Metabolic engineering of Pichia pastoris to produce ricinoleic acid, a hydroxy fatty acid of industrial importance

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DOIResolve DOI: http://doi.org/10.1194/jlr.M060954
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TypeArticle
Journal titleJournal of Lipid Research
ISSN0022-2275
Volume56
Issue11
Pages21022109; # of pages: 8
Subjecthydroxy fatty acid; ricinoleic acid; biomass; Claviceps purpurea; conceptual framework; enzyme activity; fatty acid analysis; gene amplification; gene cassette; homologous recombination; hydroxylation; Komagataella pastoris; lipid analysis; metabolic engineering; molecular cloning; nonhuman; open reading frame; plasmid; promoter region; transesterification
AbstractRicinoleic 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 12 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.
Publication date
PublisherAmerican Society for Biochemistry and Molecular Biology
LanguageEnglish
AffiliationNational Research Council Canada; Aquatic and Crop Resource Development
Peer reviewedYes
NPARC number21277325
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Record identifier206d1d42-fe94-4085-9b32-2b200a576807
Record created2016-03-09
Record modified2016-05-09
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