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dc.contributor.authorWilliams, Ernest
dc.contributor.authorBachvaroff, Tsvetan
dc.contributor.authorPlace, Allen
dc.date.accessioned2022-04-26T12:45:30Z
dc.date.available2022-04-26T12:45:30Z
dc.date.issued2022-03-23
dc.identifier.urihttp://hdl.handle.net/10713/18658
dc.description.abstractPhotosynthetic dinoflagellates synthesize many toxic but also potential therapeutic compounds therapeutics via polyketide/non-ribosomal peptide synthesis, a common means of producing natural products in bacteria and fungi. Although canonical genes are identifiable in dinoflagellate transcriptomes, the biosynthetic pathways are obfuscated by high copy numbers and fractured synteny. This study focuses on the carrier domains that scaffold natural product synthesis (thiolation domains) and the phosphopantetheinyl transferases (PPTases) that thiolate these carriers. We replaced the thiolation domain of the indigoidine producing BpsA gene from Streptomyces lavendulae with those of three multidomain dinoflagellate transcripts and coexpressed these constructs with each of three dinoflagellate PPTases looking for specific pairings that would identify distinct pathways. Surprisingly, all three PPTases were able to activate all the thiolation domains from one transcript, although with differing levels of indigoidine produced, demonstrating an unusual lack of specificity. Unfortunately, constructs with the remaining thiolation domains produced almost no indigoidine and the thiolation domain for lipid synthesis could not be expressed in E. coli. These results combined with inconsistent protein expression for different PPTase/thiolation domain pairings present technical hurdles for future work. Despite these challenges, expression of catalytically active dinoflagellate proteins in E. coli is a novel and useful tool going forward.en_US
dc.description.urihttps://doi.org/10.3390/microorganisms10040687en_US
dc.language.isoenen_US
dc.publisherMDPI AGen_US
dc.relation.ispartofMicroorganismsen_US
dc.subjectBpsAen_US
dc.subjectPKSen_US
dc.subjectdinoflagellateen_US
dc.subjectindigoidineen_US
dc.subjectnatural producten_US
dc.subjectphosphopantetheinyl transferaseen_US
dc.subjecttoxinen_US
dc.titleDinoflagellate Phosphopantetheinyl Transferase (PPTase) and Thiolation Domain Interactions Characterized Using a Modified Indigoidine Synthesizing Reporter.en_US
dc.typeArticleen_US
dc.identifier.doi10.3390/microorganisms10040687
dc.identifier.pmid35456738
dc.source.journaltitleMicroorganisms
dc.source.volume10
dc.source.issue4
dc.source.countrySwitzerland


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