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Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production
Clostridium beijerinckii, a promising industrial microorganism for butanol production, suffers from low butanol titer and lack of high-efficiency genetical engineering toolkit. A few histidine kinases (HKs) responsible for Spo0A phosphorylation have been demonstrated as functionally important compon...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098912/ https://www.ncbi.nlm.nih.gov/pubmed/32266241 http://dx.doi.org/10.3389/fbioe.2020.00214 |
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author | Xin, Xin Cheng, Chi Du, Guangqing Chen, Lijie Xue, Chuang |
author_facet | Xin, Xin Cheng, Chi Du, Guangqing Chen, Lijie Xue, Chuang |
author_sort | Xin, Xin |
collection | PubMed |
description | Clostridium beijerinckii, a promising industrial microorganism for butanol production, suffers from low butanol titer and lack of high-efficiency genetical engineering toolkit. A few histidine kinases (HKs) responsible for Spo0A phosphorylation have been demonstrated as functionally important components in regulating butanol biosynthesis in solventogenic clostridia such as C. acetobutylicum, but no study about HKs has been conducted in C. beijerinckii. In this study, six annotated but uncharacterized candidate HK genes sharing partial homologies (no less than 30%) with those in C. acetobutylicum were selected based on sequence alignment. The encoding region of these HK genes were deleted with CRISPR-Cas9n-based genome editing technology. The deletion of cbei2073 and cbei4484 resulted in significant change in butanol biosynthesis, with butanol production increased by 40.8 and 17.3% (13.8 g/L and 11.5 g/L vs. 9.8 g/L), respectively, compared to the wild-type. Faster butanol production rates were observed, with butanol productivity greatly increased by 40.0 and 20.0%, respectively, indicating these two HKs are important in regulating cellular metabolism in C. beijerinckii. In addition, the sporulation frequencies of two HKs inactivated strains decreased by 96.9 and 77.4%, respectively. The other four HK-deletion (including cbei2087, cbei2435, cbei4925, and cbei1553) mutant strains showed few phenotypic changes compared with the wild-type. This study demonstrated the role of HKs on sporulation and solventogenesis in C. beijerinckii, and provided a novel engineering strategy of HKs for improving metabolite production. The hyper-butanol-producing strains generated in this study have great potentials in industrial biobutanol production. |
format | Online Article Text |
id | pubmed-7098912 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-70989122020-04-07 Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production Xin, Xin Cheng, Chi Du, Guangqing Chen, Lijie Xue, Chuang Front Bioeng Biotechnol Bioengineering and Biotechnology Clostridium beijerinckii, a promising industrial microorganism for butanol production, suffers from low butanol titer and lack of high-efficiency genetical engineering toolkit. A few histidine kinases (HKs) responsible for Spo0A phosphorylation have been demonstrated as functionally important components in regulating butanol biosynthesis in solventogenic clostridia such as C. acetobutylicum, but no study about HKs has been conducted in C. beijerinckii. In this study, six annotated but uncharacterized candidate HK genes sharing partial homologies (no less than 30%) with those in C. acetobutylicum were selected based on sequence alignment. The encoding region of these HK genes were deleted with CRISPR-Cas9n-based genome editing technology. The deletion of cbei2073 and cbei4484 resulted in significant change in butanol biosynthesis, with butanol production increased by 40.8 and 17.3% (13.8 g/L and 11.5 g/L vs. 9.8 g/L), respectively, compared to the wild-type. Faster butanol production rates were observed, with butanol productivity greatly increased by 40.0 and 20.0%, respectively, indicating these two HKs are important in regulating cellular metabolism in C. beijerinckii. In addition, the sporulation frequencies of two HKs inactivated strains decreased by 96.9 and 77.4%, respectively. The other four HK-deletion (including cbei2087, cbei2435, cbei4925, and cbei1553) mutant strains showed few phenotypic changes compared with the wild-type. This study demonstrated the role of HKs on sporulation and solventogenesis in C. beijerinckii, and provided a novel engineering strategy of HKs for improving metabolite production. The hyper-butanol-producing strains generated in this study have great potentials in industrial biobutanol production. Frontiers Media S.A. 2020-03-20 /pmc/articles/PMC7098912/ /pubmed/32266241 http://dx.doi.org/10.3389/fbioe.2020.00214 Text en Copyright © 2020 Xin, Cheng, Du, Chen and Xue. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Xin, Xin Cheng, Chi Du, Guangqing Chen, Lijie Xue, Chuang Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title | Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title_full | Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title_fullStr | Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title_full_unstemmed | Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title_short | Metabolic Engineering of Histidine Kinases in Clostridium beijerinckii for Enhanced Butanol Production |
title_sort | metabolic engineering of histidine kinases in clostridium beijerinckii for enhanced butanol production |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098912/ https://www.ncbi.nlm.nih.gov/pubmed/32266241 http://dx.doi.org/10.3389/fbioe.2020.00214 |
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