Construction of an Escherichia Coli K-12 Mutant for Homoethanologenic Fermentation of Glucose or Xylose Without Foreign Genes

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2007 Jan 30

PMID 17259366

Citations 38

Authors

Youngnyun Kim

L O Ingram

K T Shanmugam

Affiliations

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Abstract

Conversion of lignocellulosic feedstocks to ethanol requires microorganisms that effectively ferment both hexose and pentose sugars. Towards this goal, recombinant organisms have been developed in which heterologous genes were added to platform organisms such as Saccharomyces cerevisiae, Zymomonas mobilis, and Escherichia coli. Using a novel approach that relies only on native enzymes, we have developed a homoethanologenic alternative, Escherichia coli strain SE2378. This mutant ferments glucose or xylose to ethanol with a yield of 82% under anaerobic conditions. An essential mutation in this mutant was mapped within the pdh operon (pdhR aceEF lpd), which encodes components of the pyruvate dehydrogenase complex. Anaerobic ethanol production by this mutant is apparently the result of a novel pathway that combines the activities of pyruvate dehydrogenase (typically active during aerobic, oxidative metabolism) with the fermentative alcohol dehydrogenase.

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References

Close T, Shanmugam K . Genetic analysis of a pleiotropic mutant of Klebsiella pneumoniae affected in nitrogen metabolism. J Gen Microbiol. 1980; 116(2):501-10. DOI: 10.1099/00221287-116-2-501. View

Kuyper M, Toirkens M, Diderich J, Winkler A, van Dijken J, Pronk J . Evolutionary engineering of mixed-sugar utilization by a xylose-fermenting Saccharomyces cerevisiae strain. FEMS Yeast Res. 2005; 5(10):925-34. DOI: 10.1016/j.femsyr.2005.04.004. View

Blattner F, Plunkett 3rd G, Bloch C, Perna N, Burland V, Riley M . The complete genome sequence of Escherichia coli K-12. Science. 1997; 277(5331):1453-62. DOI: 10.1126/science.277.5331.1453. View

Gonzalez R, Tao H, Shanmugam K, York S, Ingram L . Global gene expression differences associated with changes in glycolytic flux and growth rate in Escherichia coli during the fermentation of glucose and xylose. Biotechnol Prog. 2002; 18(1):6-20. DOI: 10.1021/bp010121i. View

Cassey B, Guest J, Attwood M . Environmental control of pyruvate dehydrogenase complex expression in Escherichia coli. FEMS Microbiol Lett. 1998; 159(2):325-9. DOI: 10.1111/j.1574-6968.1998.tb12878.x. View

Bothast R, Schlicher M . Biotechnological processes for conversion of corn into ethanol. Appl Microbiol Biotechnol. 2004; 67(1):19-25. DOI: 10.1007/s00253-004-1819-8. View

de Graef M, Alexeeva S, Snoep J, Teixeira de Mattos M . The steady-state internal redox state (NADH/NAD) reflects the external redox state and is correlated with catabolic adaptation in Escherichia coli. J Bacteriol. 1999; 181(8):2351-7. PMC: 93657. DOI: 10.1128/JB.181.8.2351-2357.1999. View

Lee J, Patel P, Sankar P, Shanmugam K . Isolation and characterization of mutant strains of Escherichia coli altered in H2 metabolism. J Bacteriol. 1985; 162(1):344-52. PMC: 218995. DOI: 10.1128/jb.162.1.344-352.1985. View

Clark D . The fermentation pathways of Escherichia coli. FEMS Microbiol Rev. 1989; 5(3):223-34. DOI: 10.1016/0168-6445(89)90033-8. View

10.

Talarico L, Ingram L, Maupin-Furlow J . Production of the Gram-positive Sarcina ventriculi pyruvate decarboxylase in Escherichia coli. Microbiology (Reading). 2001; 147(Pt 9):2425-2435. DOI: 10.1099/00221287-147-9-2425. View

11.

Hasona A, Kim Y, Healy F, Ingram L, Shanmugam K . Pyruvate formate lyase and acetate kinase are essential for anaerobic growth of Escherichia coli on xylose. J Bacteriol. 2004; 186(22):7593-600. PMC: 524897. DOI: 10.1128/JB.186.22.7593-7600.2004. View

12.

Lamed R, Zeikus J . Ethanol production by thermophilic bacteria: relationship between fermentation product yields of and catabolic enzyme activities in Clostridium thermocellum and Thermoanaerobium brockii. J Bacteriol. 1980; 144(2):569-78. PMC: 294704. DOI: 10.1128/jb.144.2.569-578.1980. View

13.

Kleckner N, Bender J, Gottesman S . Uses of transposons with emphasis on Tn10. Methods Enzymol. 1991; 204:139-80. DOI: 10.1016/0076-6879(91)04009-d. View

14.

Mohagheghi A, Dowe N, Schell D, Chou Y, Eddy C, Zhang M . Performance of a newly developed integrant of Zymomonas mobilis for ethanol production on corn stover hydrolysate. Biotechnol Lett. 2004; 26(4):321-5. DOI: 10.1023/b:bile.0000015451.96737.96. View

15.

Zaldivar J, Nielsen J, Olsson L . Fuel ethanol production from lignocellulose: a challenge for metabolic engineering and process integration. Appl Microbiol Biotechnol. 2001; 56(1-2):17-34. DOI: 10.1007/s002530100624. View

16.

Quail M, Haydon D, Guest J . The pdhR-aceEF-lpd operon of Escherichia coli expresses the pyruvate dehydrogenase complex. Mol Microbiol. 1994; 12(1):95-104. DOI: 10.1111/j.1365-2958.1994.tb00998.x. View

17.

Datta A . Characterization of the inhibition of Escherichia coli pyruvate dehydrogenase complex by pyruvate. Biochem Biophys Res Commun. 1991; 176(1):517-21. DOI: 10.1016/0006-291x(91)90955-7. View

18.

Underwood S, Zhou S, Causey T, Yomano L, Shanmugam K, Ingram L . Genetic changes to optimize carbon partitioning between ethanol and biosynthesis in ethanologenic Escherichia coli. Appl Environ Microbiol. 2002; 68(12):6263-72. PMC: 134451. DOI: 10.1128/AEM.68.12.6263-6272.2002. View

19.

Alam K, Clark D . Mutants of Escherichia coli deficient in the fermentative lactate dehydrogenase. J Bacteriol. 1989; 171(1):342-8. PMC: 209593. DOI: 10.1128/jb.171.1.342-348.1989. View

20.

Ingram L, Aldrich H, Borges A, Causey T, Martinez A, Morales F . Enteric bacterial catalysts for fuel ethanol production. Biotechnol Prog. 1999; 15(5):855-66. DOI: 10.1021/bp9901062. View