Deciphering the assembly process of PQQ dependent methanol dehydrogenase

Abstract 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...

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Main Authors: Haichuan Zhou, Junqing Sun, Jian Cheng, Min Wu, Jie Bai, Qian Li, Jie Shen, Manman Han, Chen Yang, Liangpo Li, Yuwan Liu, Qichen Cao, Weidong Liu, Haixia Xiao, Hongjun Dong, Feng Gao, Huifeng Jiang
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61958-w
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author Haichuan Zhou
Junqing Sun
Jian Cheng
Min Wu
Jie Bai
Qian Li
Jie Shen
Manman Han
Chen Yang
Liangpo Li
Yuwan Liu
Qichen Cao
Weidong Liu
Haixia Xiao
Hongjun Dong
Feng Gao
Huifeng Jiang
author_facet Haichuan Zhou
Junqing Sun
Jian Cheng
Min Wu
Jie Bai
Qian Li
Jie Shen
Manman Han
Chen Yang
Liangpo Li
Yuwan Liu
Qichen Cao
Weidong Liu
Haixia Xiao
Hongjun Dong
Feng Gao
Huifeng Jiang
author_sort Haichuan Zhou
collection DOAJ
description Abstract 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.
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spelling doaj-art-8197b3aebf1446009ce9e242097fa40e2025-08-20T03:05:10ZengNature PortfolioNature Communications2041-17232025-07-0116111110.1038/s41467-025-61958-wDeciphering the assembly process of PQQ dependent methanol dehydrogenaseHaichuan Zhou0Junqing Sun1Jian Cheng2Min Wu3Jie Bai4Qian Li5Jie Shen6Manman Han7Chen Yang8Liangpo Li9Yuwan Liu10Qichen Cao11Weidong Liu12Haixia Xiao13Hongjun Dong14Feng Gao15Huifeng Jiang16State Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesInnovative Vaccine and Immunotherapy Research Center, the Second Affiliated Hospital, Zhejiang University School of MedicineState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesLaboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesLaboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesState Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesAbstract 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.https://doi.org/10.1038/s41467-025-61958-w
spellingShingle Haichuan Zhou
Junqing Sun
Jian Cheng
Min Wu
Jie Bai
Qian Li
Jie Shen
Manman Han
Chen Yang
Liangpo Li
Yuwan Liu
Qichen Cao
Weidong Liu
Haixia Xiao
Hongjun Dong
Feng Gao
Huifeng Jiang
Deciphering the assembly process of PQQ dependent methanol dehydrogenase
Nature Communications
title Deciphering the assembly process of PQQ dependent methanol dehydrogenase
title_full Deciphering the assembly process of PQQ dependent methanol dehydrogenase
title_fullStr Deciphering the assembly process of PQQ dependent methanol dehydrogenase
title_full_unstemmed Deciphering the assembly process of PQQ dependent methanol dehydrogenase
title_short Deciphering the assembly process of PQQ dependent methanol dehydrogenase
title_sort deciphering the assembly process of pqq dependent methanol dehydrogenase
url https://doi.org/10.1038/s41467-025-61958-w
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