Repetitive Detection of Aromatic Hydrocarbon Contaminants with Bioluminescent Bioreporters Attached on Tapered Optical Fiber Elements

• Main Authors: Jakub Zajíc, Steven Ripp, Josef Trögl, Gabriela Kuncová, Marie Pospíšilová
• Format: Article
• Language: English
• Published: MDPI AG 2020-06-01
• Series: Sensors
• Subjects: whole-cell biosensor; bioluminescent bioreporter; optical fiber biosensor; toluene; <i>Pseudomonas putida</i> TVA8; <i>Escherichia coli</i> 652T7
• Online Access: https://www.mdpi.com/1424-8220/20/11/3237

In this study, we show the repetitive detection of toluene on a tapered optical fiber element (OFE) with an attached layer of <i>Pseudomonas putida</i> TVA8 bioluminescent bioreporters. The bioluminescent cell layer was attached on polished quartz modified with (3-aminopropyl)triethoxysilane (APTES). The repeatability of the preparation of the optical probe and its use was demonstrated with five differently shaped OFEs. The intensity of measured bioluminescence was minimally influenced by the OFE shape, possessing transmittances between 1.41% and 5.00%. OFE probes layered with <i>P. putida</i> TVA8 were used to monitor liquid toluene over a two-week period. It was demonstrated that OFE probes layered with positively induced <i>P. putida</i> TVA8 bioreporters were reliable detectors of toluene. A toluene concentration of 26.5 mg/L was detected after <30 min after immersion of the probe in the toluene solution. Additional experiments also immobilized constitutively bioluminescent cells of <i>E. coli</i> 652T7, on OFEs with polyethyleneimine (PEI). These OFEs were repetitively induced with Lauria-Bertani (LB) nutrient medium. Bioluminescence appeared 15 minutes after immersion of the OFE in LB. A change in pH from 7 to 6 resulted in a decrease in bioluminescence that was not restored following additional nutrient inductions at pH 7. The <i>E. coli</i> 652T7 OFE probe was therefore sensitive to negative influences but could not be repetitively used.