Attenuation of petroleum hydrocarbon fractions using rhizobacterial isolates possessing alkB, C23O, and nahR genes for degradation of n-alkane and aromatics

Abstract

This work assessed the catabolic versatility of functional genes in hydrocarbon-utilizing bacteria obtained from the rhizosphere of plants harvested in aged polluted soil sites in Ogoni and their attenuation efficacy in a bioremediation study. Rhizosphere soil was enumerated for its hydrocarbon-utilizing bacteria. The bacteria were in-vitro screened and selected through the quantification of their total protein and specific intermediate pathway enzyme (catechol 2,3-dioxygenase) activity in the metabolism of hydrocarbon. Thereafter, agarose gel electrophoresis technique was deployed to profile the genome of the selected strains for catechol 2,3-dioxygenase (C23O), 1,2-alkane monooxygenase (alkB), and naphthalene dioxygenase (nahR). Four rhizobacterial isolates namely Pseudomonas fluorescens (A3), Achromobacter agilis (A4), Bacillus thuringiensis (D2), and Staphylococcus lentus (L1) were selected based on the presence of C23O, alkB, and nahR genes. The gel electrophoresis results showed an approximate molecular weight of 200 bp for alkB, 300 bp for C23O, and 400 bp for nahR. The gas chromatogram for residual total petroleum hydrocarbon (TPH) revealed mineralization of fractions C₈–C₁₇, phytane, C₁₈–C₃₀. TPH for in-vitro bioremediation of crude oil-polluted soil was observed to have an optimal reduction/loss of 97% within the 56th day of the investigation. This study has further revealed that the microbiome of plants pre-exposed to crude oil pollution could serve as a reservoir for mining group of bacterial with broad catabolic potentials for eco-recovery and waste treatment purposes.

Keywords:

• Agarose gel electrophoresis
• catechol 2,3-dioxygenase
• 1,2-alkane monooxygenase
• naphthalene dioxygenase
• bioremediation
• attenuation

Acknowledgments

The authors are grateful to the Department of Microbiology, University of Port Harcourt, Rivers State, Nigeria for the laboratory space and Dr. Enobong Effiong for his immense contributions during the laboratory stage of this work.