skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Consolidated bioprocessing of Populus using Clostridium (Ruminiclostridium) thermocellum: a case study on the impact of lignin composition and structure

Journal Article · · Biotechnology for Biofuels

Higher ratios of syringyl-to-guaiacyl (S/G) lignin components of Populus were shown to improve sugar release by enzymatic hydrolysis using commercial blends. Cellulolytic microbes are often robust biomass hydrolyzers and may offer cost advantages, however, it is unknown whether their activity can also be significantly influenced by the ratio of different monolignol types in Populus biomass. Hydrolysis and fermentation of autoclaved but otherwise not pretreated Populus trichocarpa by Clostridium thermocellum ATCC 27405 was compared using feedstocks that had similar carbohydrate and total lignin contents but differed in S/G ratios. Populus with an S/G ratio of 2.1 was converted more rapidly and to a greater extent compared to similar biomass that had a ratio of 1.2. For either microbes or commercial enzymes, an approximate 50% relative difference in total solids solubilization was measured for both biomasses, which suggests that the differences and limitations in the microbial breakdown of lignocellulose may be largely from the enzymatic hydrolytic process. Unexpectedly, the reduction in glucan content per gram solid in the residual microbially processed biomass was similar (17 18 %) irrespective of S/G ratio, pointing to a similar mechanism of solubilization that proceeded at different rates. Fermentation metabolome testing did not reveal the release of known biomass-derived alcohol and aldehyde inhibitors that could explain observed differences in microbial hydrolytic activity. Biomass-derived p-hydroxybenzoic acid was up to nine-fold higher in low S/G ratio biomass fermentations, but was not found to be inhibitory in subsequent test fermentations. Cellulose crystallinity and degree of polymerization did not vary between Populus lines and had minor changes after fermentation. However, lignin molecular weights and cellulose accessibility determined by Simons staining were positively correlated to the S/G content. Higher S/G ratios in Populus biomass lead to longer and more linear lignin chains and greater access to surface cellulosic content by microbe-bound enzymatic complexes. Substrate access limitation is suggested as a primary bottleneck in solubilization of minimally processed Populus, which has important implications for microbial deconstruction of lignocellulose biomass. Lastly, our findings will allow others to examine different Populus lines and to test if similar observations are possible for other plant species.

Research Organization:
National Renewable Energy Laboratory (NREL), Golden, CO (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
Sponsoring Organization:
USDOE Office of Science (SC), Office of Biological and Environmental Research; USDOE Office of Science (SC)
Grant/Contract Number:
Bioenergy Science Center (BESC) which is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.; AC36-08GO28308; AC05-00OR22725
OSTI ID:
1618635
Alternate ID(s):
OSTI ID: 1240075; OSTI ID: 1240560
Report Number(s):
NREL/JA-5100-65961; 31; PII: 445
Journal Information:
Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Vol. 9 Journal Issue: 1; ISSN 1754-6834
Publisher:
Springer Science + Business MediaCopyright Statement
Country of Publication:
Netherlands
Language:
English
Citation Metrics:
Cited by: 49 works
Citation information provided by
Web of Science

References (40)

p -Hydroxyphenyl, Guaiacyl, and Syringyl Lignins Have Similar Inhibitory Effects on Wall Degradability journal July 1997
The characterization of pretreated lignocellulosic substrates prior to enzymatic hydrolysis, part 1: A modified Simons' staining technique journal September 2008
Lignin monomer composition affects Arabidopsis cell-wall degradability after liquid hot water pretreatment journal January 2010
Lignin Structural Variation in Hardwood Species journal May 2012
Jatropha cultivation in southern India: assessing farmers' experiences journal February 2012
The degradation pattern of cellulose by extracellular cellulases of aerobic and anaerobic microorganisms journal January 1991
Syringyl lignin production in conifers: Proof of concept in a Pine tracheary element system journal April 2015
Application of optimized alkaline pretreatment for enhancing the anaerobic digestion of different sunflower stalks varieties journal July 2013
Fermentation of soybean hulls to ethanol while preserving protein value journal July 2009
Lignin content in natural Populus variants affects sugar release journal March 2011
Do Enzymatic Hydrolyzability and Simons' Stain Reflect the Changes in the Accessibility of Lignocellulosic Substrates to Cellulase Enzymes? journal December 2001
Influence of Syringyl to Guaiacyl Ratio on the Structure of Natural and Synthetic Lignins journal January 2010
Degradation of high loads of crystalline cellulose and of unpretreated plant biomass by the thermophilic bacterium Caldicellulosiruptor bescii journal January 2014
Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass journal February 2011
Lignin modification improves fermentable sugar yields for biofuel production journal June 2007
Effect of Lignin Chemistry on the Enzymatic Hydrolysis of Woody Biomass journal June 2014
Variation in Cell Wall Composition among Forage Maize ( Zea mays L.) Inbred Lines and Its Impact on Digestibility:  Analysis of Neutral Detergent Fiber Composition by Pyrolysis-Gas Chromatography-Mass Spectrometry journal December 2003
Significant Increases in Pulping Efficiency in C4H-F5H-Transformed Poplars:  Improved Chemical Savings and Reduced Environmental Toxins journal October 2003
Enzyme-microbe synergy during cellulose hydrolysis by Clostridium thermocellum journal October 2006
High-Throughput Screening of Plant Cell-Wall Composition Using Pyrolysis Molecular Beam Mass Spectroscopy book January 2009
Industrial Robustness: Understanding the Mechanism of Tolerance for the Populus Hydrolysate-Tolerant Mutant Strain of Clostridium thermocellum journal October 2013
Elimination of metabolic pathways to all traditional fermentation products increases ethanol yields in Clostridium thermocellum journal November 2015
Use of the Simons' Staining Technique to Assess Cellulose Accessibility in Pretreated Substrates journal August 2012
Using Populus as a lignocellulosic feedstock for bioethanol journal February 2015
The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications journal August 2014
Evaluation of the bioconversion of genetically modified switchgrass using simultaneous saccharification and fermentation and a consolidated bioprocessing approach journal January 2012
Influence of initial cellulose concentration on the carbon flow distribution during batch fermentation by Clostridium thermocellum ATCC 27405 journal November 2008
Cellulose crystallinity - a key predictor of the enzymatic hydrolysis rate: Cellulose crystallinity journal February 2010
Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance journal January 2010
Conversion for Avicel and AFEX pretreated corn stover by Clostridium thermocellum and simultaneous saccharification and fermentation: Insights into microbial conversion of pretreated cellulosic biomass journal September 2011
Syringyl-Rich Lignin Renders Poplars More Resistant to Degradation by Wood Decay Fungi journal February 2013
High-resolution genetic mapping of allelic variants associated with cell wall chemistry in Populus journal January 2015
Improved saccharification and ethanol yield from field-grown transgenic poplar deficient in cinnamoyl-CoA reductase journal December 2013
Effect of lignin content on changes occurring in poplar cellulose ultrastructure during dilute acid pretreatment journal October 2014
Poplar as a feedstock for biofuels: A review of compositional characteristics journal March 2010
Enhanced biodegradation of sugarcane bagasse by Clostridium thermocellum with surfactant addition journal January 2014
Pretreatment of Rice Straw for the Improvement of Biogas Production journal May 2015
Variation of S/G Ratio and Lignin Content in a <i>Populus</i> Family Influences the Release of Xylose by Dilute Acid Hydrolysis journal January 2006
Clean Fractionation Pretreatment Reduces Enzyme Loadings for Biomass Saccharification and Reveals the Mechanism of Free and Cellulosomal Enzyme Synergy journal May 2014
Consolidated bioprocessing of transgenic switchgrass by an engineered and evolved Clostridium thermocellum strain journal January 2014

Cited By (11)

Advances in understanding the surface chemistry of lignocellulosic biomass via time-of-flight secondary ion mass spectrometry journal December 2016
Specialized activities and expression differences for Clostridium thermocellum biofilm and planktonic cells journal February 2017
Engineering microbes for direct fermentation of cellulose to bioethanol journal February 2018
Study of traits and recalcitrance reduction of field-grown COMT down-regulated switchgrass journal January 2017
Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass journal January 2017
Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring journal March 2017
Biomass Recalcitrance in Willow Under Two Biological Conversion Paradigms: Enzymatic Hydrolysis and Anaerobic Digestion journal December 2019
Enhanced depolymerization and utilization of raw lignocellulosic material by co-cultures of Ruminiclostridium thermocellum with hemicellulose-utilizing partners journal April 2019
Clostridium thermocellum LL1210 pH homeostasis mechanisms informed by transcriptomics and metabolomics journal April 2018
Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar journal March 2020
High Throughput Screening Technologies in Biomass Characterization journal November 2018


Figures / Tables (9)