Isolation and characterization of a novel strain, Bacillus sp KJ629314, with a high potential to aerobically degrade diesel

Aislamiento y caracterización de una nueva cepa, Bacillus sp KJ629314, con un alto potencial en la degradación aeróbica de diésel

S. Cisneros-de La Cueva, M.A. Martínez-Prado*, J.A. Rojas-Contreras, H. Medrano-Roldán, and M.A. Murillo-Martínez

Technological Institute of Durango, Felipe Pescador Blvd. # 1830 East. Nueva Vizcaya, Zip Code 34080, Durango, Dgo., Mexico. *Corresponding author. E-mail: adriana.martinez@orst.edu.

Abstract

In this research, a diesel-degrading bacterium (strain KJ629314) was isolated from a mining soil contaminated with total petroleum hydrocarbons (TPH) and properly characterized using the polymerase chain reaction (PCR) molecular technique. The 16s rDNA sequence analysis allowed to identify KJ629314 as a strain of Bacillus sp. Experimental phase was conducted to assess the aerobic biodegradation of diesel; to determine the removal effciency and the corresponding microbial growth; diesel was used as a substrate - electron donor - carbon source; and oxygen (via aeration) as the electron acceptor. Tests were conducted in microcosms with sterile sand with nutrients according to the Nitrogen:Phosphorus ratio of 15:1 at different diesel concentrations (10,000; 20,000; 30,000; 40,000 and 50,000 mg/kg soil). Results showed that the strain of Bacillus sp KJ629314 has a high potential in the biodegradation of diesel at the evaluated concentrations, and it was demonstrated that the removal efficiency was greater at low concentrations of diesel obtaining higher values for the microbial growth and diesel biodegradation rate constants. These promising results support the fact that Bacillus sp KJ629314 may be used as a novel biological resource to develop a bioprocess for the bioremediation of diesel-contaminated soil.

Keywords: aerobic degradation, Bacillus sp KJ629314, bioremediation, contaminated soils, mining industry, TPH.

Resumen

En esta investigación, una bacteria degradadora de diésel (cepa KJ629314) fue aislada de un suelo minero contaminado con hidrocarburos totales de petróleo (HTP) y debidamente caracterizada utilizando la técnica molecular de la reacción en cadena de la polimerasa (RCP). El análisis de la secuencia de 16s rDNA permitió identificar KJ629314 como una cepa de Bacillus sp. La fase experimental se llevó a cabo para evaluar la biodegradación aeróbica del diésel; para determinar la eficiencia de remoción y el crecimiento microbiano correspondiente; el diésel se utilizó como sustrato - donador de electrones - fuente de carbono, y el oxígeno (a través de la aireación) como aceptor de electrones. Las pruebas se realizaron en microcosmos empleando arena estéril con nutrientes de acuerdo con la relación Nitrógeno:Fósforo de 15:1 a diferentes concentraciones de diésel (10,000; 20,000; 30,000; 40,000 y 50,000 mg/kg de suelo). Los resultados mostraron que la cepa de Bacillus sp KJ629314 tiene un alto potencial en la biodegradación del diésel en las concentraciones evaluadas, y se demostró que la eficiencia de degradación fue mayor a bajas concentraciones de diésel; obteniéndose valores más altos para el crecimiento microbiano así como para las constantes de velocidad de degradación. Estos prometedores resultados apoyan el hecho de que el Bacillus sp KJ629314 puede ser utilizado como un recurso biológico novedoso para desarrollar un bioproceso para la biorremediación de suelos contaminados con diésel.

Palabras clave: Bacillus sp KJ629314, biorremediación, degradación aeróbica, HTP, industria minera, suelos contaminados.

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