, 13:327 | Cite as

Structural studies of bacterial cellulose through the solid-phase nitration and acetylation by CP/MAS 13C NMR spectroscopy

  • Hiroyuki Yamamoto
  • Fumitaka Horii
  • Asako Hirai


The solid-phase nitration and acetylation processes of bacterial cellulose have been investigated mainly by CP/MAS 13C NMR spectroscopy to clarify the features of these reactions in relation to the characterization of the disordered component included in the microfibrils. CP/MAS 13C NMR spectra of bacterial and Valonia cellulose samples are markedly changed as the nitration progresses, in a similar way to the case of cotton linters previously reported; and the relative reactivity of the OH groups in the glucose residues is found to decrease in the order of O(6)H>O(2)H>O(3)H. Moreover, the nitration rate and mode greatly depend on the concentration of nitric acid in the reaction media. At dilute and medium concentrations, the O(6)H groups in the crystalline and disordered components are subjected to nitration at nearly the same rate, indicating that these two components are distributed almost at random in the entire region of each microfibril. The preferential penetration of nitric acid into each microfibril also occurs prior to nitration at the medium concentration, resulting in an increase in the mole fraction of the disordered component. In contrast, all OH groups undergo nitration very rapidly at the higher concentration, although nitration levels off to a certain extent for O(3)H groups. In solid-phase acetylation, no regio-selective reactivity is observed among the three kinds of OH groups, which may be due to the characteristic reaction that proceeds in a very thin layer between the acetylated and nonacetylated regions in each microfibril. The almost random distribution of the disordered component in the entire region of the microfibrils is also confirmed in this solid-phase acetylation. On the basis of these results, the mechanism of the solid-phase reactions and the microfibril structure are discussed.


Bacterial cellulose CP/MAS 13C NMR Disordered structure Microfibril structure Solid-phase acetylation Solid-phase nitration 



The authors thank Professor Masaki Tsuji, Institute for Chemical Research, Kyoto University, for kindly allowing us to use a 200 kV JEOL JEM-200CS transmission electron microscope.


  1. Atalla R.H. and VanderHart D.L. (1984). Native cellulose: a composite of two distinct crystalline forms. Science 223:283–285CrossRefGoogle Scholar
  2. Chanzy H., Henrissat B., Vincendon M., Tanner S.F. and Belton P.S. (1987). Solid-state 13C N.M.R. and electron microscopy study on the reversible transformation cellulose I→cellulose IIII in Valonia. Carbohydr. Res. 160:1–11CrossRefGoogle Scholar
  3. Earl W.L. and VanderHart D.L. (1981). Observation by high-resolution carbon-13 nuclear magnetic resonance of cellulose I related to morphology and crystal structure. Macromolecules 14:570–574CrossRefGoogle Scholar
  4. Hayashi N., Sugiyama J., Okano T. and Ishihara M. (1998a). Selective degradation of the cellulose Iα component in Cladophora cellulose with Trichoderma Viride cellulase. Carbohydr. Res. 305:109–116CrossRefGoogle Scholar
  5. Hayashi N., Sugiyama J., Okano T. and Ishihara M. (1998b). The enzymatic susceptibility of cellulose microfibrils of the algal-bacterial type and the cotton-ramie type. Carbohydr. Res. 305:261–269CrossRefGoogle Scholar
  6. Hestrin S. and Schramm M. (1954). Synthesis of cellulose by Acetobacter xylinum. Biochem. J. 58:345–352Google Scholar
  7. Hirai A., Horii F. and Kitamaru R. (1987). Transformation of native cellulose crystals from cellulose Ib to Ia through solid-state chemical reactions. Macromolecules 20:1440–1442CrossRefGoogle Scholar
  8. Hirai A., Tsuji M., Yamamoto H. and Horii F. (1998). In situ crystallization of bacterial cellulose III. Influences of different polymeric additives on the formation of microfibrils as revealed by transmission electron microscopy. Cellulose 5:201–213CrossRefGoogle Scholar
  9. Horii F. (1989). The Structure of Cellulose as Studied by CP/MAS 13C NMR Spectroscopy. In: Pfeffer P.E. and Gerasimowicz W.V. (eds), Nuclear Magnetic Resonance in Agriculture. CRC Press, Boca Raton, pp. 311–335Google Scholar
  10. Horii F. (2001). Structure of Cellulose: Recent Developments in its Characterization. In: Hon D.N.-S., and Shiraishi N. (eds), Wood and Cellulosic Chemistry. Dekker, New York, pp. 83–107Google Scholar
  11. Horii F., Hirai A. and Kitamaru R. (1982). Solid-state high-resolution 13C-NMR studies of regenerated cellulose samples with different crystallinites. Polym. Bull. 8:163–170CrossRefGoogle Scholar
  12. Horii F., Hirai A. and Kitamaru R. (1984). CP/MAS CARBON-13 NMR study of spin relaxation phenomena of cellulose containing crystalline and noncrystalline components. J. Carbohydr. Chem. 3:641–662CrossRefGoogle Scholar
  13. Horii F., Hirai A., Suzuki F. and Tsujitani K. (2005a). Possible origin of disordered structure in bacterial cellulose: structure and structural evolution of sub-elementary fibrils. Abstr. Am. Chem. Soc. Nat. Meet. 229:128Google Scholar
  14. Horii F., Hirai A., Suzuki F. and Tsujitani K. (2005b). Hierarchical structure of bacterial cellulose: structure and structural evolution of the sub-elementary fibrils. Preprint Cellulose Soc. Jpn. 12:16–17Google Scholar
  15. Horii F., Yamamoto H. and Hirai A. (1997). Microstructural analysis of microfibrils of bacterial cellulose. Macromol. Symp. 120:197–205Google Scholar
  16. Horii F., Yamamoto H., Kitamaru R., Tanahashi M. and Higuchi T. (1987). Transformation of native cellulose crystals induced by saturated steam at high temperatures. Macromolecules 20:2946–2949CrossRefGoogle Scholar
  17. Kita G., Sakurada I. and Nakajima T. 1936. Sen-iso Kogyo (Cellulose Industries) 3:70Google Scholar
  18. Kono H., Yunoki S., Shikano T., Fujiwara M., Erata T., Takai M. (2002). CP/MAS 13C NMR study of cellulose and cellulose derivatives. 1. Complete assignment of the CP/MAS 13C NMR spectrum of the native cellulose. J. Am. Chem. Soc. 124:7506–7511CrossRefGoogle Scholar
  19. Masuda K., Adachi M., Hirai A., Yamamoto H., Kaji H. and Horii F. (2003). Solid-State 13C and 1H spin diffusion NMR analyses of the microfibril structure for bacterial cellulose. Solid State Nucl. Magn. Reson 23:198–212CrossRefGoogle Scholar
  20. Newman R.H. (1998). Evidence for assignment of 13C NMR signals to cellulose crystalline surfaces in wood, pulp and isolated celluloses. Holzforschung 52:157–159CrossRefGoogle Scholar
  21. Nishiyama Y., Langan P. and Chanzy H. (2002). Crystal structure and hydrogen-bonding system in cellulose Iβ from synchrotron X-ray and neutron fiber diffraction. J. Am. Chem. Soc.124:9074–9082CrossRefGoogle Scholar
  22. Nishiyama Y., Sugiyama J., Chanzy H. and Langan P. (2003). Crystal structure and hydrogen bonding system in cellulose Iα from synchrotron X-ray and neutron fiber diffraction. J. Am. Chem. Soc. 125:14300–14306CrossRefGoogle Scholar
  23. Patterson P.M., Patterson D.J., Blackwell J., Koenig J.L., Jamieson A.M., Garignan Y.P. and Turngren E.V. (1985). High-resolution solid-state 13C-NMR spectroscopy of cellulose nitrates. J. Polym. Sci. 23:483–492Google Scholar
  24. Rassow B. and Dörr E. (1924). Zur Kenntnis der Nitrocellulose. Z. Prak. Chem. 108:113–186CrossRefGoogle Scholar
  25. Sassi J.-F. and Chanzy H. (1995). Ultrastructural aspects of the acetylation of cellulose. Cellulose 2:111–128CrossRefGoogle Scholar
  26. Sassi J.-F., Tekely P. and Chanzy H. (2000). Relative susceptibility of the Iα and Iβ phase of cellulose towards acetylation. Cellulose 7:119–132CrossRefGoogle Scholar
  27. Sprague B.S., Riley J.L. and Noether H.D. (1958). Factors influencing the crystal structure of cellulose triacetate. Text. Res. J. 28:275–287CrossRefGoogle Scholar
  28. Stipanovic A. and Sarko A. (1978). Molecular and crystal structure of cellulose triacetate. I. A parallel chain structure. Polymer 19:3–8CrossRefGoogle Scholar
  29. VanderHart D.L. and Atalla R.H. (1984). Studies of microstructure in native celluloses using solid-state 13C NMR. Macromolecules 17:1465–1472CrossRefGoogle Scholar
  30. Watanabe S., Imai K. and Hayashi J. (1971). The presence of cellulose trinitrate (II). Kogyo Kagaku Zasshi 74:1420–1426Google Scholar
  31. Wu T.K. (1980). Carbon-13 and proton nuclear magnetic resonance studies of cellulose nitrates. Macromolecules 13:74–79CrossRefGoogle Scholar
  32. Yamamoto H. and Horii F. (1993). CP/MAS 13C NMR analysis of the crystal transformation induced for Valonia cellulose by annealing at high temperatures. Macromolecules 26:1313–1317CrossRefGoogle Scholar
  33. Yamamoto H. and Horii F. (1994). In situ crystallization of bacterial cellulose. I. Influences of polymeric additives, stirring and temperatures on the formation cellulose Iα and Iβ as revealed by cross polarization/magic angle spinning (CP/MAS) 13C NMR spectroscopy. Cellulose 1:57–66CrossRefGoogle Scholar
  34. Yamamoto H., Horii F. and Hirai A. (1996). In situ crystallization of bacterial cellulose. II. Influences of different polymeric additives on the formation of cellulose Iα and Iβ at the early stage of incubation. Cellulose 3:229–242CrossRefGoogle Scholar
  35. Yamamoto H., Horii F. and Odani H. (1989). Structural change of native cellulose crystals induced by annealing in aqueous alkaline and acidic solutions at high temperatures. Macromolecules 22:4130–4132CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Hiroyuki Yamamoto
    • 1
  • Fumitaka Horii
    • 2
  • Asako Hirai
    • 2
  1. 1.Fukui National College of TechnologySabaeJapan
  2. 2.Institute for Chemical ResearchKyoto UniversityUjiJapan

Personalised recommendations