Rewiring Escherichia coli to Transform Formate into Methyl Groups

Abstract

BackgroundBiotechnological applications are steadily growing and have become an important tool to reinvent the synthesis of chemicals and pharmaceuticals for lower dependence on fossil resources. In order to sustain this progression, new feedstocks for biotechnological hosts have to be explored. One-carbon (C1-)compounds, including formate, derived from CO2 or organic waste are accessible in large quantities with renewable energy, making them promising candidates. Previous studies showed that introduction of the formate assimilation machinery from Methylorubrum extorquens into Escherichia coli allows assimilation of formate into the established biotechnological host. Applying this established route for formate assimilation, we here investigated utilisation of formate for the production of value-added building blocks in E. coli using S-adenosylmethionine (SAM)-dependent methyltransferases. ResultsWe first analysed methylation activity in E. coli BL21 with a two-vector system to produce three different methyltransferases together with the formate assimilation machinery. Feeding isotopically labelled formate, products with 51 - 81% 13C-labelling could be obtained by maintaining in vivo methylation activity. Focussing on improvement of in vivo methylation, we analysed two further E. coli strains with an engineered C1-metabolism and, following condition optimisation, achieved a doubled methylation activity with a share of more than 70% formate-derived methyl groups. ConclusionsThis study demonstrates the efficient transformation of formate into methyl groups in E. coli. Our findings support that feeding formate can improve the availability of usable C1-compounds and, as a result, increase in vivo methylation activity in engineered E. coli.