Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail

Mercedes V Del Pozo, Lucía Fernández-Arrojo, Jorge Gil-Martínez, Alejandro Montesinos, Tatyana N Chernikova, Taras Y Nechitaylo, Agnes Waliszek, Marta Tortajada, Antonia Rojas, Sharon A Huws, Olga V Golyshina, Charles J Newbold, Julio Polaina, Manuel Ferrer, Peter N Golyshin

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Abstract

BACKGROUND: A complete saccharification of plant polymers is the critical step in the efficient production of bio-alcohols. Beta-glucosidases acting in the degradation of intermediate gluco-oligosaccharides produced by cellulases limit the yield of the final product.

RESULTS: In the present work, we have identified and then successfully cloned, expressed, purified and characterised 4 highly active beta-glucosidases from fibre-adherent microbial community from the cow rumen. The enzymes were most active at temperatures 45-55°C and pH 4.0-7.0 and exhibited high affinity and activity towards synthetic substrates such as p-nitrophenyl-beta-D-glucopyranoside (pNPbetaG) and pNP-beta-cellobiose, as well as to natural cello-oligosaccharides ranging from cellobiose to cellopentaose. The apparent capability of the most active beta-glucosidase, herein named LAB25g2, was tested for its ability to improve, at low dosage (31.25 units g-1 dry biomass, using pNPbetaG as substrate), the hydrolysis of pre-treated corn stover (dry matter content of 20%; 350 g glucan kg-1 dry biomass) in combination with a beta-glucosidase-deficient commercial Trichoderma reseei cellulase cocktail (5 units g-1 dry biomass in the basis of pNPbetaG). LAB25g2 increased the final hydrolysis yield by a factor of 20% (44.5 ± 1.7% vs. 34.5 ± 1.5% in control conditions) after 96-120 h as compared to control reactions in its absence or in the presence of other commercial beta-glucosidase preparations. The high stability (half-life higher than 5 days at 50°C and pH 5.2) and 2-38000 fold higher (as compared with reported beta-glucosidases) activity towards cello-oligosaccharides may account for its performance in supplementation assays.

CONCLUSIONS: The results suggest that beta-glucosidases from yet uncultured bacteria from animal digestomes may be of a potential interest for biotechnological processes related to the effective bio-ethanol production in combination with low dosage of commercial cellulases.

Original languageEnglish
Pages (from-to)73
JournalBiotechnology for Biofuels
Volume5
Issue number1
DOIs
Publication statusPrint publication - 21 Sep 2012
Externally publishedYes

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  • Cite this

    Del Pozo, M. V., Fernández-Arrojo, L., Gil-Martínez, J., Montesinos, A., Chernikova, T. N., Nechitaylo, T. Y., Waliszek, A., Tortajada, M., Rojas, A., Huws, S. A., Golyshina, O. V., Newbold, C. J., Polaina, J., Ferrer, M., & Golyshin, P. N. (2012). Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail. Biotechnology for Biofuels, 5(1), 73. https://doi.org/10.1186/1754-6834-5-73