Engineering of glycoside hydrolase family 7 cellobiohydrolases directed by natural diversity screening

Brunecky, Roman; Knott, Brandon C.; Subramanian, Venkataramanan; Linger, Jeffrey G.; Beckham, Gregg T.; Amore, Antonella; Taylor, II, Larry E.; Vander Wall, Todd A.; Lunin, Vladimir V.; Zheng, Fei; Garrido, Mercedes; Schuster, Logan; Fulk, Emily M.; Farmer, Samuel; Himmel, Michael E.; Decker, Stephen R.

doi:10.1016/j.jbc.2024.105749

Title: Engineering of glycoside hydrolase family 7 cellobiohydrolases directed by natural diversity screening

Journal Article · Fri Mar 01 00:00:00 EST 2024 · Journal of Biological Chemistry

DOI:https://doi.org/10.1016/j.jbc.2024.105749· OSTI ID:2301755

Brunecky, Roman; Knott, Brandon C.; Subramanian, Venkataramanan; Linger, Jeffrey G.; Beckham, Gregg T.; Amore, Antonella; Taylor, II, Larry E.; Vander Wall, Todd A.; Lunin, Vladimir V.; Zheng, Fei;

; Schuster, Logan;

; Farmer, Samuel; Himmel, Michael E.;

Protein engineering and screening of processive fungal cellobiohydrolases (CBHs) remain challenging due to limited expression hosts, synergy-dependency, and recalcitrant substrates. In particular, GH7 CBHs are a family critically important for the bioeconomy as well as a methodology for improving these and other difficult to engineer enzymes. Here, we target the discovery of highly active natural GH7 CBHs and engineering of variants with improved activity. Using experimentally assayed activities of genome mined CBHs, we applied sequence and structural alignments to top performers to identify key point mutations linked to improved activity. From ~1500 known GH7 sequences, an evolutionarily diverse subset of 57 GH7 CBH genes was expressed in Trichoderma reesei and screened using a multiplexed activity screening assay. Ten catalytically enhanced natural variants were identified, produced, purified, and tested for efficacy using industrially-relevant conditions and substrates. Three key amino acids in CBHs with performance comparable or superior to Penicillium funiculosum Cel7A were identified and combinatorically engineered into P. funiculosum cel7a, expressed in T. reesei, and assayed on lignocellulosic biomass. The top performer generated using this combined approach of natural diversity genome mining, experimental assays, and computational modeling produced a 41% increase in conversion extent over native P. funiculosum Cel7A, a 55% increase over the current industrial standard T. reesei Cel7A, and 10% improvement over Aspergillus oryzae Cel7C, the best natural GH7 CBH previously identified in our laboratory.

View Journal Article

Cite

Export

Save

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE; USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 2301755

Alternate ID(s):: OSTI ID: 2316135

Report Number(s):: NREL/JA-2800-87398; S002192582400125X; 105749; PII: S002192582400125X

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 300 Journal Issue: 3; ISSN 0021-9258

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

References (71)

Clustal Omega for making accurate alignments of many protein sequences: Clustal Omega for Many Protein Sequences Sievers, Fabian; Higgins, Desmond G. Protein Science, Vol. 27, Issue 1 https://doi.org/10.1002/pro.3290	journal	October 2017
Trichoderma reesei has no true exo-cellulase: all intact and truncated cellulases produce new reducing end groups on cellulose Ståhlberg, Jerry; Johansson, Gunnar; Pettersson, Göran Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1157, Issue 1 https://doi.org/10.1016/0304-4165(93)90085-M	journal	May 1993
Fungal Cellulases Payne, Christina M.; Knott, Brandon C.; Mayes, Heather B. Chemical Reviews, Vol. 115, Issue 3 https://doi.org/10.1021/cr500351c	journal	January 2015
Binding Site Dynamics and Aromatic–Carbohydrate Interactions in Processive and Non-Processive Family 7 Glycoside Hydrolases Taylor, Courtney B.; Payne, Christina M.; Himmel, Michael E. The Journal of Physical Chemistry B, Vol. 117, Issue 17 https://doi.org/10.1021/jp401410h	journal	April 2013
Kinetics of Cellobiohydrolase (Cel7A) Variants with Lowered Substrate Affinity Kari, Jeppe; Olsen, Johan; Borch, Kim Journal of Biological Chemistry, Vol. 289, Issue 47 https://doi.org/10.1074/jbc.M114.604264	journal	September 2014
High-resolution crystal structures reveal how a cellulose chain is bound in the 50 Å long tunnel of cellobiohydrolase I from Trichoderma reesei Divne, Christina; Ståhlberg, Jerry; Teeri, Tuula T. Journal of Molecular Biology, Vol. 275, Issue 2, p. 309-325 https://doi.org/10.1006/jmbi.1997.1437	journal	January 1998
Machine learning reveals sequence-function relationships in family 7 glycoside hydrolases Gado, Japheth E.; Harrison, Brent E.; Sandgren, Mats Journal of Biological Chemistry, Vol. 297, Issue 2 https://doi.org/10.1016/j.jbc.2021.100931	journal	August 2021
Heterologous protein expression in Hypocrea jecorina: A historical perspective and new developments Singh, Arjun; Taylor, Larry E.; Vander Wall, Todd A. Biotechnology Advances, Vol. 33, Issue 1 https://doi.org/10.1016/j.biotechadv.2014.11.009	journal	January 2015
Comparative insights into the saccharification potentials of a relatively unexplored but robust Penicillium funiculosum glycoside hydrolase 7 cellobiohydrolase Ogunmolu, Funso Emmanuel; Jagadeesha, Navya Bhatt Kammachi; Kumar, Rakesh Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0752-x	journal	March 2017
Effect of Single Active-Site Cleft Mutation on Product Specificity in a Thermostable Bacterial Cellulase Rignall, Tauna R.; Baker, John O.; McCarter, Suzanne L. Applied Biochemistry and Biotechnology, Vol. 98-100, Issue 1-9 https://doi.org/10.1385/ABAB:98-100:1-9:383	journal	January 2002
A Transition from Cellulose Swelling to Cellulose Dissolution by o -Phosphoric Acid: Evidence from Enzymatic Hydrolysis and Supramolecular Structure Zhang, Y. -H. Percival; Cui, Jingbiao; Lynd, Lee R. Biomacromolecules, Vol. 7, Issue 2 https://doi.org/10.1021/bm050799c	journal	February 2006
TtCel7A: A Native Thermophilic Bifunctional Cellulose/Xylanase Exogluclanase from the Thermophilic Biomass-Degrading Fungus Thielavia terrestris Co3Bag1, and Its Application in Enzymatic Hydrolysis of Agroindustrial Derivatives López-López, Azucena; Santiago-Hernández, Alejandro; Cayetano-Cruz, Maribel Journal of Fungi, Vol. 9, Issue 2 https://doi.org/10.3390/jof9020152	journal	January 2023
An update on enzymatic cocktails for lignocellulose breakdown Lopes, A. M.; Ferreira Filho, E. X.; Moreira, L. R. S. Journal of Applied Microbiology, Vol. 125, Issue 3 https://doi.org/10.1111/jam.13923	journal	July 2018
A versatile toolkit for high throughput functional genomics with Trichoderma reesei Schuster, Andre; Bruno, Kenneth S.; Collett, James R. Biotechnology for Biofuels, Vol. 5, Issue 1 https://doi.org/10.1186/1754-6834-5-1	journal	January 2012
Hypocrea jecorina CEL6A protein engineering Lantz, Suzanne E.; Goedegebuur, Frits; Hommes, Ronald Biotechnology for Biofuels, Vol. 3, Issue 1, Article No. 20 https://doi.org/10.1186/1754-6834-3-20	journal	September 2010
The Mechanism of Cellulose Hydrolysis by a Two-Step, Retaining Cellobiohydrolase Elucidated by Structural and Transition Path Sampling Studies Knott, Brandon C.; Haddad Momeni, Majid; Crowley, Michael F. Journal of the American Chemical Society, Vol. 136, Issue 1 https://doi.org/10.1021/ja410291u	journal	December 2013
SignalP 4.0: discriminating signal peptides from transmembrane regions Petersen, Thomas Nordahl; Brunak, Søren; von Heijne, Gunnar Nature Methods, Vol. 8, Issue 10 https://doi.org/10.1038/nmeth.1701	journal	September 2011
Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production Himmel, M. E.; Ding, S.-Y.; Johnson, D. K. Science, Vol. 315, Issue 5813, p. 804-807 https://doi.org/10.1126/science.1137016	journal	February 2007
Improving the thermal stability of cellobiohydrolase Cel7A from Hypocrea jecorina by directed evolution Goedegebuur, Frits; Dankmeyer, Lydia; Gualfetti, Peter Journal of Biological Chemistry, Vol. 292, Issue 42 https://doi.org/10.1074/jbc.M117.803270	journal	August 2017
Hypocrea jecorina Cellobiohydrolase I Stabilizing Mutations Identified Using Noncontiguous Recombination Smith, Matthew A.; Bedbrook, Claire N.; Wu, Timothy ACS Synthetic Biology, Vol. 2, Issue 12 https://doi.org/10.1021/sb400010m	journal	June 2013
HMMER web server: 2018 update Potter, Simon C.; Luciani, Aurélien; Eddy, Sean R. Nucleic Acids Research, Vol. 46, Issue W1 https://doi.org/10.1093/nar/gky448	journal	June 2018
ESPript: analysis of multiple sequence alignments in PostScript Gouet, P.; Courcelle, E.; Stuart, D. Bioinformatics, Vol. 15, Issue 4 https://doi.org/10.1093/bioinformatics/15.4.305	journal	April 1999
Highly Thermostable Xylanase of the Thermophilic Fungus Talaromyces thermophilus: Purification and Characterization Maalej, Ines; Belhaj, Ines; Masmoudi, Najla Fourati Applied Biochemistry and Biotechnology, Vol. 158, Issue 1 https://doi.org/10.1007/s12010-008-8317-x	journal	July 2008
Virtual Bioprospecting of Interfacial Enzymes: Relating Sequence and Kinetics Schaller, Kay S.; Molina, Gustavo Avelar; Kari, Jeppe ACS Catalysis, Vol. 12, Issue 12 https://doi.org/10.1021/acscatal.2c02305	journal	June 2022
Sabatier Principle for Interfacial (Heterogeneous) Enzyme Catalysis Kari, Jeppe; Olsen, Johan P.; Jensen, Kenneth ACS Catalysis, Vol. 8, Issue 12 https://doi.org/10.1021/acscatal.8b03547	journal	October 2018
Transient Kinetics and Rate-Limiting Steps for the Processive Cellobiohydrolase Cel7A: Effects of Substrate Structure and Carbohydrate Binding Domain Cruys-Bagger, Nicolaj; Tatsumi, Hirosuke; Ren, Guilin Robin Biochemistry, Vol. 52, Issue 49 https://doi.org/10.1021/bi401210n	journal	November 2013
Cellulase Linkers Are Optimized Based on Domain Type and Function: Insights from Sequence Analysis, Biophysical Measurements, and Molecular Simulation Sammond, Deanne W.; Payne, Christina M.; Brunecky, Roman PLoS ONE, Vol. 7, Issue 11 https://doi.org/10.1371/journal.pone.0048615	journal	November 2012
The relationship between thermal stability and pH optimum studied with wild-type and mutant Trichoderma reesei cellobiohydrolase Cel7A: Stability of cellobiohydrolase Cel7A Boer, Harry; Koivula, Anu European Journal of Biochemistry, Vol. 270, Issue 5 https://doi.org/10.1046/j.1432-1033.2003.03431.x	journal	March 2003
Status of filamentous fungi in integrated biorefineries Troiano, D.; Orsat, V.; Dumont, M. J. Renewable and Sustainable Energy Reviews, Vol. 117 https://doi.org/10.1016/j.rser.2019.109472	journal	January 2020
Fungal cellulases: protein engineering and post-translational modifications Zhang, Ruiqin; Cao, Chenghao; Bi, Jiahua Applied Microbiology and Biotechnology, Vol. 106, Issue 1 https://doi.org/10.1007/s00253-021-11723-y	journal	December 2021
A constitutive expression system for glycosyl hydrolase family 7 cellobiohydrolases in Hypocrea jecorina Linger, Jeffrey G.; Taylor, Larry E.; Baker, John O. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0230-2	journal	March 2015
Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast Alahuhta, Markus; Xu, Qi; Knoshaug, Eric P. Biotechnology for Biofuels, Vol. 14, Issue 1 https://doi.org/10.1186/s13068-020-01856-z	journal	January 2021
The role of carbohydrate binding module (CBM) at high substrate consistency: Comparison of Trichoderma reesei and Thermoascus aurantiacus Cel7A (CBHI) and Cel5A (EGII) Le Costaouëc, Tinaïg; Pakarinen, Annukka; Várnai, Anikó Bioresource Technology, Vol. 143 https://doi.org/10.1016/j.biortech.2013.05.079	journal	September 2013
Neutral and alkaline cellulases: Production, engineering, and applications Ben Hmad, Ines; Gargouri, Ali Journal of Basic Microbiology, Vol. 57, Issue 8 https://doi.org/10.1002/jobm.201700111	journal	May 2017
Engineering enhanced cellobiohydrolase activity Taylor, Larry E.; Knott, Brandon C.; Baker, John O. Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-03501-8	journal	March 2018
Physical constraints and functional plasticity of cellulases Kari, Jeppe; Molina, Gustavo A.; Schaller, Kay S. Nature Communications, Vol. 12, Issue 1 https://doi.org/10.1038/s41467-021-24075-y	journal	June 2021
A versatile 2A peptide-based bicistronic protein expressing platform for the industrial cellulase producing fungus, Trichoderma reesei Subramanian, Venkataramanan; Schuster, Logan A.; Moore, Kyle T. Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0710-7	journal	February 2017
Cloning, expression, and characterization of novel thermostable family 7 cellobiohydrolases Voutilainen, Sanni P.; Puranen, Terhi; Siika-aho, Matti Biotechnology and Bioengineering, Vol. 101, Issue 3, p. 515-528 https://doi.org/10.1002/bit.21940	journal	October 2008
The dissociation mechanism of processive cellulases Vermaas, Josh V.; Kont, Riin; Beckham, Gregg T. Proceedings of the National Academy of Sciences, Vol. 116, Issue 46 https://doi.org/10.1073/pnas.1913398116	journal	October 2019
Deciphering key features in protein structures with the new ENDscript server Robert, Xavier; Gouet, Patrice Nucleic Acids Research, Vol. 42, Issue W1 https://doi.org/10.1093/nar/gku316	journal	April 2014
Probing Substrate Interactions in the Active Tunnel of a Catalytically Deficient Cellobiohydrolase (Cel7) Colussi, Francieli; Sørensen, Trine H.; Alasepp, Kadri Journal of Biological Chemistry, Vol. 290, Issue 4 https://doi.org/10.1074/jbc.M114.624163	journal	January 2015
Expression of Talaromyces emersonii cellobiohydrolase Cel7A in Saccharomyces cerevisiae and rational mutagenesis to improve its thermostability and activity Voutilainen, S. P.; Murray, P. G.; Tuohy, M. G. Protein Engineering Design and Selection, Vol. 23, Issue 2 https://doi.org/10.1093/protein/gzp072	journal	December 2009
Compositional Analysis of Lignocellulosic Feedstocks. 1. Review and Description of Methods Sluiter, Justin B.; Ruiz, Raymond O.; Scarlata, Christopher J. Journal of Agricultural and Food Chemistry, Vol. 58, Issue 16, p. 9043-9053 https://doi.org/10.1021/jf1008023	journal	August 2010
Systematic deletions in the cellobiohydrolase (CBH) Cel7A from the fungus Trichoderma reesei reveal flexible loops critical for CBH activity Schiano-di-Cola, Corinna; Røjel, Nanna; Jensen, Kenneth Journal of Biological Chemistry, Vol. 294, Issue 6 https://doi.org/10.1074/jbc.RA118.006699	journal	December 2018
Catalytically Enhanced Endocellulase Cel5A from <I>Acidothermus cellulolyticus</I> Baker, John O.; McCarley, James R.; Lovett, Rebecca Applied Biochemistry and Biotechnology, Vol. 121, Issue 1-3 https://doi.org/10.1385/ABAB:121:1-3:0129	journal	January 2005
Improved ethanol yield and reduced Minimum Ethanol Selling Price (MESP) by modifying low severity dilute acid pretreatment with deacetylation and mechanical refining: 1) Experimental Chen, Xiaowen; Tao, Ling; Shekiro, Joseph Biotechnology for Biofuels, Vol. 5, Issue 1 https://doi.org/10.1186/1754-6834-5-60	journal	January 2012
Costs and benefits of processivity in enzymatic degradation of recalcitrant polysaccharides Horn, S. J.; Sikorski, P.; Cederkvist, J. B. Proceedings of the National Academy of Sciences, Vol. 103, Issue 48 https://doi.org/10.1073/pnas.0608909103	journal	November 2006
DMR (deacetylation and mechanical refining) processing of corn stover achieves high monomeric sugar concentrations (230 g L ⁻¹ ) during enzymatic hydrolysis and high ethanol concentrations (>10% v/v) during fermentation without hydrolysate purification or concentration Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W. Energy & Environmental Science, Vol. 9, Issue 4 https://doi.org/10.1039/C5EE03718B	journal	January 2016
Loop variants of the thermophile Rasamsonia emersonii Cel7A with improved activity against cellulose : Loop Variants of the Thermophile Sørensen, Trine Holst; Windahl, Michael Skovbo; McBrayer, Brett Biotechnology and Bioengineering, Vol. 114, Issue 1 https://doi.org/10.1002/bit.26050	journal	September 2016
The carbohydrate-active enzyme database: functions and literature Drula, Elodie; Garron, Marie-Line; Dogan, Suzan Nucleic Acids Research, Vol. 50, Issue D1 https://doi.org/10.1093/nar/gkab1045	journal	November 2021
Carbohydrate–Protein Interactions That Drive Processive Polysaccharide Translocation in Enzymes Revealed from a Computational Study of Cellobiohydrolase Processivity Knott, Brandon C.; Crowley, Michael F.; Himmel, Michael E. Journal of the American Chemical Society, Vol. 136, Issue 24 https://doi.org/10.1021/ja504074g	journal	June 2014
Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega Sievers, Fabian; Wilm, Andreas; Dineen, David Molecular Systems Biology, Vol. 7, Issue 1 https://doi.org/10.1038/msb.2011.75	journal	January 2011
A family of thermostable fungal cellulases created by structure-guided recombination Heinzelman, P.; Snow, C. D.; Wu, I. Proceedings of the National Academy of Sciences, Vol. 106, Issue 14, p. 5610-5615 https://doi.org/10.1073/pnas.0901417106	journal	March 2009
Rate of Threading a Cellulose Chain into the Binding Tunnel of a Cellulase Cruys-Bagger, Nicolaj; Alasepp, Kadri; Andersen, Morten The Journal of Physical Chemistry B, Vol. 120, Issue 25 https://doi.org/10.1021/acs.jpcb.6b01877	journal	June 2016
SWISS-MODEL: homology modelling of protein structures and complexes Waterhouse, Andrew; Bertoni, Martino; Bienert, Stefan Nucleic Acids Research, Vol. 46, Issue W1 https://doi.org/10.1093/nar/gky427	journal	May 2018
Endo-exo Synergism in Cellulose Hydrolysis Revisited Jalak, Jürgen; Kurašin, Mihhail; Teugjas, Hele Journal of Biological Chemistry, Vol. 287, Issue 34 https://doi.org/10.1074/jbc.M112.381624	journal	June 2012
Nomenclature for sugar-binding subsites in glycosyl hydrolases Davies, Gideon J.; Wilson, Keith S.; Henrissat, Bernard Biochemical Journal, Vol. 321, Issue 2 https://doi.org/10.1042/bj3210557	journal	January 1997
Engineering of a glycosidase Family 7 cellobiohydrolase to more alkaline pH optimum: the pH behaviour of Trichoderma reesei Cel7A and its E223S/ A224H/L225V/T226A/D262G mutant Becker, Dieter; Braet, Christophe; Brumer, Harry Biochemical Journal, Vol. 356, Issue 1, p. 19-30 https://doi.org/10.1042/bj3560019	journal	May 2001
Rational design and structure insights for thermostability improvement of Penicillium verruculosum Cel7A cellobiohydrolase Dotsenko, Anna S.; Dotsenko, Gleb S.; Rozhkova, Aleksandra M. Biochimie, Vol. 176 https://doi.org/10.1016/j.biochi.2020.06.007	journal	September 2020
Trichoderma reesei RUT-C30 - thirty years of strain improvement Peterson, R.; Nevalainen, H. Microbiology, Vol. 158, Issue 1, p. 58-68 https://doi.org/10.1099/mic.0.054031-0	journal	October 2011
The Sabatier principle as a tool for discovery and engineering of industrial enzymes Kari, Jeppe; Schaller, Kay; Molina, Gustavo A. Current Opinion in Biotechnology, Vol. 78 https://doi.org/10.1016/j.copbio.2022.102843	journal	December 2022
Pre-steady-state Kinetics for Hydrolysis of Insoluble Cellulose by Cellobiohydrolase Cel7A Cruys-Bagger, Nicolaj; Elmerdahl, Jens; Praestgaard, Eigil Journal of Biological Chemistry, Vol. 287, Issue 22 https://doi.org/10.1074/jbc.M111.334946	journal	April 2012
Improving Sugar Yields and Reducing Enzyme Loadings in the Deacetylation and Mechanical Refining (DMR) Process through Multistage Disk and Szego Refining and Corresponding Techno-Economic Analysis Chen, Xiaowen; Wang, Wei; Ciesielski, Peter ACS Sustainable Chemistry & Engineering, Vol. 4, Issue 1 https://doi.org/10.1021/acssuschemeng.5b01242	journal	December 2015
Removal of N-linked glycans in cellobiohydrolase Cel7A from Trichoderma reesei reveals higher activity and binding affinity on crystalline cellulose Kołaczkowski, Bartłomiej M.; Schaller, Kay S.; Sørensen, Trine Holst Biotechnology for Biofuels, Vol. 13, Issue 1 https://doi.org/10.1186/s13068-020-01779-9	journal	August 2020
Construction of thermostable cellobiohydrolase I from the fungus Talaromyces cellulolyticus by protein engineering Nakabayashi, Makoto; Kamachi, Saori; Malle, Dominggus Protein Engineering, Design and Selection, Vol. 32, Issue 1 https://doi.org/10.1093/protein/gzz001	journal	January 2019
Developments and opportunities in fungal strain engineering for the production of novel enzymes and enzyme cocktails for plant biomass degradation Kun, Roland S.; Gomes, Ana Carolina S.; Hildén, Kristiina S. Biotechnology Advances, Vol. 37, Issue 6 https://doi.org/10.1016/j.biotechadv.2019.02.017	journal	November 2019
Processivity of Cellobiohydrolases Is Limited by the Substrate Kurašin, Mihhail; Väljamäe, Priit Journal of Biological Chemistry, Vol. 286, Issue 1 https://doi.org/10.1074/jbc.M110.161059	journal	November 2010
Computing Cellulase Kinetics with a Two-Domain Linear Interaction Energy Approach Schaller, Kay S.; Kari, Jeppe; Molina, Gustavo A. ACS Omega, Vol. 6, Issue 2 https://doi.org/10.1021/acsomega.0c05361	journal	January 2021
Engineered thermostable fungal Cel6A and Cel7A cellobiohydrolases hydrolyze cellulose efficiently at elevated temperatures Wu, Indira; Arnold, Frances H. Biotechnology and Bioengineering, Vol. 110, Issue 7 https://doi.org/10.1002/bit.24864	journal	March 2013
Traffic Jams Reduce Hydrolytic Efficiency of Cellulase on Cellulose Surface Igarashi, K.; Uchihashi, T.; Koivula, A. Science, Vol. 333, Issue 6047 https://doi.org/10.1126/science.1208386	journal	September 2011
Distinct roles of N- and O-glycans in cellulase activity and stability Amore, Antonella; Knott, Brandon C.; Supekar, Nitin T. Proceedings of the National Academy of Sciences, Vol. 114, Issue 52 https://doi.org/10.1073/pnas.1714249114	journal	December 2017

Similar Records

Engineering enhanced cellobiohydrolase activity

Journal Article · Thu Mar 22 00:00:00 EDT 2018 · Nature Communications · OSTI ID:2301755

Taylor, Larry E.; Knott, Brandon C.; Baker, John O.; +16 more

Probing the Role of N-Linked Glycans in the Stability and Activity of Fungal Cellobiohydrolases by Mutational Analysis

Journal Article · Thu Jan 01 00:00:00 EST 2009 · Cellulose · OSTI ID:2301755

Adney, W S; Jeoh, T; Beckham, G T; +5 more

Biochemical and structural characterizations of two Dictyostelium cellobiohydrolases from the amoebozoa kingdom reveal a high level of conservation between distant phylogenetic trees of life

Journal Article · Fri Apr 01 00:00:00 EDT 2016 · Applied and Environmental Microbiology · OSTI ID:2301755

Hobdey, Sarah E.; Knott, Brandon C.; Momeni, Majid Haddad; +8 more

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
cellobiohydrolase
cellulase
cellulose
enzyme improvement
genomics
protein engineering
rational design

Title: Engineering of glycoside hydrolase family 7 cellobiohydrolases directed by natural diversity screening

Citation Formats

References (71)

Similar Records

Related Subjects