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bioRxiv · 10.1101/2024.08.08.607227

A synthetic cell-free pathway for biocatalytic upgrading of one-carbon substrates

Landwehr, G. M.·Vogeli, B.·Tian, C.·Singal, B.·Gupta, A.·Lion, R.·Sargent, E. H.·Karim, A. S.·Jewett, M. C.
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Abstract

Biotechnological processes hold tremendous potential for the efficient and sustainable conversion of one-carbon (C1) substrates into complex multi-carbon products. However, the development of robust and versatile biocatalytic systems for this purpose remains a significant challenge. In this study, we report a hybrid electrochemical-biochemical cell-free system for the conversion of C1 substrates into the universal biological building block acetyl-CoA. The synthetic reductive formate pathway (ReForm) consists of five core enzymes catalyzing non-natural reactions that were established through a cell-free enzyme engineering platform. We demonstrate that ReForm works in a plug-and-play manner to accept diverse C1 substrates including CO2 equivalents. We anticipate that ReForm will facilitate efforts to build and improve synthetic C1 utilization pathways for a formate-based bioeconomy.

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Landwehr, G. M., Vogeli, B., Tian, C., Singal, B., Gupta, A., Lion, R., Sargent, E. H., Karim, A. S., Jewett, M. C.. 2024-08-08. A synthetic cell-free pathway for biocatalytic upgrading of one-carbon substrates. https://doi.org/10.1101/2024.08.08.607227

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