Establishing an Autonomous Cascaded Artificial Dynamic (AutoCAD) regulation system for improved pathway performance

Area of Science:

• Synthetic biology
• Metabolic engineering
• Microbial biotechnology

Background:

• Microbial metabolic pathways require precise regulation for efficient biosynthesis.
• Introducing heterologous pathways often leads to unbalanced enzyme expression and carbon flux.
• This imbalance hinders the development of highly efficient microbial biosynthesis systems.

Purpose of the Study:

• To develop an Autonomous Cascaded Artificial Dynamic (AutoCAD) regulation system.
• To automatically coordinate heterologous pathway expression and redirect carbon fluxes.
• To enhance naringenin production in microbial hosts.

Main Methods:

• Designed an AutoCAD regulation system with intermediate-based feedforward and product-based feedback genetic circuits.
• Implemented the AutoCAD system in microbial hosts for naringenin biosynthesis.
• Conducted fed-batch fermentation to evaluate naringenin production.

Main Results:

• The AutoCAD system significantly enhanced naringenin production.
• Achieved a 16.5-fold increase in naringenin titer compared to static controls.
• Reached a final naringenin titer of 277.2 mg/L through fed-batch fermentation.

Conclusions:

• The AutoCAD regulation system provides effective dynamic control for heterologous pathways.
• This approach offers a new paradigm for engineering complex cascade dynamic control systems.
• Enables enhanced production of target compounds like naringenin in microbial systems.