Dynamic changes of substrate reactivity and enzyme adsorption on partially hydrolyzed cellulose

Jian Shi; Dong Wu; Libing Zhang; Blake A. Simmons; Seema Singh; Bin Yang; Charles E. Wyman

doi:10.1002/bit.26180

Dynamic changes of substrate reactivity and enzyme adsorption on partially hydrolyzed cellulose

Jian Shi, Dong Wu, Libing Zhang, Blake A. Simmons, Seema Singh, Bin Yang, Charles E. Wyman

Biosystems and Agricultural Engineering

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

The enzymatic hydrolysis of cellulose is a thermodynamically challenging catalytic process that is influenced by both substrate-related and enzyme-related factors. In this study, a proteolysis approach was applied to recover and clean the partially converted cellulose at the different stages of enzymatic hydrolysis to monitor the hydrolysis rate as a function of substrate reactivity/accessibility and investigate surface characteristics of the partially converted cellulose. Enzyme-substrate interactions between individual key cellulase components from wild-type Trichoderma reesei and partially converted cellulose were followed and correlated to the enzyme adsorption capacity and dynamic sugar release. Results suggest that cellobiohydrolase CBH1 (Cel7A) and endoglucanases EG2 (Cel5A) adsorption capacities decreased as cellulose was progressively hydrolyzed, likely due to the “depletion” of binding sites. Furthermore, the degree of synergism between CBH1 and EG2 varied depending on the enzyme loading and the substrates. The results provide a better understanding of the relationship between dynamic change of substrate features and the functionality of various cellulase components during enzymatic hydrolysis. Biotechnol. Bioeng. 2017;114: 503–515.

Original language	English
Pages (from-to)	503-515
Number of pages	13
Journal	Biotechnology and Bioengineering
Volume	114
Issue number	3
DOIs	https://doi.org/10.1002/bit.26180
State	Published - Mar 1 2017

Bibliographical note

Funding Information:
Research conducted by the BioEnergy Science Center (BESC) and Joint BioEnergy Institute (JBEI) was supported by Office of Biological and Environmental in the Office of Science of the U.S. Department of Energy under Contract Numbers DE-PS02-06ER64304 and DE-AC02-05CH11231, respectively. We also acknowledge the Center for Environmental Research and Technology (CE-CERT) of the Bourns College of Engineering for providing the facilities and the Ford Motor Company for funding the Chair in Environmental Engineering that facilitates projects such as this one. Work at WSU was partially funded by DARPA Young Faculty Award contract # N66001-11-1-414, and the authors acknowledge the Bioproducts, Sciences and Engineering Laboratory and Department of Biosystems Engineering at Washington State University for providing facilities and equipment to perform our research. We acknowledge the National Science Foundation under Cooperative Agreement No. 1355438 for partial support of the effort at University of Kentucky. We acknowledge Xiaodi Gao for assisting enzyme adsorption experiments. We thank Drs. Edmund Larenas and Colin Mitchinson from Genencor, a Danisco Division (now DuPont Industrial Biosciences) for generously providing purified CBH1 and EG2 enzymes and offering valuable suggestions to this research.

Publisher Copyright:
© 2016 Wiley Periodicals, Inc.

Keywords

cellulase
cellulose
enzyme–substrate interactions
proteolysis
reactivity

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1002/bit.26180

Cite this

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title = "Dynamic changes of substrate reactivity and enzyme adsorption on partially hydrolyzed cellulose",

abstract = "The enzymatic hydrolysis of cellulose is a thermodynamically challenging catalytic process that is influenced by both substrate-related and enzyme-related factors. In this study, a proteolysis approach was applied to recover and clean the partially converted cellulose at the different stages of enzymatic hydrolysis to monitor the hydrolysis rate as a function of substrate reactivity/accessibility and investigate surface characteristics of the partially converted cellulose. Enzyme-substrate interactions between individual key cellulase components from wild-type Trichoderma reesei and partially converted cellulose were followed and correlated to the enzyme adsorption capacity and dynamic sugar release. Results suggest that cellobiohydrolase CBH1 (Cel7A) and endoglucanases EG2 (Cel5A) adsorption capacities decreased as cellulose was progressively hydrolyzed, likely due to the “depletion” of binding sites. Furthermore, the degree of synergism between CBH1 and EG2 varied depending on the enzyme loading and the substrates. The results provide a better understanding of the relationship between dynamic change of substrate features and the functionality of various cellulase components during enzymatic hydrolysis. Biotechnol. Bioeng. 2017;114: 503–515.",

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author = "Jian Shi and Dong Wu and Libing Zhang and Simmons, {Blake A.} and Seema Singh and Bin Yang and Wyman, {Charles E.}",

note = "Funding Information: Research conducted by the BioEnergy Science Center (BESC) and Joint BioEnergy Institute (JBEI) was supported by Office of Biological and Environmental in the Office of Science of the U.S. Department of Energy under Contract Numbers DE-PS02-06ER64304 and DE-AC02-05CH11231, respectively. We also acknowledge the Center for Environmental Research and Technology (CE-CERT) of the Bourns College of Engineering for providing the facilities and the Ford Motor Company for funding the Chair in Environmental Engineering that facilitates projects such as this one. Work at WSU was partially funded by DARPA Young Faculty Award contract # N66001-11-1-414, and the authors acknowledge the Bioproducts, Sciences and Engineering Laboratory and Department of Biosystems Engineering at Washington State University for providing facilities and equipment to perform our research. We acknowledge the National Science Foundation under Cooperative Agreement No. 1355438 for partial support of the effort at University of Kentucky. We acknowledge Xiaodi Gao for assisting enzyme adsorption experiments. We thank Drs. Edmund Larenas and Colin Mitchinson from Genencor, a Danisco Division (now DuPont Industrial Biosciences) for generously providing purified CBH1 and EG2 enzymes and offering valuable suggestions to this research. Publisher Copyright: {\textcopyright} 2016 Wiley Periodicals, Inc.",

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AU - Singh, Seema

AU - Yang, Bin

AU - Wyman, Charles E.

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N2 - The enzymatic hydrolysis of cellulose is a thermodynamically challenging catalytic process that is influenced by both substrate-related and enzyme-related factors. In this study, a proteolysis approach was applied to recover and clean the partially converted cellulose at the different stages of enzymatic hydrolysis to monitor the hydrolysis rate as a function of substrate reactivity/accessibility and investigate surface characteristics of the partially converted cellulose. Enzyme-substrate interactions between individual key cellulase components from wild-type Trichoderma reesei and partially converted cellulose were followed and correlated to the enzyme adsorption capacity and dynamic sugar release. Results suggest that cellobiohydrolase CBH1 (Cel7A) and endoglucanases EG2 (Cel5A) adsorption capacities decreased as cellulose was progressively hydrolyzed, likely due to the “depletion” of binding sites. Furthermore, the degree of synergism between CBH1 and EG2 varied depending on the enzyme loading and the substrates. The results provide a better understanding of the relationship between dynamic change of substrate features and the functionality of various cellulase components during enzymatic hydrolysis. Biotechnol. Bioeng. 2017;114: 503–515.

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Dynamic changes of substrate reactivity and enzyme adsorption on partially hydrolyzed cellulose

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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