Optimization of Ethylene Glycol Production from (D)-xylose Via a Synthetic Pathway Implemented in Escherichia Coli

Overview

Journal Microb Cell Fact

Publisher Biomed Central

Specialties Biotechnology
Microbiology

Date 2015 Sep 5

PMID 26336892

Citations 20

Authors

Ceren Alkim

Yvan Cam

Debora Trichez

Clement Auriol

Lucie Spina

Amelie Vax

Francois Bartolo

Philippe Besse

Jean Marie Francois

Thomas Walther

Affiliations

Soon will be listed here.

Abstract

Background: Ethylene glycol (EG) is a bulk chemical that is mainly used as an anti-freezing agent and a raw material in the synthesis of plastics. Production of commercial EG currently exclusively relies on chemical synthesis using fossil resources. Biochemical production of ethylene glycol from renewable resources may be more sustainable.

Results: Herein, a synthetic pathway is described that produces EG in Escherichia coli through the action of (D)-xylose isomerase, (D)-xylulose-1-kinase, (D)-xylulose-1-phosphate aldolase, and glycolaldehyde reductase. These reactions were successively catalyzed by the endogenous xylose isomerase (XylA), the heterologously expressed human hexokinase (Khk-C) and aldolase (Aldo-B), and an endogenous glycolaldehyde reductase activity, respectively, which we showed to be encoded by yqhD. The production strain was optimized by deleting the genes encoding for (D)-xylulose-5 kinase (xylB) and glycolaldehyde dehydrogenase (aldA), and by overexpressing the candidate glycolaldehyde reductases YqhD, GldA, and FucO. The strain overproducing FucO was the best EG producer reaching a molar yield of 0.94 in shake flasks, and accumulating 20 g/L EG with a molar yield and productivity of 0.91 and 0.37 g/(L.h), respectively, in a controlled bioreactor under aerobic conditions.

Conclusions: We have demonstrated the feasibility to produce EG from (D)-xylose via a synthetic pathway in E. coli at approximately 90 % of the theoretical yield.

Citing Articles

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References

Turner P, Miller E, Jarboe L, Baggett C, Shanmugam K, Ingram L . YqhC regulates transcription of the adjacent Escherichia coli genes yqhD and dkgA that are involved in furfural tolerance. J Ind Microbiol Biotechnol. 2010; 38(3):431-9. DOI: 10.1007/s10295-010-0787-5. View

Eisen M, Spellman P, Brown P, Botstein D . Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci U S A. 1998; 95(25):14863-8. PMC: 24541. DOI: 10.1073/pnas.95.25.14863. View

Lord J . Glycolate oxidoreductase in Escherichia coli. Biochim Biophys Acta. 1972; 267(2):227-37. DOI: 10.1016/0005-2728(72)90111-9. View

Subedi K, Kim I, Kim J, Min B, Park C . Role of GldA in dihydroxyacetone and methylglyoxal metabolism of Escherichia coli K12. FEMS Microbiol Lett. 2008; 279(2):180-7. DOI: 10.1111/j.1574-6968.2007.01032.x. View

Boronat A, Aguilar J . Rhamnose-induced propanediol oxidoreductase in Escherichia coli: purification, properties, and comparison with the fucose-induced enzyme. J Bacteriol. 1979; 140(2):320-6. PMC: 216652. DOI: 10.1128/jb.140.2.320-326.1979. View

Dykxhoorn D, St Pierre R, Linn T . A set of compatible tac promoter expression vectors. Gene. 1996; 177(1-2):133-6. DOI: 10.1016/0378-1119(96)00289-2. View

Lee C, Kim I, Park C . Glyoxal detoxification in Escherichia coli K-12 by NADPH dependent aldo-keto reductases. J Microbiol. 2013; 51(4):527-30. DOI: 10.1007/s12275-013-3087-8. View

Bolstad B, Irizarry R, Astrand M, Speed T . A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 2003; 19(2):185-93. DOI: 10.1093/bioinformatics/19.2.185. View

Cherepanov P, Wackernagel W . Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene. 1995; 158(1):9-14. DOI: 10.1016/0378-1119(95)00193-a. View

10.

Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M . Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol. 2006; 2:2006.0008. PMC: 1681482. DOI: 10.1038/msb4100050. View

11.

Tang C, Ruch Jr F, Lin C . Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism. J Bacteriol. 1979; 140(1):182-7. PMC: 216794. DOI: 10.1128/jb.140.1.182-187.1979. View

12.

Liu H, Ramos K, Valdehuesa K, Nisola G, Lee W, Chung W . Biosynthesis of ethylene glycol in Escherichia coli. Appl Microbiol Biotechnol. 2012; 97(8):3409-17. DOI: 10.1007/s00253-012-4618-7. View

13.

Jarboe L . YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals. Appl Microbiol Biotechnol. 2010; 89(2):249-57. DOI: 10.1007/s00253-010-2912-9. View

14.

Child J, Willetts A . Microbial metabolism of aliphatic glycols. Bacterial metabolism of ethylene glycol. Biochim Biophys Acta. 1978; 538(2):316-27. DOI: 10.1016/0304-4165(78)90359-8. View

15.

Ritchie M, Silver J, Oshlack A, Holmes M, Diyagama D, Holloway A . A comparison of background correction methods for two-colour microarrays. Bioinformatics. 2007; 23(20):2700-7. DOI: 10.1093/bioinformatics/btm412. View

16.

Cox D . The biodegradation of polyethylene glycols. Adv Appl Microbiol. 1978; 23:173-94. DOI: 10.1016/s0065-2164(08)70068-6. View

17.

Cam Y, Alkim C, Trichez D, Trebosc V, Vax A, Bartolo F . Engineering of a Synthetic Metabolic Pathway for the Assimilation of (d)-Xylose into Value-Added Chemicals. ACS Synth Biol. 2015; 5(7):607-18. DOI: 10.1021/acssynbio.5b00103. View

18.

Chen Y, Lin E . Dual control of a common L-1,2-propanediol oxidoreductase by L-fucose and L-rhamnose in Escherichia coli. J Bacteriol. 1984; 157(3):828-32. PMC: 215334. DOI: 10.1128/jb.157.3.828-832.1984. View

19.

Curran K, Alper H . Expanding the chemical palate of cells by combining systems biology and metabolic engineering. Metab Eng. 2012; 14(4):289-97. DOI: 10.1016/j.ymben.2012.04.006. View

20.

Pellicer M, Badia J, Aguilar J, Baldoma L . glc locus of Escherichia coli: characterization of genes encoding the subunits of glycolate oxidase and the glc regulator protein. J Bacteriol. 1996; 178(7):2051-9. PMC: 177904. DOI: 10.1128/jb.178.7.2051-2059.1996. View