Testing alternative kinetic models for utilization of crystalline cellulose (Avicel) by batch cultures of Clostridium thermocellum

Descriptive kinetics of batch cellulose (Avicel) and cellobiose fermentation by Clostridium thermocellum were examined with residual substrate and biosynthate concentrations inferred based on elemental analysis. Biosynthate was formed in constant proportion to substrate consumption until substrate was exhausted for cellobiose fermentation, and until near the point of substrate exhaustion for cellulose fermentation. Cell yields (g pellet biosynthate carbon/g substrate carbon) of 0.214 and 0.200 were obtained for cellulose and cellobiose, respectively. For cellulose fermentation a sigmoidal curve fit was applied to substrate and biosynthate concentrations over time, which was then differentiated to calculate instantaneous rates of growth and substrate consumption. Three models were tested to describe the kinetics of Avicel utilization by C. thermocellum: (A) first order in cells, (B) first order in substrate, and (C) first order in cells and substrate, and second order overall. Models (A) and (B) have been proposed in the literature to describe cultures of cellulolytic microorganisms, whereas model (C) has not. Of the three models tested, model (c) provided by far the best fit to batch culture data. A second order rate constant equal to 0.735 L g C(-1) h(-1) was found for utilization of Avicel by C. thermocellum. Adding an endogenous metabolism term improved the descriptive quality of the model as substrate exhaustion was approached. Such rate constants may in the future find utility for describing and comparing cellulose fermentation involving other microbes and other substrates.