Creation of a functional hyperthermostable designer cellulosome

Amaranta Kahn, Sarah Moraïs, Anastasia P. Galanopoulou, Daehwan Chung, Nicholas S. Sarai, Neal Hengge, Dimitris G. Hatzinikolaou, Michael E. Himmel, Yannick J. Bomble, Edward A. Bayer

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

Background: Renewable energy has become a field of high interest over the past decade, and production of biofuels from cellulosic substrates has a particularly high potential as an alternative source of energy. Industrial deconstruction of biomass, however, is an onerous, exothermic process, the cost of which could be decreased significantly by use of hyperthermophilic enzymes. An efficient way of breaking down cellulosic substrates can also be achieved by highly efficient enzymatic complexes called cellulosomes. The modular architecture of these multi-enzyme complexes results in substrate targeting and proximity-based synergy among the resident enzymes. However, cellulosomes have not been observed in hyperthermophilic bacteria. Results: Here, we report the design and function of a novel hyperthermostable "designer cellulosome" system, which is stable and active at 75 °C. Enzymes from Caldicellulosiruptor bescii, a highly cellulolytic hyperthermophilic anaerobic bacterium, were selected and successfully converted to the cellulosomal mode by grafting onto them divergent dockerin modules that can be inserted in a precise manner into a thermostable chimaeric scaffoldin by virtue of their matching cohesins. Three pairs of cohesins and dockerins, selected from thermophilic microbes, were examined for their stability at extreme temperatures and were determined stable at 75 °C for at least 72 h. The resultant hyperthermostable cellulosome complex exhibited the highest levels of enzymatic activity on microcrystalline cellulose at 75 °C, compared to those of previously reported designer cellulosome systems and the native cellulosome from Clostridium thermocellum. Conclusion: The functional hyperthermophilic platform fulfills the appropriate physico-chemical properties required for exothermic processes. This system can thus be adapted for other types of thermostable enzyme systems and could serve as a basis for a variety of cellulolytic and non-cellulolytic industrial objectives at high temperatures.

Original languageEnglish
Article number44
JournalBiotechnology for Biofuels
Volume12
Issue number1
DOIs
StatePublished - 28 Feb 2019

Keywords

  • Caldicellulosiruptor bescii
  • Cellulases
  • Cohesin
  • Dockerin
  • Multi-enzyme complex
  • Scaffoldin
  • Thermostability

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • General Energy
  • Management, Monitoring, Policy and Law

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