Contribution of Scaffoldins to Biomass Degradation by Clostridium Thermocellum: The Effect of Scaffoldin-Deletions on Expression of Other Genes

Qi Xu, Michael Resch, Bryon Donohoe, Lauren Magnusson, Pin-Ching Maness, Steve Decker, Yannick Bomble, Michael Himmel, Daniel Olson, Steven Brown, Dawn Klingeman, Mustafa Syed, Lee Lynd, Charlotte Wilson, John Baker, Shihui Yang

Research output: NRELPoster

Abstract

The cellulosome system contributes greatly to the extreme efficiency of C. thermocellum cellulose degradation. In order to further understand the cellulosome working mechanism, we have knocked out C. thermocellum scaffoldin genes to generate a variety of deletion mutants. The knockout most detrimental to enzymatic hydrolysis by the secretome is that of the primary scaffoldin CipA. Deletion of multiple secondary scaffoldins results in secretome activities intermediate between those of the parent strain and the CipA-knockout mutants. The order of relative secretome activities is the same, whether the cellulosic substrate is microcrystalline cellulose (Avicel) or deacetylated acid-pretreated corn stover (DACS), but the relative magnitudes of the deletion effects are strongly substrate-dependent. Similar trends are observed in fermentation studies of the abilities of the parent and knockout strains themselves to utilize Avicel and DACS. Data from transcriptomic and proteomic studies of these strains when grown on both substrates are used to relate the activity and growth effects of the deletions to their effects on the overall expression of lignocellulose-degrading enzymes by C. thermocellum.
Original languageAmerican English
StatePublished - 2014

Publication series

NamePresented at the 36th Symposium on Biotechnology for Fuels and Chemicals, 28 April - 1 May 2014, Clearwater Beach, Florida

NREL Publication Number

  • NREL/PO-2700-60879

Keywords

  • clostridium thermocellum
  • deletion
  • gene
  • scaffoldin

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