Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

Cellulases play a key role in enzymatic routes for degradation of plant cell-wall polysaccharides into simple and economically-relevant sugars. However, their low performance on complex substrates and reduced stability under industrial conditions remain the main obstacle for the large-scale production of cellulose-derived products and biofuels. Thus, in this study a novel cellulase with unusual catalytic properties from sugarcane soil metagenome (CelE1) was isolated and characterized. The polypeptide deduced from the celE1 gene encodes a unique glycoside hydrolase domain belonging to GH5 family. The recombinant enzyme was active on both carboxymethyl cellulose and β-glucan with an endo-acting mode according to capillary electrophoretic analysis of cleavage products. CelE1 showed optimum hydrolytic activity at pH 7.0 and 50 °C with remarkable activity at alkaline conditions that is attractive for industrial applications in which conventional acidic cellulases are not suitable. Moreover, its three-dimensional structure was determined at 1.8 Å resolution that allowed the identification of an insertion of eight residues in the β8-α8 loop of the catalytic domain of CelE1, which is not conserved in its psychrophilic orthologs. This 8-residue-long segment is a prominent and distinguishing feature of thermotolerant cellulases 5 suggesting that it might be involved with thermal stability. Based on its unconventional characteristics, CelE1 could be potentially employed in biotechnological processes that require thermotolerant and alkaline cellulases.

• Publication year

  2013

• Venue

  PLoS ONE

• Publication date

  2013-12-17

• Fields of study

  Biology, Materials Science, Chemistry, Environmental Science, Medicine

• Identifiers
  DOI 10.1371/journal.pone.0083635PMID 24358302PMCID 3866126
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• The 1.8 Å structure of CelE1 reveals an eight-residue insertion in the β8-α8 loop that is absent from psychrophilic orthologs.

  Confidence 0.95

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• CelE1 shows optimum hydrolytic activity at pH 7.0 and 50 °C and retains notable activity under alkaline conditions.

  Confidence 0.97

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• CelE1 acts on carboxymethyl cellulose and β-glucan with an endo-acting cleavage pattern.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• CelE1 is a novel GH5 cellulase isolated from sugarcane soil metagenome.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review

• alkaline conditions

  condition, environment

  Reaction conditions at higher pH values used to assess enzyme activity in this work.

  Aliases: alkaline pH, basic conditions

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• carboxymethyl cellulose

  substrate, polysaccharide derivative

  A soluble cellulose derivative used here as a substrate for cellulase activity testing.

  Aliases: CMC

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• cele1

  enzyme

  A cellulase enzyme encoded by the celE1 gene and characterized from sugarcane soil metagenome.

  Aliases: CelE1 cellulase

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• gh5 family

  enzyme family

  A family of glycoside hydrolase catalytic domains that includes the cellulase domain encoded by celE1.

  Aliases: glycoside hydrolase family 5, GH5

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• psychrophilic orthologs

  protein group, comparison set

  Cold-adapted homologous proteins used here for comparison with CelE1's catalytic-domain structure.

  Aliases: psychrophilic homologs

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• sugarcane soil metagenome

  source, metagenome

  Environmental genetic material recovered from sugarcane-associated soil and used as the source for celE1.

  Aliases: sugarcane soil metagenomic DNA

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• three-dimensional structure

  structure, measurement result

  The resolved atomic structure of CelE1 determined at 1.8 Å resolution.

  Aliases: 3D structure, structure

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• β8-α8 loop

  structural region, loop

  A loop region in the catalytic domain of CelE1 that contains an eight-residue insertion.

  Aliases: beta8-alpha8 loop

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review
• β-glucan

  substrate, polysaccharide

  A glucose polymer with β-linkages that serves as a cellulase substrate in the assays.

  Aliases: beta-glucan

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewq (76h6bfydm6) review

• Development and Biotechnological Application of a Novel Endoxylanase Family GH10 Identified from Sugarcane Soil Metagenome

  2013cited by this paper
• Deciphering the synergism of endogenous glycoside hydrolase families 1 and 9 from Coptotermes gestroi.

  2013cited by this paper
• Identification and Characterization of a Cellulase-Encoding Gene from the Buffalo Rumen Metagenomic Library

  2012cited by this paper
• Dissecting structure-function-stability relationships of a thermostable GH5-CBM3 cellulase from Bacillus subtilis 168.

  2012influential reference
• A salt-bridge controlled by ligand binding modulates the hydrolysis reaction in a GH5 endoglucanase.

  2012cited by this paper
• Enzymatic hydrolysis of cellulose by the cellobiohydrolase domain of CelB from the hyperthermophilic bacterium Caldicellulosiruptor saccharolyticus.

  2011cited by this paper
• REFMAC5 for the refinement of macromolecular crystal structures

  2011cited by this paper
• Mode of operation and low-resolution structure of a multi-domain and hyperthermophilic endo-β-1,3-glucanase from Thermotoga petrophila.

  2011cited by this paper
• Cloning and functional characterization of a novel endo-β-1,4-glucanase gene from a soil-derived metagenomic library

  2011cited by this paper
• Cloning and characterization of a thermostable and halo-tolerant endoglucanase from Thermoanaerobacter tengcongensis MB4

  2010cited by this paper
• Molecular replacement with MOLREP.

  2010cited by this paper
• New improvements for lignocellulosic ethanol.

  2009cited by this paper
• Structure of endoglucanase Cel9A from the thermoacidophilic Alicyclobacillus acidocaldarius.

  2009cited by this paper
• Protein engineering in designing tailored enzymes and microorganisms for biofuels production.

  2009cited by this paper
• Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases.

  2008cited by this paper
• The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium.

  2007cited by this paper
• Inference of macromolecular assemblies from crystalline state.

  2007cited by this paper
• Structure, organization, and functions of cellulose synthase complexes in higher plants

  2007cited by this paper
• Purification and characterization of a family 5 endoglucanase from a moderately thermophilic strain of Bacillus licheniformis

  2006cited by this paper
• Material for : Ethanol Can Contribute To Energy and Environmental Goals

  2006cited by this paper
• Processive Endoglucanase Active in Crystalline Cellulose Hydrolysis by the Brown Rot Basidiomycete Gloeophyllum trabeum

  2005cited by this paper
• Metagenomics and industrial applications

  2005cited by this paper
• Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering.

  2005cited by this paper
• DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data

  2004cited by this paper
• Coot: model-building tools for molecular graphics.

  2004cited by this paper
• A novel combination of two classic catalytic schemes.

  2002cited by this paper
• Type II protein secretion in gram-negative pathogenic bacteria: the study of the structure/secretion relationships of the cellulase Cel5 (formerly EGZ) from Erwinia chrysanthemi.

  2001cited by this paper
• Crystal structure of alkaline cellulase K: insight into the alkaline adaptation of an industrial enzyme.

  2001cited by this paper
• Insights into ligand-induced conformational change in Cel5A from Bacillus agaradhaerens revealed by a catalytically active crystal form.

  2000cited by this paper
• Structure of the Bacillus agaradherans family 5 endoglucanase at 1.6 A and its cellobiose complex at 2.0 A resolution.

  1998cited by this paper
• Crystal structure of thermostable family 5 endocellulase E1 from Acidothermus cellulolyticus in complex with cellotetraose.

  1996cited by this paper
• A common protein fold and similar active site in two distinct families of β-glycanases

  1996cited by this paper
• Crystal structure of the catalytic domain of a bacterial cellulase belonging to family 5.

  1995cited by this paper
• New families in the classification of glycosyl hydrolases based on amino acid sequence similarities.

  1993cited by this paper
• Stereochemistry of the hydrolysis reaction catalyzed by endoglucanase Z from Erwinia chrysanthemi

  1992cited by this paper
• Atomic features of protein-carbohydrate interactions

  1991cited by this paper
• The TIM barrel—the most frequently occurring folding motif in proteins

  1991cited by this paper
• Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen

  1982cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper
• Solvent content of protein crystals.

  1968cited by this paper
• Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar

  1959cited by this paper
• A BUFFER SOLUTION FOR COLORIMETRIC COMPARISON

  1921cited by this paper

• Cellulases: key properties, natural sources, and industrial applications

  2025cites this paper
• Discovery, structural characterization, and functional insights into a novel apiosidase from the GH140 family, isolated from a lignocellulolytic-enriched mangrove microbial community

  2024cites this paper
• Cellulolytic enzymes in Microbulbifer sp. Strain GL-2, a marine fish intestinal bacterium, with emphasis on endo-1,4-β-glucanases Cel5A and Cel8.

  2024cites this paper
• Enhanced crystalline cellulose degradation by a novel metagenome-derived cellulase enzyme

  2024cites this paper
• Structural and molecular insights into a bifunctional glycoside hydrolase 30 xylanase specific to glucuronoxylan

  2024cites this paper
• Metagenomic mining and molecular characterization of a psychrophilic GH5-CBM5 cellulase from Krossfjorden sediments, Arctic

  2024cites this paper
• Influence of elevated pressure and pressurized fluids on microenvironment and activity of enzymes.

  2023cites this paper
• Structural and functional insights of the catalytic GH5 and Calx-β domains from the metagenome-derived endoglucanase CelE2.

  2023cites this paper
• Biochemical and structural characterisation of a family GH5 cellulase from endosymbiont of shipworm P. megotara

  2023cites this paper
• Designing microbial cellulases using genetic engineering approach: A promising strategy towards zero-waste cellulosic biorefinery

  2023cites this paper
• Soil microbiome engineering for sustainability in a changing environment

  2023cites this paper
• Metagenomics for mining of thermoalkalophilic enzymes

  2022cites this paper
• Metagenomic Approaches as a Tool to Unravel Promising Biocatalysts from Natural Resources: Soil and Water

  2022cites this paper
• Role of metagenomics in prospecting novel endoglucanases, accentuating functional metagenomics approach in second-generation biofuel production: a review

  2021cites this paper
• Effects of Herbicides on Soil Enzymes and Their Regulatory Factors in Agroecosystem: A Review

  2021cites this paper
• Investigation of cold-active and mesophilic cellulases: opportunities awaited

  2021cites this paper
• In silico characterization of the GH5-cellulase family from uncultured microorganisms: physicochemical and structural studies

  2021cites this paper
• Biodiversity and Industrial Applications of Genus Chaetomium

  2021cites this paper
• Diversity of cellulolytic microorganisms and microbial cellulases

  2021cites this paper
• Microbial Cellulolytic-Based Biofuel Production

  2021cites this paper
• Metagenomic Discovery and Characterization of Multi-Functional and Monomodular Processive Endoglucanases as Biocatalysts

  2021cites this paper
• Cellulases: From Bioactivity to a Variety of Industrial Applications

  2021cites this paper
• Xylanases of glycoside hydrolase family 30 - An overview.

  2021cites this paper
• A snapshot of microbial diversity and function in an undisturbed sugarcane bagasse pile

  2020cites this paper
• Characterization and efficient production of a thermostable, halostable and organic solvent-stable cellulase from an oil reservoir.

  2020cites this paper
• Industrial Applications of Glycoside Hydrolases

  2020cites this paper
• Role of Metagenomics in Discovery of Industrially Important Cellulase

  2020cites this paper
• A processive endoglucanase with multi-substrate specificity is characterized from porcine gut microbiota

  2019cites this paper
• Methods in Metagenomics and Environmental Biotechnology

  2019cites this paper
• Function Profiling of Microbial Community

  2019cites this paper
• Characterization of the cellulase-secretome produced by the Antarctic bacterium Flavobacterium sp. AUG42.

  2019cites this paper
• Cellulolytic thermophilic microorganisms in white biotechnology: a review

  2019cites this paper
• Biochemical Characterization of a Novel Monospecific Endo-β-1,4-Glucanase Belonging to GH Family 5 From a Rhizosphere Metagenomic Library

  2019cites this paper
• A Novel Subfamily of Endo-β-1,4-Glucanases in Glycoside Hydrolase Family 10

  2019cites this paper
• Nanoscience and Biotechnology for Environmental Applications

  2019cites this paper
• Chaetomium Enzymes and Their Applications

  2019cites this paper
• Cellulases from Thermophiles Found by Metagenomics

  2018cites this paper
• Overexpression and biochemical characterization of a recombinant psychrophilic endocellulase from Pseudoalteromonas sp. DY3.

  2018cites this paper
• A cold-adapted endoglucanase from camel rumen with high catalytic activity at moderate and low temperatures: an anomaly of truly cold-adapted evolution in a mesophilic environment

  2018cites this paper
• Recombinant thermo-alkali-stable endoglucanase of Myceliopthora thermophila BJA (rMt-egl): Biochemical characteristics and applicability in enzymatic saccharification of agro-residues.

  2017cites this paper
• Metagenomics of Plant Microbiomes

  2017cites this paper
• An Expedition to the Mechanism of Plant–Microbe Interaction by Utilization of Different Molecular Biology Tools

  2017cites this paper
• Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture.

  2017cites this paper
• META-pipe – Distributed Pipeline Analysis of Marine Metagenomic Sequence Data

  2017cites this paper
• Functional Metagenomics: Tools and Applications

  2017cites this paper
• Diversity of Microbial Carbohydrate-Active enZYmes (CAZYmes) Associated with Freshwater and Soil Samples from Caatinga Biome

  2017cites this paper
• Applied Metagenomics for Biofuel Development and Environmental Sustainability

  2017cites this paper
• A Novel Carbohydrate-binding Module from Sugar Cane Soil Metagenome Featuring Unique Structural and Carbohydrate Affinity Properties*

  2016influential citation
• From lignocellulosic metagenomes to lignocellulolytic genes: trends, challenges and future prospects

  2016cites this paper
• Screening, identification, and characterization of a novel saccharide-stimulated β-glycosidase from a soil metagenomic library

  2016cites this paper
• Discovery and Characterization of a Thermostable and Highly Halotolerant GH5 Cellulase from an Icelandic Hot Spring Isolate

  2016influential citation
• Functional and structural characterization of a novel putative cysteine protease cell wall-modifying multi-domain enzyme selected from a microbial metagenome

  2016cites this paper
• Bifunctional recombinant cellulase–xylanase (rBhcell-xyl) from the polyextremophilic bacterium Bacillus halodurans TSLV1 and its utility in valorization of renewable agro-residues

  2016cites this paper
• Synergistic function of four novel thermostable glycoside hydrolases from a long-term enriched thermophilic methanogenic digester

  2015cites this paper
• Heterologous expression and characterization of detergent stable endoglucanase EG5B from Paenibacillus sp. IHB B 3084

  2015cites this paper
• Cloning, purification, crystallization and preliminary X-ray studies of a carbohydrate-binding module from family 64 (StX).

  2015cites this paper
• Screening, identification, and characterization of a GH43 family β-xylosidase/α-arabinofuranosidase from a compost microbial metagenome

  2015cites this paper
• Chemical stability of a cold-active cellulase with high tolerance toward surfactants and chaotropic agent

  2015cites this paper
• Structural Insight of a Trimodular Halophilic Cellulase with a Family 46 Carbohydrate-Binding Module

  2015influential citation
• A Novel Member of GH16 Family Derived from Sugarcane Soil Metagenome

  2015cites this paper
• Metagenomics: An Application Based Perspective

  2014cites this paper
• Cloning, purification, crystallization and preliminary X-ray studies of a carbohydrate-binding module (CBM_E1) derived from sugarcane soil metagenome.

  2014cites this paper