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bioRxiv · 10.1101/2023.12.03.569792

High yield, low magnesium flexizyme reactions in a water-ice eutectic phase

Davisson, J.·Alejo, J.·Blank, M.·Kalb, E.·Prasad, A.·Knudson, I.·Schepartz, A.·Engelhart, A. E.·Adamala, K. P.
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Abstract

Flexizymes enable the stoichiometric acylation of tRNAs with a variety of compounds, enabling the in vitro translation of peptides with both non-natural backbones and side chains. However, flexizyme reactions have several drawbacks, including single-turnover kinetics, high Mg(II) carryover inhibiting in vitro translation, and rapid product hydrolysis. Here we present flexizyme reactions utilizing an ice-eutectic phase, with high yields, 30X lower Mg(II), and long-term product stability. The eutectic flexizyme reactions increase the ease of use and flexibility of flexizyme aminoacylation, and increase the in vitro protein production.

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Davisson, J., Alejo, J., Blank, M., Kalb, E., Prasad, A., Knudson, I., Schepartz, A., Engelhart, A. E., Adamala, K. P.. 2023-12-03. High yield, low magnesium flexizyme reactions in a water-ice eutectic phase. https://doi.org/10.1101/2023.12.03.569792

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