Encapsulating genetically modified Saccharomyces cerevisiae cells in a flow-through device towards the detection of diclofenac in wastewater

• Published in: Journal of biotechnology 2018-10, Vol.284, p.75-83
• Main Authors: Schirmer, C., Posseckardt, J., Kick, A., Rebatschek, K., Fichtner, W., Ostermann, K., Schuller, A., Rödel, G., Mertig, M.
• Format: Article
• Language: English
• Subjects: Diclofenac; Fluorescent protein; Genetically engineered whole-cell biosensor; Microfluidic device; PDMS microchannel; Saccharomyces cerevisiae (yeast)
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2018.08.003

•Novel flow-through biosensor with inclosed genetically modified yeast cells.•Safe enclosure of the sensor cells prevents them from leaving the sensor chambers.•Microfluidic system supplies the cells with nutrients and analyte.•Generation of concentration dependent fluorescence signal upon diclofenac addition.•Only the biologically active fraction of diclofenac is detected. Recently it has been proposed to use sensors based on genetically engineered reporter cells to perform continuous online water monitoring. Here we describe the design, assembly and performance of a novel flow-through device with immobilized genetically modified yeast cells that produce a fluorescent protein upon stimulation with diclofenac whose intensity is then detected by fluorescence microscopy. Although other devices employing immobilized cells for the detection of various analytes have already been described before, as novelty our system allows safe enclosure of the sensor cells, and thus, to obtain fluorescent signals that are not falsified by a loss of cells. Furthermore, the yeast cells are prevented from being released into the environment. Despite the safe containment, the immobilized reporter cells are accessible to nutrients and analytes. They thus have both the ability to grow and respond to the analyte. Both in cell culture medium and standardized synthetic wastewater, we are able to differentiate between diclofenac concentrations in a range from 10 to 100 μM. As particularly interesting feature, we show that only the biologically active fraction of diclofenac is detected. Nowadays, contamination of wastewater with diclofenac and other pharmaceutical residues is becoming a severe problem. Our investigations may pave the way for an easy-to-use and cost-efficient wastewater monitoring method.