Assessment of Anabaena sp strain PCC 7120 as a heterologous expression host for cyanobacterial natural products: production of lyngbyatoxin A

TitleAssessment of Anabaena sp strain PCC 7120 as a heterologous expression host for cyanobacterial natural products: production of lyngbyatoxin A
Publication TypeJournal Article
Year of Publication2016
AuthorsVideau P., Wells K.N, Singh A.J, Gerwick WH, Philmus B.
JournalACS Synthetic Biology
Volume5
Pagination978-988
Date Published2016/09
Type of ArticleArticle
ISBN Number2161-5063
Accession NumberWOS:000383641400010
Keywords(-)-indolactam; activator; assembly-line; biosynthetic gene-cluster; drug discovery; kinase-c; majuscula; marine cyanobacterium; nitrogen; nonribosomal peptide synthetase; promoter fusion; Secondary metabolites; streptomyces-coelicolor a3(2); v
Abstract

Cyanobacteria are well-known producers of natural products of highly varied structure and biological properties. However, the long doubling times, difficulty in establishing genetic methods for marine cyanobacteria, and low compound titers have hindered research into the biosynthesis of their secondary metabolites. While a few attempts to heterologously express cyanobacterial natural products have occurred, the results have been of varied success. Here, we report the first steps in developing the model freshwater cyanobacterium Anabaena sp. strain PCC 7120 (Anabaena 7120) as a general heterologous expression host for cyanobacterial secondary metabolites. We show that Anabaena 7120 can heterologously synthesize lyngbyatoxin A in yields comparable to those of the native producer, Moorea producens, and detail the design and use of replicative plasmids for compound production. We also demonstrate that Anabaena 7120 recognizes promoters from various biosynthetic gene clusters from both free-living and obligate symbiotic marine cyanobacteria. Through simple genetic manipulations, the titer of lyngbyatoxin A can be improved up to 13-fold. The development of Anabaena 7120 as a general heterologous expression host enables investigation of interesting cyanobacterial biosynthetic reactions and genetic engineering of their biosynthetic pathways.

DOI10.1021/acssynbio.6b00038
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