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Liquid-Liquid Phase Separation of Oil Bodies from Seeds

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Recombinant Proteins from Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1385))

Abstract

Fundamentally, oil bodies are discrete storage organelles found in oilseeds, comprising a hydrophobic triacylglycerol core surrounded by a half-unit phospholipid membrane and an outer shell of specialized proteins known as oleosins. Oil bodies possess a number of attributes that were exploited by SemBioSys Genetics to isolate highly enriched fractions of oil bodies through liquid-liquid phase separation for a number of commercial applications. The current chapter provides a general guide for the isolation of oil bodies from Arabidopsis and/or safflower seed, from which protocols can be refined for different oilseed sources. For SemBioSys Genetic’s recombinant technology, therapeutic proteins were covalently attached to oleosins or fused in-frame with ligands which bound oil bodies, facilitating their recovery to high levels of purity during “upstream processing” of transformed seed. Core to this technology was oil body isolation consisting of simple manipulation including homogenization of seeds to free the oil bodies, followed by the removal of insoluble fractions, and phase separation to recover the oil bodies. During oil body enrichment (an increase in oil body content concomitant with removal of impurities), a number of options and tips are provided to aid researchers in the manipulation and monitoring of these robust organelles.

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Author information

Authors and Affiliations

  1. SemBioSys Genetics Inc., 110 2985 23rd Avenue N.E., Calgary, AB, Canada, T1Y 7L3

    Cory L. Nykiforuk

Authors
  1. Cory L. Nykiforuk
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Correspondence to Cory L. Nykiforuk .

Editor information

Editors and Affiliations

  1. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, Ontario, Canada

    Jacqueline MacDonald

  2. Department of Biology, Western University, London, Ontario, Canada

    Igor Kolotilin

  3. London, Ontario, Canada

    Rima Menassa

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Nykiforuk, C.L. (2016). Liquid-Liquid Phase Separation of Oil Bodies from Seeds. In: MacDonald, J., Kolotilin, I., Menassa, R. (eds) Recombinant Proteins from Plants. Methods in Molecular Biology, vol 1385. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3289-4_13

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  • DOI: https://doi.org/10.1007/978-1-4939-3289-4_13

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3288-7

  • Online ISBN: 978-1-4939-3289-4

  • eBook Packages: Springer Protocols

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