Journal Article

Engineering the stambomycin modular polyketide synthase yields 37-membered mini-stambomycins

European Project: 742739,ERC-2016-ADG,SynPlex, Dynamique des Génomes et Adaptation Microbienne (DynAMic) ; Université de Lorraine (UL)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Brachmann, Alexander, Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Paris, Cédric, Piel, Jörn, Hôtel, Laurence, Chepkirui, Clara, Aigle, Bertrand, Su, Li, Laboratoire d'Ingénierie des Biomolécules (LIBio) ; Université de Lorraine (UL), Jacob, Christophe, Weissman, Kira J, ANR-15-IDEX-0004,LUE,Isite LUE, ANR-16-CE92-0006,PKS STRUCTURE,Caractérisation structurale des polykétide synthases par microscopie électronique, diffusion des rayons X aux petits angles et des approches de biophysique et de synthèse chimique alliées, IMPACT Biomolécules, Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA) ; Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)

2022

Overview

Abstract The modular organization of the type I polyketide synthases (PKSs) would seem propitious for rational engineering of desirable analogous. However, despite decades of efforts, such experiments remain largely inefficient. Here, we combine multiple, state-of-the-art approaches to reprogram the stambomycin PKS by deleting seven internal modules. One system produces the target 37-membered mini-stambomycin metabolites − a reduction in chain length of 14 carbons relative to the 51-membered parental compounds − but also substantial quantities of shunt metabolites. Our data also support an unprecedented off-loading mechanism of such stalled intermediates involving the C-terminal thioesterase domain of the PKS. The mini-stambomycin yields are reduced relative to wild type, likely reflecting the poor tolerance of the modules downstream of the modified interfaces to the non-native substrates. Overall, we identify factors contributing to the productivity of engineered whole assembly lines, but our findings also highlight the need for further research to increase production titers.

• Publisher: Nature Publishing Group,CCSD,Nature Publishing Group UK,Nature Portfolio
• Subject: 38/70 / 631/326/2522 / 631/61/318 / 631/92/552 / 631/92/60/1167 / 82/58 / 82/80 / 82/83 / Amino Acid Sequence / Assembly lines / Biochemistry / Biochemistry, Molecular Biology / Genetic engineering / Humanities and Social Sciences / Interfaces / Intermediates / Life Sciences / Macrolides - chemistry / Macrolides - metabolism / Metabolic Engineering / Metabolites / Modular engineering / Modules / multidisciplinary / Multienzyme Complexes / Polyketide synthase / Polyketide Synthases - genetics / Polyketide Synthases - metabolism / Science / Science (multidisciplinary) / Structural Biology / Substrate Specificity / Substrates / Synthetic Biology
• ISBN: 0007474071000