, Volume 17, Issue 3, pp 607–615 | Cite as

Temperature and pH stability of cellouronic acid

  • Shuji Fujisawa
  • Takuya Isogai
  • Akira Isogai


Cellouronic acid (CUA), (1 → 4)-β-d-polyglucuronate sodium salt, was prepared from regenerated cellulose by 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation in water at pH 10. Changes in chemical structure and degree of polymerization (DP) of CUA by treatment in water under various pH and temperature conditions were studied to evaluate the stability of CUA. No depolymerization occurred on CUA in water at pH 1.0–7.0 and room temperature, while clear depolymerization took place at pH 10 and 13 by β-elimination. When heated in water at >50 °C, CUA was depolymerized by hydrolysis at pH 1.0 and 4.8, and by both hydrolysis and β-elimination at pH 7.0. Kinetic studies showed that CUA depolymerization rate constant was roughly increased with increasing the pH or temperature. Especially, the depolymerization rate constant at pH 13 was approximately 128 and 55 times greater than those at pH 1.0 and 10, respectively, at 60 °C. Activation energies of hydrolysis and β-elimination of CUA were approximately 100 and 20 kJ mol−1, respectively.


TEMPO Cellouronate Cellouronic acid β-elimination Hydrolysis Activation energy Molecular mass SEC-MALLS 



This research was supported by a Grand-in-Aid for Scientific Research (S) (Grant number 21228007) from the Japan Society for the Promotion of Science (JSPS). We thank Professor Emeritus Atsushi Ishizu for valuable discussions.


  1. Bailey WF, Bobbitt JM, Wiber KB (2007) Mechanism of alcohols by oxoammonium cations. J Org Chem 72:4504–4509CrossRefGoogle Scholar
  2. Dadach ZE, Kaliaguine S (1993) Acid hydrolysis of cellulose. Part 1. Experimental kinetic analysis. Can J Chem Eng 71:880–891CrossRefGoogle Scholar
  3. de Nooy AEJ, Basemer AC, van Bekkum H (1994) Highly selective TEMPO mediated oxidation of primary alcohol groups in polysaccharides. Carbohydr Res 113:165–166Google Scholar
  4. de Nooy AEJ, Besemer AC, van Bekkum H (1995) Selective oxidation of primary alcohols mediated by nitroxyl radical in aqueous solution. Kinetics and mechanism. Tetrahedron 51:8023–8032CrossRefGoogle Scholar
  5. Delattre C, Michaud P, Lion JM, Courtois B, Courtois J (2005) Production of glucuronan oligosaccharides using a new glucuronan lyase activity from a Trichoderma sp. strain. J Biotechnol 118:448–457CrossRefGoogle Scholar
  6. Fukuzumi H, Saito T, Iwata T, Kumamoto Y, Isogai A (2008) Transparent and high gas barrier films of cellulose nanofibers prepared by TEMPO-mediated oxidation. Biomacromolecules 10:162–165CrossRefGoogle Scholar
  7. Hollabaugh CB, Burt LH, Walsh AP (1945) Carboxymethylcellulose—uses and applications. Ind Eng Chem 37:943–947CrossRefGoogle Scholar
  8. Isogai A, Kato Y (1998) Preparation of polyuronic acid from cellulose by TEMPO-mediated oxidation. Cellulose 5:153–164CrossRefGoogle Scholar
  9. Isogai T, Yanagisawa M, Isogai A (2009) Degrees of polymerization (DP) and DP distribution of cellouronic acids prepared from alkali-treated celluloses and ball-milled native celluloses by TEMPO-mediated oxidation. Cellulose 16:117–127CrossRefGoogle Scholar
  10. Johansson I, Lindberg B, Theander O (1963) Pseudo cellobiouronic acid, synthesis and acid hydrolysis. Acta Chem Scand 17:2019–2024CrossRefGoogle Scholar
  11. Kato Y, Habu N, Yamaguchi J, Iobayashi Y, Shibata I, Isogai A, Samejima M (2002) Biodegradation of β-1, 4-linked polyglucuronic acid (cellouronic acid). Cellulose 9:75–81CrossRefGoogle Scholar
  12. Kiss J (1969) A mild β-elimination reaction of some d-glucopyranosiduronate 4-sulfates. Carbohydr Res 10:328–330CrossRefGoogle Scholar
  13. Konno N, Habu N, Maeda I, Azuma N, Isogai A (2006) Cellouronate (β-1, 4-linked polyglucuronate) lyase from Brevundimonus sp. SH203: purification and characterization. Carbohydr Polym 64:589–596CrossRefGoogle Scholar
  14. Konno N, Habu N, Iihashi N, Isogai A (2008) Purification and characterization of exo-type cellouronate lyase. Cellulose 15:453–463CrossRefGoogle Scholar
  15. Konno N, Igarashi K, Habu N, Samejima M, Isogai A (2009a) Cloning of the Trichoderma reesei cDNA encoding a glucuronan lyase belonging to a novel polysaccharide lyase family. Appl Environ Microbiol 75:101–107CrossRefGoogle Scholar
  16. Konno N, Ishida T, Igarashi K, Fushinobu S, Samejima M, Habu N, Isogai A (2009b) Crystal structure of polysaccharide lyase family 20 endo-β-1, 4-glucuronan lyase from the filamentous fungus Trichoderma reesei. FEBS Lett 583:1323–1326CrossRefGoogle Scholar
  17. Lindberg B, Lönngren J, Thompson JL (1973) Degradation of polysaccharides containing uronic acid residues. Carbohydr Res 31:93–100CrossRefGoogle Scholar
  18. Marx VM, Schulz CV (1941) Methodisches zur bestimmung der viskositätszahl von cellulosen und cellulosenitraten. Makromol Chem 31:140–153CrossRefGoogle Scholar
  19. Preiss J, Ashwell G (1962) Alginic acid metabolism in bacteria. J Biol Chem 237:309–316Google Scholar
  20. Saito T, Isogai A (2004) TEMPO-mediated oxidation of native cellulose. The effect of oxidation conditions on chemical and crystal structures of the water insoluble fractions. Biomacromolecules 5:1983–1989CrossRefGoogle Scholar
  21. Saito T, Nishiyama Y, Putaux JL, Vignon M, Isogai A (2006) Homogeneous suspensions of individualized microfibrils from TEMPO-catalyzed oxidation of native cellulose. Biomacromolecules 7:1687–1691CrossRefGoogle Scholar
  22. Saito T, Kimura S, Nishiyama Y, Isogai A (2007) Cellulose nanofibers prepared by TEMPO-mediated oxidation of native cellulose. Biomacromolecules 8:2492–2496CrossRefGoogle Scholar
  23. Saito T, Hirota M, Tamura N, Kimura S, Fukuzumi H, Heux L, Isogai A (2009) Individualization of nano-sized plant cellulose fibrils by direct surface carboxylation using TEMPO catalyst under neutral conditions. Biomacromolecules 10:1992–1996CrossRefGoogle Scholar
  24. Schulz GV, Blaschke F (1941) Eine Gleichung zur Berechnung der Viscostätszahl für sehr klleine Konzentrationen. J Prak Chem 158:130–135CrossRefGoogle Scholar
  25. Sharples A (1971) Chapter XVIII, A. Acid hydrolysis and alcoholysis. In: Bikales NM, Segal L (eds) Cellulose and cellulose derivatives part V. Wiley, USA, pp 991–1006Google Scholar
  26. Shevchik VE, Kester HCM, Benen JAE, Visser J, Robert-Baudouy J, Hugouvieux-Cotte-Pattat N (1999a) Characterization of the exopolygalacturonate lyase PelX of Erwinia chrysanthemi 3937. J Bacteriol 181:1652–1663Google Scholar
  27. Shevchik VE, Condemine G, Robert-Baudouy J, Hugouvieux-Cotte-Pattat N (1999b) The exopolygalacturonate lyase PelW and the oligogalacturonate lyase Ogl, two cytoplasmic enzymes of pectin catabolism in Erwinia chrysanthemi 3937. J Bacteriol 181:3912–3919Google Scholar
  28. Shibata I, Yanagisawa M, Tsuguyuki S, Isogai A (2006) SEC-MALS analysis of cellouronic acid prepared from regenerated cellulose by TEMPO-mediated oxidation. Cellulose 13:72–80CrossRefGoogle Scholar
  29. Sihtola H, Kyrklund B, Laamanen L, Palenius I (1963) Comparison and conversion of viscosity and DP-values determined by different methods. Paperi ja puu 45:225–232Google Scholar
  30. TAPPI Test Methods T230 om-99 (1999) Viscosity of pulp (capillary viscometer method)Google Scholar
  31. Yoshimura T, Okutsu M, Hiroshima M (2009) Method for manufacturing water-soluble polyuronate having good biodegradability. WO-2009122953Google Scholar

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© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Graduate School of Agricultural and Life SciencesThe University of TokyoBunkyo-ku, TokyoJapan

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