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

Research output: Contribution to journalArticle

52 Citations (Scopus)

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

Fingerprint

Metagenome
Glucosidases
Cellulases
Saccharification
Cellulase
Rumen
beta-Glucosidase
hydrolysis
biomass
Oligosaccharides
Biomass
Cellobiose
substrate
half life
dry matter
alcohol
microbial community
ethanol
Hydrolysis
polymer

Cite this

Del Pozo, M. V., Fernández-Arrojo, L., Gil-Martínez, J., Montesinos, A., Chernikova, T. N., Nechitaylo, T. Y., ... 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
Del Pozo, Mercedes V ; Fernández-Arrojo, Lucía ; Gil-Martínez, Jorge ; Montesinos, Alejandro ; Chernikova, Tatyana N ; Nechitaylo, Taras Y ; Waliszek, Agnes ; Tortajada, Marta ; Rojas, Antonia ; Huws, Sharon A ; Golyshina, Olga V ; Newbold, Charles J ; Polaina, Julio ; Ferrer, Manuel ; Golyshin, Peter N. / Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail. In: Biotechnology for Biofuels. 2012 ; Vol. 5, No. 1. pp. 73.
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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.",
author = "{Del Pozo}, {Mercedes V} and Luc{\'i}a Fern{\'a}ndez-Arrojo and Jorge Gil-Mart{\'i}nez and Alejandro Montesinos and Chernikova, {Tatyana N} and Nechitaylo, {Taras Y} and Agnes Waliszek and Marta Tortajada and Antonia Rojas and Huws, {Sharon A} and Golyshina, {Olga V} and Newbold, {Charles J} and Julio Polaina and Manuel Ferrer and Golyshin, {Peter N}",
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Del Pozo, MV, Fernández-Arrojo, L, Gil-Martínez, J, Montesinos, A, Chernikova, TN, Nechitaylo, TY, Waliszek, A, Tortajada, M, Rojas, A, Huws, SA, Golyshina, OV, Newbold, CJ, Polaina, J, Ferrer, M & Golyshin, PN 2012, 'Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail', Biotechnology for Biofuels, vol. 5, no. 1, pp. 73. https://doi.org/10.1186/1754-6834-5-73

Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail. / Del Pozo, Mercedes V; Fernández-Arrojo, Lucía; Gil-Martínez, Jorge; Montesinos, Alejandro; Chernikova, Tatyana N; Nechitaylo, Taras Y; Waliszek, Agnes; Tortajada, Marta; Rojas, Antonia; Huws, Sharon A; Golyshina, Olga V; Newbold, Charles J; Polaina, Julio; Ferrer, Manuel; Golyshin, Peter N.

In: Biotechnology for Biofuels, Vol. 5, No. 1, 21.09.2012, p. 73.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail

AU - Del Pozo, Mercedes V

AU - Fernández-Arrojo, Lucía

AU - Gil-Martínez, Jorge

AU - Montesinos, Alejandro

AU - Chernikova, Tatyana N

AU - Nechitaylo, Taras Y

AU - Waliszek, Agnes

AU - Tortajada, Marta

AU - Rojas, Antonia

AU - Huws, Sharon A

AU - Golyshina, Olga V

AU - Newbold, Charles J

AU - Polaina, Julio

AU - Ferrer, Manuel

AU - Golyshin, Peter N

PY - 2012/9/21

Y1 - 2012/9/21

N2 - 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.

AB - 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.

U2 - 10.1186/1754-6834-5-73

DO - 10.1186/1754-6834-5-73

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JO - Biotechnology for Biofuels

JF - Biotechnology for Biofuels

SN - 1754-6834

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