The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies

Brandon C. Knott, Majid Haddad Momeni, Michael F. Crowley, Lloyd F. MacKenzie, Andreas W. Götz, Mats Sandgren, Stephen G. Withers, Jerry Ståhlberg, Gregg T. Beckham

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 
Original languageEnglish
JournalJournal of the American Chemical Society
Volume136
Issue number1
Pages (from-to)321-329
Number of pages9
ISSN0002-7863
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • Chemistry (all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry
  • Cellobiohydrolase Cel7A
  • Cellulose hydrolysis
  • Conformational change
  • Glycoside hydrolases
  • Glycosyl-enzyme intermediate
  • Glycosylation reactions
  • Hydrolytic reactions
  • Transition path sampling
  • Esterification
  • Glycosylation
  • Enzymes
  • Cellulose
  • Cellulose 1,4 beta cellobiosidase
  • Conformational transition
  • Deglycosylation
  • Enzyme active site
  • Enzyme structure
  • Hydrolysis
  • Hypocrea jecorina
  • Sampling

Cite this

Knott, Brandon C. ; Momeni, Majid Haddad ; Crowley, Michael F. ; MacKenzie, Lloyd F. ; Götz, Andreas W. ; Sandgren, Mats ; Withers, Stephen G. ; Ståhlberg, Jerry ; Beckham, Gregg T. / The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. In: Journal of the American Chemical Society. 2014 ; Vol. 136, No. 1. pp. 321-329.
@article{5defcd6a980745d6b7f052888483ee84,
title = "The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies",
abstract = "Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. ",
keywords = "Chemistry (all), Catalysis, Biochemistry, Colloid and Surface Chemistry, Cellobiohydrolase Cel7A, Cellulose hydrolysis, Conformational change, Glycoside hydrolases, Glycosyl-enzyme intermediate, Glycosylation reactions, Hydrolytic reactions, Transition path sampling, Esterification, Glycosylation, Enzymes, Cellulose, Cellulose 1,4 beta cellobiosidase, Conformational transition, Deglycosylation, Enzyme active site, Enzyme structure, Hydrolysis, Hypocrea jecorina, Sampling",
author = "Knott, {Brandon C.} and Momeni, {Majid Haddad} and Crowley, {Michael F.} and MacKenzie, {Lloyd F.} and G{\"o}tz, {Andreas W.} and Mats Sandgren and Withers, {Stephen G.} and Jerry St{\aa}hlberg and Beckham, {Gregg T.}",
year = "2014",
doi = "10.1021/ja410291u",
language = "English",
volume = "136",
pages = "321--329",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "ACS Publications",
number = "1",

}

The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. / Knott, Brandon C.; Momeni, Majid Haddad; Crowley, Michael F.; MacKenzie, Lloyd F.; Götz, Andreas W.; Sandgren, Mats; Withers, Stephen G.; Ståhlberg, Jerry; Beckham, Gregg T.

In: Journal of the American Chemical Society, Vol. 136, No. 1, 2014, p. 321-329.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies

AU - Knott, Brandon C.

AU - Momeni, Majid Haddad

AU - Crowley, Michael F.

AU - MacKenzie, Lloyd F.

AU - Götz, Andreas W.

AU - Sandgren, Mats

AU - Withers, Stephen G.

AU - Ståhlberg, Jerry

AU - Beckham, Gregg T.

PY - 2014

Y1 - 2014

N2 - Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 

AB - Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 

KW - Chemistry (all)

KW - Catalysis

KW - Biochemistry

KW - Colloid and Surface Chemistry

KW - Cellobiohydrolase Cel7A

KW - Cellulose hydrolysis

KW - Conformational change

KW - Glycoside hydrolases

KW - Glycosyl-enzyme intermediate

KW - Glycosylation reactions

KW - Hydrolytic reactions

KW - Transition path sampling

KW - Esterification

KW - Glycosylation

KW - Enzymes

KW - Cellulose

KW - Cellulose 1,4 beta cellobiosidase

KW - Conformational transition

KW - Deglycosylation

KW - Enzyme active site

KW - Enzyme structure

KW - Hydrolysis

KW - Hypocrea jecorina

KW - Sampling

U2 - 10.1021/ja410291u

DO - 10.1021/ja410291u

M3 - Journal article

C2 - 24341799

VL - 136

SP - 321

EP - 329

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 1

ER -