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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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Journal of Industrial Microbiology & Biotechnology

Abstract

An oleaginous and psychrotrophic strain (F38-3) of Sporobolomyces roseus Kluyver & 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.

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Acknowledgments

The authors would like to thank the Joint Cooperation Program of the National Research Council Canada, National Science Council of Taiwan and the Industrial Technology Research Institute Taiwan for financial support to undertake this work. The authors state they have no conflict of interest and are in agreement to publish in the Journal of Industrial Microbiology and Biotechnology. All experiments described in this manuscript were in accordance with the laws of Canada.

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Authors and Affiliations

  1. Institute for National Measurement Standards, National Research Council Canada, Building M-12, 1200 Montreal Rd., Ottawa, ON, K1A 0R6, Canada

    Yi Cui, Catharine Fraser, Graeme Gardner & Anthony J. Windust

  2. Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan

    Ching-jang Huang

  3. Institute for Marine Biosciences, National Research Council Canada, 1411 Oxford St., Halifax, NS, Canada

    Michael Reith

Authors
  1. Yi Cui
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  2. Catharine Fraser
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  3. Graeme Gardner
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  4. Ching-jang Huang
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  5. Michael Reith
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  6. Anthony J. Windust
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Correspondence to Anthony J. Windust.

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Cui, Y., Fraser, C., Gardner, G. et al. 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. J Ind Microbiol Biotechnol 39, 153–161 (2012). https://doi.org/10.1007/s10295-011-1010-z

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  • DOI: https://doi.org/10.1007/s10295-011-1010-z

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