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Optimization of phosphate-limited autoinduction broth for two-stage heterologous protein expression in Escherichia coli

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Protein expression is a routine methodology used in many areas of biological research. Autoinducible, 2-stage protein expression leveraging phosphate inducible promoters has been recently shown to enable high protein titers as well as consistent expression across scales from screening systems (microtiter plates) to instrumented bioreactors. However, to date small scale production relies on a complex autoinduction broth (AB) that requires making numerous media components. In this report, we develop a simpler media formulation (AB-2) with just a few 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. 

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