Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ

William Scott, View ORCID ProfileBrian Lowrance, View ORCID ProfileAlexander C. Anderson, View ORCID ProfileJoel T. Weadge
doi: https://doi.org/10.1101/837344
William Scott
Department of Biology, Wilfrid Laurier University 75 University Ave W. Waterloo, ON N2L 3C5
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Brian Lowrance
Department of Biology, Wilfrid Laurier University 75 University Ave W. Waterloo, ON N2L 3C5
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Brian Lowrance
Alexander C. Anderson
Department of Biology, Wilfrid Laurier University 75 University Ave W. Waterloo, ON N2L 3C5
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Alexander C. Anderson
Joel T. Weadge
Department of Biology, Wilfrid Laurier University 75 University Ave W. Waterloo, ON N2L 3C5
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Joel T. Weadge
  • For correspondence: jweadge@wlu.ca
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

Abstract

Biofilms are community structures of bacteria enmeshed in a self-produced matrix of exopolysaccharides. The biofilm matrix serves numerous roles, including resilience and persistence, making biofilms a subject of research interest among persistent clinical pathogens of global health importance. Our current understanding of the underlying biochemical pathways responsible for biosynthesis of these exopolysaccharides is largely limited to Gram-negative bacteria. Clostridia are a class of Gram-positive, anaerobic and spore-forming bacteria, and include the important human pathogens Clostridium perfringens, Clostridium botulinum, and Clostridioides difficile, among numerous others. Clostridia have been reported to form biofilms composed of cellulose, although the specific loci which encode the cellulose synthase have not been identified. Here, we report the discovery of a gene cluster, which we named ccsABZCD, among selected bacteria within class Clostridia that appears to encode a synthase complex responsible for polymerization, modification, and export of an O-acetylated cellulose exopolysaccharide. To test this hypothesis, we subcloned the putative glycoside hydrolase CcsZ and solved the X-ray crystal structure of both apo- and product-bound CcsZ. Our results demonstrate that CcsZ is in fact an endo-acting cellulase belonging to glycoside hydrolase family 5 (GH-5). This is in contrast to the Gram-negative cellulose synthase, which instead encodes BcsZ, a structurally distinct GH-8. We further show CcsZ is capable of hydrolysis of the soluble mock substrate carboxymethylcellulose (CMC) with a pH optimum of 4.5. The data we present here serves as an entry point to an understanding of biofilm formation among class Clostridia and allowed us to predict a model for Clostridial cellulose synthesis.

Author summary Biofilms are communities of microorganisms that enmesh themselves in a protective matrix of elf-produced polysaccharide materials. Biofilms have demonstrated roles in both virulence and persistence among bacterial pathogens of global health importance. The class Clostridia are a polyphyletic grouping of primarily Gram-positive, anaerobic and spore-forming bacteria which contain the important and well-studied human pathogens Clostridioides difficile, Clostridium botulinum, and Clostridium perfringens, among others. Bacteria belonging to class Clostridia have been anecdotally reported to form biofilms made of cellulose, although the molecular mechanisms governing their production has not before been described. In this work, we identify a gene cluster, which we name ccsABZHI, for the Clostridial cellulose synthase, which bears remarkable similarity to molecular machinery required for the production of cellulose biofilms in other Gram-negative bacteria. We further biochemically characterize one of these enzymes, CcsZ, a predicted endoglucanase which we predicted from our model should cleave cellulose exopolysaccharides. We show that CcsZ is in fact capable of this activity and belongs to a broader family of glycoside hydrolases with unexpected taxonomic diversity. Our work represents an entry point towards an understanding of the molecular mechanisms governing cellulose biofilm formation in Gram-positive bacteria.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Back to top
PreviousNext
Posted November 10, 2019.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ
William Scott, Brian Lowrance, Alexander C. Anderson, Joel T. Weadge
bioRxiv 837344; doi: https://doi.org/10.1101/837344
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ
William Scott, Brian Lowrance, Alexander C. Anderson, Joel T. Weadge
bioRxiv 837344; doi: https://doi.org/10.1101/837344

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (4866)
  • Biochemistry (10821)
  • Bioengineering (8068)
  • Bioinformatics (27381)
  • Biophysics (14030)
  • Cancer Biology (11166)
  • Cell Biology (16106)
  • Clinical Trials (138)
  • Developmental Biology (8808)
  • Ecology (13332)
  • Epidemiology (2067)
  • Evolutionary Biology (17399)
  • Genetics (11705)
  • Genomics (15964)
  • Immunology (11061)
  • Microbiology (26169)
  • Molecular Biology (10681)
  • Neuroscience (56748)
  • Paleontology (422)
  • Pathology (1737)
  • Pharmacology and Toxicology (3012)
  • Physiology (4570)
  • Plant Biology (9670)
  • Scientific Communication and Education (1617)
  • Synthetic Biology (2699)
  • Systems Biology (6997)
  • Zoology (1515)