Creation of a functional hyperthermostable designer cellulosome

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.

doi:10.1186/s13068-019-1386-y

Title: Creation of a functional hyperthermostable designer cellulosome

Journal Article · Thu Feb 28 00:00:00 EST 2019 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-019-1386-y· OSTI ID:1502788

Kahn, Amaranta ^[1]; Moraïs, Sarah ^[2]; Galanopoulou, Anastasia P. ^[3]; Chung, Daehwan ^[4]; Sarai, Nicholas S. ^[4]; Hengge, Neal ^[4]; Hatzinikolaou, Dimitris G. ^[3]; Himmel, Michael E. ^[4]; Bomble, Yannick J. ^[4];

^[1]

Weizmann Inst. of Science, Rehovot (Israel). Dept. of Biomolecular Sciences
Weizmann Inst. of Science, Rehovot (Israel). Dept. of Biomolecular Sciences; Ben-Gurion Univ. of the Negev, Beersheba (Israel). Faculty of Natural Sciences
National and Kapodistrian Univ. of Athens (Greece). Microbiology Group. Faculty of Biology
National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center

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.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States); Weizmann Institute of Science, Rehovot (Israel); National and Kapodistrian Univ. of Athens (Greece)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); United States—Israel Binational Science Foundation (BSF); Israel Science Foundation (ISF); European Union (EU)

Grant/Contract Number:: AC36-08GO28308; 2013284; 1349/13; 604530

OSTI ID:: 1502788

Report Number(s):: NREL/JA-2700-73261

Journal Information:: Biotechnology for Biofuels, Vol. 12; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 28 works

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09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
multi-enzyme complex
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MOESM1 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790486.v1 The current record is supplemented by this text	text	January 2019
MOESM2 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790492.v1 The current record is supplemented by this text	text	January 2019
MOESM3 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790501.v1 The current record is supplemented by this text	text	January 2019
MOESM4 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790510.v1 The current record is supplemented by this text	text	January 2019
MOESM5 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; Moraïs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790519.v1 The current record is supplemented by this text	text	January 2019
MOESM6 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790528 The current record is supplemented by this text	text	January 2019
Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; Moraïs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.c.4419314 The current record is supplemented by this collection	collection	January 2019
Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; Moraïs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.c.4419314.v1 The current record is supplemented by this collection	collection	January 2019
MOESM1 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790486 The current record is supplemented by this text	text	January 2019
MOESM5 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; Moraïs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790519 The current record is supplemented by this text	text	January 2019
MOESM6 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790528.v1 The current record is supplemented by this text	text	January 2019
MOESM3 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790501 The current record is supplemented by this text	text	January 2019
MOESM2 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790492 The current record is supplemented by this text	text	January 2019
MOESM4 of Creation of a functional hyperthermostable designer cellulosome Kahn, Amaranta; MoraĂŻs, Sarah; Galanopoulou, Anastasia https://doi.org/10.6084/m9.figshare.7790510 The current record is supplemented by this text	text	January 2019