Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene
Monoterpene α-pinene exhibits significant potential as an alternative fuel, widely recognized for its affordability and eco-friendly nature. It demonstrates multiple biological activities and has a wide range of applications. However, the limited supply of pinene extracted from plants poses a challe...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1527113/full |
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| author | Muhammad Hammad Hussain Lu Han Yanlong Wei Muhammad Javid Kamran Ashraf Maria Martuscelli Waleed Aldahmash Meijin Guo Ali Mohsin Zhanxia Li |
| author_facet | Muhammad Hammad Hussain Lu Han Yanlong Wei Muhammad Javid Kamran Ashraf Maria Martuscelli Waleed Aldahmash Meijin Guo Ali Mohsin Zhanxia Li |
| author_sort | Muhammad Hammad Hussain |
| collection | DOAJ |
| description | Monoterpene α-pinene exhibits significant potential as an alternative fuel, widely recognized for its affordability and eco-friendly nature. It demonstrates multiple biological activities and has a wide range of applications. However, the limited supply of pinene extracted from plants poses a challenge in meeting the needs of the aviation industry and other sectors. Considering this, the microbial cell factory is the only viable option for achieving sustainable pinene production. This study employed a rational design model to optimize the copy number and integration site for the heterogenous pinene expression pathway in Escherichia coli (E. coli). The integrated strain with the best pinene pathway PG1 was selected. Subsequently, the resulting strain, E. coli HSY009, accumulated 49.01 mg/L of pinene after 24 h fermentation in the flask culture. To further enhance production, pinene expression cassette PG1 was sequentially integrated into three non-essential regions (44th, 58th, 23rd), resulting in an improved pinene titer. Then, the fermentation process under optimized conditions enhanced the production of pinene to 436.68 mg/L in a 5 L batch fermenter with a mean productivity of 14.55 mg/L/h. To the best of our knowledge, this work represents the maximum mean pinene productivity based on the currently available literature. The findings of this work provide valuable insights for optimizing E. coli to produce other valuable terpenoids that share the same intermediates, IPP and DMAPP. Conclusively, this research validates the model’s universality and highlights its potential for application as cutting-edge biofuel precursors. |
| format | Article |
| id | doaj-art-b167ca548911459bb55688825324192a |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-b167ca548911459bb55688825324192a2025-01-07T06:48:34ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011510.3389/fmicb.2024.15271131527113Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pineneMuhammad Hammad Hussain0Lu Han1Yanlong Wei2Muhammad Javid3Kamran Ashraf4Maria Martuscelli5Waleed Aldahmash6Meijin Guo7Ali Mohsin8Zhanxia Li9State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaDepartment of Microbiology, Faculty of Chemical and Life Sciences, Abdul Wali Khan University Mardan, Mardan, PakistanState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaDepartment of Bioscience and Food, Agricultural and Environmental Technology, University of the Studies of Teramo, Teramo, ItalyZoology Department, College of Science, King Saud University, Riyadh, Saudi ArabiaState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaDepartment of Pulmonary and Critical Care Medicine, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, ChinaMonoterpene α-pinene exhibits significant potential as an alternative fuel, widely recognized for its affordability and eco-friendly nature. It demonstrates multiple biological activities and has a wide range of applications. However, the limited supply of pinene extracted from plants poses a challenge in meeting the needs of the aviation industry and other sectors. Considering this, the microbial cell factory is the only viable option for achieving sustainable pinene production. This study employed a rational design model to optimize the copy number and integration site for the heterogenous pinene expression pathway in Escherichia coli (E. coli). The integrated strain with the best pinene pathway PG1 was selected. Subsequently, the resulting strain, E. coli HSY009, accumulated 49.01 mg/L of pinene after 24 h fermentation in the flask culture. To further enhance production, pinene expression cassette PG1 was sequentially integrated into three non-essential regions (44th, 58th, 23rd), resulting in an improved pinene titer. Then, the fermentation process under optimized conditions enhanced the production of pinene to 436.68 mg/L in a 5 L batch fermenter with a mean productivity of 14.55 mg/L/h. To the best of our knowledge, this work represents the maximum mean pinene productivity based on the currently available literature. The findings of this work provide valuable insights for optimizing E. coli to produce other valuable terpenoids that share the same intermediates, IPP and DMAPP. Conclusively, this research validates the model’s universality and highlights its potential for application as cutting-edge biofuel precursors.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1527113/fullbiofuelchromosomal integrationrational design modelpinenefermentationEscherichia coli |
| spellingShingle | Muhammad Hammad Hussain Lu Han Yanlong Wei Muhammad Javid Kamran Ashraf Maria Martuscelli Waleed Aldahmash Meijin Guo Ali Mohsin Zhanxia Li Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene Frontiers in Microbiology biofuel chromosomal integration rational design model pinene fermentation Escherichia coli |
| title | Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene |
| title_full | Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene |
| title_fullStr | Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene |
| title_full_unstemmed | Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene |
| title_short | Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene |
| title_sort | rational engineering of escherichia coli strain for stable and enhanced biosynthesis of pinene |
| topic | biofuel chromosomal integration rational design model pinene fermentation Escherichia coli |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1527113/full |
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