Developing an immobilized enzyme technology for enrichment of Omega-3 PUFAs

Author: Mahejbin Karia

  • Thesis download: available for open access on 14 Jun 2027.

Karia, Mahejbin, 2024 Developing an immobilized enzyme technology for enrichment of Omega-3 PUFAs, Flinders University, College of Medicine and Public Health

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Majority of the population is consuming food that contains low amounts of polyunsaturated fatty acids (PUFAs) such as Omega- 3 compared to Omega-6 fatty acids. Diet intake of low omega-3 fatty acid levels has been connected to increased prevalence of various chronic illnesses. To meet the demands of omega-3 PUFAs, various approaches are employed by pharmaceutical companies for obtaining concentrated form of Omega 3 PUFAs that mostly use either expensive distillation technology or hazardous solvents that can have negative impacts on the environment. The present study focuses on employing enzymatic enrichment technology by improving the stability and activity of lipase enzyme by immobilising it on Poly-oxazoline monomer deposited through cold-plasma polymerisation technique. In this research work, two thraustochytrid in-house strains S4 and DT3 were compared for their fatty acid profile and dry cell weight. Thraustochytrid S4 strain gave better results compared to DT3 with biomass production of 16.05g/L after 96 h of fermentation and showed better EPA (0.47%) and DHA (17.21%) content. Polyoxazoline coating with thickness of 20 nm was achieved on 24 well plates and was further utilised for lipase immobilisation. The optimised conditions for enzymatic activity of free and immobilised lipase were carried out using enzyme concentration, pH and temperature as variables. Optimum enzyme concentration of 2mg/ml, pH 8.5, and a temperature of 650C was determined for both free and immobilised enzyme. Lineweaver burk plot was plotted to determine Km and Vmax values for free and immobilised enzyme. The free and immobilised enzyme were further utilised for hydrolysis of thraustochytrid S4 oil. The levels of EPA (14.4%) and DHA (21.5%) were enriched when using immobilised lipases, thus, meeting objectives of the proposed research work.

Keywords: Omega-3, Docosahexaenoic acid, eicosapentaenoic acid, cold plasma polymerisation, polyoxazoline, enzymatic enrichment.

Subject: Medical Biotechnology thesis

Thesis type: Masters
Completed: 2024
School: College of Medicine and Public Health
Supervisor: Associate Prof. Munish Puri