Cellulase Adsorption and Reactivity on a Cellulose Surface from Flow Ellipsometry

Enzymatic deconstruction of cellulose occurs at the aqueous/cellulose interface. Most assays to explore cellulase activity, however, are performed in bulk solution and, hence, fail to elucidate surface-reaction kinetics. We use flow ellipsometry to quantify the adsorption and surface reactivity of aqueous cellulase on a model cellulose film substrate. The rate of cellulose digestion at the aqueous/solid interface increases with increasing bulk concentration of enzyme, but only up to a plateau corresponding to the maximum adsorption density of cellulase. Kinetic data are analyzed according to a modified Langmuir–Michaelis–Menten framework including both reversible adsorption of cellulase to the cellulose surface and complexation of surface cellulose chains with adsorbed cellulase. At ambient temperature, the molar turnover number is 0.57 ± 0.08 s–1, commensurate with literature values, and the Langmuir adsorption equilibrium constant, characterizing the binding strength of the cellulase, is 0.086 ± 0.026 p...

[1]  J. Aubert,et al.  The biological degradation of cellulose. , 1994, FEMS microbiology reviews.

[2]  L. Wågberg,et al.  Model films of cellulose ID – improved preparation method and characterization of the cellulose film , 2004 .

[3]  T. Vuorinen,et al.  Quantitative assessment of the enzymatic degradation of amorphous cellulose by using a quartz crystal microbalance with dissipation monitoring. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[4]  N. Labbe,et al.  Compatible Ionic liquid‐cellulases system for hydrolysis of lignocellulosic biomass , 2011, Biotechnology and bioengineering.

[5]  P. Väljamäe,et al.  Mechanism of initial rapid rate retardation in cellobiohydrolase catalyzed cellulose hydrolysis , 2010, Biotechnology and bioengineering.

[6]  Andreas S Bommarius,et al.  Cellulase kinetics as a function of cellulose pretreatment. , 2008, Metabolic engineering.

[7]  Markus Pauly,et al.  Cell-wall carbohydrates and their modification as a resource for biofuels. , 2008, The Plant journal : for cell and molecular biology.

[8]  Patrizia R. Mussini,et al.  Microcrystalline cellulose powders: structure, surface features and water sorption capability , 1999 .

[9]  David K. Johnson,et al.  Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production , 2007, Science.

[10]  J. Howell,et al.  Kinetics of solka floc cellulose hydrolysis by trichoderma viride cellulase , 1975 .

[11]  M. Wada,et al.  Surface density of cellobiohydrolase on crystalline celluloses , 2006, The FEBS journal.

[12]  F. Tjerneld,et al.  Effect of poly(ethylene glycol) on enzymatic hydrolysis and adsorption of cellulase enzymes to pretreated lignocellulose , 2007 .

[13]  L. Wågberg,et al.  Model films of cellulose: I. Method development and initial results , 2002 .

[14]  S. Allen,et al.  Kinetic dynamics in heterogeneous enzymatic hydrolysis of cellulose: an overview, an experimental study and mathematical modelling , 2003 .

[15]  Jay H. Lee,et al.  Modeling cellulase kinetics on lignocellulosic substrates. , 2009, Biotechnology advances.

[16]  Tuula T. Teeri,et al.  Crystalline cellulose degradation : new insight into the function of cellobiohydrolases , 1997 .

[17]  M. Malmsten,et al.  Enzymatic degradation of model cellulose films. , 2005, Journal of colloid and interface science.

[18]  Rui M. F. Bezerra,et al.  Discrimination among eight modified michaelis-menten kinetics models of cellulose hydrolysis with a large range of substrate/enzyme ratios , 2004, Applied biochemistry and biotechnology.

[19]  Ruihong Zhang,et al.  Kinetic modeling for enzymatic hydrolysis of pretreated creeping wild ryegrass , 2009, Biotechnology and bioengineering.

[20]  L. Lynd,et al.  A functionally based model for hydrolysis of cellulose by fungal cellulase , 2006, Biotechnology and bioengineering.

[21]  M. Moo-young,et al.  Kinetics of enzymatic hydrolysis of cellulose: Analytical description of a mechanistic model , 1978, Biotechnology and bioengineering.

[22]  Sun Bok Lee,et al.  Adsorption of cellulase on cellulose: Effect of physicochemical properties of cellulose on adsorption and rate of hydrolysis , 1982, Biotechnology and bioengineering.

[23]  James D. McMillan,et al.  Bioethanol production: Status and prospects , 1997 .

[24]  A comparison of the Michaelis-Menten and HCH-1 models. , 1990, Biotechnology and bioengineering.

[25]  G. Henriksson,et al.  The physical action of cellulases revealed by a quartz crystal microbalance study using ultrathin cellulose films and pure cellulases. , 2008, Biomacromolecules.

[26]  A. C. O'sullivan.  Cellulose: the structure slowly unravels , 1997, Cellulose.

[27]  M. Himmel,et al.  Outlook for cellulase improvement: screening and selection strategies. , 2006, Biotechnology advances.

[28]  Mark Holtzapple,et al.  Inhibition of Trichoderma reesei cellulase by sugars and solvents , 1990, Biotechnology and bioengineering.

[29]  H. Blanch,et al.  A mechanistic model of the enzymatic hydrolysis of cellulose , 2010, Biotechnology and bioengineering.

[30]  T. Wood,et al.  METHODS FOR MEASURING CELLULASE ACTIVITIES , 1988 .

[31]  Computational and experimental studies of the catalytic mechanism of Thermobifida fusca cellulase Cel6A (E2). , 2003, Protein engineering.

[32]  Stephen E. Wald,et al.  Kinetics of the enzymatic hydrolysis of cellulose , 1984, Biotechnology and bioengineering.

[33]  Z. Bai,et al.  The enzymatic hydrolysis rate of cellulose decreases with irreversible adsorption of cellobiohydrolase I , 2008 .

[34]  Xavier Turon,et al.  Enzymatic kinetics of cellulose hydrolysis: a QCM-D study. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[35]  O. Rojas,et al.  In situ monitoring of cellulase activity by microgravimetry with a quartz crystal microbalance. , 2009, The journal of physical chemistry. B.

[36]  E. Bon,et al.  Endocellulase and exocellulase activities of two Streptomyces strains isolated from a forest soil , 2000, Applied biochemistry and biotechnology.

[37]  Carol L. Kozina,et al.  Structure of endoglucanase Cel9A from the thermoacidophilic Alicyclobacillus acidocaldarius. , 2009, Acta crystallographica. Section D, Biological crystallography.

[38]  P Béguin,et al.  Molecular biology of cellulose degradation. , 1990, Annual review of microbiology.

[39]  L. Lynd,et al.  How biotech can transform biofuels , 2008, Nature Biotechnology.

[40]  Jay H. Lee,et al.  Cellulose crystallinity – a key predictor of the enzymatic hydrolysis rate , 2010, The FEBS journal.

[41]  Eero Kontturi,et al.  Cellulose--model films and the fundamental approach. , 2006, Chemical Society reviews.

[42]  C. Wyman,et al.  Pretreatment: the key to unlocking low‐cost cellulosic ethanol , 2008 .

[43]  Rajeev Kumar,et al.  Cellulase adsorption and relationship to features of corn stover solids produced by leading pretreatments , 2009, Biotechnology and bioengineering.

[44]  O. Rojas,et al.  Enzymatic hydrolysis of native cellulose nanofibrils and other cellulose model films: effect of surface structure. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[45]  Charles E. Wyman,et al.  Ethanol from lignocellulosic biomass: Technology, economics, and opportunities , 1994 .

[46]  Harvey W Blanch,et al.  Initial- and processive-cut products reveal cellobiohydrolase rate limitations and the role of companion enzymes. , 2012, Biochemistry.

[47]  T. Teeri,et al.  The cellulose-binding domain of the major cellobiohydrolase of Trichoderma reesei exhibits true reversibility and a high exchange rate on crystalline cellulose. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[48]  Srinivas Karra,et al.  Modeling Intrinsic Kinetics of Enzymatic Cellulose Hydrolysis , 2008, Biotechnology progress.

[49]  B. Henrissat,et al.  Dimension, Shape, and Conformational Flexibility of a Two Domain Fungal Cellulase in Solution Probed by Small Angle X-ray Scattering* , 2002, The Journal of Biological Chemistry.

[50]  B. K. Lok,et al.  Protein adsorption on crosslinked polydimethylsiloxane using total internal reflection fluorescence , 1983 .

[51]  S. Kaddour,et al.  Adsorption of cellulase Aspergillus niger on a commercial activated carbon: kinetics and equilibrium studies. , 2010, Colloids and surfaces. B, Biointerfaces.

[52]  F. Veer,et al.  Ellipsometry as a tool to study the adsorption behavior of synthetic and biopolymers at the air–water interface , 1978 .

[53]  Bruce E Dale,et al.  Predicting digestibility of ammonia fiber explosion (AFEX)-treated rice straw , 2002, Applied biochemistry and biotechnology.

[54]  C. Radke,et al.  Kinetics of adsorption and proteolytic cleavage of a multilayer ovalbumin film by subtilisin Carlsberg. , 2008, Langmuir : the ACS journal of surfaces and colloids.