Mutational analysis in Corynebacterium stationis MFS transporters for improving nucleotide bioproduction

Product secretion from an engineered cell can be advantageous for microbial cell factories. Extensive work on nucleotide manufacturing, one of the most successful microbial fermentation processes, has enabled Corynebacterium stationis to transport nucleotides outside the cell by random mutagenesis;...

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Veröffentlicht in:Applied microbiology and biotechnology 2024-12, Vol.108 (1), p.251-251, Article 251
Hauptverfasser: Kinose, Keita, Shinoda, Keiko, Konishi, Tomoyuki, Kawasaki, Hisashi
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creator Kinose, Keita
Shinoda, Keiko
Konishi, Tomoyuki
Kawasaki, Hisashi
description Product secretion from an engineered cell can be advantageous for microbial cell factories. Extensive work on nucleotide manufacturing, one of the most successful microbial fermentation processes, has enabled Corynebacterium stationis to transport nucleotides outside the cell by random mutagenesis; however, the underlying mechanism has not been elucidated, hindering its applications in transporter engineering. Herein, we report the nucleotide-exporting major facilitator superfamily (MFS) transporter from the C. stationis genome and its hyperactive mutation at the G64 residue. Structural estimation and molecular dynamics simulations suggested that the activity of this transporter improved via two mechanisms: (1) enhancing interactions between transmembrane helices through the conserved “RxxQG” motif along with substrate binding and (2) trapping substrate-interacting residue for easier release from the cavity. Our results provide novel insights into how MFS transporters change their conformation from inward- to outward-facing states upon substrate binding to facilitate efflux and can contribute to the development of rational design approaches for efflux improvements in microbial cell factories. Keypoints • An MFS transporter from C. stationis genome and its mutation at residue G64 were assessed • It enhanced the transporter activity by strengthening transmembrane helix interactions and trapped substrate-interacting residues • Our results contribute to rational design approach development for efflux improvement Graphical Abstract
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subjects Binding
Biological Transport
Biomedical and Life Sciences
Biotechnologically Relevant Enzymes and Proteins
Biotechnology
Conformation
Corynebacterium
Corynebacterium - genetics
Efflux
Factories
Fermentation
Genomes
Helices
Industrial engineering
Life Sciences
Manufacturing engineering
Membrane Transport Proteins - genetics
Microbial Genetics and Genomics
Microbiology
Microorganisms
Molecular dynamics
Mutation
Nucleotides
Random mutagenesis
Residues
Substrates
title Mutational analysis in Corynebacterium stationis MFS transporters for improving nucleotide bioproduction
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