There is a wealth of scientific evidence associating the dietary intake of omega 3 long-chain polyunsaturated fatty acids with beneficial health properties. In this study, alternative natural sources of these polyunsaturated fatty acids are sought from novel low temperature isolated fungi. Over 100 low temperature isolated fungi were screened for very long chain polyunsaturated fatty acids (VLCPUF As), such as C20:5 n3 and C22:6 n3. Of those screened, only ten fungi were capable of VLCPUFA production, with Mortierella the predominant VLCPUF A producing species. Four Oomycete species were also capable of VLCPUF A production. It is thought that only basal fungal lineages, such as species from the Chytridiomycota and Zygomycota, are capable of VLCPUF A production. It was also found that VLCPUF As are not essential for growth at low temperatures, as Penicillium rugulosum, capable of producing 'fatty acids no longer or more unsaturated than C18:3 n3, demonstrated over 2 g of biomass per 100 ml of broth when grown at 5°C. This indicates that trienoic fatty acids are sufficient for maintaining membrane fluidity, although other factors may play a role in P. rugulosum's low temperature growth. Comparatively, VLCPUFA producing Mortierella species produced 200-250 mg of biomass, whereas the majority of non- VLCPUF A producing isolates produced 106-115 mg of biomass per 100 ml of broth. The total lipid unsaturation indices of nine isolates grown under three temperature regimes showed that the lowest growth temperature, 5°C, produced the highest un saturation index value III SIX of the organisms. 15°C produced the highest unsaturation index value in two of the isolates. This suggests that temperature has an effect on fungal lipid composition, and that lower temperatures may increase lipid unsaturation levels. It was also found that the ~6 elongase, initially identified from Mortierella alpina, is indicative for VLCPUF A producing fungi. The genomic conserved sequence found within ~6 elongases was used to develop primer sets that could be used with a PCR based methodology to screen fungal isolates for VLCPUF A production. The method successfully identified VLCPUF A producing Mortierella and Allomyces species, and was not found to amplify non-Ad elongases. Finally, recombinantPhaffia rhodozyma strains were developed using the ~5 desaturase and ~6 elongase from Mortierella alpina. The fatty acid profiles of the recombinant strains displayed novel fatty acids such as C20:2 n6 and C20:3 n3, and putatively, C18:2 ~5, 9 and C18:3 ~5, 9, 12 which correlated with the inserted genes.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:553832 |
| Date | January 2012 |
| Creators | Broughton, Richard |
| Contributors | Bramley, Peter : Fraser, Paul : Ryan, Matthew |
| Publisher | Royal Holloway, University of London |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://repository.royalholloway.ac.uk/items/38797691-c5ac-ef52-3a6c-12c3097e2765/6/ |
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