title: High-titer heterologous production in E. coli of lyngbyatoxin, a protein kinase C activator from an uncultured marine cyanobacterium.
abstract: Many chemically complex cyanobacterial polyketides and nonribosomal peptides are of great pharmaceutical interest, but the levels required for exploitation are difficult to achieve from native sources. Here we develop a framework for the expression of these multifunctional cyanobacterial assembly lines in Escherichia coli using the lyngbyatoxin biosynthetic pathway, derived from a marine microbial assemblage dominated by the cyanobacterium Moorea producens. Heterologous expression of this pathway afforded high titers of both lyngbyatoxin A (25.6 mg L(-1)) and its precursor indolactam-V (150 mg L(-1)). Production, isolation, and identification of all expected chemical intermediates of lyngbyatoxin biosynthesis in E. coli also confirmed the previously proposed biosynthetic route, setting a solid chemical foundation for future pathway engineering. The successful production of the nonribosomal peptide lyngbyatoxin A in E. coli also opens the possibility for future heterologous expression, characterization, and exploitation of other cyanobacterial natural product pathways.
authors.affiliation: School of Biotechnology and Biomolecular Sciences, The University of New South Wales , Sydney 2052, Australia.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Ongley
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Bian
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.forename: Rocky
authors.initials: R
authors.lastname: Chau
authors.forename: William H
authors.initials: WH
authors.lastname: Gerwick
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
authors.forename: Brett A
authors.initials: BA
authors.lastname: Neilan
mesh_terms: D053898:Biosynthetic Pathways; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D007218:Industrial Microbiology / Q000379:methods; D008235:Lyngbya Toxins / Q000235:genetics / Q000378:metabolism*; D008387:Marine Toxins / Q000235:genetics / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism
chemical_list: D008235:Lyngbya Toxins; D008387:Marine Toxins; D011493:Protein Kinase C; C019457:lyngbyatoxin A