High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it>
<p>Abstract</p> <p>Background</p> <p>The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and p...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2011-09-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://www.biotechnologyforbiofuels.com/content/4/1/30 |
| _version_ | 1818547333172822016 |
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| author | Ahlgren Simon Thorngren Naomi la Grange Daniël C Siika-aho Matti Voutilainen Sanni P Koivula Anu Froehlich Allan Wiswall Erin McBride John Brevnova Elena den Haan Riaan Ilmén Marja Mellon Mark Deleault Kristen Rajgarhia Vineet van Zyl Willem H Penttilä Merja |
| author_facet | Ahlgren Simon Thorngren Naomi la Grange Daniël C Siika-aho Matti Voutilainen Sanni P Koivula Anu Froehlich Allan Wiswall Erin McBride John Brevnova Elena den Haan Riaan Ilmén Marja Mellon Mark Deleault Kristen Rajgarhia Vineet van Zyl Willem H Penttilä Merja |
| author_sort | Ahlgren Simon |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and produce a valuable product such as ethanol at a high rate and titer could significantly reduce the costs of biomass conversion technologies, and will allow separate conversion steps to be combined in a consolidated bioprocess (CBP). Development of <it>Saccharomyces cerevisiae </it>for CBP requires the high level secretion of cellulases, particularly cellobiohydrolases.</p> <p>Results</p> <p>We expressed various cellobiohydrolases to identify enzymes that were efficiently secreted by <it>S. cerevisiae</it>. For enhanced cellulose hydrolysis, we engineered bimodular derivatives of a well secreted enzyme that naturally lacks the carbohydrate-binding module, and constructed strains expressing combinations of <it>cbh1 </it>and <it>cbh2 </it>genes. Though there was significant variability in the enzyme levels produced, up to approximately 0.3 g/L CBH1 and approximately 1 g/L CBH2 could be produced in high cell density fermentations. Furthermore, we could show activation of the unfolded protein response as a result of cellobiohydrolase production. Finally, we report fermentation of microcrystalline cellulose (Avicel™) to ethanol by CBH-producing <it>S. cerevisiae </it>strains with the addition of beta-glucosidase.</p> <p>Conclusions</p> <p>Gene or protein specific features and compatibility with the host are important for efficient cellobiohydrolase secretion in yeast. The present work demonstrated that production of both CBH1 and CBH2 could be improved to levels where the barrier to CBH sufficiency in the hydrolysis of cellulose was overcome.</p> |
| first_indexed | 2024-12-12T08:05:21Z |
| format | Article |
| id | doaj.art-3071f5110d624ddd8d68fcffcc1afb49 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T08:05:21Z |
| publishDate | 2011-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-3071f5110d624ddd8d68fcffcc1afb492022-12-22T00:32:00ZengBMCBiotechnology for Biofuels1754-68342011-09-01413010.1186/1754-6834-4-30High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it>Ahlgren SimonThorngren Naomila Grange Daniël CSiika-aho MattiVoutilainen Sanni PKoivula AnuFroehlich AllanWiswall ErinMcBride JohnBrevnova Elenaden Haan RiaanIlmén MarjaMellon MarkDeleault KristenRajgarhia Vineetvan Zyl Willem HPenttilä Merja<p>Abstract</p> <p>Background</p> <p>The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and produce a valuable product such as ethanol at a high rate and titer could significantly reduce the costs of biomass conversion technologies, and will allow separate conversion steps to be combined in a consolidated bioprocess (CBP). Development of <it>Saccharomyces cerevisiae </it>for CBP requires the high level secretion of cellulases, particularly cellobiohydrolases.</p> <p>Results</p> <p>We expressed various cellobiohydrolases to identify enzymes that were efficiently secreted by <it>S. cerevisiae</it>. For enhanced cellulose hydrolysis, we engineered bimodular derivatives of a well secreted enzyme that naturally lacks the carbohydrate-binding module, and constructed strains expressing combinations of <it>cbh1 </it>and <it>cbh2 </it>genes. Though there was significant variability in the enzyme levels produced, up to approximately 0.3 g/L CBH1 and approximately 1 g/L CBH2 could be produced in high cell density fermentations. Furthermore, we could show activation of the unfolded protein response as a result of cellobiohydrolase production. Finally, we report fermentation of microcrystalline cellulose (Avicel™) to ethanol by CBH-producing <it>S. cerevisiae </it>strains with the addition of beta-glucosidase.</p> <p>Conclusions</p> <p>Gene or protein specific features and compatibility with the host are important for efficient cellobiohydrolase secretion in yeast. The present work demonstrated that production of both CBH1 and CBH2 could be improved to levels where the barrier to CBH sufficiency in the hydrolysis of cellulose was overcome.</p>http://www.biotechnologyforbiofuels.com/content/4/1/30biofuelscellulolytic yeastUPR |
| spellingShingle | Ahlgren Simon Thorngren Naomi la Grange Daniël C Siika-aho Matti Voutilainen Sanni P Koivula Anu Froehlich Allan Wiswall Erin McBride John Brevnova Elena den Haan Riaan Ilmén Marja Mellon Mark Deleault Kristen Rajgarhia Vineet van Zyl Willem H Penttilä Merja High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> Biotechnology for Biofuels biofuels cellulolytic yeast UPR |
| title | High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> |
| title_full | High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> |
| title_fullStr | High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> |
| title_full_unstemmed | High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> |
| title_short | High level secretion of cellobiohydrolases by <it>Saccharomyces cerevisiae</it> |
| title_sort | high level secretion of cellobiohydrolases by it saccharomyces cerevisiae it |
| topic | biofuels cellulolytic yeast UPR |
| url | http://www.biotechnologyforbiofuels.com/content/4/1/30 |
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