Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply
Beta (β)-carotene (C40H56; a provitamin) is a particularly important carotenoid for human health. Many studies have focused on engineering Escherichia coli as an efficient heterologous producer of β-carotene. Moreover, several strains with potential for use in the industrial production of this provi...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2020-06-01
|
Series: | Frontiers in Bioengineering and Biotechnology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00585/full |
id |
doaj-381d6d19b150453fb53bc8421f84c5a8 |
---|---|
record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuanqing Wu Yuanqing Wu Panpan Yan Panpan Yan Yang Li Yang Li Yang Li Xuewei Liu Xuewei Liu Zhiwen Wang Zhiwen Wang Tao Chen Tao Chen Xueming Zhao Xueming Zhao |
spellingShingle |
Yuanqing Wu Yuanqing Wu Panpan Yan Panpan Yan Yang Li Yang Li Yang Li Xuewei Liu Xuewei Liu Zhiwen Wang Zhiwen Wang Tao Chen Tao Chen Xueming Zhao Xueming Zhao Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply Frontiers in Bioengineering and Biotechnology Escherichia coli metabolic engineering β-carotene phosphotransferase system inactivation NADPH supply |
author_facet |
Yuanqing Wu Yuanqing Wu Panpan Yan Panpan Yan Yang Li Yang Li Yang Li Xuewei Liu Xuewei Liu Zhiwen Wang Zhiwen Wang Tao Chen Tao Chen Xueming Zhao Xueming Zhao |
author_sort |
Yuanqing Wu |
title |
Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply |
title_short |
Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply |
title_full |
Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply |
title_fullStr |
Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply |
title_full_unstemmed |
Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH Supply |
title_sort |
enhancing β-carotene production in escherichia coli by perturbing central carbon metabolism and improving the nadph supply |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Bioengineering and Biotechnology |
issn |
2296-4185 |
publishDate |
2020-06-01 |
description |
Beta (β)-carotene (C40H56; a provitamin) is a particularly important carotenoid for human health. Many studies have focused on engineering Escherichia coli as an efficient heterologous producer of β-carotene. Moreover, several strains with potential for use in the industrial production of this provitamin have already been constructed via different metabolic engineering strategies. In this study, we aimed to improve the β-carotene-producing capacity of our previously engineered E. coli strain ZF43ΔgdhA through further gene deletion and metabolic pathway manipulations. Deletion of the zwf gene increased the resultant strain's β-carotene production and content by 5.1 and 32.5%, respectively, relative to the values of strain ZF43ΔgdhA, but decreased the biomass by 26.2%. Deletion of the ptsHIcrr operon further increased the β-carotene production titer from 122.0 to 197.4 mg/L, but the provitamin content was decreased. Subsequently, comparative transcriptomic analysis was used to explore the dynamic transcriptional responses of the strains to the blockade of the pentose phosphate pathway and inactivation of the phosphotransferase system. Lastly, based on the analyses of comparative transcriptome and reduction cofactor, several strategies to increase the NADPH supply were evaluated for enhancement of the β-carotene content. The combination of yjgB gene deletion and nadK overexpression led to increased β-carotene production and content. The best strain, ECW4/p5C-nadK, produced 266.4 mg/L of β-carotene in flask culture and 2,579.1 mg/L in a 5-L bioreactor. The latter value is the highest reported from production via the methylerythritol phosphate pathway in E. coli. Although the strategies applied is routine in this study, the combinations reported were first implemented, are simple but efficient and will be helpful for the production of many other natural products, especially isoprenoids. Importantly, we demonstrated that the use of the methylerythritol phosphate pathway alone for efficient β-carotene biosynthesis could be achieved via appropriate modifications of the cell metabolic functions. |
topic |
Escherichia coli metabolic engineering β-carotene phosphotransferase system inactivation NADPH supply |
url |
https://www.frontiersin.org/article/10.3389/fbioe.2020.00585/full |
work_keys_str_mv |
AT yuanqingwu enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT yuanqingwu enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT panpanyan enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT panpanyan enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT yangli enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT yangli enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT yangli enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT xueweiliu enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT xueweiliu enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT zhiwenwang enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT zhiwenwang enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT taochen enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT taochen enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT xuemingzhao enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply AT xuemingzhao enhancingbcaroteneproductioninescherichiacolibyperturbingcentralcarbonmetabolismandimprovingthenadphsupply |
_version_ |
1724497066037608448 |
spelling |
doaj-381d6d19b150453fb53bc8421f84c5a82020-11-25T03:48:48ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-06-01810.3389/fbioe.2020.00585535957Enhancing β-Carotene Production in Escherichia coli by Perturbing Central Carbon Metabolism and Improving the NADPH SupplyYuanqing Wu0Yuanqing Wu1Panpan Yan2Panpan Yan3Yang Li4Yang Li5Yang Li6Xuewei Liu7Xuewei Liu8Zhiwen Wang9Zhiwen Wang10Tao Chen11Tao Chen12Xueming Zhao13Xueming Zhao14Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaCollege of Life Science, Shihezi University, Shihezi, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSynthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaBeta (β)-carotene (C40H56; a provitamin) is a particularly important carotenoid for human health. Many studies have focused on engineering Escherichia coli as an efficient heterologous producer of β-carotene. Moreover, several strains with potential for use in the industrial production of this provitamin have already been constructed via different metabolic engineering strategies. In this study, we aimed to improve the β-carotene-producing capacity of our previously engineered E. coli strain ZF43ΔgdhA through further gene deletion and metabolic pathway manipulations. Deletion of the zwf gene increased the resultant strain's β-carotene production and content by 5.1 and 32.5%, respectively, relative to the values of strain ZF43ΔgdhA, but decreased the biomass by 26.2%. Deletion of the ptsHIcrr operon further increased the β-carotene production titer from 122.0 to 197.4 mg/L, but the provitamin content was decreased. Subsequently, comparative transcriptomic analysis was used to explore the dynamic transcriptional responses of the strains to the blockade of the pentose phosphate pathway and inactivation of the phosphotransferase system. Lastly, based on the analyses of comparative transcriptome and reduction cofactor, several strategies to increase the NADPH supply were evaluated for enhancement of the β-carotene content. The combination of yjgB gene deletion and nadK overexpression led to increased β-carotene production and content. The best strain, ECW4/p5C-nadK, produced 266.4 mg/L of β-carotene in flask culture and 2,579.1 mg/L in a 5-L bioreactor. The latter value is the highest reported from production via the methylerythritol phosphate pathway in E. coli. Although the strategies applied is routine in this study, the combinations reported were first implemented, are simple but efficient and will be helpful for the production of many other natural products, especially isoprenoids. Importantly, we demonstrated that the use of the methylerythritol phosphate pathway alone for efficient β-carotene biosynthesis could be achieved via appropriate modifications of the cell metabolic functions.https://www.frontiersin.org/article/10.3389/fbioe.2020.00585/fullEscherichia colimetabolic engineeringβ-carotenephosphotransferase system inactivationNADPH supply |