Functional analysis of the glucan degradation locus in Caldicellulosiruptor bescii reveals essential roles of component glycoside hydrolases in plant biomass deconstruction

Jonathan M. Conway, Bennett S. McKinley, Nathaniel L. Seals, Diana Hernandez, Piyum A. Khatibi, Suresh Poudel, Richard J. Giannone, Robert L. Hettich, Amanda M. Williams-Rhaesa, Gina L. Lipscomb, Michael W.W. Adams, Robert M. Kelly

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36 Scopus citations


The ability to hydrolyze microcrystalline cellulose is an uncommon feature in the microbial world, but it can be exploited for conversion of lignocellulosic feedstocks into biobased fuels and chemicals. Understanding the physiological and biochemical mechanisms by which microorganisms deconstruct cellulosic material is key to achieving this objective. The glucan degradation locus (GDL) in the genomes of extremely thermophilic Caldicellulosiruptor species encodes polysaccharide lyases (PLs), unique cellulose binding proteins (tāpirins), and putative posttranslational modifying enzymes, in addition to multidomain, multifunctional glycoside hydrolases (GHs), thereby representing an alternative paradigm for plant biomass degradation compared to fungal or cellulosomal systems. To examine the individual and collective in vivo roles of the glycolytic enzymes, the six GH genes in the GDL of Caldicellulosiruptor bescii were systematically deleted, and the extents to which the resulting mutant strains could solubilize microcrystalline cellulose (Avicel) and plant biomass (switchgrass or poplar) were examined. Three of the GDL enzymes, Athe_1867 (CelA) (GH9-CBM3-CBM3-CBM3-GH48), Athe_1859 (GH5-CBM3-CBM3-GH44), and Athe_1857 (GH10-CBM3-CBM3-GH48), acted synergistically in vivo and accounted for 92% of naked microcrystalline cellulose (Avicel) degradation. However, the relative importance of the GDL GHs varied for the plant biomass substrates tested. Furthermore, mixed cultures of mutant strains showed that switchgrass solubilization depended on the secretome-bound enzymes collectively produced by the culture, not on the specific strain from which they came. These results demonstrate that certain GDL GHs are primarily responsible for the degradation of microcrystalline cellulose-containing substrates by C. bescii and provide new insights into the workings of a novel microbial mechanism for lignocellulose utilization.

Original languageEnglish (US)
Article numbere01828-17
JournalApplied and Environmental Microbiology
Issue number24
StatePublished - Dec 1 2017
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology


  • Caldicellulosiruptor
  • Cellulase
  • Cellulose degradation
  • Extreme thermophile
  • Glycoside hydrolase
  • Lignocellulose


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