In vivo and in vitro trans-acylation by BryP, the putative bryostatin pathway acyltransferase derived from an uncultured marine symbiont.
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.
Lopanik
Nicole B
NB
Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA.
Shields
Jennifer A
JA
Buchholz
Tonia J
TJ
Rath
Christopher M
CM
Hothersall
Joanne
J
Haygood
Margo G
MG
Håkansson
Kristina
K
Thomas
Christopher M
CM
Sherman
David H
DH
D000213:Acyl Carrier Protein / Q000737:chemistry / Q000378:metabolism; D000215:Acylation; D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D055162:Biocatalysis; D001688:Biological Products / Q000096:biosynthesis; D054713:Bryostatins / Q000096:biosynthesis*; D002013:Bryozoa / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D004917:Erythromycin / Q000378:metabolism; D018942:Macrolides / Q000378:metabolism; D008314:Malonates / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016712:Mupirocin / Q000096:biosynthesis; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000378:metabolism; D010802:Phylogeny; D048630:Polyketide Synthases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011551:Pseudomonas fluorescens / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D013379:Substrate Specificity; D013559:Symbiosis
D000213:Acyl Carrier Protein; D001688:Biological Products; D054713:Bryostatins; D018942:Macrolides; D008314:Malonates; D004917:Erythromycin; D048630:Polyketide Synthases; C030290:malonic acid; D016712:Mupirocin; D000217:Acyltransferases; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; C003145:picromycin