Conversion of cellulose to glucose is the rate-limiting step in converting biomass into fuel. In this paper, we explore the effects of micro-and macro-mixing on the enzymatic hydrolysis of various cellulosic substrates to glucose by using a spatially averaged low-dimensional CSTR model. We quantify the effects of mixing on glucose yield, cellulose depolymerization rate and the synergy between the enzymes. We conclude that micromixing limitations provide an important mechanism to increase yield, reduce the dominance of synergy and guide optimum process design by offsetting inhibitory effects by preventing the inhibitors from coming in contact with the enzymes. On quantifying the effects of inhibition type (competitive vs noncompetitive) and mixing type (macro-vs micro-), we find that noncompetitive inhibition inhibits glucose yield more strongly than competitive inhibition, and that unlike micromixing limitations, macromixing limitations reduce glucose yield. On the basis of our analysis, we recommend 2 tank reactors in series with minimal local mixing in each tank and glucose removal at the exit of the first tank as the optimal reactor configuration for maximizing glucose yield from enzymatic hydrolysis of cellulose.
This
article presents a tightly coupled experimental and theoretical
study to explore the effects of mixing and mass transfer on the kinetics
and dynamics of cellulase-mediated cellulose (Avicel) hydrolysis for
bioethanol production in batch reactors. The kinetic parameters (K
M and V
Max) for
the three enzymes (endoglucanase, exoglucanase, β-glucosidase)
that constitute cellulase are determined at various mixing speeds:
0 (no mixing), 40, 80, and 150 rpm (high mixing). The experimental
values of K
M and V
Max are fitted to algebraic expressions that quantify them
as functions of mixing speed, and that, in the asymptotic limit of
complete mixing, give their purely kinetic values (without any mass
transfer disguise). The glucose and reducing sugar yields as well
as the degree of polymerization (DP) for Avicel are measured at all
four mixing speeds for 45 h of incubation. Maximum yields of 76% for
glucose and 87% for reducing sugar are obtained at no mixing condition
(i.e., mass transfer controlled regime), and the DP was found to reduce
from 300 to 41. An unsteady-state multistep three-enzyme kinetic model
incorporating competitive and noncompetitive inhibition caused by
the products glucose (monomer) and cellobiose (dimer) is simulated
using the experimentally obtained K
M and V
Max values at various mixing speeds, and our
model simulations are validated with our experiments. Our analysis
shows that the K
M and V
Max for the three cellulase enzymes remain mass-transfer
disguised kinetic parameters even at high mixing speeds, and we quantify
the purely kinetic values they attain in the limit of perfect mixing.
Our experiments and simulations show that lower mixing speeds increase
glucose and reducing sugar yields and decrease DP by preventing the
products glucose and cellobiose from coming in contact with the active
sites of the cellulase, thus reducing product inhibition, an observation
that may significantly reduce the energy costs for bioethanol production.
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