4.7 Article

Engineering of a Synthetic Metabolic Pathway for the Assimilation of (D)-Xylose into Value-Added Chemicals

期刊

ACS SYNTHETIC BIOLOGY
卷 5, 期 7, 页码 607-618

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.5b00103

关键词

synthetic pathway; xylose utilization; glycolic acid production; Escherichia coli

资金

  1. Toulouse White Biotechnology (TWB) consortium (project PENTOSYS)
  2. CAPES foundation (Ministry of Education, Brazil)
  3. Institut National de la Recherche Agronomique (INRA, France)
  4. Region Midi Pyrenees

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A synthetic pathway for (D)-xylose assimilation was stoichiometrically evaluated and implemented in Escherichia coli strains. The pathway proceeds via isomerization of (D)-xylose to (D)-xylulose, phosphorylation of (D)-xylulose to obtain (D)-xylulose-l-phosphate (XIP), and aldolytic cleavage of the latter to yield glycolaldehyde and DHAP. Stoichiometric analyses showed that this pathway provides access to ethylene glycol with a theoretical molar yield of 1. Alternatively, both glycolaldehyde and DHAP can be converted to glycolic acid with a theoretical yield that is 20% higher than for the exclusive production of this acid via the glyoxylate shunt. Simultaneous expression of xylulose-1 kinase and X1P aldolase activities, provided by human ketohexokinase-C and human aldolase-B, respectively, restored growth of a (D)-xylulose-5-kinase mutant on xylose. This strain produced ethylene glycol as the major metabolic endproduct. Metabolic engineering provided strains that assimilated the entire C2 fraction into the central metabolism or that produced 4.3 g/L glycolic acid at a molar yield of 0.9 in shake flasks.

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