Inhibitory effects of lignin on enzymatic hydrolysis: The role of lignin chemistry and molecular weight
Abstract
Lignocellulose is a promising feedstock for biofuel production, while lignin poses a grand challenge on the entire process, especially enzymatic hydrolysis. In this study, different types of lignin inhibited enzymatic hydrolysis by different mechanisms. Organosolv lignin from Loblolly pine adsorbed enzyme nonproductively and reduced the available enzyme for cellulose, therefore decreasing hydrolysis rate and ultimate sugar yield. Kraft pine lignin precipitated on the surface of cellulose, preventing it from contacting with enzyme. The molecular weight influenced the inhibition effect of lignin. Lignin of lower molecular weight could bind enzyme nonproductively and when the molecular weight increased, the steric repulsion caused by lignin deposition on cellulose became more significant. The NMR analysis revealed that lignin structural features, e.g., functional groups, S/G ratio, determined the behaviors of lignin in enzymatic hydrolysis. High content of aliphatic hydroxyl groups, or low content of carboxylic groups led to high surface hydrophobicity, increasing the adsorption between lignin and enzyme. In addition, the substrate reactivity is also an important factor that affects enzymatic hydrolysis. Cellulose with higher crystallinity exhibited slower hydrolysis rate and lower conversion. When the crystallinity index increased from 0.43 to 0.72 and 0.81, the ultimate conversion decreased from 80 to 68% and 57%, respectively.
- Authors:
-
- Clemson Univ., SC (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1474540
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Renewable Energy
- Additional Journal Information:
- Journal Volume: 123; Journal Issue: C; Journal ID: ISSN 0960-1481
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Li, Xiang, Li, Mi, Pu, Yunqiao, Ragauskas, Arthur J., Klett, Adam S., Thies, Mark, and Zheng, Yi. Inhibitory effects of lignin on enzymatic hydrolysis: The role of lignin chemistry and molecular weight. United States: N. p., 2018.
Web. doi:10.1016/j.renene.2018.02.079.
Li, Xiang, Li, Mi, Pu, Yunqiao, Ragauskas, Arthur J., Klett, Adam S., Thies, Mark, & Zheng, Yi. Inhibitory effects of lignin on enzymatic hydrolysis: The role of lignin chemistry and molecular weight. United States. https://doi.org/10.1016/j.renene.2018.02.079
Li, Xiang, Li, Mi, Pu, Yunqiao, Ragauskas, Arthur J., Klett, Adam S., Thies, Mark, and Zheng, Yi. Tue .
"Inhibitory effects of lignin on enzymatic hydrolysis: The role of lignin chemistry and molecular weight". United States. https://doi.org/10.1016/j.renene.2018.02.079. https://www.osti.gov/servlets/purl/1474540.
@article{osti_1474540,
title = {Inhibitory effects of lignin on enzymatic hydrolysis: The role of lignin chemistry and molecular weight},
author = {Li, Xiang and Li, Mi and Pu, Yunqiao and Ragauskas, Arthur J. and Klett, Adam S. and Thies, Mark and Zheng, Yi},
abstractNote = {Lignocellulose is a promising feedstock for biofuel production, while lignin poses a grand challenge on the entire process, especially enzymatic hydrolysis. In this study, different types of lignin inhibited enzymatic hydrolysis by different mechanisms. Organosolv lignin from Loblolly pine adsorbed enzyme nonproductively and reduced the available enzyme for cellulose, therefore decreasing hydrolysis rate and ultimate sugar yield. Kraft pine lignin precipitated on the surface of cellulose, preventing it from contacting with enzyme. The molecular weight influenced the inhibition effect of lignin. Lignin of lower molecular weight could bind enzyme nonproductively and when the molecular weight increased, the steric repulsion caused by lignin deposition on cellulose became more significant. The NMR analysis revealed that lignin structural features, e.g., functional groups, S/G ratio, determined the behaviors of lignin in enzymatic hydrolysis. High content of aliphatic hydroxyl groups, or low content of carboxylic groups led to high surface hydrophobicity, increasing the adsorption between lignin and enzyme. In addition, the substrate reactivity is also an important factor that affects enzymatic hydrolysis. Cellulose with higher crystallinity exhibited slower hydrolysis rate and lower conversion. When the crystallinity index increased from 0.43 to 0.72 and 0.81, the ultimate conversion decreased from 80 to 68% and 57%, respectively.},
doi = {10.1016/j.renene.2018.02.079},
journal = {Renewable Energy},
number = C,
volume = 123,
place = {United States},
year = {Tue Feb 20 00:00:00 EST 2018},
month = {Tue Feb 20 00:00:00 EST 2018}
}
Web of Science
Works referenced in this record:
Pretreatment: The key to efficient utilization of lignocellulosic materials
journal, November 2012
- Galbe, Mats; Zacchi, Guido
- Biomass and Bioenergy, Vol. 46
Inhibition of cellulase, xylanase and β-glucosidase activities by softwood lignin preparations
journal, September 2006
- Berlin, Alex; Balakshin, Mikhail; Gilkes, Neil
- Journal of Biotechnology, Vol. 125, Issue 2, p. 198-209
Differences in the adsorption of enzymes onto lignins from diverse types of lignocellulosic biomass and the underlying mechanism
journal, January 2014
- Guo, Fenfen; Shi, Wenjing; Sun, Wan
- Biotechnology for Biofuels, Vol. 7, Issue 1
Inhibitory effect of lignin during cellulose bioconversion: The effect of lignin chemistry on non-productive enzyme adsorption
journal, April 2013
- Rahikainen, Jenni L.; Martin-Sampedro, Raquel; Heikkinen, Harri
- Bioresource Technology, Vol. 133
Contrasting effects of hardwood and softwood organosolv lignins on enzymatic hydrolysis of lignocellulose
journal, July 2014
- Lai, Chenhuan; Tu, Maobing; Shi, Zhiqiang
- Bioresource Technology, Vol. 163
Adsorption of proteins involved in hydrolysis of lignocellulose on lignins and hemicelluloses
journal, November 2013
- Pareek, Nidhi; Gillgren, Thomas; Jönsson, Leif J.
- Bioresource Technology, Vol. 148
Effects of Lignin−Metal Complexation on Enzymatic Hydrolysis of Cellulose
journal, June 2010
- Liu, H.; Zhu, J. Y.; Fu, S. Y.
- Journal of Agricultural and Food Chemistry, Vol. 58, Issue 12
Inhibition of enzymatic hydrolysis by residual lignins from softwood-study of enzyme binding and inactivation on lignin-rich surface
journal, June 2011
- Rahikainen, Jenni; Mikander, Saara; Marjamaa, Kaisa
- Biotechnology and Bioengineering, Vol. 108, Issue 12
Enhancing the enzymatic hydrolysis of lignocellulosic biomass by increasing the carboxylic acid content of the associated lignin
journal, November 2010
- Nakagame, Seiji; Chandra, Richard P.; Kadla, John F.
- Biotechnology and Bioengineering, Vol. 108, Issue 3
Characterization and analysis of the molecular weight of lignin for biorefining studies
journal, June 2014
- Tolbert, Allison; Akinosho, Hannah; Khunsupat, Ratayakorn
- Biofuels, Bioproducts and Biorefining, Vol. 8, Issue 6
Disparate roles of solvent extractable lignin and residual bulk lignin in enzymatic hydrolysis of pretreated sweetgum
journal, January 2015
- Lai, Chenhuan; Tu, Maobing; Yong, Qiang
- RSC Advances, Vol. 5, Issue 119
Current Understanding of the Correlation of Lignin Structure with Biomass Recalcitrance
journal, November 2016
- Li, Mi; Pu, Yunqiao; Ragauskas, Arthur J.
- Frontiers in Chemistry, Vol. 4
Practical screening of purified cellobiohydrolases and endoglucanases with alpha-cellulose and specification of hydrodynamics
journal, January 2010
- Jager, Gernot; Wu, Zhuojun; Garschhammer, Kerstin
- Biotechnology for Biofuels, Vol. 3, Issue 1
Continuous-Flow Process for the Purification and Fractionation of Alkali and Organosolv Lignins
journal, September 2016
- Klett, Adam S.; Payne, A. Mark; Thies, Mark C.
- ACS Sustainable Chemistry & Engineering, Vol. 4, Issue 12
Recovering ultraclean lignins of controlled molecular weight from Kraft black-liquor lignins
journal, January 2015
- Klett, A. S.; Chappell, P. V.; Thies, M. C.
- Chemical Communications, Vol. 51, Issue 64
Elucidation of Lignin Structure by Quantitative 2D NMR
journal, June 2011
- Sette, Marco; Wechselberger, Rainer; Crestini, Claudia
- Chemistry - A European Journal, Vol. 17, Issue 34
Elucidating Structural Characteristics of Biomass using Solution-State 2 D NMR with a Mixture of Deuterated Dimethylsulfoxide and Hexamethylphosphoramide
journal, April 2016
- Yoo, Chang Geun; Pu, Yunqiao; Li, Mi
- ChemSusChem, Vol. 9, Issue 10
Lignin Modification during Eucalyptus globulus Kraft Pulping Followed by Totally Chlorine-Free Bleaching: A Two-Dimensional Nuclear Magnetic Resonance, Fourier Transform Infrared, and Pyrolysis−Gas Chromatography/Mass Spectrometry Study
journal, May 2007
- Ibarra, David; Chávez, María Isabel; Rencoret, Jorge
- Journal of Agricultural and Food Chemistry, Vol. 55, Issue 9
Structural characterization of Kraft lignins from different spent cooking liquors by 1D and 2D Nuclear Magnetic Resonance spectroscopy
journal, April 2014
- Fernández-Costas, C.; Gouveia, S.; Sanromán, M. A.
- Biomass and Bioenergy, Vol. 63
2-Chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane, a Reagent for the Accurate Determination of the Uncondensed and Condensed Phenolic Moieties in Lignins
journal, June 1995
- Granata, Alessandro; Argyropoulos, Dimitris S.
- Journal of Agricultural and Food Chemistry, Vol. 43, Issue 6
Application of quantitative 31P NMR in biomass lignin and biofuel precursors characterization
journal, January 2011
- Pu, Yunqiao; Cao, Shilin; Ragauskas, Arthur J.
- Energy & Environmental Science, Vol. 4, Issue 9
Comparison of dilute acid and ionic liquid pretreatment of switchgrass: Biomass recalcitrance, delignification and enzymatic saccharification
journal, July 2010
- Li, Chenlin; Knierim, Bernhard; Manisseri, Chithra
- Bioresource Technology, Vol. 101, Issue 13
Enzyme Adsorption and Cellulose Conversion during Hydrolysis of Dilute-Acid-Pretreated Corn Stover
journal, February 2014
- Tai, Chao; Keshwani, Deepak R.
- Energy & Fuels, Vol. 28, Issue 3
Effects of dilute acid pretreatment conditions on enzymatic hydrolysis monomer and oligomer sugar yields for aspen, balsam, and switchgrass
journal, April 2010
- Jensen, Jill R.; Morinelly, Juan E.; Gossen, Kelsey R.
- Bioresource Technology, Vol. 101, Issue 7
Comparison of enzymatic reactivity of corn stover solids prepared by dilute acid, AFEX™, and ionic liquid pretreatments
journal, January 2014
- Gao, Xiadi; Kumar, Rajeev; Singh, Seema
- Biotechnology for Biofuels, Vol. 7, Issue 1
The Structure of Lignin of Corn Stover and its Changes Induced by Mild Sodium Hydroxide Treatment
journal, February 2014
- Min, Dou-yong; Chang, Hou-min; Jameel, Hasan
- BioResources, Vol. 9, Issue 2
Structural Characterization and Comparison of Switchgrass Ball-milled Lignin Before and After Dilute Acid Pretreatment
journal, August 2009
- Samuel, Reichel; Pu, Yunqiao; Raman, Babu
- Applied Biochemistry and Biotechnology, Vol. 162, Issue 1
Exploring the effect of different plant lignin content and composition on ionic liquid pretreatment efficiency and enzymatic saccharification of Eucalyptus globulus L. mutants
journal, August 2012
- Papa, G.; Varanasi, P.; Sun, L.
- Bioresource Technology, Vol. 117
Alkaline Pretreatment of Switchgrass
journal, May 2015
- Karp, Eric M.; Resch, Michael G.; Donohoe, Bryon S.
- ACS Sustainable Chemistry & Engineering, Vol. 3, Issue 7
Remarkable solvent and extractable lignin effects on enzymatic digestibility of organosolv pretreated hardwood
journal, March 2014
- Lai, Chenhuan; Tu, Maobing; Li, Mi
- Bioresource Technology, Vol. 156
Characterization of milled wood lignin and ethanol organosolv lignin from miscanthus
journal, October 2009
- El Hage, Roland; Brosse, Nicolas; Chrusciel, Laurent
- Polymer Degradation and Stability, Vol. 94, Issue 10
New insights into the structure and composition of technical lignins: a comparative characterisation study
journal, January 2016
- Constant, Sandra; Wienk, Hans L. J.; Frissen, Augustinus E.
- Green Chemistry, Vol. 18, Issue 9
Structural Characterization of Pine Kraft Lignin: BioChoice Lignin vs Indulin AT
journal, May 2016
- Hu, Zhoujian; Du, Xueyu; Liu, Jie
- Journal of Wood Chemistry and Technology, Vol. 36, Issue 6
Adsorption of cellulase on cellulose: Effect of physicochemical properties of cellulose on adsorption and rate of hydrolysis
journal, October 1982
- Lee, Sun Bok; Shin, H. S.; Ryu, Dewey D. Y.
- Biotechnology and Bioengineering, Vol. 24, Issue 10
Effects of cellulase and xylanase enzymes on the deconstruction of solids from pretreatment of poplar by leading technologies
journal, March 2009
- Kumar, Rajeev; Wyman, Charles E.
- Biotechnology Progress, Vol. 25, Issue 2
Distinct Roles of Residual Xylan and Lignin in Limiting Enzymatic Hydrolysis of Organosolv Pretreated Loblolly Pine and Sweetgum
journal, January 2013
- Li, Mi; Tu, Maobing; Cao, Dongxu
- Journal of Agricultural and Food Chemistry, Vol. 61, Issue 3
Cellulose crystallinity - a key predictor of the enzymatic hydrolysis rate: Cellulose crystallinity
journal, February 2010
- Hall, Mélanie; Bansal, Prabuddha; Lee, Jay H.
- FEBS Journal, Vol. 277, Issue 6
Effects of Cellulose Crystallinity, Hemicellulose, and Lignin on the Enzymatic Hydrolysis of Miscanthus sinensis to Monosaccharides
journal, March 2008
- Yoshida, Makoto; Liu, Yuan; Uchida, Satoshi
- Bioscience, Biotechnology, and Biochemistry, Vol. 72, Issue 3
The initial kinetics of hydrolysis by cellobiohydrolases I and II is consistent with a cellulose surface - erosion model
journal, April 1998
- Valjamae, Priit; Sild, Veljo; Pettersson, Goran
- European Journal of Biochemistry, Vol. 253, Issue 2
Surface density of cellobiohydrolase on crystalline celluloses.: A critical parameter to evaluate enzymatic kinetics at a solid-liquid interface
journal, July 2006
- Igarashi, Kiyohiko; Wada, Masahisa; Hori, Ritsuko
- FEBS Journal, Vol. 273, Issue 13
Investigation of lignin deposition on cellulose during hydrothermal pretreatment, its effect on cellulose hydrolysis, and underlying mechanisms: Investigation of Lignin Deposition on Cellulose
journal, October 2013
- Li, Hongjia; Pu, Yunqiao; Kumar, Rajeev
- Biotechnology and Bioengineering, Vol. 111, Issue 3
Solvent-Driven Preferential Association of Lignin with Regions of Crystalline Cellulose in Molecular Dynamics Simulation
journal, September 2013
- Lindner, Benjamin; Petridis, Loukas; Schulz, Roland
- Biomacromolecules, Vol. 14, Issue 10
Effects of lignin and surfactant on adsorption and hydrolysis of cellulases on cellulose
journal, January 2016
- Li, Yanfei; Sun, Zongping; Ge, Xiaoyan
- Biotechnology for Biofuels, Vol. 9, Issue 1
Lignin-enzyme interaction: Mechanism, mitigation approach, modeling, and research prospects
journal, July 2017
- Li, Xiang; Zheng, Yi
- Biotechnology Advances, Vol. 35, Issue 4
Scanning probe microscopy of pine and birch kraft pulp fibres
journal, March 2000
- Simola, J.; Malkavaara, P.; Alén, R.
- Polymer, Vol. 41, Issue 6
Effect of alkali lignins with different molecular weights from alkali pretreated rice straw hydrolyzate on enzymatic hydrolysis
journal, January 2016
- Li, Yun; Qi, Benkun; Luo, Jianquan
- Bioresource Technology, Vol. 200
Cellulase–lignin interactions—The role of carbohydrate-binding module and pH in non-productive binding
journal, October 2013
- Rahikainen, Jenni Liisa; Evans, James David; Mikander, Saara
- Enzyme and Microbial Technology, Vol. 53, Issue 5
Deactivation of cellulases by phenols
journal, January 2011
- Ximenes, Eduardo; Kim, Youngmi; Mosier, Nathan
- Enzyme and Microbial Technology, Vol. 48, Issue 1
Role of Functional Groups in Lignin Inhibition of Enzymatic Hydrolysis of Cellulose to Glucose
journal, March 2008
- Pan, Xuejun
- Journal of Biobased Materials and Bioenergy, Vol. 2, Issue 1
Works referencing / citing this record:
Effect of cellulolytic enzyme binding on lignin isolated from alkali and acid pretreated switchgrass on enzymatic hydrolysis
journal, November 2019
- Jung, Woochul; Sharma-Shivappa, Ratna; Park, Sunkyu
- 3 Biotech, Vol. 10, Issue 1
Overcoming the drying-induced pore closure of APMP poplar fibers in old newsprint by surfactant treatment to promote enzymatic hydrolysis of the cellulose
journal, May 2019
- Mo, Wenxuan; Li, Bo; Li, Yaping
- Cellulose, Vol. 26, Issue 9
The Use of Lignin as a Microbial Carrier in the Co-Digestion of Cheese and Wafer Waste
journal, December 2019
- Pilarska, Agnieszka A.; Wolna-Maruwka, Agnieszka; Pilarski, Krzysztof
- Polymers, Vol. 11, Issue 12
Ethanol production by simultaneous saccharification and cofermentation of pretreated corn stalk
journal, June 2019
- Zhao, Wenxuan; Zhao, Fuguang; Zhang, Sitong
- Journal of Basic Microbiology, Vol. 59, Issue 7
Short Ozonation of Lignocellulosic Waste as Energetically Favorable Pretreatment
journal, February 2019
- Rosen, Yan; Mamane, Hadas; Gerchman, Yoram
- BioEnergy Research, Vol. 12, Issue 2
Preparation of Syringaldehyde from Lignin by Catalytic Oxidation of Perovskite-Type Oxides
journal, January 2020
- Li, Ying-xia; Zhu, Jun-peng; Zhang, Zhong-jun
- ACS Omega, Vol. 5, Issue 5
Lignin-Based Hollow Nanoparticles for Controlled Drug Delivery: Grafting Preparation Using β-Cyclodextrin/Enzymatic-Hydrolysis Lignin
journal, July 2019
- Zhou, Yu; Han, Yanming; Li, Gaiyun
- Nanomaterials, Vol. 9, Issue 7
The Use of Lignin as a Microbial Carrier in the Co-Digestion of Cheese and Wafer Waste
journal, December 2019
- Pilarska, Agnieszka A.; Wolna-Maruwka, Agnieszka; Pilarski, Krzysztof
- Polymers, Vol. 11, Issue 12