Construction of a synthetic pathway for the production of 1,3-propanediol from glucose.

Autor: Frazão CJR; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France., Trichez D; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France., Serrano-Bataille H; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France., Dagkesamanskaia A; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France., Topham CM; Molecular Forces Consulting, 40 rue Boyssonne, F-31400, Toulouse, France., Walther T; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France.; TWB, 3 Rue des Satellites, Canal Biotech Building 2, F-31400, Toulouse, France.; TU Dresden, Institute of Natural Materials Technology, 01062, Dresden, Germany., François JM; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, 135 Avenue de Rangueil, F-31077, Toulouse, France. fran_jm@insa-toulouse.fr.; TWB, 3 Rue des Satellites, Canal Biotech Building 2, F-31400, Toulouse, France. fran_jm@insa-toulouse.fr.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2019 Aug 09; Vol. 9 (1), pp. 11576. Date of Electronic Publication: 2019 Aug 09.
DOI: 10.1038/s41598-019-48091-7
Abstrakt: 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: MEDLINE
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