Construction of a synthetic pathway for the production of 1,3-propanediol from glucose
| Autor: | Frazão, Cláudio J. R., Trichez, Débora, Serrano-Bataille, Hélène, Dagkesamanskaia, Adilia, Topham, Christopher M., Walther, Thomas, François, Jean Marie |
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| Přispěvatelé: | Toulouse Biotechnology Institute (TBI), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Molecular Forces Consulting, Toulouse White Biotechnology (TWB), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), ANR14-CE06-0024, Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
propan-1
[SDV.BIO]Life Sciences [q-bio]/Biotechnology gene overexpression Citric Acid Cycle lcsh:Medicine Biotechnologies Article Industrial Microbiology propanediol lcsh:Science surexpression génique Zymomonas lcsh:R Biosynthetic Pathways Lactococcus lactis enzyme Glucose Metabolic Engineering Propylene Glycols 2-diol criblage protéine lcsh:Q escherichia coli protein size grading Pyruvate Decarboxylase Biotechnology |
| Zdroj: | Scientific Reports Scientific Reports, Nature Publishing Group, 2019, 9 (1), 12 p. ⟨10.1038/s41598-019-48091-7⟩ Scientific Reports, Vol 9, Iss 1, Pp 1-12 (2019) Scientific Reports, 2019, 9 (1), 12 p. ⟨10.1038/s41598-019-48091-7⟩ Scientific Reports 1 (9), 12 p.. (2019) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-019-48091-7⟩ |
| Popis: | In this work, we describe the construction of a synthetic metabolic pathway enabling direct biosynthesis of 1,3-propanediol (PDO) from glucose via the Krebs cycle intermediate malate. This non-natural pathway extends a previously published synthetic pathway for the synthesis of (L)-2,4-dihydroxybutyrate (L-DHB) from malate by three additional reaction steps catalyzed respectively, by a DHB dehydrogenase, a 2-keto-4-hydroxybutyrate (OHB) dehydrogenase and a PDO oxidoreductase. Screening and structure-guided protein engineering provided a (L)-DHB dehydrogenase from the membrane-associated (L)-lactate dehydrogenase of E. coli and OHB decarboxylase variants derived from the branched-chain keto-acid decarboxylase encoded by kdcA from Lactococcus lactis or pyruvate decarboxylase from Zymomonas mobilis. The simultaneous overexpression of the genes encoding these enzymes together with the endogenous ydhD-encoded aldehyde reductase enabled PDO biosynthesis from (L)-DHB. While the simultaneous expression of the six enzymatic activities in a single engineered E. coli strain resulted in a low production of 0.1 mM PDO from 110 mM glucose, a 40-fold increased PDO titer was obtained by co-cultivation of an E. coli strain expressing the malate-DHB pathway with another strain harboring the DHB-to-PDO pathway. |
| Databáze: | OpenAIRE |
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