Cargando…

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....

Descripción completa

Detalles Bibliográficos
Autores principales: Long, Mengfei, Xu, Meijuan, Ma, Zhenfeng, Pan, Xuewei, You, Jiajia, Hu, Mengkai, Shao, Yu, Yang, Taowei, Zhang, Xian, Rao, Zhiming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2020
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
_version_ 1783537546265886720
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
work_keys_str_mv AT longmengfei significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT xumeijuan significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT mazhenfeng significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT panxuewei significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT youjiajia significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT humengkai significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT shaoyu significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT yangtaowei significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT zhangxian significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli
AT raozhiming significantlyenhancingproductionoftrans4hydroxylprolinebyintegratedsystemengineeringinescherichiacoli