Deciphering the assembly process of PQQ dependent methanol dehydrogenase
Pyrroloquinoline quinone (PQQ)-dependent methanol dehydrogenases (MDHs), the periplasmic metalloenzymes in Gram-negative methylotrophic bacteria, play a pivotal role in methane and methanol bio-utilization. Although the structures of many PQQ-dependent MDHs have been resolved, including the canonical heterotetrameric enzymes composed of two MxaF and two MxaI subunits with a molecule of PQQ and a calcium ion in the active site in MxaF, the biogenesis of these enzymes remains elusive. Here, we characterize a chaperone, MxaJ, responsible for PQQ incorporation by reconstructing a PQQ-dependent MDH assembly system in Escherichia coli. Using cryo-electron microscopy, we capture the structures of the intermediate complexes formed by the chaperone MxaJ and catalytic subunit MxaF during PQQ-dependent MDH maturation, revealing a chaperone-mediated molecular mechanism of cofactor incorporation. These findings not only advance our understanding on the biogenesis of PQQ-dependent MDH, but also provide an alternative engineering way for methane and methanol bioconversion.
Authors
- Jian Cheng (ORCID: https://orcid.org/0009-0008-7059-9764)
- Qichen Cao (ORCID: https://orcid.org/0000-0001-8157-5882)
- Haichuan Zhou
- Huifeng Jiang (ORCID: https://orcid.org/0000-0002-3757-7896)
- Yuwan Liu (ORCID: https://orcid.org/0000-0001-8200-3077)
- Mian Wu (ORCID: https://orcid.org/0000-0002-2714-0500)
- Liangpo Li
- Hongjun Dong (ORCID: https://orcid.org/0000-0003-2708-0947)
- Haixia Xiao (ORCID: https://orcid.org/0000-0003-0032-5430)
- Junqing Sun
- Feng Gao (ORCID: https://orcid.org/0000-0002-0670-010X)
- Chen Yang (ORCID: https://orcid.org/0009-0002-1447-9617)
- Weidong Liu (ORCID: https://orcid.org/0000-0001-7954-7700)
- Jie Bai (ORCID: https://orcid.org/0000-0002-5051-7169)
- Qian Li
- Manman Han
- Jie Shen (ORCID: https://orcid.org/0000-0003-1768-8748)
Institutions
- Zhejiang A & F University (CN)
- Chinese Academy of Sciences (CN)
- Intelligent Synthetic Biology Center (KR)
- Shanxi Academy of Building Research (CN)
- Tianjin Institute of Industrial Biotechnology (CN)
- Second Affiliated Hospital of Zhejiang University (CN)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-07-19
- DOI
- https://doi.org/10.1038/s41467-025-61958-w
- Citations
- 3
- Primary Topic
- Microbial metabolism and enzyme function
- Type
- article
- Field-Weighted Citation Impact
- 2.42
Funders
- Qingdao Agricultural University
- Chinese Academy of Sciences
- Tianjin Science and Technology Committee
- Dalian Institute of Chemical Physics