Methane monooxygenase (MMO), which exists in particulate (pMMO) or soluble forms (sMMO) in methanotrophic bacteria, is an industrially promising enzyme that catalyses oxidation of low-reactive methane and other carbon feedstocks into methanol and their corresponding oxidation products. However, the simple, fast and high-yield production of functionally active MMO, which has so far been unsuccessful despite diverse approaches based on either native methanotroph culture or recombinant expression systems, remains a major challenge for its industrial applications. Here we developed pMMO-mimetic catalytic protein constructs by genetically encoding the beneficial reassembly of catalytic domains of pMMO on apoferritin as a biosynthetic scaffold. This approach resulted in high-yield synthesis of stable and soluble protein constructs in Escherichia coli, which successfully retain enzymatic activity for methanol production with a turnover number comparable to that of native pMMO.
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This study was supported by the 2015 NLRL (National Leading Research Lab.) Project (grant no. NRF (National Research Foundation Korea)-2015R1A2A1A05001861), Bio & Medical Technology Development Program (grant no. NRF‐2017M3A9F5032628), and also partly by NRF-Korea (NRF-2016R1A6A3A11933393).
The authors declare no competing interests.
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Supplementary Discussion, Supplementary Methods, Supplementary Figures 1–21, Supplementary Tables 1 and 2, Supplementary Note 1 and Supplementary References
Cartesian coordinates of Figure 1b
MD simulations of pmoB with duroquinol and hydroquinone
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Kim, H.J., Huh, J., Kwon, Y.W. et al. Biological conversion of methane to methanol through genetic reassembly of native catalytic domains. Nat Catal 2, 342–353 (2019). https://doi.org/10.1038/s41929-019-0255-1
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