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Synthesis of C‐Glucosylated Octaketide Anthraquinones in Nicotiana benthamiana by Using a Multispecies‐Based Biosynthetic Pathway
Authors:Dr. Johan Andersen‐Ranberg  Dr. Kenneth Thermann Kongstad  Dr. Majse Nafisi  Prof. Dan Staerk  Finn Thyge Okkels  Prof. Uffe Hasbro Mortensen  Prof. Birger Lindberg Møller  Dr. Rasmus John Normand Frandsen  Dr. Rubini Kannangara
Affiliation:1. Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg, Denmark;2. Present address: Department of Plant Microbial Biology, University of California Berkeley, Berkeley, CA, USA;3. Faculty of Health and Medical Sciences, Department of Drug Design and Pharmacology, University of Copenhagen, Copenhagen, Denmark;4. Chr. Hansen A/S, H?rsholm, Denmark;5. Present address: ActaBio ApS, Roskilde, Denmark;6. Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark
Abstract:Carminic acid is a C‐glucosylated octaketide anthraquinone and the main constituent of the natural dye carmine (E120), possessing unique coloring, stability, and solubility properties. Despite being used since ancient times, longstanding efforts to elucidate its route of biosynthesis have been unsuccessful. Herein, a novel combination of enzymes derived from a plant (Aloe arborescens, Aa), a bacterium (Streptomyces sp. R1128, St), and an insect (Dactylopius coccus, Dc) that allows for the biosynthesis of the C‐glucosylated anthraquinone, dcII, a precursor for carminic acid, is reported. The pathway, which consists of AaOKS, StZhuI, StZhuJ, and DcUGT2, presents an alternative biosynthetic approach for the production of polyketides by using a type III polyketide synthase (PKS) and tailoring enzymes originating from a type II PKS system. The current study showcases the power of using transient expression in Nicotiana benthamiana for efficient and rapid identification of functional biosynthetic pathways, including both soluble and membrane‐bound enzymes.
Keywords:biosynthesis  carmine  enzymes  polyketides  synthetic biology
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