Nanoscale dynamics of cellulose digestion by the cellobiohydrolase TrCel7A

Zachary K. Haviland, Daguan Nong, Kate L.Vasquez Kuntz, Thomas J. Starr, Dengbo Ma, Ming Tien, Charles T. Anderson, William O. Hancock

Research output: Contribution to journalArticlepeer-review

Abstract

Understanding the mechanism by which cellulases from bacteria, fungi, and protozoans catalyze the digestion of lignocellulose is important for developing cost-effective strategies for bioethanol production. Cel7A from the fungus Trichoderma reesei is a model exoglucanase that degrades cellulose strands from their reducing ends by processively cleaving individual cellobiose units. Despite being one of the most studied cellulases, the binding and hydrolysis mechanisms of Cel7A are still debated. Here, we used single-molecule tracking to analyze the dynamics of 11,116 quantum dot-labeled TrCel7A molecules binding to and moving processively along immobilized cellulose. Individual enzyme molecules were localized with a spatial precision of a few nanometers and followed for hundreds of seconds. Most enzyme molecules bound to cellulose in a static state and dissociated without detectable movement, whereas a minority of molecules moved processively for an average distance of 39 nm at an average speed of 3.2 nm/s. These data were integrated into a three-state model in which TrCel7A molecules can bind from solution into either static or processive states and can reversibly switch between states before dissociating. From these results, we conclude that the rate-limiting step for cellulose degradation by Cel7A is the transition out of the static state, either by dissociation from the cellulose surface or by initiation of a processive run. Thus, accelerating the transition of Cel7A out of its static state is a potential avenue for improving cellulase efficiency.

Original languageEnglish (US)
Article number101029
JournalJournal of Biological Chemistry
Volume297
Issue number3
DOIs
StatePublished - Sep 1 2021

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Nanoscale dynamics of cellulose digestion by the cellobiohydrolase TrCel7A'. Together they form a unique fingerprint.

Cite this