Pathway Engineering for Phenethylamine Production in Escherichia coli

Area of Science:

• Metabolic Engineering
• Synthetic Biology
• Biotechnology

Background:

• Phenethylamine is a valuable bioproduct with diverse applications.
• Efficient microbial production of phenethylamine is crucial for industrial viability.

Purpose of the Study:

• To engineer a robust Escherichia coli (E. coli) strain for high-yield phenethylamine production.
• To reconstruct and optimize the phenethylamine biosynthetic pathway in E. coli.

Main Methods:

• Chromosomal integration and overexpression of key genes, including Enterococcus faecium pdc (phenylalanine decarboxylase).
• Sequential overexpression of phenethylamine pathway genes (aroG, aroL, pheA, tyrB).
• Modification of the phosphotransferase system (ptsH, ptsI, crr deletion; galP, glk overexpression) and pentose phosphate pathway (zwf overexpression).

Main Results:

• The engineered E. coli strain AUD9 achieved 2.65 g/L phenethylamine in batch fermentation with a yield of 0.27 g/g glucose.
• Fed-batch fermentation of AUD9 resulted in 38.82 g/L phenethylamine with a productivity of 1.08 g/L/h.

Conclusions:

• The engineered E. coli strain AUD9 demonstrates significant potential for industrial-scale phenethylamine production.
• Metabolic engineering strategies successfully optimized phenethylamine biosynthesis in E. coli.