Genome-wide host-pathway interactions affecting cis-cis-muconic acid production in yeast.

Autor: Cachera P; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby, Denmark., Kurt NC; Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark., Røpke A; Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark., Strucko T; Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark., Mortensen UH; Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark., Jensen MK; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby, Denmark. Electronic address: mije@biosustain.dtu.dk.
Jazyk: angličtina
Zdroj: Metabolic engineering [Metab Eng] 2024 May; Vol. 83, pp. 75-85. Date of Electronic Publication: 2024 Feb 28.
DOI: 10.1016/j.ymben.2024.02.015
Abstrakt: The success of forward metabolic engineering depends on a thorough understanding of the behaviour of a heterologous metabolic pathway within its host. We have recently described CRI-SPA, a high-throughput gene editing method enabling the delivery of a metabolic pathway to all strains of the Saccharomyces cerevisiae knock-out library. CRI-SPA systematically quantifies the effect of each modified gene present in the library on product synthesis, providing a complete map of host:pathway interactions. In its first version, CRI-SPA relied on the colour of the product betaxanthins to quantify strains synthesis ability. However, only a few compounds produce a visible or fluorescent phenotype limiting the scope of our approach. Here, we adapt CRI-SPA to onboard a biosensor reporting the interactions between host genes and the synthesis of the colourless product cis-cis-muconic acid (CCM). We phenotype >9,000 genotypes, including both gene knock-out and overexpression, by quantifying the fluorescence of yeast colonies growing in high-density agar arrays. We identify novel metabolic targets belonging to a broad range of cellular functions and confirm their positive impact on CCM biosynthesis. In particular, our data suggests a new interplay between CCM biosynthesis and cytosolic redox through their common interaction with the oxidative pentose phosphate pathway. Our genome-wide exploration of host:pathway interaction opens novel strategies for improved production of CCM in yeast cell factories.
Competing Interests: Declaration of competing interest The authors declare no financial interest.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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