Applied Biochemistry and Biotechnology

, Volume 144, Issue 1, pp 47–58 | Cite as

Development of a New Bioprocess for Production of 1,3-propanediol I.: Modeling of Glycerol Bioconversion to 1,3-propanediol with Klebsiella pneumoniae Enzymes



Glycerol is a renewable resource for it is formed as a byproduct during biodiesel production. Because of its large volume production, it seems to be a good idea to develop a technology that converts this waste into products of high value, for example, to 1,3-propanediol (1,3-PD). We suggested an enzymatic bioconversion in a membrane reactor in which the NAD coenzyme can be regenerated, and three key enzymes are retained by a 10-kDa ultrafilter membrane. Unfortunately, some byproducts also formed during successful glycerol to 1,3-PD bioconversion runs, as we used crude enzyme solution of Klebsiella pneumoniae. To study the possibilities to avoid this byproduct formation, we built a mathematical description of this system. The model was also used for simulation bioconversions of high glycerol concentration with and without elimination of byproduct formation and of continuous operation.


1,3-Propanediol Klebsiella pneumoniae Enzymatic bioconversion Modelling 







glycerol dehydrogenase enzyme


glycerol dehydratase enzyme


1,3-propanediol oxydoreductase


dihydroxyacetone kinase


triose-phosphate isomerase


glyceraldehyde-3-phosphate dehydrogenase


phosphoglycerate kinase


phosphoglycerate mutase


phosphopyruvate hydratase


pyruvate kinase


pyruvate synthase


phosphate acetyltransferase


phospholipid-translocating ATPase




lactate 2-monooxygenase







We are thankful to OTKA T032015 and NKFP-3/A/0035/2002 for the financial support of our work.


  1. 1.
    Dunn-Coleman, N. S., Gatenby, A. A., Valle, F. (1998). Methods for the production of 1,3-propanediol by recombinant organisms. World Patent WO9821339.Google Scholar
  2. 2.
    Németh, Á., Kupcsulik, B., & Sevella, B. (2003). 1,3-Propanediol oxidoreductase production with Klebsiella pneumoniae DSM2026. World Journal of Microbiology and Biotechnology, 19(7), 659–663.CrossRefGoogle Scholar
  3. 3.
    Johnson, E. A., & Lin, C. C. (1987). Klebsiella pneumoniae 1,3-Propanediol:NAD+ Oxidoreductase. Journal of Bacteriology, 169(5), 2050–2054.Google Scholar
  4. 4.
    Toraya, T., Ushio, K., Fukui, S., & Hogenkamp, P. C. (1977). Studies on the mechanism of the adenosylcobalamin-dependent diol dehydrase reaction by the use of analogs of coenzyme. Journal of Biological Chemistry, 252(3), 963–970.Google Scholar
  5. 5.
    Biebl, H., Menzel, K., Zeng, A. P., & Deckwer, W. D. (1999). Microbial production of 1,3-propanediol. Applied Microbiology and Biotechnology, 52(3), 289–297.CrossRefGoogle Scholar
  6. 6.
    Mori, K., Tobimatsu, T., Hara, T., & Toraya, T. (1997). Characterization, sequencing, and expression of the genes encoding a reactivating factor for glycerol-inactivated adenosylcobalamin-dependent diol dehydratase. Journal of Biological Chemistry 272(51), 32034–32041.CrossRefGoogle Scholar
  7. 7.
    McGregor, W. J., Phillips, J., Suelter, C. H. (1974). Purification and kinetic characterization of a monovalent cation-activated glycerol dehydrogenase from Aerobacter aerogenes. Journal of Biological Chemistry, 249(10), 3132–3139.Google Scholar
  8. 8.
    Garcia-Alles, L. F., Siebold, C., Nyffeler, T. L., Flukiger-Bruhwiler, K., Schneider, P., Burgi, H. B., et al. (2004). Phosphoenolpyruvate and ATP-dependent dihydroxyacetone kinases: Covalent substrate-binding and kinetic mechanism. Biochemistry, 43, 13037–13045.CrossRefGoogle Scholar
  9. 9.
    Yamanishi, M., Yunoki, M., Tobimatsu, T., Sato, H., Matsui, J., Dokiya, A., et al. (2002). The crystal structure of coenzyme B12-dependent glycerol dehydratase in complex with cobalamin and propane-1,2-diol. European Journal of Biochemistry, 269(18), 4484–4494.CrossRefGoogle Scholar
  10. 10.
    Johnson, E. A., Burke, S. K., Forage, R. G., & Lin, E. C. (1984). Purification and properties of dihydroxyacetone kinase from Klebsiella pneumoniae. Journal of Bacteriology, 160(1), 55–60.Google Scholar
  11. 11.
    Cornish Bowden, A. (2004). Fundamentals of Enzyme Kinetics, 3rd edn. London, UK: Portland.Google Scholar
  12. 12.
    Berglund, O., & Eckstein, F. (1972). ATP and dATP-substituted agaroses and the purification of ribonucleotide reductase. Journal of Biological Chemistry, 224(7276), 253–261.Google Scholar
  13. 13.
    Bückmann, A. F. (1981). An efficient synthesis of High-Molecular-Weight NAD(H) derivatives suitable for continuous operation with coenzyme-dependent enzyme systems. Journal of Applied Biochemistry, 3, 301–315.Google Scholar
  14. 14.
    Obon, J. M., Manjon, A., & Iborra, J. L. (1998). Retention and regeneration of native NAD(H) in noncharged ultrafiltration membrane reactors: application to l-lactate and gluconate production. Biotechnology and Bioengineering, 57(5), 510–517.CrossRefGoogle Scholar
  15. 15.
    Reynaud, C., Sarçabal, P., Meynial-Salles, I., Croux, C., & Soucaille, P. (2003). Molecular characterization of the 1,3-propanediol (1,3-PD) operon of Clostridium butyricum. Applied Biological Sciences, 100(9), 5010–5015.Google Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  1. 1.Department of Agricultural Chemical TechnologyBudapest University of Technology and EconomicsBudapestHungary

Personalised recommendations