title: Regioselective Dichlorination of a Non-Activated Aliphatic Carbon Atom and Phenolic Bismethylation by a Multifunctional Fungal Flavoenzyme.
abstract: The regioselective functionalization of non-activated carbon atoms such as aliphatic halogenation is a major synthetic challenge. A novel multifunctional enzyme catalyzing the geminal dichlorination of a methyl group was discovered in Aspergillus oryzae (Koji mold), an important fungus that is widely used for Asian food fermentation. A biosynthetic pathway encoded on two different chromosomes yields mono- and dichlorinated polyketides (diaporthin derivatives), including the cytotoxic dichlorodiaporthin as the main product. Bioinformatic analyses and functional genetics revealed an unprecedented hybrid enzyme (AoiQ) with two functional domains, one for halogenation and one for O-methylation. AoiQ was successfully reconstituted in vivo and in vitro, unequivocally showing that this FADH2 -dependent enzyme is uniquely capable of the stepwise gem-dichlorination of a non-activated carbon atom on a freestanding substrate. Genome mining indicated that related hybrid enzymes are encoded in cryptic gene clusters in numerous ecologically relevant fungi.
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Pranatchareeya
authors: P
authors: Chankhamjon
authors: Department of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, 422-8526, Japan.
authors: Yuta
authors: Y
authors: Tsunematsu
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Mie
authors: M
authors: Ishida-Ito
authors: Department of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, 422-8526, Japan.
authors: Yuzuka
authors: Y
authors: Sasa
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Florian
authors: F
authors: Meyer
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Daniela
authors: D
authors: Boettger-Schmidt
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Barbara
authors: B
authors: Urbansky
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Klaus-Dieter
authors: KD
authors: Menzel
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany.
authors: Kirstin
authors: K
authors: Scherlach
authors: Department of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, 422-8526, Japan.
authors: Kenji
authors: K
authors: Watanabe
authors: Department of Biomolecular Chemistry and BioPilotPlant, Leibniz Institute for Natural Product Chemistry and Infection Biology (HKI), Beutenbergstrasse 11a, 07745, Jena, Germany. christian.hertweck@leibniz-hki.de.
authors: Christian
authors: C
authors: Hertweck
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D053898:Biosynthetic Pathways; D005285:Fermentation; D005800:Genes, Fungal; D054879:Halogenation; D008745:Methylation; D010636:Phenols / Q000737:chemistry / Q000378:metabolism*; D061065:Polyketides / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism
chemical_list: D010636:Phenols; D061065:Polyketides
keywords: Aspergillus oryzae; chlorination; enzyme catalysis; genome mining; halogenase