Iterative assembly of two separate polyketide chains by the same single-module bacterial polyketide synthase in the biosynthesis of HSAF

Yaoyao Li, Haotong Chen, Yanjiao Ding, Yunxuan Xie, Haoxin Wang, Ronald Cerny, Yuemao Shen, Liangcheng Du

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Antifungal HSAF (heat-stable antifungal factor, dihydromaltophilin) is a polycyclic tetramate macrolactam from the biocontrol agent Lysobacter enzymogenes. Its biosynthetic gene cluster contains only a single-module polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS), although two separate hexaketide chains are required to assemble the skeleton. To address the unusual biosynthetic mechanism, we expressed the biosynthetic genes in two "clean" strains of Streptomyces and showed the production of HSAF analogues and a polyene tetramate intermediate. We then expressed the PKS module in Escherichia coli and purified the enzyme. Upon incubation of the enzyme with acyl-coenzyme A and reduced nicotinamide adenine dinucleotide phosphate (NADPH), a polyene was detected in the tryptic acyl carrier protein (ACP). Finally, we incubated the polyene-PKS with the NRPS module in the presence of ornithine and adenosine triphosphate (ATP), and we detected the same polyene tetramate as that in Streptomyces transformed with the PKS-NRPS alone. Together, our results provide evidence for an unusual iterative biosynthetic mechanism for bacterial polyketide-peptide natural products.

Original languageEnglish (US)
Pages (from-to)7524-7530
Number of pages7
JournalAngewandte Chemie - International Edition
Volume53
Issue number29
DOIs
StatePublished - Jul 14 2014

Fingerprint

Polyketide Synthases
Polyketides
Polyenes
Biosynthesis
Peptide Synthases
Peptides
Enzymes
Genes
Biocontrol
Acyl Carrier Protein
Coenzymes
Acyl Coenzyme A
Ornithine
Biological Products
NADP
Escherichia coli
Phosphates
Adenosine Triphosphate
Hot Temperature
dihydromaltophilin

Keywords

  • biosynthesis
  • enzymes
  • macrocycles
  • natural products
  • polyketides

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

Cite this

Iterative assembly of two separate polyketide chains by the same single-module bacterial polyketide synthase in the biosynthesis of HSAF. / Li, Yaoyao; Chen, Haotong; Ding, Yanjiao; Xie, Yunxuan; Wang, Haoxin; Cerny, Ronald; Shen, Yuemao; Du, Liangcheng.

In: Angewandte Chemie - International Edition, Vol. 53, No. 29, 14.07.2014, p. 7524-7530.

Research output: Contribution to journalArticle

@article{22186861992940f197cad4bc4baa7987,
title = "Iterative assembly of two separate polyketide chains by the same single-module bacterial polyketide synthase in the biosynthesis of HSAF",
abstract = "Antifungal HSAF (heat-stable antifungal factor, dihydromaltophilin) is a polycyclic tetramate macrolactam from the biocontrol agent Lysobacter enzymogenes. Its biosynthetic gene cluster contains only a single-module polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS), although two separate hexaketide chains are required to assemble the skeleton. To address the unusual biosynthetic mechanism, we expressed the biosynthetic genes in two {"}clean{"} strains of Streptomyces and showed the production of HSAF analogues and a polyene tetramate intermediate. We then expressed the PKS module in Escherichia coli and purified the enzyme. Upon incubation of the enzyme with acyl-coenzyme A and reduced nicotinamide adenine dinucleotide phosphate (NADPH), a polyene was detected in the tryptic acyl carrier protein (ACP). Finally, we incubated the polyene-PKS with the NRPS module in the presence of ornithine and adenosine triphosphate (ATP), and we detected the same polyene tetramate as that in Streptomyces transformed with the PKS-NRPS alone. Together, our results provide evidence for an unusual iterative biosynthetic mechanism for bacterial polyketide-peptide natural products.",
keywords = "biosynthesis, enzymes, macrocycles, natural products, polyketides",
author = "Yaoyao Li and Haotong Chen and Yanjiao Ding and Yunxuan Xie and Haoxin Wang and Ronald Cerny and Yuemao Shen and Liangcheng Du",
year = "2014",
month = "7",
day = "14",
doi = "10.1002/anie.201403500",
language = "English (US)",
volume = "53",
pages = "7524--7530",
journal = "Angewandte Chemie - International Edition",
issn = "1433-7851",
publisher = "John Wiley and Sons Ltd",
number = "29",

}

TY - JOUR

T1 - Iterative assembly of two separate polyketide chains by the same single-module bacterial polyketide synthase in the biosynthesis of HSAF

AU - Li, Yaoyao

AU - Chen, Haotong

AU - Ding, Yanjiao

AU - Xie, Yunxuan

AU - Wang, Haoxin

AU - Cerny, Ronald

AU - Shen, Yuemao

AU - Du, Liangcheng

PY - 2014/7/14

Y1 - 2014/7/14

N2 - Antifungal HSAF (heat-stable antifungal factor, dihydromaltophilin) is a polycyclic tetramate macrolactam from the biocontrol agent Lysobacter enzymogenes. Its biosynthetic gene cluster contains only a single-module polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS), although two separate hexaketide chains are required to assemble the skeleton. To address the unusual biosynthetic mechanism, we expressed the biosynthetic genes in two "clean" strains of Streptomyces and showed the production of HSAF analogues and a polyene tetramate intermediate. We then expressed the PKS module in Escherichia coli and purified the enzyme. Upon incubation of the enzyme with acyl-coenzyme A and reduced nicotinamide adenine dinucleotide phosphate (NADPH), a polyene was detected in the tryptic acyl carrier protein (ACP). Finally, we incubated the polyene-PKS with the NRPS module in the presence of ornithine and adenosine triphosphate (ATP), and we detected the same polyene tetramate as that in Streptomyces transformed with the PKS-NRPS alone. Together, our results provide evidence for an unusual iterative biosynthetic mechanism for bacterial polyketide-peptide natural products.

AB - Antifungal HSAF (heat-stable antifungal factor, dihydromaltophilin) is a polycyclic tetramate macrolactam from the biocontrol agent Lysobacter enzymogenes. Its biosynthetic gene cluster contains only a single-module polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS), although two separate hexaketide chains are required to assemble the skeleton. To address the unusual biosynthetic mechanism, we expressed the biosynthetic genes in two "clean" strains of Streptomyces and showed the production of HSAF analogues and a polyene tetramate intermediate. We then expressed the PKS module in Escherichia coli and purified the enzyme. Upon incubation of the enzyme with acyl-coenzyme A and reduced nicotinamide adenine dinucleotide phosphate (NADPH), a polyene was detected in the tryptic acyl carrier protein (ACP). Finally, we incubated the polyene-PKS with the NRPS module in the presence of ornithine and adenosine triphosphate (ATP), and we detected the same polyene tetramate as that in Streptomyces transformed with the PKS-NRPS alone. Together, our results provide evidence for an unusual iterative biosynthetic mechanism for bacterial polyketide-peptide natural products.

KW - biosynthesis

KW - enzymes

KW - macrocycles

KW - natural products

KW - polyketides

UR - http://www.scopus.com/inward/record.url?scp=84904515170&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84904515170&partnerID=8YFLogxK

U2 - 10.1002/anie.201403500

DO - 10.1002/anie.201403500

M3 - Article

VL - 53

SP - 7524

EP - 7530

JO - Angewandte Chemie - International Edition

JF - Angewandte Chemie - International Edition

SN - 1433-7851

IS - 29

ER -