Development of a biosensor for on-line detection of tributyltin with a recombinant bioluminescent Escherichia coli strain

• Published in: Applied microbiology and biotechnology 2003-08, Vol.62 (2-3), p.218-225
• Main Authors: THOUAND, G, HORRY, H, DURAND, M. J, PICART, P, BENDRIAA, L, DANIEL, P, DUBOW, M. S
• Format: Article
• Language: English
• Subjects: Analysis; Antifouling substances; ATP; Bacteria; Bacterial physiology; Biological and medical sciences; Bioluminescence; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biosensors; Biotechnology; Dissolved oxygen; E coli; Environmental Engineering; Environmental Sciences; Escherichia coli; Escherichia coli - genetics; Escherichia coli - growth & development; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Life Sciences; Luciferases - genetics; Luminescent Measurements; Methods. Procedures. Technologies; Microbial activity; Molecular microbiology; Observations; Online Systems; Recombination, Genetic; Trialkyltin Compounds - analysis; Tributyltin; Various methods and equipments; Vibrio - enzymology; Vibrio - genetics
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-003-1279-6

A biosensor was developed for the detection of tributyltin (TBT), using a bioluminescent recombinant Escherichia coli:: luxAB strain. Dedicated devices allowed the on-line measurement of bioluminescence, pH and dissolved oxygen values and the feed-back regulation of temperature. Bacterial physiology was monitored by the measurement of the cellular density, respiratory activity and the intracellular level of ATP, glucose and acetate levels. Our results showed that a synthetic glucose medium gave a better TBT detection limit than LB medium (respectively 0.02 micro M and 1.5 micro M TBT). High growth and dilution rates ( D=0.9 h(-1)) allowed maximum light emission from the bacterium. Moreover, simple atmospheric air bubbling was sufficient to provide oxygen for growth and the bioluminescence reaction. Real-time monitoring of bioluminescence after TBT induction occurred with continuous addition of decanal up to 300 micro M, which was not toxic throughout a 7-day experiment. The design of our biosensor and the optimization of the main parameters that influence microbial activity led to the capacity for the detection of TBT.