iodure (PI), ribonuclease A from bovine pancreas (RNase A), bovine serum albumin (BSA), phosphate-buffered saline(PBS), trypsin, 3-(4,5-dimethyl-2-thiazolyl)-2,5 diphenyltetrazolium bromide (MTT), and DMEM (Dubelcco’s Modified Eagle's medium) and RPMI-1640 (Roswell Park Memorial Institute) media were purchased in Sigma-Aldrich (Saint Louis, MO, USA). Acetonitrile HPLC- and MS-grade (ACN), acetone, and n-hexane were from Scharlau Chemie (Barcelona, Spain). Peptide LLPSY was synthesized by GenScript Corp. (Piscataway, NJ, USA) with a purity >95%. Raw olives of Picual variety were collected in Toledo (Spain). | Get A Quote |
Peptides with molecular weights below 3 kDa from the hydrolysis of olive seed proteins with Thermolysin (OS-3 kDa) have demonstrated a high in vitro hypotensive effect. This fraction has been further fractionated by semipreparative RP-HPLC to obtain 8 fractions. ACE inhibitor capacity of fractions was evaluated observing the highest capacity in fraction F5. Peptides in fraction F5 were identified by RP-HPLC- and HILIC-ESI-Q-ToF and cytotoxic effect was assessed in different cell lines. Peptide LLPSY, present in this faction, was synthesized and characterized. Despite antihypertensive capacity was not as high as in fraction F5, a significant anti-proliferative capacity on two different cancer cell lines wa... More
