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Rapid construction of E.coli chassis with genome multi-position integration of isopentenol utilization pathway for efficient and stable terpenoid accumulation
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  • Yu-zhou Wang,
  • Hong-yan Jing,
  • Xin Li,
  • Feng Zhang,
  • Xiaoman Sun,
  • He Huang
Yu-zhou Wang
Nanjing Normal University
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Hong-yan Jing
Nanjing Normal University
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Xin Li
Nanjing Normal University
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Feng Zhang
Nanjing Normal University
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Xiaoman Sun
Nanjing Normal University

Corresponding Author:[email protected]

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He Huang
Nanjing Normal University
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The isopentenol utilization pathway (IUP) is potential in terpenoids synthesis. This study aimed to construct IUP-employed E. coli chassis for stably synthesizing terpenoids. As to effectiveness, promotor engineering strategy was employed to regulate IUP expression level, while ribosome-binding site (RBS) library of the key enzyme was constructed for screening the optimal RBS, followed by optimization of concentration of inducer and substrates, the titer of reporting production, lycopene, from 0.087 to 8.67 mg/OD600. As about stability, the IUP expression cassette was integrated into the genome through transposition tool based on CRISPR-associated transposases. Results showed that the strain with 13 copies produced 1.78-fold lycopene titer that of the controlled strain with IUP-harbored plasmid, and it exhibited stable expression after ten successions while the plasmid loss was observed in the controlled strain in the 3rd succession. This strategy provides valuable information for rapid construction of highly effective and stable chassis employing IUP for terpenoids production.
12 Jun 2023Submitted to Biotechnology Journal
12 Jun 2023Submission Checks Completed
12 Jun 2023Assigned to Editor
13 Jun 2023Reviewer(s) Assigned
29 Jun 2023Review(s) Completed, Editorial Evaluation Pending
30 Jun 2023Editorial Decision: Revise Minor
03 Jul 20231st Revision Received
03 Jul 2023Submission Checks Completed
03 Jul 2023Assigned to Editor
03 Jul 2023Reviewer(s) Assigned
17 Jul 2023Review(s) Completed, Editorial Evaluation Pending
19 Jul 2023Editorial Decision: Accept