Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Characterisation of a glucose phosphotransferase system in Clostridium acetobutylicum ATCC 824


The transport of glucose by the solventogenic anaerobe Clostridium acetobutylicum was investigated. Glucose phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) activity was detected in extracts prepared from cultures grown on glucose and extract fractionation revealed that both soluble and membrane components are required for activity. Glucose PTS activity was inhibited by the analogue methyl α-glucoside, indicating that the PTS enzyme II belongs to the glucose-glucoside (Glc) family of proteins. Consistent with this conclusion, labelled methyl α-glucoside was phosphorylated by PEP in cell-free extracts and this activity was inhibited by glucose. A single gene encoding a putative enzyme II of the glucose family, which we have designated glcG, was identified from the C. acetobutylicum ATCC 824 genome sequence. In common with certain other low-GC gram-positive bacteria, including Bacillus subtilis, the C. acetobutylicumglcG gene appears to be associated with a BglG-type regulator mechanism, as it is preceded by a transcription terminator that is partially overlapped by a typical ribonucleic antiterminator (RAT) sequence, and is downstream of an open reading frame that appears to encode a transcription antiterminator protein. This is the first report of a glucose transport mechanism in this industrially important organism.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

  2. Aymerich S, Steinmetz M (1992) Specificity determinants and structural features in the RNA target of the bacterial antiterminator proteins of the BglG/SacY family. Proc Natl Acad Sci USA 89:10410–10414

  3. Barabote RD, Saier MH Jr (2005) Comparative genome analyses of the bacterial phosphotransferase system. Microbiol Mol Biol Rev 69:608–634

  4. Behrens S, Mitchell WJ, Bahl H (2001) Molecular analysis of the mannitol operon of Clostridium acetobutylicum encoding a phosphotransferase system and a putative PTS-modulated regulator. Microbiology 147:75–86

  5. Binet MRB, Bouvet OM (1998) Transport of glucose by a phosphoenolpyruvate:mannose phosphotransferase system in Pasteurella multocida. Res Microbiol 149:83–94

  6. Brückner R, Titgemeyer F (2002) Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization. FEMS Microbiol Lett 209:141–148

  7. Christiansen I, Hengstenberg W (1999) Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system—two highly similar glucose permeases in Staphylococcus carnosus with different glucoside specificity: protein engineering in vivo? Microbiology 145:2881–2889

  8. Crutz AM, Steinmetz M (1992) Transcription of the Bacillus subtilis sacX and sacY genes, encoding regulators of sucrose metabolism, is both inducible by sucrose and controlled by the DegS–DegU signaling system. J Bacteriol 174:6087–6095

  9. Debarbouillé M, Arnaud M, Fouet A, Klier A, Rapoport G (1990) The sacT gene regulating the sacPA operon in Bacillus subtilis shares strong homology with transcriptional antiterminators. J Bacteriol 172:3966–3973

  10. Dürre P (1998) New insights and novel developments in clostridial acetone/butanol/isopropanol fermentation. Appl Microbiol Biotechnol 49:639–648

  11. Knezevic I, Bachem S, Sickmann A, Meyer HE, Stulke J, Hengstenberg W (2000) Regulation of the glucose-specific phosphotransferase system (PTS) of Staphylococcus carnosus by the antiterminator protein GlcT. Microbiology 146:2333–2342

  12. Krüger S, Hecker M (1995) Regulation of the putative bglPH operon for aryl-beta-glucoside utilization in Bacillus subtilis. J Bacteriol 177:5590–5597

  13. Langbein I, Bachem S, Stülke J (1999) Specific interaction of the RNA-binding domain of the Bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT. J Mol Biol 293:795–805

  14. Lee J, Mitchell WJ, Blaschek HP (2001) Glucose uptake in Clostridium beijerinckii NCIMB 8052 and the solvent-hyperproducing mutant BA101. Appl Environ Microbiol 67:5025–5031

  15. Lengeler JW, Jahreis K, Wehmeier UF (1994) Enzymes II of the phosphoenolpyruvate-dependent phosphotransferase systems: their structure and function in carbohydrate transport. Biochim Biophys Acta 1188:1–28

  16. Mathews DH, Sabina J, Zuker M, Turner DH (1999) Expanded sequence dependence of thermodynamic parameters provides robust prediction of RNA secondary structure. J Mol Biol 288:911–940

  17. Mitchell WJ (1996) Carbohydrate uptake and utilization by Clostridium beijerinckii NCIMB 8052. Anaerobe 2:379–384

  18. Mitchell WJ (1998) Physiology of carbohydrate to solvent conversion by clostridia. Adv Microb Physiol 39:31–130

  19. Mitchell WJ, Booth IR (1984) Characterization of the Clostridium pasteurianum phosphotransferase system. J Gen Microbiol 130:2193–2200

  20. Mitchell WJ, Tangney M (2005) Carbohydrate uptake by the phosphotransferase system and other mechanisms. In: Dürre P (ed) Handbook on Clostridia. CRC Press pp165–185

  21. Mitchell WJ, Shaw JE, Andrews L (1991) Properties of the glucose phosphotransferase system of Clostridium acetobutylicum NCIB 8052. Appl Environ Microbiol 57:2534–2539

  22. Nölling J et al (2001) Genome sequencing and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum. J Bacteriol 183:4823–4838

  23. O’Brien RW, Morris JG (1971) Oxygen and the growth and metabolism of Clostridium acetobutylicum. J Gen Microbiol 68:307–318

  24. Postma PW, Lengeler JW, Jacobson GR (1993) Phosphoenolpyruvate: carbohydrate phosphotransferase systems of bacteria. Microbiol Rev 57:543–594

  25. Robillard GT, Broos J (1999) Structure/function studies on the bacterial carbohydrate transporters, enzymes II, of the phosphoenolpyruvate-dependent phosphotransferase system. Biochem Biophys Acta 1422:73–104

  26. Rutberg B (1997) Antitermination of transcription of catabolic operons. Mol Microbiol 23:413–421

  27. Saier MH Jr, Reizer J (1992) Proposed uniform nomenclature for the proteins and protein domains of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. J Bacteriol 174:1433–1438

  28. Schmalisch MH, Bachem S, Stülke J (2003) Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation. Elucidation of the phosphorylation chain leading to inactivation of GlcT. J Biol Chem 278:51108–51115

  29. Schnetz K, Stülke J, Gertz S, Krüger S, Krieg M, Hecker M, Rak B (1996) LicT, a Bacillus subtilis transcriptional antiterminator protein of the BglG family. J Bacteriol 178:1971–1979

  30. Stülke J, Martin-Verstraete I, Zagorec M, Rose M, Klier A, Rapoport G (1997) Induction of the Bacillus subtilis ptsGHI operon by glucose is controlled by a novel antiterminator, GlcT. Mol Microbiol 25:65–78

  31. Stülke J, Arnaud M, Rapoport G, Martin-Verstraete I (1998) PRD—a protein domain involved in PTS-dependent induction and carbon catabolite repression of catabolic operons in bacteria. Mol Microbiol 28:865–874

  32. Tangney M, Mitchell WJ (2000) Analysis of a catabolic operon for sucrose transport and metabolism in Clostridium acetobutylicum ATCC 824. J Mol Microbiol Biotechnol 2:71–80

  33. Tangney M, Mitchell WJ (2004) Clostridium tetani encodes a phosphocarrier protein, HPr. Microbiology 150:525–526

  34. Tangney M, Mitchell WJ (2005) Regulation of catabolic gene systems. In: Dürre P (ed) Handbook on clostridia. CRC Press pp583–605

  35. Tangney M, Brehm J, Minton N, Mitchell WJ (1998) A gene system for glucitol transport and metabolism in Clostridium beijerinckii NCIMB 8052. Appl Environ Microbiol 64:1612–1619

  36. Tangney M, Winters GT, Mitchell WJ (2001) Characterization of a maltose transport system in Clostridium acetobutylicum ATCC 824. J Ind Microbiol Biotechnol 27:298–306

  37. Tangney M, Galinier A, Deutscher J, Mitchell WJ (2003) Analysis of the elements of catabolite repression in Clostridium acetobutylicum ATCC 824. J Mol Microbiol Biotechnol 6:6–11

  38. Zagorec M, Postma PW (1992) Cloning and nucleotide sequence of the ptsG gene of Bacillus subtilis. Mol Gen Genet 234:325–328

Download references

Author information

Correspondence to Martin Tangney.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tangney, M., Mitchell, W.J. Characterisation of a glucose phosphotransferase system in Clostridium acetobutylicum ATCC 824. Appl Microbiol Biotechnol 74, 398–405 (2007). https://doi.org/10.1007/s00253-006-0679-9

Download citation


  • Solventogenic clostridia
  • Sugar transport
  • PTS
  • Enzyme II
  • Antiterminator