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Lookup NU author(s): Dr James StachORCiD, Professor Paul RaceORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© The Author(s) 2024.Methanogenic archaea are chemolithotrophic prokaryotes that can reduce carbon dioxide with hydrogen gas to form methane. These microorganisms make a significant contribution to the global carbon cycle, with methanogenic archaea from anoxic environments estimated to contribute > 500 million tons of global methane annually. Archaeal methanogenesis is dependent on the methanofurans; aminomethylfuran containing coenzymes that act as the primary C1 acceptor molecule during carbon dioxide fixation. Although the biosynthetic pathway to the methanofurans has been elucidated, structural adaptations which confer thermotolerance to Mfn enzymes from extremophilic archaea are yet to be investigated. Here we focus on the methanofuran biosynthetic enzyme MfnB, which catalyses the condensation of two molecules of glyceralde-3-phosphate to form 4‑(hydroxymethyl)-2-furancarboxaldehyde-phosphate. In this study, MfnB enzymes from the hyperthermophile Methanocaldococcus jannaschii and the mesophile Methanococcus maripaludis have been recombinantly overexpressed and purified to homogeneity. Thermal unfolding studies, together with steady-state kinetic assays, demonstrate thermoadaptation in the M. jannaschii enzyme. Molecular dynamics simulations have been used to provide a structural explanation for the observed properties. These reveal a greater number of side chain interactions in the M. jannaschii enzyme, which may confer protection from heating effects by enforcing spatial residue constraints.
Author(s): Maddock RMA, Marsh CO, Johns ST, Rooms LD, Duke PW, van der Kamp MW, Stach JEM, Race PR
Publication type: Article
Publication status: Published
Journal: Extremophiles
Year: 2024
Volume: 28
Issue: 3
Print publication date: 01/12/2024
Online publication date: 30/08/2024
Acceptance date: 20/08/2024
Date deposited: 09/09/2024
ISSN (print): 1431-0651
ISSN (electronic): 1433-4909
Publisher: Springer
URL: https://doi.org/10.1007/s00792-024-01359-x
DOI: 10.1007/s00792-024-01359-x
Data Access Statement: All data supporting the findings of this study are available within the paper and its Supplementary Information.
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