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Dehalogenase gene detection and microbial diversity of a chlorinated hydrocarbon contaminated site

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Abstract

In recent years large quantities of mixtures of chlorinated hydrocarbons have accumulated in the environment due to the widespread use and production of these compounds. Microbes have been found to demonstrate a widespread and diverse potential to adapt to the dechlorination of such compounds. Therefore the aim of this study was to investigate the presence and diversity of reductive and hydrolytic dehalogenase genes in a site contaminated with a mixture of chlorinated hydrocarbons. Primers targeting reductive and hydrolytic bacterial dehalogenase genes were designed. In addition, DGGE analysis was performed in order to determine the presence of any known dehalogenase-producing organisms. Total DNA isolated from borehole water samples was used as the template for the amplification reactions. All PCR products obtained with the reductive and hydrolytic gene primers, as well as the dominant bands present on the DGGE gel were cloned and sequenced. Sequencing of the individual amplicons revealed significant identities to the tceA gene of Dehalococcoides ethenogenes 195, the vcrA gene of Dehalococcoides sp. VS as well as the dhlA and dhlB genes of Xanthobacter autotrophicus GJ10. DGGE analysis indicated a high level of commonality with the different sampling times and depths. However, sequence analysis revealed that 66% of the cloned fragments showed significant (95–99%) identity with uncultured microorganisms. Phylogenetic analysis of the sequences revealed that the DGGE clones clustered into two groups when compared to known bacteria having hydrocarbon degradative capabilities. This indicated that the sequences of the clones were diverse when compared to known microorganisms. This diversity represents a largely untapped genetic pool that can be exploited for the discovery of novel biocatalysts that can be employed in bioremediation. In addition, the presence of both hydrolytic and reductive dehalogenases provided strong evidence that bacteria capable of dehalogenation of chlorinated hydrocarbons may be present in sites contaminated with these compounds.

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Acknowledgments

The authors wish to thank the National Research Foundation of South Africa for financial support and Inqaba Biotechnical Industries (Pty) Ltd (Pretoria, South Africa) for performing the sequencing reactions.

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Authors and Affiliations

  1. Discipline of Microbiology, Faculty of Science and Agriculture, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban, 4000, Republic of South Africa

    Algasan Govender, Rehana Shaik, Nathlee Samantha Abbai & Balakrishna Pillay

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  1. Algasan Govender
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  2. Rehana Shaik
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  3. Nathlee Samantha Abbai
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  4. Balakrishna Pillay
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Correspondence to Balakrishna Pillay.

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Govender, A., Shaik, R., Abbai, N.S. et al. Dehalogenase gene detection and microbial diversity of a chlorinated hydrocarbon contaminated site. World J Microbiol Biotechnol 27, 2407–2414 (2011). https://doi.org/10.1007/s11274-011-0713-7

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  • DOI: https://doi.org/10.1007/s11274-011-0713-7

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