Učinak izolata i hidrolizata proteina iz pogače konoplje na rast i oksidacijski stres normalnih i tumorskih stanica

master's thesis

• Title (english): Hempseed protein isolates' and hydrolysates' effects on the proliferation and induced oxidative stress in normal and cancer cell lines
• Author: Tena Dominko
• Mentor: Kristina Radošević (mentor)
• Committee member: Kristina Radošević (predsjednik povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2020-09-28, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Najčešći proizvodi industrijske konoplje (Cannabis sativa L.) su brašno i ulje koje se proizvodi hladnim prešanjem sjemenki. Prilikom tog postupka kao nusproizvod zaostaje uljna pogača konoplje koja je bogata proteinima. Enzimskom hidrolizom proteina konoplje dobivaju se peptidi iznimnog biološkog potencijala te se zato koriste u proizvodnji hrane i farmaceutika. Zato je cilj ovog rada bio ispitati antiproliferacijsko djelovanje proteinskih hidrolizata uljne pogače konoplje pripremljenih s tri različita enzima (alkalaza, neutraza i protamex) in vitro na tumorskoj HeLa i netumorskoj HaCaT staničnoj liniji, protektivni učinak na HaCaT stanicama u kojima je induciran oksidativni stres te induciranje stanične smrti u HeLa stanicama. Dobiveni rezultati pokazuju kako svi pripremljeni hidrolizati i izolat posjeduju antiproliferacijsku aktivnost. HeLa stanice su osjetljivije na učinak hidrolizata te je najjači antiproliferativni učinak imao hidrolizat dobiven pomoću enzima alkalaze. Dokazan je i antioksidativni učinak hidrolizata pripravljenog enzimom neutrazom na HaCaT stanice u kojima je induciran oksidativni stres. Također, dokazano je i kako hidrolizat dobiven pomoću enzima alkalaze inducira staničnu smrt u HeLa stanicama.

Abstract (english)

The most common products of industrial hempseed (Cannabis sativa L.) are flour and oil, which is produced by cold pressing of seeds. Byproduct of this process is hempseed oilcake that is rich in protein. Product of enzymatic hydrolysis of hemp proteins are peptides with biological potential and therefore are used in the production of food and pharmaceuticals. In this thesis the antiproliferative effects of hempseed protein isolate and hydrolysates prepared using three proteases (Alcalase, Neutrase and Protamex) were tested in vitro on cancer (HeLa) and normal (HaCaT) cell lines. Protective effects were evaluated on H2O2-induced oxidative stress in HaCaT cells. In addition, it was tested whether hempseed protein hydrolysate induces cell death in HeLa cells. The obtained results showed that hempseed protein isolate and hydrolysates exhibit antiproliferative effects. The strongest antiproliferative effect was observed in hydrolysates obtained by Alcalase. The study has confirmed the protective effects of hydrolysate obtained by Neutrase during induced oxidative stress in HaCaT cell line. Furthermore, it is confirmed that hydrolysate obtained by Alcalase induces cell death in HeLa cell line.

• Keywords: citotoksičnost; oksidativni stres; proteinski hidrolizati; stanična smrt; uljna pogača konoplje
• Keywords (english): cytotoxicity; oxidative stress; protein hydrolysate; cell death; hempseed oilcake
• Language: croatian
• URN:NBN: urn:nbn:hr:159:725116
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Repository: Repository of the Faculty of Food Technology and Biotechnology
• Created on: 2021-01-20 12:09:15