A rationally designed miniature of soluble methane monooxygenase enables rapid and high-yield methanol production in Escherichia coli

Yeonhwa Yu, Yongfan Shi, Young Wan Kwon, Yoobin Choi, Yusik Kim, Jeong-Geol Na, June Huh () and Jeewon Lee ()
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Yeonhwa Yu: Korea University
Yongfan Shi: Sogang University
Young Wan Kwon: Korea University
Yoobin Choi: Korea University
Yusik Kim: Korea University
Jeong-Geol Na: Sogang University
June Huh: Korea University
Jeewon Lee: Korea University

Nature Communications, 2024, vol. 15, issue 1, 1-16

Abstract: Abstract Soluble methane monooxygenase (sMMO) oxidizes a wide range of carbon feedstocks (C1 to C8) directly using intracellular NADH and is a useful means in developing green routes for industrial manufacturing of chemicals. However, the high-throughput biosynthesis of active recombinant sMMO and the ensuing catalytic oxidation have so far been unsuccessful due to the structural and functional complexity of sMMO, comprised of three functionally complementary components, which remains a major challenge for its industrial applications. Here we develop a catalytically active miniature of sMMO (mini-sMMO), with a turnover frequency of 0.32 s−1, through an optimal reassembly of minimal and modified components of sMMO on catalytically inert and stable apoferritin scaffold. We characterise the molecular characteristics in detail through in silico and experimental analyses and verifications. Notably, in-situ methanol production in a high-cell-density culture of mini-sMMO-expressing recombinant Escherichia coli resulted in higher yield and productivity (~ 3.0 g/L and 0.11 g/L/h, respectively) compared to traditional methanotrophic production.

Date: 2024
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DOI: 10.1038/s41467-024-48671-w

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