Journal of Lipid Research
Volume 56, Issue 11, November 2015, Pages 2102-2109
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Research Articles
Metabolic engineering of Pichia pastoris to produce ricinoleic acid, a hydroxy fatty acid of industrial importance[S]

https://doi.org/10.1194/jlr.M060954Get rights and content
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Ricinoleic acid (12-hydroxyoctadec-cis-9-enoic acid) has many specialized uses in bioproduct industries, while castor bean is currently the only commercial source for the fatty acid. This report describes metabolic engineering of a microbial system (Pichia pastoris) to produce ricinoleic acid using a “push” (synthesis) and “pull” (assembly) strategy. CpFAH, a fatty acid hydroxylase from Claviceps purpurea, was used for synthesis of ricinoleic acid, and CpDGAT1, a diacylglycerol acyl transferase for the triacylglycerol synthesis from the same species, was used for assembly of the fatty acid. Coexpression of CpFAH and CpDGAT1 produced higher lipid contents and ricinoleic acid levels than expression of CpFAH alone. Coexpression in a mutant haploid strain defective in the Δ12 desaturase activity resulted in a higher level of ricinoleic acid than that in the diploid strain. Intriguingly, the ricinoleic acid produced was mainly distributed in the neutral lipid fractions, particularly the free fatty acid form, but with little in the polar lipids. This work demonstrates the effectiveness of the metabolic engineering strategy and excellent capacity of the microbial system for production of ricinoleic acid as an alternative to plant sources for industrial uses.

Claviceps purpurea
fatty acid hydroxylase
diacylglycerol acyltransferase

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The data discussed in this publication have been deposited in the National Institutes of Health database GenBank (Meesapyodsuk et al., 2015) and are accessible through the accession number for C. purpurea CpDGAT1, KT581979.

    Abbreviations:

    CpDGAT1

    diacylglycerol acyltransferase from Claviceps purpurea

    CpFAH

    fatty acid hydroxylase from Claviceps purpurea

    DGAT

    diacylglycerol acyltransferase

    FAME

    fatty acid methyl ester

    FRA

    free ricinoleic acid

    TAG

    triacylglycerol

[S]

The online version of this article (available at http://www.jlr.org) contains a supplement.