Using a Riboswitch Sensor to Examine Coenzyme B12 Metabolism and Transport in E. coli

SummarySmall molecules play crucial roles in every major cellular process. Despite this, detecting their levels within cells remains a significant challenge. Here, we describe intracellular sensors of coenzyme B12 that make use of the exquisite molecular detection capabilities of a naturally occurring riboswitch. These probes sensitively detect their target using colorimetric, fluorescent, or luminescent reporters. To assess their utility in the study of biological systems, the sensors were applied to examine the synthesis and the import of coenzyme B12. The sensors were able to monitor the effects of genetic deletions, recombinant expression of foreign genes, and varied growth conditions on both of these processes. These results indicate that riboswitch-based sensors can provide valuable information on intracellular small molecule concentrations that can be employed in the study of related cellular processes.

► Intracellular coenzyme B12 sensors constructed using riboswitches ►Sensors are flexible, specific, and sensitive at physiological range ►Sensors applied to study coenzyme B12 transport and metabolism ►Genetic factors and growth conditions affecting these processes effectively detected