International Journal of Modern Biological Research
ISSN: 2053-180X
Vol. 2(4), pp. 31-38, November 2014
doi.org/10.33500/ijmbr.2014.2.005



Effect of oxygen on bioluminescence emitted by the biosensor Pseudomonas putida HK-44 in presence of phenanthrene

J. Meléndez-Estrada1* and and R. Rodríguez-Vázquez2

1Escuela Superior de Ingeniería y Arquitectura- Unidad Zacatenco del IPN. Av. Juan de Dios Bátiz S/N, Edificio de Posgrado e Investigación. Unidad Profesional “Adolfo López Mateos” Zacatenco. Z. C. 07738. Mexico City.
2Departmento de Biotecnología y Bioingeniería, CINVESTAV-IPN, Instituto Politécnico Nacional 2508, Apartado Postal 14-740, Col. San Pedro Zacatenco Z. C. 07360, Mexico City.

*To whom correspondence should be addressed. E-mail: jmelende31@hotmail.com.

Received 11 September, 2014; Received in revised form 03 October, 2014; Accepted 08 October, 2014.

Abstract


Key words:
Pseudomonas putida HK-44, Oxygen tension, Phenanthrene, Biosensor, Bioluminescence.


Pseudomonas putida HK-44, a bioluminescent reporter strain with a nah-lux fusion, can be used to assess the presence of polynuclear aromatic hydrocarbons by stimulating the light production upon inducing the naphthalene dioxygenase system. However, this assay can be affected by the presence of oxygen. The aim of this work was to evaluate the effect of different oxygen tensions on luminescence emission by HK-44 in presence of phenanthrene. The relationship between light emission and bacterial cell density was positively correlated (P<0.05). Light emission produced by the strain HK-44 was enhanced in the presence of 2 ppm of phenanthrene and a maximum light emission was observed at the highest oxygen tension, this is because the bioluminescence reaction requires oxygen. Data fitted to a Stern-Volmer model at longer periods of time (starting from 150 min) with phenanthrene were observed to have decreased bioluminescence (Kq) at 2 to 10% oxygen tensions, which indicated a possible luminescence quenching by the oxygen. However, at 20% of oxygen saturation, bioluminescence of biosensor was not affected, therefore this biosensor is reliable for polycyclic aromatic hydrocarbons (PAHs) detection under this oxygen saturation conditions.

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