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Relationships Between Cellobiose Catabolism, Enzyme Levels, and Metabolic Intermediates in Clostridium cellulolyticum Grown in a Synthetic Medium

Abstract : Continuous cultures, under cellobiose sufficient concentrations (14.62 mM) using a chemically defined medium, were examined to determine the carbon regulation selected by Clostridium cellulolyticum. Using a synthetic medium, a q cellobiose of 2.57 mmol g cells −1 h −1 was attained whereas the highest value obtained on complex media was 0.68 mmol g cells −1 h −1 (Payot et al. 1998. Microbiology 144:375-384). On a synthetic medium at D = 0.035 h −1 under cellobiose excess, lactate and etha-nol biosynthesis were able to use the reducing equivalents supplied by acetic acid formation and the H 2 /CO 2 ratio was found equal to 1. At a higher dilution rate (D = 0.115 h −1), there was no lactate production and the pathways toward ethanol and NADH-ferredoxin-hydrogenase contributed to balance the reducing equivalents; in this case a H 2 /CO 2 ratio of 1.54 was found. With increasing D, there was a progressive increase (i) in the steady-state concentration of NADH and NAD + pools from 11.8 to 22.1 µmol (g cells) −1 , (ii) in the intracellular NADH/NAD + ratios from 0.43 to 1.51. On synthetic media, under cello-biose excess the carbon flow was also equilibrated by three overflows: exopolysaccharide, extracellular protein , and amino acid excretions. At D = 0.115 h −1 , 34% of the cellobiose consumed was converted into exopolysac-charides; this deviation of the carbon flow and the increase of the phosphoroclastic activity decreased dramatically the pyruvate excretion and explained the break in lactate production. Whatever the dilution rate, C. cel-lulolyticum, using ammonium and cellobiose excess, always spilled usual amino acids accompanied by other amino compounds. In vitro, GAPDH, phosphoroclastic reaction, alcohol dehydrogenase, and acetate kinase activities were high under conditions giving high in vivo specific production rates. There were also correlations between the in vitro lactate dehydrogenase activity and in vivo lactate production, but in contrast with the preceding activities, these two parameters decreased with D. All the results demonstrate that C. cellulolyticum was able to optimize carbon catabolism from cellulosic sub-strates in a synthetic medium.
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https://hal.inrae.fr/hal-02910822
Déposant : Stéphanie Dutilloy <>
Soumis le : lundi 3 août 2020 - 10:10:03
Dernière modification le : mercredi 19 août 2020 - 03:31:32

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  • HAL Id : hal-02910822, version 1

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Emmanuel Guedon, S. Payot, M Desvaux, H. Petitdemange. Relationships Between Cellobiose Catabolism, Enzyme Levels, and Metabolic Intermediates in Clostridium cellulolyticum Grown in a Synthetic Medium. Inc. Biotechnol Bioeng, 2000, 67, pp.327 - 335. ⟨hal-02910822⟩

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