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Title: Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

Abstract

To efficiently deconstruct recalcitrant plant biomass to fermentable sugars in industrial processes, biocatalysts of higher performance and lower cost are required. The genetic diversity found in the metagenomes of natural microbial biomass decay communities may harbor such enzymes. Our goal was to discover and characterize new glycoside hydrolases (GHases) from microbial biomass decay communities, especially those from unknown or never previously cultivated microorganisms. From the metagenome sequences of an anaerobic microbial community actively decaying poplar biomass, we identified approximately 4,000 GHase homologs. Based on homology to GHase families/activities of interest and the quality of the sequences, candidates were selected for full-length cloning and subsequent expression. As an alternative strategy, a metagenome expression library was constructed and screened for GHase activities. These combined efforts resulted in the cloning of four novel GHases that could be successfully expressed in Escherichia coli. Further characterization showed that two enzymes showed significant activity on p-nitrophenyl-{alpha}-L-arabinofuranoside, one enzyme had significant activity against p-nitrophenyl-{beta}-D-glucopyranoside, and one enzyme showed significant activity against p-nitrophenyl-{beta}-D-xylopyranoside. Enzymes were also tested in the presence of ionic liquids. Metagenomics provides a good resource for mining novel biomass degrading enzymes and for screening of cellulolytic enzyme activities. The four GHases that were cloned maymore » have potential application for deconstruction of biomass pretreated with ionic liquids, as they remain active in the presence of up to 20% ionic liquid (except for 1-ethyl-3-methylimidazolium diethyl phosphate). Alternatively, ionic liquids might be used to immobilize or stabilize these enzymes for minimal solvent processing of biomass.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
BROOKHAVEN NATIONAL LABORATORY (BNL)
Sponsoring Org.:
USDOE SC OFFICE OF SCIENCE (SC)
OSTI Identifier:
1044001
Report Number(s):
BNL-96107-2012-JA
R&D Project: BOW-139; YN1901000; TRN: US201214%%256
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 4; Journal Issue: 23
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 54 ENVIRONMENTAL SCIENCES; BIOMASS; CLONING; COMMUNITIES; DECAY; ENZYMES; ESCHERICHIA COLI; GENETICS; GLYCOSIDES; HYDROLASES; MICROORGANISMS; MINING; MOLTEN SALTS; PERFORMANCE; POPLARS; PROCESSING; SACCHARIDES; SOLVENTS; WOOD

Citation Formats

Li L. L., van der Lelie D., Taghavi, S., McCorkle, S. M., Zhang, Y.-B., Blewitt, M. G., Brunecky, R., Adney, W. S., Himmel, M. E., Brumm, P., Drinkwater, C., Mead, D. A., and Tringe, S. G. Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases. United States: N. p., 2011. Web.
Li L. L., van der Lelie D., Taghavi, S., McCorkle, S. M., Zhang, Y.-B., Blewitt, M. G., Brunecky, R., Adney, W. S., Himmel, M. E., Brumm, P., Drinkwater, C., Mead, D. A., & Tringe, S. G. Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases. United States.
Li L. L., van der Lelie D., Taghavi, S., McCorkle, S. M., Zhang, Y.-B., Blewitt, M. G., Brunecky, R., Adney, W. S., Himmel, M. E., Brumm, P., Drinkwater, C., Mead, D. A., and Tringe, S. G. Mon . "Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases". United States.
@article{osti_1044001,
title = {Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases},
author = {Li L. L. and van der Lelie D. and Taghavi, S. and McCorkle, S. M. and Zhang, Y.-B. and Blewitt, M. G. and Brunecky, R. and Adney, W. S. and Himmel, M. E. and Brumm, P. and Drinkwater, C. and Mead, D. A. and Tringe, S. G.},
abstractNote = {To efficiently deconstruct recalcitrant plant biomass to fermentable sugars in industrial processes, biocatalysts of higher performance and lower cost are required. The genetic diversity found in the metagenomes of natural microbial biomass decay communities may harbor such enzymes. Our goal was to discover and characterize new glycoside hydrolases (GHases) from microbial biomass decay communities, especially those from unknown or never previously cultivated microorganisms. From the metagenome sequences of an anaerobic microbial community actively decaying poplar biomass, we identified approximately 4,000 GHase homologs. Based on homology to GHase families/activities of interest and the quality of the sequences, candidates were selected for full-length cloning and subsequent expression. As an alternative strategy, a metagenome expression library was constructed and screened for GHase activities. These combined efforts resulted in the cloning of four novel GHases that could be successfully expressed in Escherichia coli. Further characterization showed that two enzymes showed significant activity on p-nitrophenyl-{alpha}-L-arabinofuranoside, one enzyme had significant activity against p-nitrophenyl-{beta}-D-glucopyranoside, and one enzyme showed significant activity against p-nitrophenyl-{beta}-D-xylopyranoside. Enzymes were also tested in the presence of ionic liquids. Metagenomics provides a good resource for mining novel biomass degrading enzymes and for screening of cellulolytic enzyme activities. The four GHases that were cloned may have potential application for deconstruction of biomass pretreated with ionic liquids, as they remain active in the presence of up to 20% ionic liquid (except for 1-ethyl-3-methylimidazolium diethyl phosphate). Alternatively, ionic liquids might be used to immobilize or stabilize these enzymes for minimal solvent processing of biomass.},
doi = {},
journal = {Biotechnology for Biofuels},
number = 23,
volume = 4,
place = {United States},
year = {2011},
month = {8}
}