Skip to main content

Main menu

  • Home
  • Articles
    • Current
    • Special Feature Articles - Most Recent
    • Special Features
    • Colloquia
    • Collected Articles
    • PNAS Classics
    • List of Issues
  • Front Matter
    • Front Matter Portal
    • Journal Club
  • News
    • For the Press
    • This Week In PNAS
    • PNAS in the News
  • Podcasts
  • Authors
    • Information for Authors
    • Editorial and Journal Policies
    • Submission Procedures
    • Fees and Licenses
  • Submit
  • Submit
  • About
    • Editorial Board
    • PNAS Staff
    • FAQ
    • Accessibility Statement
    • Rights and Permissions
    • Site Map
  • Contact
  • Journal Club
  • Subscribe
    • Subscription Rates
    • Subscriptions FAQ
    • Open Access
    • Recommend PNAS to Your Librarian

User menu

  • Log in
  • My Cart

Search

  • Advanced search
Home
Home
  • Log in
  • My Cart

Advanced Search

  • Home
  • Articles
    • Current
    • Special Feature Articles - Most Recent
    • Special Features
    • Colloquia
    • Collected Articles
    • PNAS Classics
    • List of Issues
  • Front Matter
    • Front Matter Portal
    • Journal Club
  • News
    • For the Press
    • This Week In PNAS
    • PNAS in the News
  • Podcasts
  • Authors
    • Information for Authors
    • Editorial and Journal Policies
    • Submission Procedures
    • Fees and Licenses
  • Submit
Research Article

Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance

Marcelo Kern, John E. McGeehan, Simon D. Streeter, Richard N. A. Martin, Katrin Besser, Luisa Elias, Will Eborall, Graham P. Malyon, Christina M. Payne, Michael E. Himmel, Kirk Schnorr, Gregg T. Beckham, Simon M. Cragg, Neil C. Bruce, and Simon J. McQueen-Mason
  1. aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  2. bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  3. cDepartment of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506;
  4. dBiosciences Center and
  5. fNational Bioenergy Center, National Renewable Energy Laboratory, Golden, CO 80401; and
  6. eNovozymes A/S, 2880 Bagsvaerd, Denmark

See allHide authors and affiliations

PNAS first published June 3, 2013; https://doi.org/10.1073/pnas.1301502110
Marcelo Kern
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John E. McGeehan
bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Simon D. Streeter
bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard N. A. Martin
bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Katrin Besser
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Luisa Elias
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Will Eborall
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Graham P. Malyon
bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Christina M. Payne
cDepartment of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506;
dBiosciences Center and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael E. Himmel
dBiosciences Center and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kirk Schnorr
eNovozymes A/S, 2880 Bagsvaerd, Denmark
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Gregg T. Beckham
fNational Bioenergy Center, National Renewable Energy Laboratory, Golden, CO 80401; and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: simon.mcqueenmason@york.ac.uk gregg.beckham@nrel.gov Simon.Cragg@port.ac.uk neil.bruce@york.ac.uk
Simon M. Cragg
bSchool of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: simon.mcqueenmason@york.ac.uk gregg.beckham@nrel.gov Simon.Cragg@port.ac.uk neil.bruce@york.ac.uk
Neil C. Bruce
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: simon.mcqueenmason@york.ac.uk gregg.beckham@nrel.gov Simon.Cragg@port.ac.uk neil.bruce@york.ac.uk
Simon J. McQueen-Mason
aCentre for Novel Agricultural Products, Department of Biology, University of York, York YO10 5DD, United Kingdom;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: simon.mcqueenmason@york.ac.uk gregg.beckham@nrel.gov Simon.Cragg@port.ac.uk neil.bruce@york.ac.uk
  1. Edited by Alexis T. Bell, University of California, Berkeley, CA, and approved May 8, 2013 (received for review January 24, 2013)

  • Article
  • Figures & SI
  • Info & Metrics
  • PDF
Loading

Abstract

Nature uses a diversity of glycoside hydrolase (GH) enzymes to convert polysaccharides to sugars. As lignocellulosic biomass deconstruction for biofuel production remains costly, natural GH diversity offers a starting point for developing industrial enzymes, and fungal GH family 7 (GH7) cellobiohydrolases, in particular, provide significant hydrolytic potential in industrial mixtures. Recently, GH7 enzymes have been found in other kingdoms of life besides fungi, including in animals and protists. Here, we describe the in vivo spatial expression distribution, properties, and structure of a unique endogenous GH7 cellulase from an animal, the marine wood borer Limnoria quadripunctata (LqCel7B). RT-quantitative PCR and Western blot studies show that LqCel7B is expressed in the hepatopancreas and secreted into the gut for wood degradation. We produced recombinant LqCel7B, with which we demonstrate that LqCel7B is a cellobiohydrolase and obtained four high-resolution crystal structures. Based on a crystallographic and computational comparison of LqCel7B to the well-characterized Hypocrea jecorina GH7 cellobiohydrolase, LqCel7B exhibits an extended substrate-binding motif at the tunnel entrance, which may aid in substrate acquisition and processivity. Interestingly, LqCel7B exhibits striking surface charges relative to fungal GH7 enzymes, which likely results from evolution in marine environments. We demonstrate that LqCel7B stability and activity remain unchanged, or increase at high salt concentration, and that the L. quadripunctata GH mixture generally contains cellulolytic enzymes with highly acidic surface charge compared with enzymes derived from terrestrial microbes. Overall, this study suggests that marine cellulases offer significant potential for utilization in high-solids industrial biomass conversion processes.

  • gribble
  • carbohydrate degrading enzymes

Footnotes

  • ↵1M.K. and J.E.M. contributed equally to this work.

  • ↵2To whom correspondence may be addressed. E-mail: simon.mcqueenmason{at}york.ac.uk, gregg.beckham{at}nrel.gov, Simon.Cragg{at}port.ac.uk, or neil.bruce{at}york.ac.uk.
  • Author contributions: M.K., J.E.M., K.S., G.T.B., N.C.B., and S.J.M.-M. designed research; M.K., J.E.M., S.D.S., R.N.A.M., K.B., L.E., W.E., G.P.M., C.M.P., K.S., and G.T.B. performed research; M.K., J.E.M., S.D.S., C.M.P., M.E.H., G.T.B., S.M.C., N.C.B., and S.J.M.-M. analyzed data; and M.K., J.E.M., C.M.P., G.T.B., S.M.C., N.C.B., and S.J.M.-M. wrote the paper.

  • The authors declare no conflict of interest.

  • This article is a PNAS Direct Submission.

  • Data deposition: The atomic coordinates and structure factors have been deposited in the Protein Data Bank, www.pdb.org (PDB ID codes 4GWA, 4HAP, 4HAQ, and 4IPM) and the sequence reported in this paper has been deposited in the GenBank database (accession no. KC776193).

  • This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1301502110/-/DCSupplemental.

Next
Back to top
Article Alerts
Email Article

Thank you for your interest in spreading the word on PNAS.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance
(Your Name) has sent you a message from PNAS
(Your Name) thought you would like to see the PNAS web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Salt-tolerant marine cellulase
Marcelo Kern, John E. McGeehan, Simon D. Streeter, Richard N. A. Martin, Katrin Besser, Luisa Elias, Will Eborall, Graham P. Malyon, Christina M. Payne, Michael E. Himmel, Kirk Schnorr, Gregg T. Beckham, Simon M. Cragg, Neil C. Bruce, Simon J. McQueen-Mason
Proceedings of the National Academy of Sciences Jun 2013, 201301502; DOI: 10.1073/pnas.1301502110

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Request Permissions
Share
Salt-tolerant marine cellulase
Marcelo Kern, John E. McGeehan, Simon D. Streeter, Richard N. A. Martin, Katrin Besser, Luisa Elias, Will Eborall, Graham P. Malyon, Christina M. Payne, Michael E. Himmel, Kirk Schnorr, Gregg T. Beckham, Simon M. Cragg, Neil C. Bruce, Simon J. McQueen-Mason
Proceedings of the National Academy of Sciences Jun 2013, 201301502; DOI: 10.1073/pnas.1301502110
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Mendeley logo Mendeley
Proceedings of the National Academy of Sciences: 118 (9)
Current Issue

Submit

Sign up for Article Alerts

Jump to section

  • Article
  • Figures & SI
  • Info & Metrics
  • PDF

You May Also be Interested in

Setting sun over a sun-baked dirt landscape
Core Concept: Popular integrated assessment climate policy models have key caveats
Better explicating the strengths and shortcomings of these models will help refine projections and improve transparency in the years ahead.
Image credit: Witsawat.S.
Model of the Amazon forest
News Feature: A sea in the Amazon
Did the Caribbean sweep into the western Amazon millions of years ago, shaping the region’s rich biodiversity?
Image credit: Tacio Cordeiro Bicudo (University of São Paulo, São Paulo, Brazil), Victor Sacek (University of São Paulo, São Paulo, Brazil), and Lucy Reading-Ikkanda (artist).
Syrian archaeological site
Journal Club: In Mesopotamia, early cities may have faltered before climate-driven collapse
Settlements 4,200 years ago may have suffered from overpopulation before drought and lower temperatures ultimately made them unsustainable.
Image credit: Andrea Ricci.
Steamboat Geyser eruption.
Eruption of Steamboat Geyser
Mara Reed and Michael Manga explore why Yellowstone's Steamboat Geyser resumed erupting in 2018.
Listen
Past PodcastsSubscribe
Birds nestling on tree branches
Parent–offspring conflict in songbird fledging
Some songbird parents might improve their own fitness by manipulating their offspring into leaving the nest early, at the cost of fledgling survival, a study finds.
Image credit: Gil Eckrich (photographer).

Similar Articles

Site Logo
Powered by HighWire
  • Submit Manuscript
  • Twitter
  • Facebook
  • RSS Feeds
  • Email Alerts

Articles

  • Current Issue
  • Special Feature Articles – Most Recent
  • List of Issues

PNAS Portals

  • Anthropology
  • Chemistry
  • Classics
  • Front Matter
  • Physics
  • Sustainability Science
  • Teaching Resources

Information

  • Authors
  • Editorial Board
  • Reviewers
  • Subscribers
  • Librarians
  • Press
  • Site Map
  • PNAS Updates
  • FAQs
  • Accessibility Statement
  • Rights & Permissions
  • About
  • Contact

Feedback    Privacy/Legal

Copyright © 2021 National Academy of Sciences. Online ISSN 1091-6490