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Caffeine addicted bacteria—an iGEM project

Quandt EM, Hammerling MJ, et al. (2013) Decaffeination and measurement of caffeine content by addicted Escherichia coli with a refactored N-demethylation operon from Pseudomonas putida CBB5. ACS Synth Biol. DOI: 10.1021/sb4000146.

Citation summary: These researchers use IDT gBlocks® Gene Fragments in Gibson Assembly® reactions to generate some of the plasmid constructs in this work. See how these custom, double-stranded DNA fragments can streamline your research.

As part of an undergraduate project for the International Genetically Engineered Machine (iGEM) competition, this research group from the University of Texas at Austin imported a refactored N-demethylation operon that provides a caffeine degradation pathway from Pseudomonas putida CBB5 to E. coli, an organism that is more amenable to genetic engineering and industrial applications. The E. coli strain was a guaB knockout that required xanthine, a precursor of guanine, to grow. Xanthine is produced by N-demethylation of caffeine; thus, these cells required caffeine to survive.

IDT gBlocks Gene Fragments were used in Gibson Isothermal Assembly reactions to generate some of the plasmid constructs within this work. In addition to characterizing the sequence elements regulating the caffeine degradation pathway, the researchers were able to use the growth yield of the “addicted” E. coli strain to precisely measure caffeine concentrations, for example, in sodas and energy drinks. The synthetic N-demethylation operon has applications for cheaply producing pharmaceuticals or their precursor molecules, and for detoxifying waste that could subsequently be recycled into animal feed and biofuels.

For years, IDT has supported iGEM participants in their efforts to create solutions to all kinds environmental, medical, and technical problems. Support comes in the form of custom, high quality, double-stranded DNA fragments—gBlocks Gene Fragments. In 2016, over 250 teams took advantage of our offer to receive up to 20 kb of gBlocks Gene Fragments for free. Find out more at

Published Nov 9, 2017