ATP biosensor reveals microbial energetic dynamics and facilitates bioproduction

Area of Science:

• Metabolic Engineering and Synthetic Biology
• Microbial Biotechnology
• Cellular Energetics

Background:

• Adenosine-5'-triphosphate (ATP) is the central energy currency in cells, crucial for metabolic activities and biosynthesis.
• The role of intracellular ATP dynamics in microbial bioproduction and strategies for its enhancement are not well understood.
• Optimizing ATP levels could significantly improve the efficiency of producing valuable biomaterials and chemicals.

Purpose of the Study:

• To investigate the impact of intracellular ATP dynamics on bioproduction across different microbial strains and conditions.
• To identify specific growth phases and carbon sources that influence ATP levels and correlate with product formation.
• To utilize ATP dynamics as a diagnostic tool for metabolic burden assessment and bioproduction optimization.

Main Methods:

• Utilized a novel ATP biosensor to monitor intracellular ATP dynamics in real-time.
• Analyzed ATP fluctuations across various growth phases (exponential, stationary) and with different carbon sources (acetate, oleate).
• Quantified the production of key biomolecules, such as fatty acids (FA) and polyhydroxyalkanoates (PHA), in relation to ATP levels.
• Assessed metabolic burden and identified bottlenecks in limonene bioproduction using ATP dynamics.

Main Results:

• Observed transient ATP accumulations during the transition from exponential to stationary growth phases.
• Correlated ATP accumulation with increased production of fatty acids in Escherichia coli and polyhydroxyalkanoates in Pseudomonas putida.
• Identified acetate (for E. coli) and oleate (for P. putida) as carbon sources that elevate steady-state ATP levels and enhance FA/PHA production.
• Demonstrated the utility of ATP dynamics in diagnosing metabolic bottlenecks limiting limonene bioproduction.

Conclusions:

• Intracellular ATP dynamics are intrinsically linked to the efficiency of microbial bioproduction.
• Specific carbon sources and growth phase transitions can be manipulated to optimize ATP levels for enhanced production of valuable compounds.
• ATP dynamics serve as a valuable indicator for assessing metabolic burden and guiding metabolic engineering strategies for improved bioproduction.