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Cell-Free Optimized Production of Protoporphyrins

Garcia DC , Davies JP , Rhea K, Lee MF, Lux MM. bioRxiv, 2023.

Porphyrins are molecules composed of one or more cyclic tetrapyrroles whose aromaticity enable semiconductor-like properties, making them useful in a broad range of applications, including artificial photosynthesis and light harvesting, catalysis, single-molecule electronics, sensors, nonlinear optics, and chemical warfare agent degradation. Due to its utility in this work we take advantage of porphyrins as both molecules of interest and as an easily detectable products in a cell-free extract to i) show our ability to produce porphyrins using enriched cell-free extracts, ii) explore consolidating the extract source cells into a single co-culture fermentation in order to limit the need for multiple extract productions, and iii) rapidly generate ideal cofactor and substrate mixtures using DBTL-cycles powered by Design of Experiments (DOE).

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