• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. Dec 1996; 62(12): 4465–4470.
PMCID: PMC168273

Development of an arabinose-fermenting Zymomonas mobilis strain by metabolic pathway engineering.

Abstract

The substrate fermentation range of the ethanologenic bacterium Zymomonas mobilis was expanded to include the pentose sugar, L-arabinose, which is commonly found in agricultural residues and other lignocellulosic biomass. Five genes, encoding L-arabinose isomerase (araA), L-ribulokinase (araB), L-ribulose-5-phosphate-4-epimerase (araD), transaldolase (talB), and transketolase (tktA), were isolated from Escherichia coli and introduced into Z. mobilis under the control of constitutive promoters that permitted their expression even in the presence of glucose. The engineered strain grew on and produced ethanol from L-arabinose as a sole C source at 98% of the maximum theoretical ethanol yield, based on the amount of consumed sugar. This indicates that arabinose was metabolized almost exclusively to ethanol as the sole fermentation product, with little by-product formation. Although no diauxic growth pattern was evident, the microorganism preferentially utilized glucose before arabinose, apparently reflecting the specificity of the indigenous facilitated diffusion transport system. This microorganism may be useful, along with the previously developed xylose-fermenting Z. mobilis (M. Zhang, C. Eddy, K. Deanda, M. Finkelstein, and S. Picataggio, Science 267:240-243, 1995), in a mixed culture for efficient fermentation of the predominant hexose and pentose sugars in agricultural residues and other lignocellulosic feedstocks to ethanol.

Full Text

The Full Text of this article is available as a PDF (225K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnell WO, Yi KC, Conway T. Sequence and genetic organization of a Zymomonas mobilis gene cluster that encodes several enzymes of glucose metabolism. J Bacteriol. 1990 Dec;172(12):7227–7240. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Conway T, Sewell GW, Ingram LO. Glyceraldehyde-3-phosphate dehydrogenase gene from Zymomonas mobilis: cloning, sequencing, and identification of promoter region. J Bacteriol. 1987 Dec;169(12):5653–5662. [PMC free article] [PubMed]
  • Dimarco AA, Romano AH. d-Glucose Transport System of Zymomonas mobilis. Appl Environ Microbiol. 1985 Jan;49(1):151–157. [PMC free article] [PubMed]
  • Feldmann SD, Sahm H, Sprenger GA. Cloning and expression of the genes for xylose isomerase and xylulokinase from Klebsiella pneumoniae 1033 in Escherichia coli K12. Mol Gen Genet. 1992 Aug;234(2):201–210. [PubMed]
  • Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. [PubMed]
  • Ingram LO, Conway T, Clark DP, Sewell GW, Preston JF. Genetic engineering of ethanol production in Escherichia coli. Appl Environ Microbiol. 1987 Oct;53(10):2420–2425. [PMC free article] [PubMed]
  • LEE N, ENGLESBERG E. Dual effects of structural genes in Escherichia coli. Proc Natl Acad Sci U S A. 1962 Mar 15;48:335–348. [PMC free article] [PubMed]
  • Lee N, Gielow W, Martin R, Hamilton E, Fowler A. The organization of the araBAD operon of Escherichia coli. Gene. 1986;47(2-3):231–244. [PubMed]
  • McMillan JD, Boynton BL. Arbinose utilization by xylose-fermenting yeasts and fungi. Appl Biochem Biotechnol. 1994 Spring;45-46:569–584. [PubMed]
  • Parker C, Barnell WO, Snoep JL, Ingram LO, Conway T. Characterization of the Zymomonas mobilis glucose facilitator gene product (glf) in recombinant Escherichia coli: examination of transport mechanism, kinetics and the role of glucokinase in glucose transport. Mol Microbiol. 1995 Mar;15(5):795–802. [PubMed]
  • Patrick JW, Lee N. Purification and properties of an L-arabinose isomerase from Escherichia coli. J Biol Chem. 1968 Aug 25;243(16):4312–4318. [PubMed]
  • Swings J, De Ley J. The biology of Zymomonas. Bacteriol Rev. 1977 Mar;41(1):1–46. [PMC free article] [PubMed]
  • Weisser P, Krämer R, Sahm H, Sprenger GA. Functional expression of the glucose transporter of Zymomonas mobilis leads to restoration of glucose and fructose uptake in Escherichia coli mutants and provides evidence for its facilitator action. J Bacteriol. 1995 Jun;177(11):3351–3354. [PMC free article] [PubMed]
  • Zhang M, Eddy C, Deanda K, Finkelstein M, Picataggio S. Metabolic Engineering of a Pentose Metabolism Pathway in Ethanologenic Zymomonas mobilis. Science. 1995 Jan 13;267(5195):240–243. [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...