The isolation, characterization and effect of lignin isolated from steam pretreated Douglas-fir on the enzymatic hydrolysis of cellulose.

Douglas-fir was SO(2)-steam pretreated at different severities (190, 200, and 210°C) to assess the possible negative effect of the residual and isolated lignins on the enzymatic hydrolysis of the steam pretreated substrates. When various isolated lignins were added to the Avicel hydrolysis reactions, the decrease in glucose yields ranged from 15.2% to 29.0% after 72 h. It was apparent that the better hydrolysis yields obtained at higher pretreatment severities were more a result of the greater accessibly of the cellulose rather than any specific change in the non-productive binding of the lignin to the enzymes. FTIR and (13)C NMR characterization indicated that the lignin in the steam pretreated substrates became more condensed with increasing severity, suggesting that the cellulases were adsorbed to the lignin by hydrophobic interactions. Electrostatic interactions were also involved as the positively charged cellulase components were preferentially adsorbed to the lignins.

[1]  John N. Saddler,et al.  Steam pretreatment of douglas-fir wood chips , 2000, Applied biochemistry and biotechnology.

[2]  H. Ooshima,et al.  Enhancement of enzymatic hydrolysis of cellulose by surfactant , 1986, Biotechnology and bioengineering.

[3]  John N. Saddler,et al.  The effect of initial pore volume and lignin content on the enzymatic hydrolysis of softwoods , 1998 .

[4]  R. Overend,et al.  Fractionation of lignocellulosics by steam-aqueous pretreatments , 1987, Philosophical Transactions of the Royal Society of London. Series A, Mathematical and Physical Sciences.

[5]  J. Saddler,et al.  Acid‐catalyzed steam pretreatment of lodgepole pine and subsequent enzymatic hydrolysis and fermentation to ethanol , 2007, Biotechnology and bioengineering.

[6]  F. Tjerneld,et al.  Ion-exchange chromatographic purification and quantitative analysis of Trichoderma reesei cellulases cellobiohydrolase I, II and endoglucanase II by fast protein liquid chromatography , 1998 .

[7]  P. Thibault,et al.  Characterization of cellobiohydrolase I (Cel7A) glycoforms from extracts of Trichoderma reesei using capillary isoelectric focusing and electrospray mass spectrometry. , 2001, Journal of chromatography. B, Biomedical sciences and applications.

[8]  W. Mabee,et al.  Comparison of methods to assess the enzyme accessibility and hydrolysis of pretreated lignocellulosic substrates , 2009, Biotechnology Letters.

[9]  Xuejun Pan Role of Functional Groups in Lignin Inhibition of Enzymatic Hydrolysis of Cellulose to Glucose , 2008 .

[10]  Linoj Kumar,et al.  Can the same steam pretreatment conditions be used for most softwoods to achieve good, enzymatic hydrolysis and sugar yields? , 2010, Bioresource technology.

[11]  John N. Saddler,et al.  Structure and properties of lignin in softwoods after SO2- catalyzed steam explosion and enzymatic hydrolysis , 2001 .

[12]  G. Gellerstedt,et al.  Steam Explosion of Aspen Wood. Characterisation of Reaction Products , 2002 .

[13]  J. Kadla,et al.  A comprehensive approach for quantitative lignin characterization by NMR spectroscopy. , 2004, Journal of agricultural and food chemistry.

[14]  Rajeev Kumar,et al.  Access of cellulase to cellulose and lignin for poplar solids produced by leading pretreatment technologies , 2009, Biotechnology progress.

[15]  Jack N Saddler,et al.  Enhancing the enzymatic hydrolysis of lignocellulosic biomass by increasing the carboxylic acid content of the associated lignin , 2011, Biotechnology and bioengineering.

[16]  J. Saddler,et al.  The bioconversion of mountain pine beetle‐killed lodgepole pine to fuel ethanol using the organosolv process , 2008, Biotechnology and bioengineering.

[17]  Johan Börjesson,et al.  Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose , 2002 .

[18]  J. Saddler,et al.  Inhibition of cellulase, xylanase and beta-glucosidase activities by softwood lignin preparations. , 2006, Journal of biotechnology.

[19]  W. G. Glasser,et al.  Lignin Impact on Fiber Degradation. 3. Reversal of Inhibition of Enzymatic Hydrolysis by Chemical Modification of Lignin and by Additives , 1997 .

[20]  J. O. Baker,et al.  Fingerprinting Trichoderma reesei hydrolases in a commercial cellulase preparation , 2001, Applied biochemistry and biotechnology.

[21]  W. Mabee,et al.  Substrate pretreatment: the key to effective enzymatic hydrolysis of lignocellulosics? , 2007, Advances in biochemical engineering/biotechnology.

[22]  Guido Zacchi,et al.  Adsorption of Trichoderma reesei CBH I and EG II and their catalytic domains on steam pretreated softwood and isolated lignin. , 2004, Journal of biotechnology.

[23]  J. Saddler,et al.  Adsorption of cellulase on cellulolytic enzyme lignin from lodgepole pine. , 2009, Journal of agricultural and food chemistry.

[24]  J. Saddler,et al.  The effect of isolated lignins, obtained from a range of pretreated lignocellulosic substrates, on enzymatic hydrolysis , 2010, Biotechnology and bioengineering.

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

[26]  A. Nada,et al.  Infrared spectroscopy of some treated lignins , 1998 .

[27]  E. Cowling,et al.  Comparative Studies on Cellulolytic Enzyme Lignin and Milled Wood Lignin of Sweetgum and Spruce , 1975 .

[28]  J. Saddler,et al.  Substrate and Enzyme Characteristics that Limit Cellulose Hydrolysis , 1999, Biotechnology progress.

[29]  Scott Renneckar,et al.  Nanocomposite-based lignocellulosic fibers 1. Thermal stability of modified fibers with clay-polyelectrolyte multilayers , 2008 .

[30]  J. Kadla,et al.  Polyoxometalate (POM) Oxidation of Milled Wood Lignin (MWL) , 2007 .

[31]  Ross D. Brown,et al.  Purification and characterization of a β‐glucosidase from Trichoderma reesei , 1987 .

[32]  Bin Yang,et al.  Cellulase adsorption and an evaluation of enzyme recycle during hydrolysis of steam-exploded softwood residues. , 2002, Applied biochemistry and biotechnology.