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Title: Heterologous expression of family 10 xylanases from Acidothermus cellulolyticus enhances the exoproteome of Caldicellulosiruptor bescii and growth on xylan substrates

Journal Article · · Biotechnology for Biofuels
 [1];  [2];  [2];  [2];  [1]
  1. Univ. of Georgia, Athens, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

The ability to deconstruct plant biomass without conventional pretreatment has made members of the genus Caldicellulosiruptor the target of investigation for the consolidated processing of lignocellulosic biomass to biofuels and bioproducts. These Gram-positive bacteria are hyperthermophilic anaerobes and the most thermophilic cellulolytic organisms so far described. They use both C5 and C6 sugars simultaneously and have the ability to grow well on xylan, a major component of plant cell walls. This is an important advantage for their use to efficiently convert biomass at yields sufficient for an industrial process. For commodity chemicals, yield from substrate is perhaps the most important economic factor. In an attempt to improve even further the ability of C. bescii to use xylan, we introduced two xylanases from Acidothermus cellulolyticus. Acel_0180 includes tandem carbohydrate-binding modules (CBM2 and CBM3) located at the C-terminus, one of which, CBM2, is not present in C. bescii. Also, the sequences of Xyn10A and Acel_0180 have very little homology with the GH10 domains present in C. bescii. For these reasons, we selected these xylanases as potential candidates for synergistic interaction with those in the C. bescii exoproteome. As a result, heterologous expression of two xylanases from Acidothermus cellulolyticus in Caldicellulosiruptor bescii resulted in a modest, but significant increase in the activity of the exoproteome of C. bescii on xylan substrates. Even though the increase in extracellular activity was modest, the ability of C. bescii to grow on these substrates was dramatically improved suggesting that the xylan substrate/microbe interaction substantially increased deconstruction over the secreted free enzymes alone. In conclusion, we anticipate that the ability to efficiently use xylan, a major component of plant cell walls for conversion of plant biomass to products of interest, will allow the conversion of renewable, sustainable, and inexpensive plant feedstocks to products at high yields.

Research Organization:
National Renewable Energy Laboratory (NREL), Golden, CO (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC36-08GO28308; AC05-00OR22725
OSTI ID:
1319281
Report Number(s):
NREL/JA-2700-67046
Journal Information:
Biotechnology for Biofuels, Vol. 9, Issue 1; ISSN 1754-6834
Publisher:
BioMed CentralCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 20 works
Citation information provided by
Web of Science

References (42)

Xyn10A, a Thermostable Endoxylanase from Acidothermus cellulolyticus 11B journal September 2010
Identification and characterization of CbeI, a novel thermostable restriction enzyme from Caldicellulosiruptor bescii DSM 6725 and a member of a new subfamily of HaeIII-like enzymes journal May 2011
Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose journal August 2015
Feature-based reappraisal of theBacillus subtilis exoproteome journal January 2007
Xylanases, xylanase families and extremophilic xylanases journal January 2005
Degradation of microcrystalline cellulose and non-pretreated plant biomass by a cell-free extracellular cellulase/hemicellulase system from the extreme thermophilic bacterium Caldicellulosiruptor bescii journal January 2013
Secretion and subcellular localizations of bacterial proteins: a semantic awareness issue journal April 2009
Substrate Specificity in Glycoside Hydrolase Family 10: TYROSINE 87 AND LEUCINE 314 PLAY A PIVOTAL ROLE IN DISCRIMINATING BETWEEN GLUCOSE AND XYLOSE BINDING IN THE PROXIMAL ACTIVE SITE OF journal April 2000
Ethanol production from xylose in engineered Saccharomyces cerevisiae strains: current state and perspectives journal July 2009
Enhancement of Synthetic Trichoderma-Based Enzyme Mixtures for Biomass Conversion with an Alternative Family 5 Glycosyl Hydrolase from Sporotrichum thermophile journal October 2014
Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii journal June 2014
Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations journal March 2009
Insights into plant biomass conversion from the genome of the anaerobic thermophilic bacterium Caldicellulosiruptor bescii DSM 6725 journal January 2011
Revealing Nature's Cellulase Diversity The Digestion Mechanism of Caldicellulosiruptor bescii CelA journal December 2013
A Proteomic View on Genome-Based Signal Peptide Predictions journal September 2001
Xylanases: from Biology to BioTechnology journal December 1996
Carbon catabolite repression in bacteria: many ways to make the most out of nutrients journal August 2008
Construction of a Stable Replicating Shuttle Vector for Caldicellulosiruptor Species: Use for Extending Genetic Methodologies to Other Members of This Genus journal May 2013
Assembly of Xylanases into Designer Cellulosomes Promotes Efficient Hydrolysis of the Xylan Component of a Natural Recalcitrant Cellulosic Substrate journal November 2011
Competition between pentoses and glucose during uptake and catabolism in recombinant Saccharomyces cerevisiae journal January 2012
Novel architecture of family-9 glycoside hydrolases identified in cellulosomal enzymes of Acetivibrio cellulolyticus and Clostridium thermocellum journal January 2006
Cellulosic ethanol production via consolidated bioprocessing at 75 °C by engineered Caldicellulosiruptor bescii journal October 2015
Hemicellulose bioconversion journal May 2003
Homologous Expression of the Caldicellulosiruptor bescii CelA Reveals that the Extracellular Protein Is Glycosylated journal March 2015
Use of Label-Free Quantitative Proteomics To Distinguish the Secreted Cellulolytic Systems of Caldicellulosiruptor bescii and Caldicellulosiruptor obsidiansis journal April 2011
Recent progress in consolidated bioprocessing journal June 2012
Strain engineering of Saccharomyces cerevisiae for enhanced xylose metabolism journal November 2013
Heterologous complementation of a pyrF deletion in Caldicellulosiruptor hydrothermalisgenerates a new host for the analysis of biomass deconstruction journal September 2014
Overcoming restriction as a barrier to DNA transformation in Caldicellulosiruptor species results in efficient marker replacement journal January 2013
Xylans from Oat Spelts and Birch Kraft Pulp journal December 2005
Improved growth media and culture techniques for genetic analysis and assessment of biomass utilization by Caldicellulosiruptor bescii journal November 2012
Crotonylation of key metabolic enzymes regulates carbon catabolite repression in Streptomyces roseosporus journal April 2020
Substrate Specificity in Glycoside Hydrolase Family 10 journal July 2000
Degradation of different [(glucurono)arabino]xylans by a combination of purified xylan-degrading enzymes journal February 1993
Relationships between activities of xylanases and xylan structures journal July 2000
Structural Insights into Serine-rich Fimbriae from Gram-positive Bacteria journal June 2010
Isolation and Characterization of Acidothermus cellulolyticus gen. nov., sp. nov., a New Genus of Thermophilic, Acidophilic, Cellulolytic Bacteria journal July 1986
Efficient Degradation of Lignocellulosic Plant Biomass, without Pretreatment, by the Thermophilic Anaerobe "Anaerocellum thermophilum" DSM 6725 journal May 2009
Reconstitution of a Thermostable Xylan-Degrading Enzyme Mixture from the Bacterium Caldicellulosiruptor bescii journal December 2012
Characterization and sequence of a Thermomonospora fusca xylanase. journal January 1994
Caldicellulosiruptor Core and Pangenomes Reveal Determinants for Noncellulosomal Thermophilic Deconstruction of Plant Biomass journal May 2012
Proteomics of Protein Secretion by Bacillus subtilis: Separating the "Secrets" of the Secretome journal June 2004

Cited By (2)

Genomic and physiological analyses reveal that extremely thermophilic Caldicellulosiruptor changbaiensis deploys uncommon cellulose attachment mechanisms journal August 2019
Genome Stability in Engineered Strains of the Extremely Thermophilic Lignocellulose-Degrading Bacterium Caldicellulosiruptor bescii journal May 2017


Figures / Tables (6)