RESUMO
1-Methylxanthine is a high-value derivative of caffeine of limited natural availability with many potential pharmaceutical applications. Unfortunately, production of 1-methylxanthine through purely chemical methods of synthesis are unfavorable due to lengthy chemical processes and the requirement of hazardous chemicals, ultimately resulting in low yields. Here, we describe a novel biosynthetic process for the production of 1-methylxanthine from theophylline using engineered Escherichia coli whole-cell biocatalysts and reaction optimization. When scaled-up to 1590â¯mL, the simple biocatalytic reaction produced approximately 1188â¯mg 1-methylxanthine from 1444â¯mg theophylline, constituting gram-scale production of 1-methylxanthine in as little as 3â¯hours. Following HPLC purification and solvent evaporation, 1163â¯mg of dried 1-methylxanthine powder was collected, resulting in a 97.9â¯wt% product recovery at a purity of 97.8%. This is the first report of a biocatalytic process designed specifically for the production and purification of the high-value biochemical 1-methylxanthine from theophylline. This process is also the most robust methylxanthine N-demethylation process featuring engineered E. coli to date, capable of gram-scale production.