Engineering Enhanced Cellobiohydrolase Activity

Steve Decker, Brandon Knott, Petri Alahuhta, Jeffrey Linger, Vladimir Lunin, Venkataramanan Subramanian, Qi Xu, Todd VanderWall, Michael Crowley, Michael Himmel, Gregg Beckham, Larry Taylor II, Sarah Hobdey, Kara Podkaminer, Logan Schuster, Yogesh Chaudhari, Wiiliam Adney, John Baker, Antonella Amore

Research output: Contribution to journalArticlepeer-review

68 Scopus Citations


Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure-activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.

Original languageAmerican English
Article number1186
Number of pages10
JournalNature Communications
Issue number1
StatePublished - 1 Dec 2018

Bibliographical note

Publisher Copyright:
© 2018 The Author(s).

NREL Publication Number

  • NREL/JA-2700-70605


  • hydrolases
  • polysaccharides
  • protein design
  • x-ray crystallography


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