Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification
Abstract Background Corn stover (CS) is evaluated as the most favorable candidate feedstock for butanol production via microbial acetone–butanol–ethanol (ABE) fermentation by Clostridium acetobutylicum. By independent acid pretreatment and enzymatic hydrolysis, fermentable sugars (mainly glucose and...
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doaj-4364a19b4a424661827a65b8a1488afa2020-11-25T04:08:09ZengBMCBiotechnology for Biofuels1754-68342019-11-0112111110.1186/s13068-019-1604-7Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxificationYouduo Wu0Yidi Bai1Daojing Zhang2Chi Cheng3Lijie Chen4Fengwu Bai5Chuang Xue6School of Bioengineering, Dalian University of TechnologySchool of Bioengineering, Dalian University of TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologySchool of Bioengineering, Dalian University of TechnologySchool of Bioengineering, Dalian University of TechnologySchool of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversitySchool of Bioengineering, Dalian University of TechnologyAbstract Background Corn stover (CS) is evaluated as the most favorable candidate feedstock for butanol production via microbial acetone–butanol–ethanol (ABE) fermentation by Clostridium acetobutylicum. By independent acid pretreatment and enzymatic hydrolysis, fermentable sugars (mainly glucose and xylose) were released, of which glucose was naturally utilized as the most preferred carbon source by C. acetobutylicum. However, the ABE fermentation using corn stover hydrolysate (CSH) without detoxification is typically limited to poor sugars utilization, butanol production and productivity. In the presence of pretreatment-derived inhibitors, the intracellular ATP and NADH, as important factors involved in cell growth, solventogenesis initiation and stress response, are exceedingly challenged owing to disrupted glucose phosphotransferase system (PTS). Therefore, there is a necessity to develop effective engineering approaches to overcome these limitations for high-efficient butanol production from CSH without detoxification. Results PTS-engineered C. acetobutylicum strains were constructed via overexpression and knockout of gene glcG encoding glucose-specific PTS IICBA, which pleiotropically regulated glucose utilization, cell growth, solventogenesis and inhibitors tolerance. The PTSGlcG-overexpressing strain exhibited high fermentation efficiency, wherein butanol production and productivity was 11.1 g/L and 0.31 g/L/h, compared to those of 11.0 g/L and 0.15 g/L/h with the PTSGlcG-deficient strain. During CSH culture without detoxification, the PTSGlcG-overexpressing strain exhibited desirable inhibitors tolerance and solventogenesis with butanol production of 10.0 g/L, increased by 300% and 400% compared to those of 2.5 and 2.0 g/L with the control and PTSGlcG-deficient strains, respectively. As a result of extra glucose and 10 g/L CaCO3 addition into CSH, butanol production and productivity were further maximized to 12.5 g/L and 0.39 g/L/h. These validated improvements on the PTSGlcG-overexpressing strain were ascribed to not only efficient glucose transport but also its cascading effects on intracellular ATP and NADH generation, solventogenesis initiation and inhibitors tolerance at the exponential growth phase. Conclusion The PTSGluG regulation could be an effective engineering approach for high-efficient ABE fermentation from lignocellulosic hydrolysates without detoxification or wastewater generation, providing fundamental information for economically sustainable butanol production with high productivity.http://link.springer.com/article/10.1186/s13068-019-1604-7Clostridium acetobutylicumCorn stoverPTS regulationInhibitor toleranceButanol productivity |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Youduo Wu Yidi Bai Daojing Zhang Chi Cheng Lijie Chen Fengwu Bai Chuang Xue |
spellingShingle |
Youduo Wu Yidi Bai Daojing Zhang Chi Cheng Lijie Chen Fengwu Bai Chuang Xue Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification Biotechnology for Biofuels Clostridium acetobutylicum Corn stover PTS regulation Inhibitor tolerance Butanol productivity |
author_facet |
Youduo Wu Yidi Bai Daojing Zhang Chi Cheng Lijie Chen Fengwu Bai Chuang Xue |
author_sort |
Youduo Wu |
title |
Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
title_short |
Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
title_full |
Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
title_fullStr |
Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
title_full_unstemmed |
Pleiotropic regulation of a glucose-specific PTS in Clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
title_sort |
pleiotropic regulation of a glucose-specific pts in clostridium acetobutylicum for high-efficient butanol production from corn stover without detoxification |
publisher |
BMC |
series |
Biotechnology for Biofuels |
issn |
1754-6834 |
publishDate |
2019-11-01 |
description |
Abstract Background Corn stover (CS) is evaluated as the most favorable candidate feedstock for butanol production via microbial acetone–butanol–ethanol (ABE) fermentation by Clostridium acetobutylicum. By independent acid pretreatment and enzymatic hydrolysis, fermentable sugars (mainly glucose and xylose) were released, of which glucose was naturally utilized as the most preferred carbon source by C. acetobutylicum. However, the ABE fermentation using corn stover hydrolysate (CSH) without detoxification is typically limited to poor sugars utilization, butanol production and productivity. In the presence of pretreatment-derived inhibitors, the intracellular ATP and NADH, as important factors involved in cell growth, solventogenesis initiation and stress response, are exceedingly challenged owing to disrupted glucose phosphotransferase system (PTS). Therefore, there is a necessity to develop effective engineering approaches to overcome these limitations for high-efficient butanol production from CSH without detoxification. Results PTS-engineered C. acetobutylicum strains were constructed via overexpression and knockout of gene glcG encoding glucose-specific PTS IICBA, which pleiotropically regulated glucose utilization, cell growth, solventogenesis and inhibitors tolerance. The PTSGlcG-overexpressing strain exhibited high fermentation efficiency, wherein butanol production and productivity was 11.1 g/L and 0.31 g/L/h, compared to those of 11.0 g/L and 0.15 g/L/h with the PTSGlcG-deficient strain. During CSH culture without detoxification, the PTSGlcG-overexpressing strain exhibited desirable inhibitors tolerance and solventogenesis with butanol production of 10.0 g/L, increased by 300% and 400% compared to those of 2.5 and 2.0 g/L with the control and PTSGlcG-deficient strains, respectively. As a result of extra glucose and 10 g/L CaCO3 addition into CSH, butanol production and productivity were further maximized to 12.5 g/L and 0.39 g/L/h. These validated improvements on the PTSGlcG-overexpressing strain were ascribed to not only efficient glucose transport but also its cascading effects on intracellular ATP and NADH generation, solventogenesis initiation and inhibitors tolerance at the exponential growth phase. Conclusion The PTSGluG regulation could be an effective engineering approach for high-efficient ABE fermentation from lignocellulosic hydrolysates without detoxification or wastewater generation, providing fundamental information for economically sustainable butanol production with high productivity. |
topic |
Clostridium acetobutylicum Corn stover PTS regulation Inhibitor tolerance Butanol productivity |
url |
http://link.springer.com/article/10.1186/s13068-019-1604-7 |
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