| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Polymerase chain reaction (PCR) amplification, purification, digestion, ligation, transformation, and other routine molecular manipulations were performed according to established protocols. The tea caffeine synthase gene TCS1 from Camellie sinensis, the coffee xanthosine methyltransferase gene CaXMT from Coffea arabica, the methionine adenosyltransferase gene SAM2 fromSaccharomyces cerevisiae, the Vitreoscilla hemoglobin gene vgb, and the guanine deaminase gene, GUD1, from Saccharomyces cerevisiae were codon optimized and then commercially synthesized (Genscript Bio Tech Co., Lid, NJ) to permit high-efficiency expression in E. coli. | Get A Quote |
Caffeine (Cf, 1,3,7-trimethylxanthine), a major secondary metabolite of many higher plants, is widely used in popular non-alcoholic beverages, and in the pharmaceutical and health industries. Currently, this valuable chemical is mainly manufactured by chemical synthesis. In this study, we developed a novel approach for de novo caffeine production in metabolically engineered Escherichia coli. Xanthine-to-caffeine conversion was first achieved by the expression of a plant-derived gene encoding tea caffeine synthase (TCS1). Caffeine accumulation was then increased using two metabolic strategies: higher-level expression of the target enzymes, and enhancement of xanthine and S-adenosyl-L-methionine biosynthesis.... More
