Exploring the role of Alk, PAH and CYP153 genes in removing of bitumen contaminants

Document Type : Research Paper

Authors

1 National Center for Laboratories and Construction Research, Wasit Construction Laboratory, AL-Kut, Wasit, Iraq.

2 Department of Biology, College of Science, Wasit University, Iraq.

10.22103/jab.2024.24493.1634

Abstract

Objective
Alkane hydroxylase is an enzyme involved in the first stage of alkane degradation the alkB gene is important for the biodegradation of bitumen because bitumen contains a lot of hydrocarbons such as alkanes, bacteria with the is alkane monooxygenase (AlkB) gene can break it down. The regulatory mechanisms can be intricate and species-specific; there are enzymes encoded by the PAH gene that initiate the breakdown of polycyclic aromatic hydrocarbons (PAHs); and PAHs are dangerous pollutants that can be detected in water and soil. An alkane hydroxylase, specifically a cytochrome P450 enzyme, is encoded by the cytochrome P450 Class I P450 (CYP153) gene. this enzyme is essential for the biodegradation of hydrocarbons, which includes bitumen. Enzymes belonging to the cytochrome P450 family play an important role in the metabolism of many different compounds, including those that are foreign to the body. One of bitumen's main components, alkanes, can be oxidized by the enzyme CYP153. Thus, the aim of this study was to explore the role of Alk, PAH and CYP153 genes in removing of bitumen contaminants.

Materials and Methods
Genomic DNA was extracted using the standard DNA extraction Kit. The quality and quantity of extracted DNA were determined using nanodrop device. The specific primers were used to amplify AlkB, PAHs and Cyp153 genes. Visualization of the amplified fragments was performed using a transilluminator under ultraviolet light and photographed.

Results
Extracted DNA had good quality and quantity (10ng/μL). Alkane monooxygenase (alkB), PAH, and CYP153 are three key enzyme-encoding genes that play an essential role in the mineralization of aliphatic and PAH chemicals, respectively. The presence of these three genes (alkB, PAH, and CYP153) was detected based on PCR amplification and visualized on agarose gel. Frequency of bitumen utilization genes was different. It was the highest for CYP152 gene and the lowest for AlkB gene.

Conclusions
The results highlight the potential for bioremediation applications, especially in bitumen-contaminated areas of employing native bacteria such as Pseudomonas aeruginosa to validate these bacteria's effectiveness in practical settings and create scalable bioremediation techniques for reducing hydrocarbon contamination field research is necessary.

Keywords

References

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Agricultural Biotechnology Journal
Volume 16, Issue 4
January 2025
Pages 377-390

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