Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Strategies to reduce end-product inhibition in family 48 glycoside hydrolases

Journal Article · · Proteins
DOI:https://doi.org/10.1002/prot.24965· OSTI ID:1240085
 [1];  [2];  [2];  [2];  [2];  [2];  [1];  [2];  [2];  [2]
  1. Cornell Univ., Ithaca, NY (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States)
+ Show Author Affiliations
Family 48 cellobiohydrolases are some of the most abundant glycoside hydrolases in nature. They are able to degrade cellulosic biomass and therefore serve as good enzyme candidates for biofuel production. Family 48 cellulases hydrolyze cellulose chains via a processive mechanism, and produce end products composed primarily of cellobiose as well as other cellooligomers (dp ≤ 4). The challenge of utilizing cellulases in biofuel production lies in their extremely slow turnover rate. A factor contributing to the low enzyme activity is suggested to be product binding to enzyme and the resulting performance inhibition. In this study, we quantitatively evaluated the product inhibitory effect of four family 48 glycoside hydrolases using molecular dynamics simulations and product expulsion free-energy calculations. We also suggested a series of single mutants of the four family 48 glycoside hydrolases with theoretically reduced level of product inhibition. As a result, the theoretical calculations provide a guide for future experimental studies designed to produce mutant cellulases with enhanced activity.
Research Organization:
NREL (National Renewable Energy Laboratory (NREL), Golden, CO (United States))
Sponsoring Organization:
USDOE Office of Science (SC)
Grant/Contract Number:
AC36-08GO28308
OSTI ID:
1240085
Report Number(s):
NREL/JA--2700-65427
Journal Information:
Proteins, Journal Name: Proteins Journal Issue: 3 Vol. 84; ISSN 0887-3585
Publisher:
WileyCopyright Statement
Country of Publication:
United States
Language:
English

References (37)

Alleviating product inhibition in cellulase enzyme Cel7A: Alleviating Product Inhibition in Cel7A journal September 2015
Extending the treatment of backbone energetics in protein force fields: Limitations of gas-phase quantum mechanics in reproducing protein conformational distributions in molecular dynamics simulations journal August 2004
Additive empirical force field for hexopyranose monosaccharides journal November 2008
CHARMM: The biomolecular simulation program journal July 2009
CHARMM: A program for macromolecular energy, minimization, and dynamics calculations journal July 1983
CHAMBER: Comprehensive support for CHARMM force fields within the AMBER software journal December 2009
Product inhibition of the recombinant CelS, an exoglucanase component of the Clostridium thermocellum cellulosome journal December 1995
Free Energy Calculations with Non-Equilibrium Methods: Applications of the Jarzynski Relationship journal January 2006
The noncellulosomal family 48 cellobiohydrolase from Clostridium phytofermentans ISDg: heterologous expression, characterization, and processivity journal October 2009
Major characteristics of the cellulolytic system of Clostridium thermocellum coincide with those of the purified cellulosome journal January 1985
The Crystal Structure and Catalytic Mechanism of Cellobiohydrolase CelS, the Major Enzymatic Component of the Clostridium thermocellum Cellulosome journal July 2002
Structures of Mutants of Cellulase Cel48F of Clostridium cellulolyticum in Complex with Long Hemithiocellooligosaccharides Give Rise to a New View of the Substrate Pathway during Processive Action journal January 2008
Crystal Structures of the Cellulase Cel48F in Complex with Inhibitors and Substrates Give Insights into Its Processive Action † , ‡ journal September 2000
Identification of Two Functionally Different Classes of Exocellulases † journal January 1996
Ricin Toxin Contains at Least Three Galactose-Binding Sites Located in B Chain Subdomains 1α, 1β, and 2γ † journal January 1996
Catalytic mechanism of enzymic glycosyl transfer journal November 1990
All-Atom Empirical Potential for Molecular Modeling and Dynamics Studies of Proteins † journal April 1998
Updating the sequence-based classification of glycosyl hydrolases journal June 1996
Cloning, expression and characterization of a Family 48 exocellulase, Cel48A, from Thermobifida fusca: Thermobifida fusca exocellulase Cel48A journal August 2000
A “fast growth” method of computing free energy differences journal April 2001
Comparison of simple potential functions for simulating liquid water journal July 1983
Deletion of the Cel48S cellulase from Clostridium thermocellum journal September 2010
Probing Carbohydrate Product Expulsion from a Processive Cellulase with Multiple Absolute Binding Free Energy Methods journal March 2011
Product Binding Varies Dramatically between Processive and Nonprocessive Cellulase Enzymes journal May 2012
The crystal structure of the processive endocellulase CelF of Clostridium cellulolyticum in complex with a thiooligosaccharide inhibitor at 2.0Aresolution journal October 1998
Equilibrium free-energy differences from nonequilibrium measurements: A master-equation approach journal November 1997
Nonequilibrium Equality for Free Energy Differences journal April 1997
Equilibrium Information from Nonequilibrium Measurements in an Experimental Test of Jarzynski's Equality journal June 2002
Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production journal February 2007
Revealing Nature's Cellulase Diversity The Digestion Mechanism of Caldicellulosiruptor bescii CelA journal December 2013
Isolation and properties of a major cellobiohydrolase from the cellulosome of Clostridium thermocellum. journal July 1991
The processive endocellulase CelF, a major component of the Clostridium cellulolyticum cellulosome: purification and characterization of the recombinant form. journal January 1997
The Cellulosomes: Multienzyme Machines for Degradation of Plant Cell Wall Polysaccharides journal October 2004
High temperature pre-digestion of corn stover biomass for improved product yields journal December 2014
An Introduction to the Bootstrap book May 1994
An Improved Strategy for Generating Forces in Steered Molecular Dynamics: The Mechanical Unfolding of Titin, e2lip3 and Ubiquitin journal September 2010
Catalytically Enhanced Endocellulase Cel5A from Acidothermus cellulolyticus journal January 2005

Cited By (2)

Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure journal November 2017
Determination of the native features of the exoglucanase Cel48S from Clostridium thermocellum journal January 2018

Figures / Tables (9)

Table I(p. 25)table Table I
Table II(p. 26)table Table II
Table III(p. 27)table Table III
Figure 1(p. 28)figure Figure 1
Figure 2(p. 29)figure Figure 2
Figure 3(p. 30)figure Figure 3
Figure 4(p. 31)figure Figure 4
Figure 5(p. 32)figure Figure 5
Figure 6(p. 33)figure Figure 6

Similar Records

Comparative Community Proteomics Demonstrates the Unexpected Importance of Actinobacterial Glycoside Hydrolase Family 12 Protein for Crystalline Cellulose Hydrolysis
Journal Article · Tue Aug 23 00:00:00 EDT 2016 · mBio (Online) · OSTI ID:1340800
Product Binding Varies Dramatically between Processive and Nonprocessive Cellulase Enzymes
Journal Article · Fri Jul 13 00:00:00 EDT 2012 · Journal of Biological Chemistry · OSTI ID:1050115

Related Subjects

09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
biofuels
cellulose
glycoside hydrolases
molecular dynamics
product inhibition