Microbial production of creatine using growth-coupled selection systems.

Area of Science:

• Biotechnology
• Metabolic Engineering
• Synthetic Biology

Background:

• Creatine is essential for energy storage and muscle development, particularly valuable as a supplement for plant-based diets.
• Chemical synthesis of creatine has limitations, necessitating sustainable alternatives.

Purpose of the Study:

• To develop a novel biosynthetic process for creatine production using an engineered Escherichia coli cell factory.
• To overcome metabolic bottlenecks and enhance creatine yield through a combination of computational biology and adaptive laboratory evolution.

Main Methods:

• Engineered Escherichia coli to express a heterologous pathway for creatine synthesis.
• Employed a model-driven growth-coupled selection approach and adaptive laboratory evolution.
• Utilized genome-scale modeling to optimize the glycine amidinotransferase step and improve creatine tolerance.

Main Results:

• Achieved a 58% increase in creatine titer compared to the baseline strain, starting from glycine and arginine.
• Successfully overcame metabolic bottlenecks in the heterologous synthesis pathway.
• Developed an engineered E. coli strain with enhanced creatine production capabilities.

Conclusions:

• The study demonstrates an efficient cell factory generation strategy by combining production with growth, driven by evolutionary engineering and computational biology.
• This biosynthetic approach offers a sustainable and effective alternative to chemical synthesis for creatine production.
• Highlights the power of integrating computational modeling and adaptive evolution for optimizing microbial cell factories.