Rational design of a bacterial import system for new-to-nature molecules.

Autor: Rodríguez-Robles E; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Müller D; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Künzl T; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Nemat SJ; Department of Chemistry, University of Basel, Basel, Switzerland., Edelmann MP; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Srivastava P; Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium., Louis D; Alderys SAS, Orsay, France., Groaz E; Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium., Tiefenbacher K; Department of Chemistry, University of Basel, Basel, Switzerland., Roberts TM; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Herdewijn P; Laboratory of Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium., Marlière P; TESSSI, Paris, France; Scientist of Fortune SA, Luxembourg. Electronic address: phmarliere@tesssi.eu., Panke S; Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland. Electronic address: sven.panke@bsse.ethz.ch.
Jazyk: angličtina
Zdroj: Metabolic engineering [Metab Eng] 2024 Sep; Vol. 85, pp. 26-34. Date of Electronic Publication: 2024 May 25.
DOI: 10.1016/j.ymben.2024.05.005
Abstrakt: Integration of novel compounds into biological processes holds significant potential for modifying or expanding existing cellular functions. However, the cellular uptake of these compounds is often hindered by selectively permeable membranes. We present a novel bacterial transport system that has been rationally designed to address this challenge. Our approach utilizes a highly promiscuous sulfonate membrane transporter, which allows the passage of cargo molecules attached as amides to a sulfobutanoate transport vector molecule into the cytoplasm of the cell. These cargoes can then be unloaded from the sulfobutanoyl amides using an engineered variant of the enzyme γ-glutamyl transferase, which hydrolyzes the amide bond and releases the cargo molecule within the cell. Here, we provide evidence for the broad substrate specificity of both components of the system by evaluating a panel of structurally diverse sulfobutanoyl amides. Furthermore, we successfully implement the synthetic uptake system in vivo and showcase its functionality by importing an impermeant non-canonical amino acid.
Competing Interests: Declaration of competing interest Tilmann Künzl, Philippe Marlière, and Sven Panke are inventors of the patent WO 2019068726 A1 which is related to this work. All other authors declare no competing interests.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: