Journal of Biotechnology 77 (2000) 37 – 47 13 C and 1 H NMR study of cellulose metabolism by Fibrobacter succinogenes S85 Xavier Bibollet a , Nathalie Bosc b , Maria Matulova a,c , Anne-Marie Delort a, *, Genevieve Gaudet b , Evelyne Forano b a Laboratoire de Synthe `se, Electrosynthe `se et Etude de Syste `mes a ` Inte ´re ˆt Biologique, UMR 6504 Uniersite ´ Blaise Pascal -CNRS, 63177 Aubie `re cedex, France b Laboratoire de Microbiologie, INRA, Centre de Recherches de Clermont -Ferrand -Theix, 63122 Saint -Gene `s -Champanelle, France c Institute of Chemistry, Sloak Academy of Sciences, Dubraska cesta 9, 842 38 Bratislaa, Sloak Republic Received 8 February 1999; received in revised form 5 July 1999; accepted 6 July 1999 Abstract Fibrobacter succinogenes S85, a cellulolytic rumen bacterium, is very efficient in degrading lignocellulosic substrates and could be used to develop a biotechnological process for the treatment of wastes. In this work, the metabolism of cellulose by F. succinogenes S85 was investigated using in vivo 13 C NMR and 13 C-filtered spin-echo difference 1 H NMR spectroscopy. The degradation of unlabelled cellulose synthesised by Acetobacter xylinum was studied indirectly, in the presence of [1- 13 C]glucose, by estimating the isotopic dilution of the final bacterial fermentation products (glycogen, succinate, acetate). During the pre-incubation period of F. succinogenes cells with cellulose fibres, some cells (‘non-adherent’) did not attach to the solid material. Results for ‘adherent’ cells showed that about one fourth of the glucose units entering F. succinogenes metabolism originated from cellulose degradation. A huge reversal of succinate metabolism pathway and production of large amounts of unlabelled acetate which was observed during incubation with glucose only, was found to be much decreased in the presence of solid substrate. The synthesis of glucose 6-phophate was slightly increased in the presence of cellulose. Results clearly showed that ‘non-adherent’ cells were able to metabolise glucose very efficiently; consequently the metabolic state of these cells was not responsible for their ‘non-adherence’ to cellulose fibre. © 2000 Elsevier Science B.V. All rights reserved. Keywords: Cellulose; 13 C and 1 H NMR; Fibrobacter ; Metabolism; Adhesion; Rumen www.elsevier.com/locate/jbiotec 1. Introduction Microbial cellulases and hemicellulases are widely used in different industrial activities, such as in textile, detergent, brewery or wood-process- ing, and also in the treatment of domestic wastes and in biological treatment of fibrous feeds in the Abbreiations: 13 C-FSED, 13 C-filtered spin-echo difference 1 H NMR spectroscopy. * Corresponding author. Fax: +33-4-73407717. E-mail address: amdelort@chimtp.univ-bpclermont.fr (A.-M. Delort) 0168-1656/00/$ - see front matter © 2000 Elsevier Science B.V. All rights reserved. PII:S0168-1656(99)00206-0