Effect of Multiple Copies of Cohesins on Cellulase and Hemicellulase Activities of Clostridium cellulovorans Mini-cellulosomes

  • Cha, Jae-Ho (Section of Molecular and Cellular Biology, University of California) ;
  • Matsuoka, Satoshi (Section of Molecular and Cellular Biology, University of California) ;
  • Chan, Helen (Section of Molecular and Cellular Biology, University of California) ;
  • Yukawa, Hideaki (Research Institute of Innovative Technology for the Earth) ;
  • Inui, Masayuki (Research Institute of Innovative Technology for the Earth) ;
  • Doi, Roy H. (Section of Molecular and Cellular Biology, University of California)
  • Published : 2007.11.30

Abstract

Cellulosomes in Clostridium cellulovorans are assembled by the interaction between the repeated cohesin domains of a scaffolding protein (CbpA) and the dockerin domain of enzyme components. In this study, we determined the synergistic effects on cellulosic and hemicellulosic substrates by three different recombinant mini-cellulosomes containing either endoglucanase EngB or endoxylanase XynA bound to mini-CbpA with one cohesin domain (mini-CbpAl), two cohesins (mini-CbpA12), or four cohesins (mini-CbpAl234). The assembly of EngB or XynA with mini-CbpA increased the activity against carboxymethyl cellulose, acid-swollen cellulose, Avicel, xylan, and com fiber 1.1-1.8-fold compared with that for the corresponding enzyme alone. A most distinct improvement was shown with com fiber, a natural substrate containing xylan, arabinan, and cellulose. However, there was little difference in activity between the three different mini-cellulosomes when the cellulosomal enzyme concentration was held constant regardless of the copy number of cohesins in the cellulosome. A synergistic effect was observed when the enzyme concentration was increased to be proportional to the number of cohesins in the mini-cellulosome. The highest degree of synergy was observed with mini-CbpAl234 (1.8-fold) and then mini-CbpAl2 (1.3-fold), and the lowest synergy was observed with mini-CbpAl (1.2-fold) when Avicel was used as the substrate. As the copy number of cohesin was increased, there was more synergy. These results indicate that the clustering effect (physical enzyme proximity) of the enzyme within the mini-cellulosome is one of the important factors for efficient degradation of plant cell walls.

Keywords

References

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