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However, attempts to engineer\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    to metabolize methanol have achieved limited success. Here, we provide a rigorous systematic analysis of several potential pathway bottlenecks. We show that regeneration of ribulose 5-phosphate in\n                    <jats:italic>E. coli<\/jats:italic>\n                    is insufficient to sustain methanol assimilation, and overcome this by activating the sedoheptulose bisphosphatase variant of the ribulose monophosphate pathway. By leveraging the kinetic isotope effect associated with deuterated methanol as a chemical probe, we further demonstrate that under these conditions overall pathway flux is kinetically limited by methanol dehydrogenase. Finally, we identify NADH as a potent kinetic inhibitor of this enzyme. These results provide direction for future engineering strategies to improve methanol utilization, and underscore the value of chemical biology methodologies in metabolic engineering.\n                  <\/jats:p>","DOI":"10.1038\/s41467-018-04795-4","type":"journal-article","created":{"date-parts":[[2018,6,15]],"date-time":"2018-06-15T10:14:30Z","timestamp":1529057670000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":130,"title":["Improving formaldehyde consumption drives methanol assimilation in engineered E. coli"],"prefix":"10.1038","volume":"9","author":[{"given":"Benjamin M.","family":"Woolston","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason R.","family":"King","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Reiter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bob","family":"Van Hove","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory","family":"Stephanopoulos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,6,19]]},"reference":[{"key":"4795_CR1","doi-asserted-by":"crossref","unstructured":"Clomburg, J. 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