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Optimization of Phosphate Limited Autoinduction Broth for 2-Stage Heterologous Protein Expression in E. coli

Romel Menacho-Melgar, Jennifer N. Hennigan, View ORCID ProfileMichael D. Lynch
doi: https://doi.org/10.1101/2021.06.08.447502
Romel Menacho-Melgar
1Department of Biomedical Engineering, Duke University
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Jennifer N. Hennigan
2Department of Chemistry, Duke University
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Michael D. Lynch
1Department of Biomedical Engineering, Duke University
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  • For correspondence: michael.lynch@duke.edu
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Abstract

Autoinducible, 2-stage protein expression leveraging phosphate inducible promoters has been recently shown to enable not only high protein titers but also consistent performance across scales from screening systems (microtiter plates) to instrumented bioreactors. However, to date small scale production using microtiter plates and shake flasks rely on a complex autoinduction broth (AB) that requires making numerous media components, not all amenable to autoclaving. In this report, we develop a simpler media formulation (AB-2) with just a few autoclavable components. We show that AB-2 is robust to small changes in its composition and performs equally, if not better, than AB across different scales. AB-2 will facilitate adoption of phosphate limited 2-stage protein expression protocols.

Competing Interest Statement

M.D. Lynch has a financial interest in DMC Biotechnologies, Inc. M.D. Lynch, J.N. Hennigan and R. Menacho-Melgar have a financial interest in Roke Biotechnologies, Inc.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted June 08, 2021.
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Optimization of Phosphate Limited Autoinduction Broth for 2-Stage Heterologous Protein Expression in E. coli
Romel Menacho-Melgar, Jennifer N. Hennigan, Michael D. Lynch
bioRxiv 2021.06.08.447502; doi: https://doi.org/10.1101/2021.06.08.447502
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Optimization of Phosphate Limited Autoinduction Broth for 2-Stage Heterologous Protein Expression in E. coli
Romel Menacho-Melgar, Jennifer N. Hennigan, Michael D. Lynch
bioRxiv 2021.06.08.447502; doi: https://doi.org/10.1101/2021.06.08.447502

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