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Reprogramming Escherichia coli pyruvate-forming reaction towards chorismate derivatives production
Institution:1. Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, AZ 85287, ECG 301, 501 E. Tyler Mall, Arizona, 85287, United States;2. School of Life Sciences, Arizona State University, 427 E. Tyler Mall, Tempe, AZ, 85287, United States;1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan;2. Center for Sustainable Resource Science, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan;3. Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan
Abstract:Microbial metabolic pathway engineering is a potent strategy used worldwide to produce aromatic compounds. We drastically rewired the primary metabolic pathway of Escherichia coli to produce aromatics and their derivatives. The metabolic pathway of E. coli was compartmentalized into the production and energy modules. We focused on the pyruvate-forming reaction in the biosynthesis pathway of some compounds as the reaction connecting those modules. E. coli strains were engineered to show no growth unless pyruvate was synthesized along with the compounds of interest production. Production of salicylate and maleate was demonstrated to confirm our strategy's versatility. In maleate production, the production, yield against the theoretical yield, and production rate reached 12.0 g L?1, 67%, and up to fourfold compared to that in previous reports, respectively; these are the highest values of maleate production in microbes to our knowledge. The results reveal that our strategy strongly promotes the production of aromatics and their derivatives.
Keywords:Metabolic engineering  Maleate  Pyruvate  Metabolic rewiring  Emden–Meyerhof pathway"}  {"#name":"keyword"  "$":{"id":"kwrd0040"}  "$$":[{"#name":"text"  "_":"EM pathway  pentose phosphate pathway"}  {"#name":"keyword"  "$":{"id":"kwrd0050"}  "$$":[{"#name":"text"  "_":"PP pathway  tricarboxylic acid (TCA) cycle"}  {"#name":"keyword"  "$":{"id":"kwrd0060"}  "$$":[{"#name":"text"  "_":"TCA cycle  pyruvate"}  {"#name":"keyword"  "$":{"id":"kwrd0070"}  "$$":[{"#name":"text"  "_":"PYR  acetyl-coenzyme A"}  {"#name":"keyword"  "$":{"id":"kwrd0080"}  "$$":[{"#name":"text"  "_":"AceCoA  phosphoenolpyruvate"}  {"#name":"keyword"  "$":{"id":"kwrd0090"}  "$$":[{"#name":"text"  "_":"PEP  L-phenylalanine"}  {"#name":"keyword"  "$":{"id":"kwrd0100"}  "$$":[{"#name":"text"  "_":"Phe  phosphotransferase system"}  {"#name":"keyword"  "$":{"id":"kwrd0110"}  "$$":[{"#name":"text"  "_":"PTS  optical density"}  {"#name":"keyword"  "$":{"id":"kwrd0120"}  "$$":[{"#name":"text"  "$$":[{"#name":"__text__"  "_":"OD"}  {"#name":"inf"  "$":{"loc":"post"}  "_":"600  L-tyrosine"}  {"#name":"keyword"  "$":{"id":"kwrd0130"}  "$$":[{"#name":"text"  "_":"Tyr  L-tryptophan"}  {"#name":"keyword"  "$":{"id":"kwrd0140"}  "$$":[{"#name":"text"  "_":"Trp  Entner–Doudoroff pathway"}  {"#name":"keyword"  "$":{"id":"kwrd0150"}  "$$":[{"#name":"text"  "_":"ED pathway  glyoxylate cycle"}  {"#name":"keyword"  "$":{"id":"kwrd0160"}  "$$":[{"#name":"text"  "_":"GO cycle  disodium succinate"}  {"#name":"keyword"  "$":{"id":"kwrd0170"}  "$$":[{"#name":"text"  "_":"SUC  L-valine"}  {"#name":"keyword"  "$":{"id":"kwrd0180"}  "$$":[{"#name":"text"  "_":"Val  citrate lyase"}  {"#name":"keyword"  "$":{"id":"kwrd0190"}  "$$":[{"#name":"text"  "_":"CTL  L-serine deaminase"}  {"#name":"keyword"  "$":{"id":"kwrd0200"}  "$$":[{"#name":"text"  "_":"LSD  3-hydroxybenzoate"}  {"#name":"keyword"  "$":{"id":"kwrd0210"}  "$$":[{"#name":"text"  "_":"3HBA  alpha-ketoglutarate"}  {"#name":"keyword"  "$":{"id":"kwrd0220"}  "$$":[{"#name":"text"  "_":"aKG  malate dehydrogenase"}  {"#name":"keyword"  "$":{"id":"kwrd0230"}  "$$":[{"#name":"text"  "_":"MDH  dissolved oxygen"}  {"#name":"keyword"  "$":{"id":"kwrd0240"}  "$$":[{"#name":"text"  "_":"DO  4-hydroxybenzoate"}  {"#name":"keyword"  "$":{"id":"kwrd0250"}  "$$":[{"#name":"text"  "_":"4HBA  L-methionine"}  {"#name":"keyword"  "$":{"id":"kwrd0260"}  "$$":[{"#name":"text"  "_":"Met  Gas chromatography–mass spectrometry"}  {"#name":"keyword"  "$":{"id":"kwrd0270"}  "$$":[{"#name":"text"  "_":"GC–MS
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