In Vivo and In Vitro Trans-Acylation by BryP, the Putative Bryostatin Pathway Acyltransferase Derived from an Uncultured Marine Symbiont

Nicole B. Lopanik, Jennifer A. Shields, Tonia J. Buchholz, Christopher M. Rath, Joanne Hothersall, Margo G. Haygood, Kristina Håkansson, Christopher M. Thomas, David H. Sherman

Research output: Contribution to journalArticle

62 Scopus citations

Abstract

The putative modular polyketide synthase (PKS) that prescribes biosynthesis of the bryostatin natural products from the uncultured bacterial symbiont of the marine bryozoan Bugula neritina possesses a discrete open reading frame (ORF) (bryP) that encodes a protein containing tandem acyltransferase (AT) domains upstream of the PKS ORFs. BryP is hypothesized to catalyze in trans acylation of the PKS modules for polyketide chain elongation. To verify conservation of function, bryP was introduced into AT-deletion mutant strains of a heterologous host containing a PKS cluster with similar architecture, and polyketide production was partially rescued. Biochemical characterization demonstrated that BryP catalyzes selective malonyl-CoA acylation of native and heterologous acyl carrier proteins and complete PKS modules in vitro. The results support the hypothesis that BryP loads malonyl-CoA onto Bry PKS modules, and provide the first biochemical evidence of the functionality of the bry cluster.

Original languageEnglish (US)
Pages (from-to)1175-1186
Number of pages12
JournalChemistry and Biology
Volume15
Issue number11
DOIs
StatePublished - Nov 24 2008

Keywords

  • CHEMBIO

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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    Lopanik, N. B., Shields, J. A., Buchholz, T. J., Rath, C. M., Hothersall, J., Haygood, M. G., Håkansson, K., Thomas, C. M., & Sherman, D. H. (2008). In Vivo and In Vitro Trans-Acylation by BryP, the Putative Bryostatin Pathway Acyltransferase Derived from an Uncultured Marine Symbiont. Chemistry and Biology, 15(11), 1175-1186. https://doi.org/10.1016/j.chembiol.2008.09.013