Coupling S-adenosylmethionine–dependent methylation to growth: Design and uses

by Hao Luo, Anne Sofie L. Hansen, Lei Yang, Konstantin Schneider, Mette Kristensen, Ulla Christensen, Hanne B. Christensen, Bin Du, Emre Özdemir, Adam M. Feist, Jay D. Keasling, Michael K. Jensen, Markus J. Herrgård, Bernhard O. Palsson

We present a selection design that couples S-adenosylmethionine–dependent methylation to growth. We demonstrate its use in improving the enzyme activities of not only N-type and O-type methyltransferases by 2-fold but also an acetyltransferase of another enzyme category when linked to a methylation pathway in Escherichia coli using adaptive laboratory evolution. We also demonstrate its application for drug discovery using a catechol O-methyltransferase and its inhibitors entacapone and tolcapone. Implementation of this design in Saccharomyces cerevisiae is also demonstrated.