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    Area of Science:

    • Plant biochemistry
    • Lipid metabolism
    • Molecular biology

    Background:

    • Fatty acid elongation (FAE) is a crucial metabolic process.
    • Standard FAE involves a four-step cycle producing very long-chain fatty acids.
    • Plant seed oils are sources of valuable lipids.

    Purpose of the Study:

    • To investigate an unusual fatty acid synthesis pathway in Orychophragmus limprichtianus.
    • To characterize novel keto-hydroxy fatty acids found in its seed oil.
    • To identify the enzymes and mechanisms responsible for this unique FAE process.

    Main Methods:

    • Analysis of seed oil composition.
    • Transcriptomic analysis to identify key enzymes.
    • Functional assays and enzyme activity studies.
    • Protein modeling and mutant analysis.

    Main Results:

    • Discovery of C24-C28 keto-hydroxy fatty acids in O. limprichtianus seed oil, comprising ~25% of total fatty acids.
    • Identification of a discontinuous elongation process involving a 3-keto hydroxy intermediate.
    • Characterization of OlFAE1-2 and a low-activity ketoreductase OlKCR1-1 as central enzymes.
    • Evidence for altered OlKCR1-1 activity due to specific amino acid substitutions.
    • Accumulation of these novel fatty acids in estolides, expanding seed oil diversity.

    Conclusions:

    • Orychophragmus limprichtianus exhibits a unique variation of plant fatty acid elongation.
    • This pathway bypasses full reduction, utilizing a polyketide synthase-like mechanism.
    • Enzyme engineering, particularly of OlKCR1-1, is key to this process.
    • Findings provide tools for engineering plants and microbes for novel renewable oils.