title: Discovery of Unclustered Fungal Indole Diterpene Biosynthetic Pathways through Combinatorial Pathway Reassembly in Engineered Yeast.
abstract: The structural diversity and biological activities of fungal indole diterpenes (IDTs) are generated in large part by the IDT cyclases (IDTCs). Identifying different IDTCs from IDT biosynthetic pathways is therefore important toward understanding how these enzymes introduce chemical diversity from a common linear precursor. However, IDTCs involved in the cyclization of the well-known aflavinine subgroup of IDTs have not been discovered. Here, using Saccharomyces cerevisiae as a heterologous host and a phylogenetically guided enzyme mining approach, we combinatorially assembled IDT biosynthetic pathways using IDTCs homologues identified from different fungal hosts. We identified the genetically standalone IDTCs involved in the cyclization of aflavinine and anominine and produced new IDTs not previously isolated. The cyclization mechanisms of the new IDTCs were proposed based on the yeast reconstitution results. Our studies demonstrate heterologous pathway assembly is a useful tool in the reconstitution of unclustered biosynthetic pathways.
authors.forename: Man-Cheng
authors.initials: MC
authors.lastname: Tang
authors.forename: Hsiao-Ching
authors.initials: HC
authors.lastname: Lin
authors.affiliation: Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China , Qingdao, Shandong 266003, P. R. China.
authors.forename: Dehai
authors.initials: D
authors.lastname: Li
authors.forename: Yi
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Stanford Genome Technology Center, Stanford University , Palo Alto, California 94304, United States.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Xu
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Cacho
authors.affiliation: Stanford Genome Technology Center, Stanford University , Palo Alto, California 94304, United States.
authors.forename: Maureen E
authors.initials: ME
authors.lastname: Hillenmeyer
authors.forename: Neil K
authors.initials: NK
authors.lastname: Garg
authors.forename: Yi
authors.initials: Y
authors.lastname: Tang
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D003500:Cyclization; D004224:Diterpenes / Q000737:chemistry / Q000378:metabolism*; D005818:Genetic Engineering; D007211:Indoles / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D019761:Phosphorus-Oxygen Lyases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism
chemical_list: D004224:Diterpenes; D007211:Indoles; D019761:Phosphorus-Oxygen Lyases