Fermentative N-Methylanthranilate Production by Engineered Corynebacterium glutamicum

Area of Science:

• Biotechnology
• Metabolic Engineering
• Synthetic Biology

Background:

• N-methylanthranilate (NMA) is a key precursor for valuable bioactive compounds.
• Current NMA synthesis methods are often costly and inefficient.
• Fermentation using Corynebacterium glutamicum offers a scalable alternative for amino acid production.

Purpose of the Study:

• To develop a metabolically engineered Corynebacterium glutamicum strain for the fermentative production of N-methylanthranilate (NMA).
• To establish a novel biocatalytic route for NMA synthesis using heterologous gene expression.

Main Methods:

• Metabolic engineering of C. glutamicum to enhance anthranilate supply via the shikimate pathway.
• Heterologous expression of anthranilate N-methyltransferase (anmt) from Ruta graveolens.
• Coexpression of adenosylhomocysteinase (sahH) to improve S-adenosyl methionine regeneration.

Main Results:

• Engineered C. glutamicum produced NMA, which accumulated in the culture medium.
• Coexpression of sahH significantly improved NMA production titers.
• A C1* strain achieved a final NMA titer of 0.5 g/L with a yield of 4.8 mg/g glucose in bioreactor culture.

Conclusions:

• Metabolic engineering and heterologous gene expression provide a viable strategy for NMA production in C. glutamicum.
• This biocatalytic approach offers a sustainable and potentially cost-effective alternative to traditional synthesis methods.
• Further optimization could lead to industrial-scale NMA production for bioactive compound synthesis.