Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose

The efficient diversion of pyruvate from normal fermentative pathways to ethanol production in Klebsiella oxytoca M5A1 requires the expression of Zymomonas mobilis genes encoding both pyruvate decarboxylase and alcohol dehydrogenase. Final ethanol concentrations obtained with the best recombinant, strain M5A1 (pLOI555), were in excess of 40 g/liter with an efficiency of 0.48 g of ethanol (xylose) and 0.50 g of ethanol (glucose) per g of sugar, as compared with a theoretical maximum of 0.51 g of ethanol per g of sugar. The maximal volumetric productivity per hour for both sugars was 2.0 g/liter. This volumetric productivity with xylose is almost twice that previously obtained with ethanologenic Escherichia coli. Succinate was also produced as a minor product during fermentation.

[1]  K. Shanmugam,et al.  Genetic improvement of Escherichia coli for ethanol production: chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase and alcohol dehydrogenase II , 1991, Applied and environmental microbiology.

[2]  S. Maloy,et al.  Regulation of proline utilization in enteric bacteria: cloning and characterization of the Klebsiella put control region , 1991, Journal of bacteriology.

[3]  N. Misawa,et al.  Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli , 1990, Journal of bacteriology.

[4]  L. Ingram,et al.  Effects of environmental conditions on xylose fermentation by recombinant Escherichia coli , 1990, Applied and environmental microbiology.

[5]  D. Clark,et al.  The fermentation pathways of Escherichia coli. , 1989, FEMS microbiology reviews.

[6]  L. Ingram,et al.  Efficient ethanol production from glucose, lactose, and xylose by recombinant Escherichia coli , 1989, Applied and environmental microbiology.

[7]  S. Slater,et al.  Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly-beta-hydroxybutyrate biosynthetic pathway , 1988, Journal of bacteriology.

[8]  T. Jeffries,et al.  Fermentation of hemicellulosic sugars and sugar mixtures by Candida shehatae. , 1988, Biotechnology and bioengineering.

[9]  L. Ingram,et al.  Expression of Different Levels of Ethanologenic Enzymes from Zymomonas mobilis in Recombinant Strains of Escherichia coli , 1988, Applied and environmental microbiology.

[10]  L. Ingram,et al.  Genetic engineering of ethanol production in Escherichia coli , 1987, Applied and environmental microbiology.

[11]  R. K. Finn,et al.  Fermentation of d-Xylose to Ethanol by Genetically Modified Klebsiella planticola , 1987, Applied and environmental microbiology.

[12]  R. K. Finn,et al.  Fermentation of d-Xylose and l-Arabinose to Ethanol by Erwinia chrysanthemi , 1987, Applied and environmental microbiology.

[13]  L. Ingram,et al.  Cloning and sequencing of the alcohol dehydrogenase II gene from Zymomonas mobilis , 1987, Journal of bacteriology.

[14]  L. Ingram,et al.  Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase , 1987, Journal of bacteriology.

[15]  H. Ikeda,et al.  Production of ‘hybrid’ antibiotics by genetic engineering , 1985, Nature.

[16]  A. Poustka,et al.  Selective isolation of cosmid clones by homologous recombination in Escherichia coli. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[17]  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.

[18]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[19]  P. W. Wilson,et al.  Nitrogen Fixation by Members of the Tribe Klebsielleae , 1965, Journal of bacteriology.

[20]  F. Neidhardt,et al.  Physiology of the bacterial cell : a molecular approach , 1990 .

[21]  John D. Wright,et al.  Xylose fermentation , 1989 .