Get access

Rational Design of Modular Polyketide Synthases: Morphing the Aureothin Pathway into a Luteoreticulin Assembly Line

Authors

  • Yuki Sugimoto,

    1. Leibniz Institute for Natural Product Research and Infection Biology, HKI, Dept. of Biomolecular Chemistry, Beutenbergstr. 11a, 07745 Jena (Germany)
    Search for more papers by this author
  • Dr. Ling Ding,

    1. Leibniz Institute for Natural Product Research and Infection Biology, HKI, Dept. of Biomolecular Chemistry, Beutenbergstr. 11a, 07745 Jena (Germany)
    Search for more papers by this author
  • Dr. Keishi Ishida,

    1. Leibniz Institute for Natural Product Research and Infection Biology, HKI, Dept. of Biomolecular Chemistry, Beutenbergstr. 11a, 07745 Jena (Germany)
    Search for more papers by this author
  • Prof. Dr. Christian Hertweck

    Corresponding author
    1. Leibniz Institute for Natural Product Research and Infection Biology, HKI, Dept. of Biomolecular Chemistry, Beutenbergstr. 11a, 07745 Jena (Germany)
    2. Chair of Natural Product Chemistry, Friedrich Schiller University, Jena (Germany)
    • Leibniz Institute for Natural Product Research and Infection Biology, HKI, Dept. of Biomolecular Chemistry, Beutenbergstr. 11a, 07745 Jena (Germany)

    Search for more papers by this author

  • We thank A. Perner for MS analysis and H. Heinecke for NMR measurements. This work was supported by the Federal Ministry of Science and Technology (BMBF (Germany)) and the German Science Foundation (DFG).

Abstract

The unusual nitro-substituted polyketides aureothin, neoaureothin (spectinabilin), and luteoreticulin, which are produced by diverse Streptomyces species, point to a joint evolution. Through rational genetic recombination and domain exchanges we have successfully reprogrammed the modular (type I) aur polyketide synthase (PKS) into a synthase that generates luteoreticulin. This is the first rational transformation of a modular PKS to produce a complex polyketide that was initially isolated from a different bacterium. A unique aspect of this synthetic biology approach is that we exclusively used genes from a single biosynthesis gene cluster to design the artificial pathway, an avenue that likely emulates natural evolutionary processes. Furthermore, an unexpected, context-dependent switch in the regiospecificity of a pyrone methyl transferase was observed. We also describe an unprecedented scenario where an AT domain iteratively loads an extender unit onto the cognate ACP and the downstream ACP. This aberrant function is a novel case of non-colinear behavior of PKS domains.

Ancillary