Isolation and optimisation of the oleaginous yeast Sporobolomyces roseus for biosynthesis of 13C isotopically labelled 18-carbon unsaturated fatty acids and trans 18:1 and 18:2 derivatives through synthesis

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DOIResolve DOI: http://doi.org/10.1007/s10295-011-1010-z
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleJournal of Industrial Microbiology & Biotechnology
ISSN1367-5435
1476-5535
Volume39
Issue1
Pages153161; # of pages: 9
SubjectTrans fats; Conjugated linoleic acid; 13C labelling; Oleaginous yeasts; Sporobolomyces roseus
AbstractAn oleaginous and psychrotrophic strain (F38-3) of Sporobolomyces roseus Kluyver and van Niel was isolated from a salt marsh environment in Nova Scotia, Canada following a screening program to select for high producers of 18-carbon unsaturated fatty acids. Fatty acid production was characterised as a function of temperature at 20 g glucose L−1, and optimal yields were obtained at 14°C, achieving 5.7 g dw biomass and 39.2% total fatty acids by dry weight, with 18:1, 18:2 and 18:3 all-cis fatty acids accounting for 49.4%, 14.3% and 6.7% of total fatty acids (TFA), respectively—the highest reported for this species. Production of 18:3 was inversely correlated to growth temperature, rising from 2% of TFA at 30°C to 8.9% at 6°C. Cultivation of isolate F38-3 on universally 13C (U-13C) labelled glucose and subsequent transesterification and isolation of the fatty acid methyl esters (FAMEs) by preparative chromatography yielded pure, highly 13C-enriched (>90%) 18:1, 18:2 and 18:3 all-cis FAMEs. The U-13C 18:1 FAME was catalytically converted to U-13C 18:1 trans-9 and purified to >99.5% purity. The U-13C 18:2 was converted by alkaline isomerisation into a 50/50 mixture of 18:2 cis-9, trans-11 and 18:2 trans-10, cis-12 isomers and purified to >95.0% purity. Overall, 10%, by weight, of labelled glucose fed to isolate F38-3 was recovered as fatty acid methyl esters and 7.5% as 18-carbon unsaturated fats, and the final isomerisation reactions resulted in yields of 80% or greater. The ultimate goal of the work is to develop methodologies to produce 13C-labelled metabolic tracers as tools to study the metabolism of trans fats.
Publication date
LanguageEnglish
AffiliationNRC Institute for National Measurement Standards; NRC Institute for Marine Biosciences; National Research Council Canada
Peer reviewedYes
NPARC number21268376
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Record identifierd908b429-60b7-4593-999b-1e38d1ec03e5
Record created2013-07-04
Record modified2016-05-09
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