DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Composition and yield of non-cellulosic and cellulosic sugars in soluble and particulate fractions during consolidated bioprocessing of poplar biomass by Clostridium thermocellum

Journal Article · · Biotechnology for Biofuels and Bioproducts

Abstract Background Terrestrial plant biomass is the primary renewable carbon feedstock for enabling transition to a sustainable bioeconomy. Consolidated bioprocessing (CBP) by the cellulolytic thermophile Clostridium thermocellum offers a single step microbial platform for production of biofuels and biochemicals via simultaneous solubilization of carbohydrates from lignocellulosic biomass and conversion to products. Here, solubilization of cell wall cellulosic, hemicellulosic, and pectic polysaccharides in the liquor and solid residues generated during CBP of poplar biomass by C. thermocellum was analyzed. Results The total amount of biomass solubilized in the C. thermocellum DSM1313 fermentation platform was 5.8, 10.3, and 13.7% of milled non-pretreated poplar after 24, 48, and 120 h, respectively. These results demonstrate solubilization of 24% cellulose and 17% non-cellulosic sugars after 120 h, consistent with prior reports. The net solubilization of non-cellulosic sugars by C. thermocellum (after correcting for the uninoculated control fermentations) was 13 to 36% of arabinose (Ara), xylose (Xyl), galactose (Gal), mannose (Man), and glucose (Glc); and 15% and 3% of fucose and glucuronic acid, respectively. No rhamnose was solubilized and 71% of the galacturonic acid (GalA) was solubilized. These results indicate that C. thermocellum may be selective for the types and/or rate of solubilization of the non-cellulosic wall polymers. Xyl, Man, and Glc were found to accumulate in the fermentation liquor at levels greater than in uninoculated control fermentations, whereas Ara and Gal did not accumulate, suggesting that C. thermocellum solubilizes both hemicelluloses and pectins but utilizes them differently. After five days of fermentation, the relative amount of Rha in the solid residues increased 21% indicating that the Rha-containing polymer rhamnogalacturonan I (RG-I) was not effectively solubilized by C. thermocellum CBP, a result confirmed by immunoassays. Comparison of the sugars in the liquor versus solid residue showed that C. thermocellum solubilized hemicellulosic xylan and mannan, but did not fully utilize them, solubilized and appeared to utilize pectic homogalacturonan, and did not solubilize RG-I. Conclusions The significant relative increase in RG-I in poplar solid residues following CBP indicates that C. thermocellum did not solubilize RG-I. These results support the hypothesis that this pectic glycan may be one barrier for efficient solubilization of poplar by C. thermocellum .

Sponsoring Organization:
USDOE
Grant/Contract Number:
SC0015662; AC36-08GO28308
OSTI ID:
1846685
Journal Information:
Biotechnology for Biofuels and Bioproducts, Journal Name: Biotechnology for Biofuels and Bioproducts Journal Issue: 1 Vol. 15; ISSN 2731-3654
Publisher:
Springer Science + Business MediaCopyright Statement
Country of Publication:
United Kingdom
Language:
English

References (65)

Poplar as a feedstock for biofuels: A review of compositional characteristics journal March 2010
Automated Synthesis of Arabinoxylan-Oligosaccharides Enables Characterization of Antibodies that Recognize Plant Cell Wall Glycans journal February 2015
Functional subgenomics ofClostridium thermocellum cellulosomal genes: Identification of the major catalytic components in the extracellular complex and detection of three new enzymes journal September 2005
Lignin Exhibits Recalcitrance‐Associated Features Following the Consolidated Bioprocessing of Populus trichocarpa Natural Variants journal November 2017
From cellulosomes to cellulosomics journal January 2008
Pectinolytic activity ofClostridium thermocellum: Its use for anaerobic fermentation of sugar beet pulp journal March 1986
Polysaccharides in some industrially important hardwood species journal October 2005
Mutant selection and phenotypic and genetic characterization of ethanol-tolerant strains of Clostridium thermocellum journal August 2011
Nucleotide sequences of two contiguous and highly homologous xylanase genes xynA and xynB and characterization of XynA from Clostridium thermocellum journal March 1999
A defined growth medium with very low background carbon for culturing Clostridium thermocellum journal February 2012
Cloning of LicB from Clostridium thermocellum and its efficient secretive expression of thermostable β-1,3-1,4-glucanase journal March 2014
A New Member of Family 11 Polysaccharide Lyase, Rhamnogalacturonan Lyase (CtRGLf) from Clostridium thermocellum journal February 2016
Clostridium thermocellum: A microbial platform for high-value chemical production from lignocellulose book August 2020
Carbohydrate analysis by a phenol–sulfuric acid method in microplate format journal April 2005
The multi-ligand binding first family 35 Carbohydrate Binding Module (CBM35) of Clostridium thermocellum targets rhamnogalacturonan I journal September 2018
Characterization of reduced carbohydrate solubilization during Clostridium thermocellum fermentation with high switchgrass concentrations journal August 2020
Effect of xylanase supplementation of cellulase on digestion of corn stover solids prepared by leading pretreatment technologies journal September 2009
Xylooligomers are strong inhibitors of cellulose hydrolysis by enzymes journal December 2010
Closing the carbon balance for fermentation by Clostridium thermocellum (ATCC 27405) journal January 2012
Xylans inhibit enzymatic hydrolysis of lignocellulosic materials by cellulases journal October 2012
The structure, function, and biosynthesis of plant cell wall pectic polysaccharides journal September 2009
Towards sustainable development through the circular economy—A review and critical assessment on current circularity metrics journal December 2019
Comprehensive Monosaccharide Composition Analysis of Insoluble Polysaccharides by Permethylation To Produce Methyl Alditol Derivatives for Gas Chromatography/Mass Spectrometry journal September 2019
Primary, Secondary, Tertiary and Quaternary Structure Levels in Linear Polysaccharides: From Random Coil, to Single Helix to Supramolecular Assembly journal February 2019
Elucidating the Structural Changes to Populus Lignin during Consolidated Bioprocessing with Clostridium thermocellum journal July 2017
Biocommodity Engineering journal October 1999
Quantitative Saccharification of Wood and Cellulose journal January 1945
Novel Xylanase from a Holstein Cattle Rumen Metagenomic Library and Its Application in Xylooligosaccharide and Ferulic Acid Production from Wheat Straw journal December 2012
A Colorimetric Method for the Determination of Sugars journal July 1951
Sugar release and growth of biofuel crops are improved by downregulation of pectin biosynthesis journal February 2018
Development of a thermophilic coculture for corn fiber conversion to ethanol journal April 2020
Identification of endoxylanase XynE from Clostridium thermocellum as the first xylanase of glycoside hydrolase family GH141 journal September 2017
Cellulose and lignin colocalization at the plant cell wall surface limits microbial hydrolysis of Populus biomass journal January 2017
Carbohydrate-binding modules: fine-tuning polysaccharide recognition journal September 2004
A family 26 mannanase produced by Clostridium thermocellum as a component of the cellulosome contains a domain which is conserved in mannanases from anaerobic fungi journal November 1999
Two new major subunits in the cellulosome of Clostridium thermocellum: xyloglucanase Xgh74A and endoxylanase Xyn10D journal October 2005
A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies journal September 2017
The Arabidopsis irregular xylem8 Mutant Is Deficient in Glucuronoxylan and Homogalacturonan, Which Are Essential for Secondary Cell Wall Integrity journal January 2007
An Arabidopsis Cell Wall Proteoglycan Consists of Pectin and Arabinoxylan Covalently Linked to an Arabinogalactan Protein journal January 2013
RUBY, a Putative Galactose Oxidase, Influences Pectin Properties and Promotes Cell-To-Cell Adhesion in the Seed Coat Epidermis of Arabidopsis journal March 2019
Rhamnogalacturonan‐I is a determinant of cell–cell adhesion in poplar wood journal October 2019
Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities journal February 2016
Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production journal February 2007
Chi18A, the Endochitinase in the Cellulosome of the Thermophilic, Cellulolytic Bacterium Clostridium thermocellum journal June 2002
Feruloyl Esterase Activity of the Clostridium thermocellum Cellulosome Can Be Attributed to Previously Unknown Domains of XynY and XynZ journal March 2000
Microbial Cellulose Utilization: Fundamentals and Biotechnology journal September 2002
Purification of Clostridium thermocellum xylanase Z expressed in Escherichia coli and identification of the corresponding product in the culture medium of C. thermocellum. journal October 1988
Sequence of xynC and properties of XynC, a major component of the Clostridium thermocellum cellulosome journal July 1997
Aspen pectate lyase PtxtPL1-27 mobilizes matrix polysaccharides from woody tissues and improves saccharification yield journal January 2014
Deletion of a gene cluster encoding pectin degrading enzymes in Caldicellulosiruptor bescii reveals an important role for pectin in plant biomass recalcitrance journal October 2014
The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading journal October 2014
Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock journal January 2015
Consolidated bioprocessing of Populus using Clostridium (Ruminiclostridium) thermocellum: a case study on the impact of lignin composition and structure journal February 2016
Comparison of four glycosyl residue composition methods for effectiveness in detecting sugars from cell walls of dicot and grass tissues journal July 2017
The effect of switchgrass loadings on feedstock solubilization and biofuel production by Clostridium thermocellum journal November 2017
Comparative characterization of all cellulosomal cellulases from Clostridium thermocellum reveals high diversity in endoglucanase product formation essential for complex activity journal October 2017
Working towards recalcitrance mechanisms: increased xylan and homogalacturonan production by overexpression of GAlactUronosylTransferase12 (GAUT12) causes increased recalcitrance and decreased growth in Populus journal January 2018
Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum journal August 2018
Multiple levers for overcoming the recalcitrance of lignocellulosic biomass journal January 2019
Elucidation of the recognition mechanisms for hemicellulose and pectin in Clostridium cellulovorans using intracellular quantitative proteome analysis journal May 2015
Sequence of the Clostridium thermocellum Mannanase Gene man26B and Characterization of the Translated Product journal January 2001
Impact of Pretreated Switchgrass and Biomass Carbohydrates on Clostridium thermocellum ATCC 27405 Cellulosome Composition: A Quantitative Proteomic Analysis journal April 2009
Role of Pectinolytic Enzymes Identified in Clostridium thermocellum Cellulosome journal February 2015
The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications journal August 2014
Loss of Arabidopsis GAUT12/IRX8 causes anther indehiscence and leads to reduced G lignin associated with altered matrix polysaccharide deposition journal July 2014