Dissociation of the cellulosome of Clostridium thermocellum under nondenaturing conditions

Ely Morag, Sima Yaron, Raphael Lamed, Rina Kenig, Yuval Shoham, Edward A. Bayer*

*Corresponding author for this work

Research output: Contribution to journalConference articlepeer-review

Abstract

The cellulosome from the anaerobic, thermophilic, cellulolytic bacterium, Clostridium thermocellum, was dissociated under nondenaturing conditions by incubation at 60°C in the combined presence of EDTA and cellulose. This approach differs from previous strategies in that detergents were not necessary for dissociation of the subunits. Selected enzymatic subunits could be isolated with relative ease. One of these is the major cellulosomal cellobiohydrolase, CelS (previously designated subunit S8 or S(s)). Both the intact cellulosome complex and the combined dissociated subunits exhibited similar levels of activities on soluble and acid-swollen cellulosic substrates. In contrast, the mixture of the free enzymatic subunits was markedly deficient in its degradation of crystalline cellulose compared with that of the intact cellulosome. The results support the critical role of divalent cations, e.g. calcium, in the stability of the cellulosome complex. In addition, the results reinforce previous indications that the cellulosome undergoes conformational changes upon binding to cellulose.

Original languageEnglish
Pages (from-to)235-242
Number of pages8
JournalJournal of Biotechnology
Volume51
Issue number3
DOIs
StatePublished - 15 Nov 1996
EventProceedings of the 1995 Workshop on Environmental Biotechnology -
Duration: 28 Nov 199528 Nov 1995

Keywords

  • Clostridium thermocellum
  • cellulases
  • cellulose degradation
  • cellulosome
  • multi-enzyme complex
  • subunit dissociation

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