@article{21b9d3915f8c450d877819f660d1c192,
title = "Glycosylation of hyperthermostable designer cellulosome components yields enhanced stability and cellulose hydrolysis",
abstract = "Biomass deconstruction remains integral for enabling second-generation biofuel production at scale. However, several steps necessary to achieve significant solubilization of biomass, notably harsh pretreatment conditions, impose economic barriers to commercialization. By employing hyperthermostable cellulase machinery, biomass deconstruction can be made more efficient, leading to milder pretreatment conditions and ultimately lower production costs. The hyperthermophilic bacterium Caldicellulosiruptor bescii produces extremely active hyperthermostable cellulases, including the hyperactive multifunctional cellulase CbCel9A/Cel48A. Recombinant CbCel9A/Cel48A components have been previously produced in Escherichia coli and integrated into synthetic hyperthermophilic designer cellulosome complexes. Since then, glycosylation has been shown to be vital for the high activity and stability of CbCel9A/Cel48A. Here, we studied the impact of glycosylation on a hyperthermostable designer cellulosome system in which two of the cellulosomal components, the scaffoldin and the GH9 domain of CbCel9A/Cel48A, were glycosylated as a consequence of employing Ca. bescii as an expression host. Inclusion of the glycosylated components yielded an active cellulosome system that exhibited long-term stability at 75 °C. The resulting glycosylated designer cellulosomes showed significantly greater synergistic activity compared to the enzymatic components alone, as well as higher thermostability than the analogous nonglycosylated designer cellulosomes. These results indicate that glycosylation can be used as an essential engineering tool to improve the properties of designer cellulosomes. Additionally, Ca. bescii was shown to be an attractive candidate for production of glycosylated designer cellulosome components, which may further promote the viability of this bacterium both as a cellulase expression host and as a potential consolidated bioprocessing platform organism.",
keywords = "Caldicellulosiruptor bescii, cellulosome, expression host, glycosylation, thermostability",
author = "Amaranta Kahn and Sarah Mora{\"i}s and Daehwan Chung and Sarai, {Nicholas S.} and Hengge, {Neal N.} and Audrey Kahn and Himmel, {Michael E.} and Bayer, {Edward A.} and Bomble, {Yannick J.}",
note = "Funding Information: AmK greatly appreciates scholarships received from the Ministry of Immigrant Absorption, Jerusalem, Israel, and from the ministry of Foreign Affairs, Paris, France. AmK is a Sustainability and Energy Weizmann Fellow. EAB is the incumbent of The Maynard I. and Elaine Wishner Chair of Bio‐organic Chemistry. This work was authored in part by Alliance for Sustainable Energy, LLC, the manager and operator of the National Renewable Energy Laboratory for the U.S. Department of Energy (DOE) under Contract No. DE‐AC36‐08GO28308. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. Funding was also provided by the Center for Bioenergy Innovation (CBI), a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. This research was also supported by the United States–Israel Binational Science Foundation (BSF grant No. 2013284), Jerusalem, Israel; the Israel Science Foundation (ISF grant no. 1349/13); and the European Union NMP.2013.1.1‐2: CellulosomePlus Project number 604530. The funding sources were not involved in the design of the study, data interpretation, report writing, or decision in submitting the article for publication. Funding Information: AmK greatly appreciates scholarships received from the Ministry of Immigrant Absorption, Jerusalem, Israel, and from the ministry of Foreign Affairs, Paris, France. AmK is a Sustainability and Energy Weizmann Fellow. EAB is the incumbent of The Maynard I. and Elaine Wishner Chair of Bio-organic Chemistry. This work was authored in part by Alliance for Sustainable Energy, LLC, the manager and operator of the National Renewable Energy Laboratory for the U.S. Department of Energy (DOE) under Contract No. DE-AC36-08GO28308. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. Funding was also provided by the Center for Bioenergy Innovation (CBI), a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. This research was also supported by the United States–Israel Binational Science Foundation (BSF grant No. 2013284), Jerusalem, Israel; the Israel Science Foundation (ISF grant no. 1349/13); and the European Union NMP.2013.1.1-2: CellulosomePlus Project number 604530. The funding sources were not involved in the design of the study, data interpretation, report writing, or decision in submitting the article for publication. Publisher Copyright: {\textcopyright} 2020 Federation of European Biochemical Societies",
year = "2020",
month = oct,
day = "1",
doi = "10.1111/febs.15251",
language = "English",
volume = "287",
pages = "4370--4388",
journal = "FEBS Journal",
issn = "1742-4658",
publisher = "wiley",
number = "20",
}