skip to main content

DOE PAGESDOE PAGES

Title: Creation of a functional hyperthermostable designer cellulosome

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. 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 hyperthermostablemore » 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. 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.« less
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [4] ;  [4] ;  [3] ;  [4] ;  [4] ; ORCiD logo [1]
  1. Weizmann Inst. of Science, Rehovot (Israel). Dept. of Biomolecular Sciences
  2. Weizmann Inst. of Science, Rehovot (Israel). Dept. of Biomolecular Sciences; Ben-Gurion Univ. of the Negev, Beersheba (Israel). Faculty of Natural Sciences
  3. National and Kapodistrian Univ. of Athens (Greece). Microbiology Group. Faculty of Biology
  4. National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center
Publication Date:
Report Number(s):
NREL/JA-2700-73261
Journal ID: ISSN 1754-6834
Grant/Contract Number:
AC36-08GO28308; 2013284; 1349/13; 604530
Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 12; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Research Org:
National Renewable Energy Lab. (NREL), Golden, CO (United States); Weizmann Inst. of Science, Rehovot (Israel); National and Kapodistrian Univ. of Athens (Greece)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); United States—Israel Binational Science Foundation (BSF); Israel Science Foundation (ISF); European Union (EU)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; multi-enzyme complex; cellulases; thermostability; Caldicellulosiruptor bescii; scaffoldin; dockerin; cohesin
OSTI Identifier:
1502788

Kahn, Amaranta, Moraïs, Sarah, Galanopoulou, Anastasia P., Chung, Daehwan, Sarai, Nicholas S., Hengge, Neal, Hatzinikolaou, Dimitris G., Himmel, Michael E., Bomble, Yannick J., and Bayer, Edward A.. Creation of a functional hyperthermostable designer cellulosome. United States: N. p., Web. doi:10.1186/s13068-019-1386-y.
Kahn, Amaranta, Moraïs, Sarah, Galanopoulou, Anastasia P., Chung, Daehwan, Sarai, Nicholas S., Hengge, Neal, Hatzinikolaou, Dimitris G., Himmel, Michael E., Bomble, Yannick J., & Bayer, Edward A.. Creation of a functional hyperthermostable designer cellulosome. United States. doi:10.1186/s13068-019-1386-y.
Kahn, Amaranta, Moraïs, Sarah, Galanopoulou, Anastasia P., Chung, Daehwan, Sarai, Nicholas S., Hengge, Neal, Hatzinikolaou, Dimitris G., Himmel, Michael E., Bomble, Yannick J., and Bayer, Edward A.. 2019. "Creation of a functional hyperthermostable designer cellulosome". United States. doi:10.1186/s13068-019-1386-y. https://www.osti.gov/servlets/purl/1502788.
@article{osti_1502788,
title = {Creation of a functional hyperthermostable designer cellulosome},
author = {Kahn, Amaranta and Moraïs, Sarah and Galanopoulou, Anastasia P. and Chung, Daehwan and Sarai, Nicholas S. and Hengge, Neal and Hatzinikolaou, Dimitris G. and Himmel, Michael E. and Bomble, Yannick J. and Bayer, Edward A.},
abstractNote = {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. 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. 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.},
doi = {10.1186/s13068-019-1386-y},
journal = {Biotechnology for Biofuels},
number = ,
volume = 12,
place = {United States},
year = {2019},
month = {2}
}

Works referenced in this record:

Hyperthermophilic Enzymes: Sources, Uses, and Molecular Mechanisms for Thermostability
journal, March 2001

Cellulases and biofuels
journal, June 2009

Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production
journal, February 2007
  • Himmel, M. E.; Ding, S.-Y.; Johnson, D. K.
  • Science, Vol. 315, Issue 5813, p. 804-807
  • DOI: 10.1126/science.1137016

How biotech can transform biofuels
journal, February 2008
  • Lynd, Lee R.; Laser, Mark S.; Bransby, David
  • Nature Biotechnology, Vol. 26, Issue 2, p. 169-172
  • DOI: 10.1038/nbt0208-169

Efficient Degradation of Lignocellulosic Plant Biomass, without Pretreatment, by the Thermophilic Anaerobe "Anaerocellum thermophilum" DSM 6725
journal, May 2009
  • Yang, Sung-Jae; Kataeva, Irina; Hamilton-Brehm, Scott D.
  • Applied and Environmental Microbiology, Vol. 75, Issue 14, p. 4762-4769
  • DOI: 10.1128/AEM.00236-09

Phylogenetic, Microbiological, and Glycoside Hydrolase Diversities within the Extremely Thermophilic, Plant Biomass-Degrading Genus Caldicellulosiruptor
journal, October 2010
  • Blumer-Schuette, S. E.; Lewis, D. L.; Kelly, R. M.
  • Applied and Environmental Microbiology, Vol. 76, Issue 24, p. 8084-8092
  • DOI: 10.1128/AEM.01400-10

Description of Caldicellulosiruptor saccharolyticus gen. nov., sp. nov: An obligately anaerobic, extremely thermophilic, cellulolytic bacterium
journal, July 1994

Revealing Nature's Cellulase Diversity The Digestion Mechanism of Caldicellulosiruptor bescii CelA
journal, December 2013