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Applied and Environmental Microbiology, September 2001, p. 3846-3851, Vol. 67, No. 9
Laboratoire de Biochimie des Bactéries
Gram +, Domaine Scientifique Victor Grignard, Université
Henri Poincaré, Faculté des Sciences, 54506 Vand
Received 20 March 2001/Accepted 31 May 2001
An investigation of cellulose degradation by the nonruminal,
cellulolytic, mesophilic bacterium Clostridium
cellulolyticum was performed in cellulose-fed chemostat
cultures with ammonium as the growth-limiting nutrient. At any
dilution rate (D), acetate was always the main product
of the catabolism, with a yield of product from substrate
ranging between 37.7 and 51.5 g per mol of hexose equivalent
fermented and an acetate/ethanol ratio always higher than 1. As
D rose, the acetyl coenzyme A was rerouted in favor of
ethanol pathways, and ethanol production could represent up to 17.7%
of the carbon consumed. Lactate was significantly produced, but with
increasing D, the specific lactate production rate
declined, as did the specific rate of production of extracellular pyruvate. The proportion of the original carbon directed towards phosphoglucomutase remained constant, and the carbon surplus was balanced mainly by exopolysaccharide and glycogen biosyntheses at high
D values, while cellodextrin excretion occurred mainly at lower ones. With increasing D, the specific rate of
carbon flowing down catabolites increased as well, but when expressed as a percentage of carbon it declined, while the percentage of carbon
directed through biosynthesis pathways was enhanced. The maximum growth
and energetic yields were lower than those obtained in
cellulose-limited chemostats and were related to an
uncoupling between catabolism and anabolism leading to an excess of
energy. Compared to growth on cellobiose in ammonium-limited chemostats (E. Guedon, M. Desvaux, and H. Petitdemange, J. Bacteriol.
182:2010-2017, 2000), (i) a specific consumption rate of carbon of as
high as 26.72 mmol of hexose equivalent g of cells
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.9.3846-3851.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Flux Analysis of the Metabolism of
Clostridium cellulolyticum Grown in Cellulose-Fed
Continuous Culture on a Chemically Defined Medium under
Ammonium-Limited Conditions
uvre-lès-Nancy Cédex, France
1
h
1 could not be reached and (ii) the proportions of
carbon directed towards cellodextrin, glycogen, and
exopolysaccharide pathways were not as high as first determined
on cellobiose. While the use of cellobiose allows highlighting of
metabolic limitation and regulation of C. cellulolyticum
under ammonium-limited conditions, some of these events should then
rather be interpreted as distortions of the metabolism. Growth of
cellulolytic bacteria on easily available carbon and nitrogen sources
represents conditions far different from those of the natural
lignocellulosic compounds.
*
Corresponding author. Mailing address: Laboratoire de
Biochimie des Bactéries Gram +, Domaine Scientifique Victor
Grignard, Université Henri Poincaré, Faculté des
Sciences, BP 239, 54506 Vand
uvre-lès-Nancy Cédex, France.
Phone: 33 3 83 91 20 53. Fax: 33 3 83 91 25 50. E-mail:
hpetitde{at}lcb.u-nancy.fr.
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