Exploiting nongenetic cell-to-cell variation for enhanced biosynthesis.

Area of Science:

• Metabolic Engineering
• Synthetic Biology
• Biotechnology

Background:

• Biosynthesis offers renewable chemical production but often requires performance improvements for economic viability.
• Nongenetic variations within cell populations naturally lead to high- and low-performing variants.
• Leveraging high-performing variants is key to enhancing overall biosynthetic efficiency.

Purpose of the Study:

• To develop and implement an in vivo method for continuous selection of high-performing, nongenetic cellular variants.
• To improve the economic feasibility of biosynthetic processes through enhanced production yields.
• To demonstrate the broad applicability of the developed method across different metabolic pathways.

Main Methods:

• Development of in vivo population quality control (PopQC) for real-time selection of superior cellular variants.
• Application of PopQC to two distinct biosynthetic pathways, free fatty acid (FFA) and tyrosine production.
• Utilizing two alternative design principles within the PopQC framework.

Main Results:

• Achieved a threefold increase in the production of both free fatty acid (FFA) and tyrosine.
• Confirmed that PopQC enhances ensemble biosynthesis by effectively selecting for nongenetic high performers.
• Demonstrated high performance in fed-batch FFA production with a titer of 21.5 g/L and productivity of 0.5 g/L/h.

Conclusions:

• In vivo population quality control (PopQC) is an effective strategy for enhancing microbial biosynthesis.
• The PopQC method significantly improves yields for target compounds by exploiting inherent nongenetic cellular variation.
• PopQC holds broad potential for optimizing diverse metabolic pathways in various biotechnological applications.