Heterologous expression of a β- d -glucosidase in Caldicellulosiruptor bescii has a surprisingly modest effect on the activity of the exoproteome and growth on crystalline cellulose

Kim, Sun-Ki; Chung, Daehwan; Himmel, Michael E.; Bomble, Yannick J.; Westpheling, Janet

doi:10.1007/s10295-017-1982-4

Heterologous expression of a β- d -glucosidase in Caldicellulosiruptor bescii has a surprisingly modest effect on the activity of the exoproteome and growth on crystalline cellulose

Journal Article · Fri Dec 01 00:00:00 EST 2017 · Journal of Industrial Microbiology and Biotechnology

DOI:https://doi.org/10.1007/s10295-017-1982-4· OSTI ID:1773520

Kim, Sun-Ki ^[1]; Chung, Daehwan ^[2]; Himmel, Michael E. ^[2]; Bomble, Yannick J. ^[2]; Westpheling, Janet ^[1]

0000 0004 1936 738X grid.213876.9 Department of Genetics University of Georgia 30602 Athens GA USA, 0000 0004 0446 2659 grid.135519.a The BioEnergy Science Center, Oak Ridge National Laboratory 37831 Oak Ridge TN USA
0000 0001 2199 3636 grid.419357.d Biosciences Center, National Renewable Energy Laboratory Golden CO USA, 0000 0004 0446 2659 grid.135519.a The BioEnergy Science Center, Oak Ridge National Laboratory 37831 Oak Ridge TN USA

Abstract

Members of the genus Caldicellulosiruptor are the most thermophilic cellulolytic bacteria so far described and are capable of efficiently utilizing complex lignocellulosic biomass without conventional pretreatment. Previous studies have shown that accumulation of high concentrations of cellobiose and, to a lesser extent, cellotriose, inhibits cellulase activity both in vivo and in vitro and high concentrations of cellobiose are present in C. bescii fermentations after 90 h of incubation. For some cellulolytic microorganisms, β-d-glucosidase is essential for the efficient utilization of cellobiose as a carbon source and is an essential enzyme in commercial preparations for efficient deconstruction of plant biomass. In spite of its ability to grow efficiently on crystalline cellulose, no extracellular β-d-glucosidase or its GH1 catalytic domain could be identified in the C. bescii genome. To investigate whether the addition of a secreted β-d-glucosidase would improve growth and cellulose utilization by C. bescii, we cloned and expressed a thermostable β-d-glucosidase from Acidothermus cellulolyticus (Acel_0133) in C. bescii using the CelA signal sequence for protein export. The effect of this addition was modest, suggesting that β-d-glucosidase is not rate limiting for cellulose deconstruction and utilization by C. bescii.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC05-00OR22725; AC36-08GO28308

OSTI ID:: 1773520

Alternate ID(s):: OSTI ID: 1399844

Report Number(s):: NREL/JA--2700-70290

Journal Information:: Journal of Industrial Microbiology and Biotechnology, Journal Name: Journal of Industrial Microbiology and Biotechnology Journal Issue: 12 Vol. 44; ISSN 1367-5435

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: Germany

Language:: English

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