Recombinant expression and subcellular targeting of the particulate methane monooxygenase (pMMO) protein components in plants.
Recombinant expression and subcellular targeting of the particulate methane monooxygenase (pMMO) protein components in plants.
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DOI：
10.1038/s41598-023-42224-9
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发表时间：
2023-09-15
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
甲烷是一种强有力的温室气体，自工业化前以来，它导致了大约五分之一的全球变暖。农业部门产生大量甲烷排放，特别是畜牧业、废物管理和水稻种植。仅稻田就产生了约9%的人为排放量。甲烷是由产甲烷古菌在淹水稻田中产生的，并通过水稻植株的通气组织输送到大气中。因此，生物工程水稻与催化剂解毒甲烷的途中可能有助于有效的减排战略。颗粒甲烷单加氧酶（pMMO）是自然界中发现的主要甲烷催化剂，并且是由甲烷氧化细菌表达的酶复合物。pMMO的重组表达一直具有挑战性，这可能是由于其膜定位、多聚体结构和多顺反子操纵子。在这里，我们展示了第一步的工程植物的甲烷解毒与模型系统烟草和拟南芥中表达的三个pMMO亚基。膜拓扑结构和蛋白质-蛋白质相互作用与植物ER上pMMO亚基的正确折叠和组装一致。此外，合成的自切割多肽导致所有三个亚基的同时表达，尽管低表达水平排除了更详细的结构研究。这项工作提出了植物细胞作为一种新的异源系统的pMMO允许蛋白质的表达和修饰。
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.
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影响因子：
14.9
作者：
Dobson L;Reményi I;Tusnády GE
通讯作者：
Tusnády GE
影响因子：
13.8
作者：
Ellgaard L;Sevier CS;Bulleid NJ
通讯作者：
Bulleid NJ
影响因子：
2
作者：
Botchway, Stanley W.;Scherer, Kathrin M.;Parker, Anthony W.
通讯作者：
Parker, Anthony W.
DOI：
10.1139/o64-190
发表时间：
1964-01-01
期刊：
CANADIAN JOURNAL OF BIOCHEMISTRY AND PHYSIOLOGY
影响因子：
--
作者：
COSSINS, EA
通讯作者：
COSSINS, EA
影响因子：
7.2
作者：
Clough, SJ;Bent, AF
通讯作者：
Bent, AF