Current Microbiology

, 63:319 | Cite as

PHB Biosynthesis in Catabolite Repression Mutant of Burkholderia sacchari

  • Mateus Schreiner Garcez Lopes
  • Guillermo Gosset
  • Rafael Costa Santos Rocha
  • José Gregório Cabrera Gomez
  • Luiziana Ferreira da Silva


Due to the effect of catabolite repression, sugar mixtures cannot be metabolized in a rapid and efficient way implicating in lower productivity in bioprocesses using lignocellulosic hydrolysates. In gram-negative bacteria, this mechanism is mediated by the phosphotransferase system (PTS), which concomitantly internalizes and phosphorylates sugars. In this study, we isolated a UV mutant of Burkholderia sacchari, called LFM828, which transports hexoses and pentoses by a non-PTS uptake system. This mutant presented released glucose catabolite repression over the pentoses. In mixtures of glucose, xylose, and arabinose, specific growth rates and the specific sugar consumption rates were, respectively, 10 and 23% higher in LFM828, resulting in a reduced time to exhaust all sugars in the medium. However, in polyhydroxybutyrate (PHB) biosynthesis experiments it was necessary the supplementation of yeast extract to maintain higher values of growth rate and sugar consumption rate. The deficient growth in mineral medium was partially recovered by replacing the ammonium nitrogen source by glutamate. It was demonstrated that the ammonium metabolism is not defective in LFM828, differently from ammonium, glutamate can also be used as carbon and energy allowing an improvement on the carbohydrates utilization for PHB production in LFM828. In contrast, higher rates of ammonia consumption and CO2 production in LFM828 indicate altered fluxes through the central metabolism in LFM828 and the parental. In conclusion, PTS plays an important role in cell physiology and the elimination of its components has a significant impact on catabolite repression, carbon flux distribution, and PHB biosynthesis in B. sacchari.


Xylose Glutamine Synthetase Catabolite Repression Carbon Catabolite Repression Sugar Mixture 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Authors would like to thank Georgina Hernández-Chávez for HPLC determinations. This study was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and from Program of International Mobility of Santander Bank.


  1. 1.
    Cordaro JC, Melton T, Stratis JP et al (1976) Fosfomycin resistance: selection method for internal and extended deletions of the phosphoenaolpyruvate:sugar phosphotransferase genes of Salmonella typhimurium. J Bacteriol 128:785–793PubMedGoogle Scholar
  2. 2.
    Dien BS, Nichols NN, Bothast RJ (2001) Recombinant Escherichia coli engineered for production of l-lactic acid from hexose and pentose sugars. J Ind Microbiol Biotechnol 27(4):259–264PubMedCrossRefGoogle Scholar
  3. 3.
    Dien BS, Nichols NN, Bothast RJ (2002) Fermentation of sugar mixtures using Escherichia coli catabolite repression mutants engineered for production of l-lactic acid. J Ind Microbiol Biotechnol 2002:221–227CrossRefGoogle Scholar
  4. 4.
    Faires N, Tobisch S, Bachem S et al (1999) The catabolite control protein CcpA controls ammonium assimilation in Bacillus subtilis. J Mol Microbiol Biotechnol 1:141–148PubMedGoogle Scholar
  5. 5.
    Farrel RE Jr (2005) RNA methodologies: a laboratory guide for isolation and characterization, vol 3rd ed. Elsevier Academic Press, New YorkGoogle Scholar
  6. 6.
    Flores N, Yong-Xiao J, Berry A et al (1996) Pathway engineering for the production of aromatic compounds in Escherichia coli. Nat Biotechnol 14:620–623PubMedCrossRefGoogle Scholar
  7. 7.
    Flores S, Gosset G, Flores N et al (2002) Analysis of carbon metabolism in Escherichia coli strains with an inactive phosphotransferase system by 13C labeling and NMR spectroscopy. Metab Eng 4:124–137PubMedCrossRefGoogle Scholar
  8. 8.
    Gomez JGC, Rodrigues MFA, Alli RCP et al (1996) Evaluation of soil gram-negative bacteria yielding polyhydroxyalkanoate acids from carbohydrates and propionic acid. Appl Microbiol Biotechnol 45:785–791CrossRefGoogle Scholar
  9. 9.
    Gosset G (2005) Improvement of Escherichia coli production strains by modification of the phosphoenolpyruvate:sugar phosphotransferase system. Microb Cell Fact 4:14PubMedCrossRefGoogle Scholar
  10. 10.
    Hernandez-Montalvo V, Valle F, Bolivar F et al (2001) Characterization of sugar mixtures utilization by an Escherichia coli mutant devoid of the phosphotransferase system. Appl Microbiol Biotechnol 57:186–191PubMedCrossRefGoogle Scholar
  11. 11.
    Kang HY, Song S, Park C (1998) Priority of pentose utilization at the level of transcription: arabinose, xylose, and ribose operons. Mol Cells 8:318–323PubMedGoogle Scholar
  12. 12.
    Li R, Chen Q, Wang PG et al (2007) A novel-designed Escherichia coli for the production of various polyhydroxyalkanoates from inexpensive substrate mixture. Appl Microbiol Biotechnol 75:1103–1109PubMedCrossRefGoogle Scholar
  13. 13.
    Lindsay SE, Bothast RJ, Ingram LO (1995) Improved strains of recombinant Escherichia coli for ethanol production from sugar mixtures. Appl Microbiol Biotechnol 43:70–75PubMedCrossRefGoogle Scholar
  14. 14.
    Lopes MSG, Rocha RCS, Zanotto SP et al (2009) Screening of bacteria to produce polyhydroxyalkanoates from xylose. World J Microbiol Biotechnol 25:1751–1756CrossRefGoogle Scholar
  15. 15.
    Mao XJ, Huo YX, Buck M et al (2007) Interplay between CRP-cAMP and PII-Ntr systems forms novel regulatory network between carbon metabolism and nitrogen assimilation in Escherichia coli. Nucleic Acids Res 35:1432–1440PubMedCrossRefGoogle Scholar
  16. 16.
    Martin C, Alriksson B, Sjöde A et al (2007) Dilute sulfuric acid pretreatment of agricultural and agro-industrial residues for ethanol production. Appl Biochem Biotechnol 137:339–352PubMedCrossRefGoogle Scholar
  17. 17.
    Nichols NN, Dien BS, Bothast RJ (2001) Use of catabolite repression mutants for fermentation of sugar mixtures to ethanol. Appl Microbiol Biotechnol 56:120–125PubMedCrossRefGoogle Scholar
  18. 18.
    Picon A, Teixeira de Mattos MJ, Postma PW (2005) Reducing the glucose uptake rate in Escherichia coli affects growth rate but not protein production. Biotechnol Bioeng 90:191–200PubMedCrossRefGoogle Scholar
  19. 19.
    Ramsay JA, Aly Hasssan MC, Ramsay BA (1995) Hemicellulose as a potential substrate for production of PHA. Can J Microbiol 41:262–266CrossRefGoogle Scholar
  20. 20.
    Riis V, Mai W (1988) Gas chromatographic determination of poly-b-hydroxybutyric acid in microbial biomass after hydrochloric acid propanolysis. J Chromatogr 445:285–289CrossRefGoogle Scholar
  21. 21.
    Rocha RCS, Silva LF, Taciro MK et al (2008) Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) P(3H-co-3HV) with a broad range of 3HV content at high yields by Burkholderia sacchari IPT 189. World J Microbiol Biotechnol 24:427–431CrossRefGoogle Scholar
  22. 22.
    Sambrook J, Fritsch EF, Maniats T (1989) Molecular cloning: a laboratory manual, 3rd ed edn. Cold Spring Harbor Lab Press, Cold Spring HarborGoogle Scholar
  23. 23.
    Sigüenza R, Flores N, Hernández G et al (1999) Kinetic characterization in batch continuous culture of Escherichia coli mutants affected in phosphoenolpyruvate metabolism: differences in acetic acid production. World J Microbiol Biotechnol 15:587–592CrossRefGoogle Scholar
  24. 24.
    Silva LF, Taciro MK, Michelin MER et al (2004) Poly-3-hydroxybutiyrate (P3HB) production by bacteria from xylose, glucose and sugarcane bagasse hydrolysate. Ind Microbiol Biotechnol 31:245–254CrossRefGoogle Scholar
  25. 25.
    Sudesh K, Abe H, Doi Y (2000) Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Prog Polym Sci 25:1503–1555CrossRefGoogle Scholar
  26. 26.
    Tian ZX, Li QS, Buck M et al (2001) The CRP–cAMP complex and downregulation of the glnAp2 promoter provides a novel regulatory linkage between carbon metabolism and nitrogen assimilation in Escherichia coli. Mol Microbiol 41:911–924PubMedCrossRefGoogle Scholar
  27. 27.
    Wacker I, Ludwig H, Reif I et al (2003) The regulatory link between carbon and nitrogen metabolism in Bacillus subtilis: regulation of the gltAB operon by the catabolite control protein CcpA. Microbiology 149:3001–3009PubMedCrossRefGoogle Scholar
  28. 28.
    White D (2000) The physiology and biochemistry of prokaryotes, vol 2nd ed. Oxford University Press, OxfordGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Mateus Schreiner Garcez Lopes
    • 1
  • Guillermo Gosset
    • 2
  • Rafael Costa Santos Rocha
    • 1
  • José Gregório Cabrera Gomez
    • 1
  • Luiziana Ferreira da Silva
    • 1
  1. 1.Departamento de Microbiologia, Instituto de Ciências BiomédicasUniversidade de São PauloSão PauloBrazil
  2. 2.Departamento de Ingeniería Celular y Biocatálisis, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico

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