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The Biosynthesis of Norsesquiterpene Aculenes Requires Three Cytochrome P450 Enzymes to Catalyze a Stepwise Demethylation Process
Author(s) -
Lee ChiFang,
Chen LiXun,
Chiang ChenYu,
Lai ChenYu,
Lin HsiaoChing
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201910200
Subject(s) - demethylation , aspergillus oryzae , biosynthesis , cyclase , enzyme , decarboxylation , biochemistry , heterologous expression , stereochemistry , chemistry , cytochrome p450 , biology , gene , catalysis , gene expression , recombinant dna , dna methylation
Aculenes are a unique class of norsequiterpenes (C 14 ) that are produced by Aspergillus aculeatus . The nordaucane skeleton in aculenes A–D may be derived from an ent ‐daucane precursor through demethylation, however, the enzymes involved remain unexplored. We identified the biosynthetic gene cluster and characterized the biosynthetic pathway based on gene inactivation, feeding experiments, and heterologous reconstitution in Saccharomyces cerevisiae and Aspergillus oryzae . We discovered that three cytochrome P450 monoxygenases are required to catalyze the stepwise demethylation process. AneF converts the 12‐methyl group into a carboxylic acid and AneD installs the 10‐hydroxy group for later tautomerization and stabilization. Finally, AneG installs an electron‐withdrawing carbonyl group at the C‐2 position, which triggers C‐12 decarboxylation to yield the nordaucane skeleton. Additionally, a terpene cyclase (AneC) was found that forms a new product (dauca‐4,7‐diene).