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Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli
Trans-4-hydroxy-l-proline is produced by trans-proline-4-hydroxylase with l-proline through glucose fermentation. Here, we designed a thorough “from A to Z” strategy to significantly improve trans-4-hydroxy-l-proline production. Through rare codon selected evolution, Escherichia coli M1 produced 18....
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Association for the Advancement of Science
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7244267/ https://www.ncbi.nlm.nih.gov/pubmed/32494747 http://dx.doi.org/10.1126/sciadv.aba2383 |
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author | Long, Mengfei Xu, Meijuan Ma, Zhenfeng Pan, Xuewei You, Jiajia Hu, Mengkai Shao, Yu Yang, Taowei Zhang, Xian Rao, Zhiming |
author_facet | Long, Mengfei Xu, Meijuan Ma, Zhenfeng Pan, Xuewei You, Jiajia Hu, Mengkai Shao, Yu Yang, Taowei Zhang, Xian Rao, Zhiming |
author_sort | Long, Mengfei |
collection | PubMed |
description | Trans-4-hydroxy-l-proline is produced by trans-proline-4-hydroxylase with l-proline through glucose fermentation. Here, we designed a thorough “from A to Z” strategy to significantly improve trans-4-hydroxy-l-proline production. Through rare codon selected evolution, Escherichia coli M1 produced 18.2 g L(−1) l-proline. Metabolically engineered M6 with the deletion of putA, proP, putP, and aceA, and proB mutation focused carbon flux to l-proline and released its feedback inhibition. It produced 15.7 g L(−1) trans-4-hydroxy-l-proline with 10 g L(−1) l-proline retained. Furthermore, a tunable circuit based on quorum sensing attenuated l-proline hydroxylation flux, resulting in 43.2 g L(−1) trans-4-hydroxy-l-proline with 4.3 g L(−1) l-proline retained. Finally, rationally designed l-proline hydroxylase gave 54.8 g L(−1) trans-4-hydroxy-l-proline in 60 hours almost without l-proline remaining—the highest production to date. The de novo engineering carbon flux through rare codon selected evolution, dynamic precursor modulation, and metabolic engineering provides a good technological platform for efficient hydroxyl amino acid synthesis. |
format | Online Article Text |
id | pubmed-7244267 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Association for the Advancement of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-72442672020-06-02 Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli Long, Mengfei Xu, Meijuan Ma, Zhenfeng Pan, Xuewei You, Jiajia Hu, Mengkai Shao, Yu Yang, Taowei Zhang, Xian Rao, Zhiming Sci Adv Research Articles Trans-4-hydroxy-l-proline is produced by trans-proline-4-hydroxylase with l-proline through glucose fermentation. Here, we designed a thorough “from A to Z” strategy to significantly improve trans-4-hydroxy-l-proline production. Through rare codon selected evolution, Escherichia coli M1 produced 18.2 g L(−1) l-proline. Metabolically engineered M6 with the deletion of putA, proP, putP, and aceA, and proB mutation focused carbon flux to l-proline and released its feedback inhibition. It produced 15.7 g L(−1) trans-4-hydroxy-l-proline with 10 g L(−1) l-proline retained. Furthermore, a tunable circuit based on quorum sensing attenuated l-proline hydroxylation flux, resulting in 43.2 g L(−1) trans-4-hydroxy-l-proline with 4.3 g L(−1) l-proline retained. Finally, rationally designed l-proline hydroxylase gave 54.8 g L(−1) trans-4-hydroxy-l-proline in 60 hours almost without l-proline remaining—the highest production to date. The de novo engineering carbon flux through rare codon selected evolution, dynamic precursor modulation, and metabolic engineering provides a good technological platform for efficient hydroxyl amino acid synthesis. American Association for the Advancement of Science 2020-05-22 /pmc/articles/PMC7244267/ /pubmed/32494747 http://dx.doi.org/10.1126/sciadv.aba2383 Text en Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (http://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited. |
spellingShingle | Research Articles Long, Mengfei Xu, Meijuan Ma, Zhenfeng Pan, Xuewei You, Jiajia Hu, Mengkai Shao, Yu Yang, Taowei Zhang, Xian Rao, Zhiming Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title | Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title_full | Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title_fullStr | Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title_full_unstemmed | Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title_short | Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli |
title_sort | significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in escherichia coli |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7244267/ https://www.ncbi.nlm.nih.gov/pubmed/32494747 http://dx.doi.org/10.1126/sciadv.aba2383 |
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