Abstract
We describe a general and simple modification to the standard M9 minimal medium recipe that leads to an approximate twofold increase in the yield of heterologously expressed proteins in Escherichia coli BL21(DE3) bacteria. We monitored the growth of bacteria transformed with plasmids for three different test proteins in five minimal media with different concentrations of buffering salts and/or initial media pH. After purification of the over-expressed proteins, we found a clear correlation between the protein yield and change in media pH over time, where the minimal media that were the most buffered and therefore most resistant to change in pH produced the most protein. And in all three test protein cases, the difference in yield was nearly twofold between the best and worst buffering media. Thus, we propose that increasing the buffering capacity of M9 minimal media will generally lead to a similar increase for most of the proteins currently produced by this standard protein expression protocol. Moreover, we have qualitatively found that this effect also extends to deuterated M9 minimal media growths, which could lead to significant cost savings in these preparations.
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Acknowledgements
We would like to thank Marella D. Canny for critically reading this manuscript, and other members of the Latham laboratory for help with bacterial growths and stimulating discussions. This work was supported by grant D-1798 from the Welch Foundation (M.P.L.).
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Azatian, S.B., Kaur, N. & Latham, M.P. Increasing the buffering capacity of minimal media leads to higher protein yield. J Biomol NMR 73, 11–17 (2019). https://doi.org/10.1007/s10858-018-00222-4
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DOI: https://doi.org/10.1007/s10858-018-00222-4