Synthesis of Methylated Anthranilate Derivatives Using Engineered Strains of Escherichia coli

Anthranilate derivatives have been used as flavoring and fragrant agents for a long time. Recently, these compounds are gaining attention due to new biological functions including antinociceptive and analgesic activities. Three anthranilate derivatives, N -methylanthranilate, methyl anthranilate, and methyl N -methylanthranilate were synthesized using metabolically engineered stains of Escherichia coli . NMT encoding N -methyltransferase from Ruta graveolens , AMAT encoding anthraniloyl-coenzyme A (CoA):methanol acyltransferase from Vitis labrusca , and pqsA encoding anthranilate coenzyme A ligase from Pseudomonas aeruginosa were cloned and E. coli strains harboring these genes were used to synthesize the three desired compounds. E. coli mutants ( metJ , rpD , yrR mutants), which provide more anthranilate and/or S -adenosyl methionine, were used to increase the production of the synthesized compounds. MS/MS analysis was used to determine the structure of the products. Approximately, 185.3 μM N -methylanthranilate and 95.2 μM methyl N -methylanthranilate were synthesized. This is the first report about the synthesis of anthranilate derivatives in E. coli .