Recombinant expression and subcellular targeting of the particulate methane monooxygenase (pMMO) protein components in plants

SPATOLA ROSSI, Tatiana, TOLMIE, A. Frances, NICHOL, Tim, PAIN, Charlotte, HARRISON, Patrick, SMITH, Thomas J., FRICKER, Mark and KRIECHBAUMER, Verena (2023). Recombinant expression and subcellular targeting of the particulate methane monooxygenase (pMMO) protein components in plants. Scientific Reports, 13 (1): 15337.

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Abstract

Methane is a potent greenhouse gas, which has contributed to approximately a fifth of global warming since pre-industrial times. The agricultural sector produces significant methane emissions, especially from livestock, waste management and rice cultivation. Rice fields alone generate around 9% of total anthropogenic emissions. Methane is produced in waterlogged paddy fields by methanogenic archaea, and transported to the atmosphere through the aerenchyma tissue of rice plants. Thus, bioengineering rice with catalysts to detoxify methane en route could contribute to an efficient emission mitigation strategy. Particulate methane monooxygenase (pMMO) is the predominant methane catalyst found in nature, and is an enzyme complex expressed by methanotrophic bacteria. Recombinant expression of pMMO has been challenging, potentially due to its membrane localization, multimeric structure, and polycistronic operon. Here we show the first steps towards the engineering of plants for methane detoxification with the three pMMO subunits expressed in the model systems tobacco and Arabidopsis. Membrane topology and protein–protein interactions were consistent with correct folding and assembly of the pMMO subunits on the plant ER. Moreover, a synthetic self-cleaving polypeptide resulted in simultaneous expression of all three subunits, although low expression levels precluded more detailed structural investigation. The work presents plant cells as a novel heterologous system for pMMO allowing for protein expression and modification.

Item Type:	Article
Additional Information:	** From Springer Nature via Jisc Publications Router ** Licence for this article: http://creativecommons.org/licenses/by/4.0/ ** Acknowledgements: This work was funded by a Leverhulme Trust fund (RGP-2015-1927) to VK, a BBSRC Engineering Biology Breakthrough Award (BB/W011166/1) to VK and TJS as well as a BBSRC Doctoral Training Program PhD studentship to TSR. **Journal IDs: eissn 2045-2322 **Article IDs: publisher-id: s41598-023-42224-9; manuscript: 42224 **History: collection 12-2023; online 15-09-2023; published_online 15-09-2023; accepted 07-09-2023; registration 07-09-2023; submitted 28-03-2023
Identification Number:	https://doi.org/10.1038/s41598-023-42224-9
SWORD Depositor:	Colin Knott
Depositing User:	Colin Knott
Date Deposited:	18 Sep 2023 13:21
Last Modified:	11 Oct 2023 11:45
URI:	https://shura.shu.ac.uk/id/eprint/32408

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