Engineering a complex, multiple enzyme-mediated synthesis of natural plant pigments in the silkworm, Bombyx mori .

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  • Additional Information
    • Source:
      Publisher: National Academy of Sciences Country of Publication: United States NLM ID: 7505876 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1091-6490 (Electronic) Linking ISSN: 00278424 NLM ISO Abbreviation: Proc Natl Acad Sci U S A Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : National Academy of Sciences
    • Subject Terms:
      Genetic Engineering* ; Bombyx*; Animals, Genetically Modified ; Animals ; Pigments, Biological/biosynthesis ; Betalains/biosynthesis ; Betalains/chemistry ; Gene Expression ; Gene Expression Regulation, Enzymologic ; Color
    • Abstract:
      Plants produce various pigments that not only appear as attractive colors but also provide valuable resources in applications in daily life and scientific research. Biosynthesis pathways for these natural plant pigments are well studied, and most have multiple enzymes that vary among plant species. However, adapting these pathways to animals remains a challenge. Here, we describe successful biosynthesis of betalains, water-soluble pigments found only in a single plant order, Caryophyllales, in transgenic silkworms by coexpressing three betalain synthesis genes, cytochrome P450 enzyme CYP76AD1 , DOPA 4,5-dioxygenase, and betanidin 5-O-glucosyltransferase . Betalains can be synthesized in various tissues under the control of the ubiquitous IE1 promoter but accumulate mainly in the hemolymph with yields as high as 274 μg/ml. Additionally, transformed larvae and pupae show a strong red color easily distinguishable from wild-type animals. In experiments in which expression is controlled by the promoter of silk gland-specific gene, fibroin heavy-chain , betalains are found predominantly in the silk glands and can be secreted into cocoons through spinning. Betalains in transformed cocoons are easily recovered from cocoon shells in water with average yields reaching 14.4 μg/mg. These data provide evidence that insects can synthesize natural plant pigments through a complex, multiple enzyme-mediated synthesis pathway. Such pigments also can serve as dominant visible markers in insect transgenesis applications. This study provides an approach to producing valuable plant-derived compounds by using genetically engineered silkworms as a bioreactor.
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    • Contributed Indexing:
      Keywords: Bombyx mori; betalains; bioreactor; silk gland
    • Accession Number:
      0 (Pigments, Biological)
      37279-84-8 (Betalains)
    • Publication Date:
      Date Created: 20230807 Date Completed: 20231016 Latest Revision: 20231016
    • Publication Date:
      20231016
    • Accession Number:
      PMC10433459
    • Accession Number:
      10.1073/pnas.2306322120
    • Accession Number:
      37549256