Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases
<p>Abstract</p> <p>Background</p> <p>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...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2011-08-01
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| Series: | Biotechnology for Biofuels |
| Online Access: | http://www.biotechnologyforbiofuels.com/content/4/1/23 |
| _version_ | 1818554277913690112 |
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| author | Li Luen-Luen Taghavi Safiyh McCorkle Sean M Zhang Yian-Biao Blewitt Michael G Brunecky Roman Adney William S Himmel Michael E Brumm Phillip Drinkwater Colleen Mead David A Tringe Susannah G Lelie Daniel van der |
| author_facet | Li Luen-Luen Taghavi Safiyh McCorkle Sean M Zhang Yian-Biao Blewitt Michael G Brunecky Roman Adney William S Himmel Michael E Brumm Phillip Drinkwater Colleen Mead David A Tringe Susannah G Lelie Daniel van der |
| author_sort | Li Luen-Luen |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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 <it>Escherichia coli</it>. Further characterization showed that two enzymes showed significant activity on <it>p</it>-nitrophenyl-α-<smcaps>L</smcaps>-arabinofuranoside, one enzyme had significant activity against <it>p</it>-nitrophenyl-β-<smcaps>D</smcaps>-glucopyranoside, and one enzyme showed significant activity against <it>p</it>-nitrophenyl-β-<smcaps>D</smcaps>-xylopyranoside. Enzymes were also tested in the presence of ionic liquids.</p> <p>Conclusions</p> <p>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.</p> |
| first_indexed | 2024-12-12T09:37:15Z |
| format | Article |
| id | doaj.art-b51f2007477d4bb590c569947e527af6 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T09:37:15Z |
| publishDate | 2011-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-b51f2007477d4bb590c569947e527af62022-12-22T00:28:41ZengBMCBiotechnology for Biofuels1754-68342011-08-01412310.1186/1754-6834-4-23Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolasesLi Luen-LuenTaghavi SafiyhMcCorkle Sean MZhang Yian-BiaoBlewitt Michael GBrunecky RomanAdney William SHimmel Michael EBrumm PhillipDrinkwater ColleenMead David ATringe Susannah GLelie Daniel van der<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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 <it>Escherichia coli</it>. Further characterization showed that two enzymes showed significant activity on <it>p</it>-nitrophenyl-α-<smcaps>L</smcaps>-arabinofuranoside, one enzyme had significant activity against <it>p</it>-nitrophenyl-β-<smcaps>D</smcaps>-glucopyranoside, and one enzyme showed significant activity against <it>p</it>-nitrophenyl-β-<smcaps>D</smcaps>-xylopyranoside. Enzymes were also tested in the presence of ionic liquids.</p> <p>Conclusions</p> <p>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.</p>http://www.biotechnologyforbiofuels.com/content/4/1/23 |
| spellingShingle | Li Luen-Luen Taghavi Safiyh McCorkle Sean M Zhang Yian-Biao Blewitt Michael G Brunecky Roman Adney William S Himmel Michael E Brumm Phillip Drinkwater Colleen Mead David A Tringe Susannah G Lelie Daniel van der Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases Biotechnology for Biofuels |
| title | Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases |
| title_full | Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases |
| title_fullStr | Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases |
| title_full_unstemmed | Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases |
| title_short | Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases |
| title_sort | bioprospecting metagenomics of decaying wood mining for new glycoside hydrolases |
| url | http://www.biotechnologyforbiofuels.com/content/4/1/23 |
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