Mutational analysis in Corynebacterium stationis MFS transporters for improving nucleotide bioproduction.
Corynebacterium
stationis
MFS transporter
Microbial cell factory
Nucleotide transport
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
04 Mar 2024
04 Mar 2024
Historique:
received:
04
10
2023
accepted:
19
02
2024
revised:
16
02
2024
medline:
4
3
2024
pubmed:
4
3
2024
entrez:
4
3
2024
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 38436751
doi: 10.1007/s00253-024-13080-y
pii: 10.1007/s00253-024-13080-y
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
251Subventions
Organisme : The research center for computational science, Okazaki
ID : 22-IMS-C089
Organisme : The research center for computational science, Okazaki
ID : 23-IMS-C077
Organisme : Japan Society for the Promotion of Science
ID : JP20H05453
Organisme : Institute for Fermentation, Osaka
ID : K-2018-009
Informations de copyright
© 2024. The Author(s).
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