Biosynthesis of ethylene glycol in Escherichia coli

Ethylene glycol (EG) is an important platform chemical with steadily expanding global demand. Its commercial production is currently limited to fossil resources; no biosynthesis route has been delineated. Herein, a biosynthesis route for EG production from d-xylose is reported. This route consists of four steps: d-xylose → d-xylonate → 2-dehydro-3-deoxy-d-pentonate → glycoaldehyde → EG. Respective enzymes, d-xylose dehydrogenase, d-xylonate dehydratase, 2-dehydro-3-deoxy-d-pentonate aldolase, and glycoaldehyde reductase, were assembled. The route was implemented in a metabolically engineered Escherichia coli, in which the d-xylose → d-xylulose reaction was prevented by disrupting the d-xylose isomerase gene. The most efficient construct produced 11.7 g L−1 of EG from 40.0 g L−1 of d-xylose. Glycolate is a carbon-competing by-product during EG production in E. coli; blockage of glycoaldehyde → glycolate reaction was also performed by disrupting the gene encoding aldehyde dehydrogenase, but from this approach, EG productivity was not improved but rather led to d-xylonate accumulation. To channel more carbon flux towards EG than the glycolate pathway, further systematic metabolic engineering and fermentation optimization studies are still required to improve EG productivity.

[1]  M. Doudoroff,et al.  A new phosphorylated intermediate in glucose oxidation. , 1954, The Journal of biological chemistry.

[2]  M. Mavrovouniotis Group contributions for estimating standard gibbs energies of formation of biochemical compounds in aqueous solution , 1990, Biotechnology and bioengineering.

[3]  O. J. Sweeting,et al.  Polyethylene: Preparation, Structure, And Properties , 1957 .

[4]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[5]  J. Aguilar,et al.  Metabolism of L-fucose and L-rhamnose in Escherichia coli: aerobic-anaerobic regulation of L-lactaldehyde dissimilation , 1988, Journal of bacteriology.

[6]  W. Wackernagel,et al.  Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. , 1995, Gene.

[7]  Xinbin Ma,et al.  Ethylene Glycol: Properties, Synthesis, and Applications , 2012 .

[8]  A. Dahms 3-Deoxy-D-pentulosonic acid aldolase and its role in a new pathway of D-xylose degradation. , 1974, Biochemical and biophysical research communications.

[9]  H. Stålbrand,et al.  Heat extraction of corn fiber hemicellulose , 2007, Applied biochemistry and biotechnology.

[10]  L. Jarboe YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals , 2010, Applied Microbiology and Biotechnology.

[11]  H. Mori,et al.  Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection , 2006, Molecular systems biology.

[12]  L. Baldomà,et al.  Involvement of lactaldehyde dehydrogenase in several metabolic pathways of Escherichia coli K12. , 1987, The Journal of biological chemistry.

[13]  Jiying Sun,et al.  Selective hydrogenolysis of biomass-derived xylitol to ethylene glycol and propylene glycol on supported Ru catalysts , 2011 .

[14]  A. Baudot,et al.  Thermal properties of ethylene glycol aqueous solutions. , 2004, Cryobiology.

[15]  G. Huber,et al.  Renewable Chemical Commodity Feedstocks from Integrated Catalytic Processing of Pyrolysis Oils , 2010, Science.

[16]  Zhiguang Zhu,et al.  Cellulose solvent- and organic solvent-based lignocellulose fractionation enabled efficient sugar release from a variety of lignocellulosic feedstocks. , 2012, Bioresource technology.

[17]  A. Zeng,et al.  Microbial production of diols as platform chemicals: recent progresses. , 2011, Current opinion in biotechnology.

[18]  Chankyu Park,et al.  Transcriptional Activation of the Aldehyde Reductase YqhD by YqhC and Its Implication in Glyoxal Metabolism of Escherichia coli K-12 , 2010, Journal of bacteriology.

[19]  W. Chung,et al.  High yield production of D-xylonic acid from D-xylose using engineered Escherichia coli. , 2012, Bioresource technology.

[20]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[21]  L. Warren,et al.  The thiobarbituric acid assay of sialic acids. , 1959, The Journal of biological chemistry.

[22]  Christine Nicole S. Santos,et al.  An Engineered Microbial Platform for Direct Biofuel Production from Brown Macroalgae , 2012, Science.

[23]  A. Burgard,et al.  Metabolic engineering of Escherichia coli for direct production of 1,4-butanediol. , 2011, Nature chemical biology.