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A Single-Component Blue Light-Induced System Based on EL222 in Yarrowia lipolytica
Zhiqian Wang1, Yunjun Yan1, Houjin Zhang1
1Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, MOE Key Laboratory of Molecular Biophysics, Wuhan 430074, China.
International Journal of Molecular Sciences
|June 10, 2022
Summary
Researchers developed a novel optogenetic system for Yarrowia lipolytica, enabling precise control over gene expression using blue light. This light-induced system successfully synthesized and verified the Bleomycin resistance protein (BleoR).
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Optogenetics
Background:
- Optogenetics offers precise control over biological systems due to its rapid response, reversibility, and high resolution.
- Metabolic engineering of microbial chassis cells benefits from advanced control strategies like optogenetics.
Purpose of the Study:
- To construct a blue light-induced gene expression system in Yarrowia lipolytica.
- To demonstrate the system's capability for synthesizing and functionally verifying proteins, such as Bleomycin resistance protein (BleoR).
Main Methods:
- Engineered a light-responsive element (TF) using EL222 and VP16 for blue light sensing.
- Integrated TF with an upstream activation sequence (C120)5 and minimal promoter CYC102 to create a light-inducible promoter.
- Utilized GFPMut3 as a reporter gene to validate system response to blue light induction.
- Assembled four copies of the responsive promoter and reporter gene for enhanced signal output.
- Investigated the impact of light dose and periodicity on system performance.
- Applied the light-controlled system for BleoR synthesis and verification.
Main Results:
- The constructed light-induced sensor responded effectively to blue light, initiating gene expression.
- A 128.5-fold increase in fluorescent signal was observed with four copies of the promoter and reporter gene after 8 hours of induction compared to dark conditions.
- The system demonstrated good spatial and temporal controllability under varying light conditions.
- Successful synthesis and functional verification of BleoR were achieved using the light-controlled system.
Conclusions:
- The developed optogenetic system provides robust transcriptional regulation in Yarrowia lipolytica.
- This system holds potential for constructing large-scale synthetic networks and overproducing target products.
- Optogenetics offers a powerful tool for precise metabolic engineering and synthetic biology applications.
