Učinci obogaćivanja hranjivoga medija proteinskim hidrolizatima sjemenki lana i konoplje na rast i produktivnost biotehnološki značajnih životinjskih staničnih linija

doctoral thesis

• Title (english): Effects of growth media supplementation with flaxseed and hempseed protein hydrolysates on growth and productivity of biotechnologically relevant animal cell lines
• Author: Marijan Logarušić
• Mentor: Igor Slivac (mentor)
• Committee member: Višnja Gaurina-Srček (predsjednik povjerenstva)
• Committee member: Marko Obranović (član povjerenstva)
• Committee member: Ana Butorac (član 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: 2023-02-28, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology
• Universal decimal classification (UDC): 57 - Biological sciences. Physical anthropology. Biodiversity

Abstract

Cilj održivog gospodarstva je što učinkovitije iskoristiti organske ostatke nastale preradom sirovina iz različitih grana poljoprivrede i proizvodnje hrane. Primjeri takvih organskih ostataka su i uljane pogače koje zaostaju nakon ekstrakcije ulja iz sjemenki uljarica poput lana i konoplje. Zbog udjela i sastava proteina, uljane pogače imaju potencijal biti iskorištene kao sekundarne sirovine u tehnologiji životinjskih stanica čiji su proizvodi visokovrijedni spojevi poput rekombinantnih proteina i cjepiva. Tijekom ovog istraživanja izolirani su proteini iz uljanih pogača lana i konoplje te su podvrgnuti zasebnim hidrolizama pomoću tri različita mikrobna enzima. Pripravljeni hidrolizati su kvantitativno i kvalitativno analizirani te korišteni kao dodatak mediju za uzgoj životinjskih stanica, uz pretpostavku da će pozitivno utjecati na stanični rast i produktivnost. Glavnina istraživanja provedena je na proizvodnoj staničnoj liniji CHO DP-12 koja stabilno proizvodi rekombinantno monoklonsko protutijelo. Utvrđeno je da je učinak hidrolizata dvojak, tj. da poboljšanje staničnog rasta i/ili produktivnosti ovisi o vrsti enzima kojim su hidrolizati pripravljeni, veličini peptidne frakcije hidrolizata te koncentraciji hidrolizata u mediju. Sličan ishod dobiven je pri uzgoju stanica HEK293T i još je ustanovljeno da prisutnost hidrolizata u mediju nepovoljno djeluje na staničnu transfekciju. Istraživanje antioksidacijskog potencijala i antiproliferacijskog djelovanja hidrolizata na tumorskim (HeLa) i normalnim (HaCaT) staničnim linijama također nije dalo jednoznačne razultate. Nešto povoljnije proliferacijsko djelovanje pokazali su hidrolizati lana dok je antitumorsko djelovanje bilo izraženije kod hidrolizata konoplje.

Abstract (english)

The goal of sustainable economy is to efficiently exploit organic residues resulting from the processing of raw materials from various domains of agriculture and food manufacturing. Examples of such organic residues are oil cakes remained after extraction of oil from flaxseeds and hempseeds. Due to their protein content and composition, oil cakes have the potential to be used as secondary raw material in animal cell culture technology, the products of which are high-value compounds such as recombinant proteins and vaccines. In this research, proteins were isolated from flaxseed and hempseed oil cakes and were subjected to three independent hydrolyses using three microbial enzymes. The produced hydrolysates were quantitatively and qualitatively analyzed and applied as supplements in cell growth media, with the assumption that they will have a positive effect on cell growth and productivity. Most of the research was performed with CHO DP-12 cells producing recombinant monoclonal antibody. It was found that the effect of hydrolysates is ambigous, i.e. that the improvement of cell growth and/or productivity depends on the type of enzyme used for protein digestion, the peptide size in hydrolysate fraction and the concentration of the hydrolysate applied in the cell growth media. Similar results were obtained during the cultivation of HEK293T cells and where the presence of hydrolysates had adverse impact on cell transfection. The investigation of the antioxidative potential and antiproliferative effect of the hydrolysates on tumor (HeLa) and normal (HaCaT) cell lines respectively, showed somewhat disparate results. However, the hempseed hydrolysates showed more favorable antitumor activity, while the flaxseed hydrolysates were more proliferative.

• Keywords: sjemenke lana i konoplje; uljana pogača; proteinski hidrolizat; antioksidacijski potencijal; hranjivi medij; životinjske stanične linije; stanični rast; stanična produktivnost; transfekcija
• Keywords (english): flaxseed; hempseed; oil cake; protein hydrolysates; antioxidative potential; growth medium; animal cell lines; cell growth; cell productivity; transfection
• Language: croatian
• URN:NBN: urn:nbn:hr:159:864925
• Promotion: 2023
• Study programme: Title: Postgraduate PhD Study in Nutrition; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje biotehničkih znanosti, polje nutricionizam (doktor/doktorica znanosti, područje biotehničkih znanosti, polje nutricionizam)
• Type of resource: Text
• Extent: 186 str. : 30 cm.
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-01-13 13:19:13