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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Microbiotechnology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1527113

Rational engineering of Escherichia coli strain for stable and enhanced biosynthesis of pinene Authors

Provisionally accepted
Muhammad Hammad Hussain Muhammad Hammad Hussain 1Lu Han Lu Han 1Yanlong Wei Yanlong Wei 1Muhammad Javid Muhammad Javid 2Kamran Ashraf Kamran Ashraf 1Maria Martuscelli Maria Martuscelli 3Waleed Aldahmash Waleed Aldahmash 4Guo Meijin Guo Meijin 1Ali Mohsin Ali Mohsin 1*Zhanxia Li Zhanxia Li 5*
  • 1 School of Biotechnology, East China University of Science and Technology, Shanghai, China
  • 2 Abdul Wali Khan University Mardan, Mardan, Khyber Pakhtunkhwa, Pakistan
  • 3 University of Teramo, Teramo, Abruzzo, Italy
  • 4 King Saud University, Riyadh, Riyadh, Saudi Arabia
  • 5 Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, Shanghai Municipality, China

The final, formatted version of the article will be published soon.

    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.

    Keywords: biofuel, Chromosomal integration, Rational design model, pinene, Fermentation

    Received: 12 Nov 2024; Accepted: 12 Dec 2024.

    Copyright: © 2024 Hussain, Han, Wei, Javid, Ashraf, Martuscelli, Aldahmash, Meijin, Mohsin and Li. 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) or licensor 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.

    * Correspondence:
    Ali Mohsin, School of Biotechnology, East China University of Science and Technology, Shanghai, China
    Zhanxia Li, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, 200233, Shanghai Municipality, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.