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Optimization of an inducible, chromosomally encoded benzo [a] pyrene (BaP) degradation pathway in Bacillus subtilis BMT4i (MTCC 9447)

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Abstract

Benzo [a] pyrene (BaP), a pentacyclic polyaromatic hydrocarbon, is 1 of the 12 target compounds defined in the new US Environmental Protection Agency strategy for controlling persistent, bioaccumulative, and toxic pollutants. We previously isolated a novel strain Bacillus subtilis BMT4i capable of utilizing BaP as sole source of carbon and energy. The present study investigated (1) whether the BaP degradation pathway is inducible, and (2) whether it is plasmid-encoded. Furthermore, physical (temperature, pH, and UV-induced photolysis of BaP) and chemical (BaP concentration, surfactant, and ionic strength) parameters for BaP degradation were determined. Our findings revealed a ten-fold enhanced degradation rate in induced vs non-induced culture in the presence of chloramphenicol, suggesting that the BaP degradation pathway is inducible. Physical methods demonstrated the lack of plasmid in BMT4i—a result further complimented by plasmid curing, which had no effect on BaP degradability thus a chromosomal localization can be inferred. Maximum BaP degradation in BMT4i was observed under the following physical and chemical conditions: 30°C, pH 8.0, UV-induced photolysis of BaP-basal salt mineral medium (BSM), 150 μg/ml BaP, 0.01% Tween-20, and 400–1,800 μM MgSO4. These conditions could be beneficial in the development and standardization of effective bioremediation protocols using B. subtilis BMT4i.

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Acknowledgements

This work was supported in part by the Modern Institute of Technology, Rishikesh, Uttarakhand, India and Uttarakhand Council of Science and Technology, Uttarakhand, India (UCOST; grant no. UCS and T/R and D/LS-46/06-07), which are gratefully acknowledged. We also thank Prof Aditya Shastri, Vice Chancellor, Banasthali University, Rajasthan, India for providing facilities in the Department of Bioscience and Biotechnology at this university. We gratefully acknowledge Dr. Yogesh Shouche, Scientist E and Mr. Jay Siddharth, Senior Research Fellow (SRF), Molecular Biology Laboratory, National Centre for Cell Science (NCCS), Pune, India for providing the pulse field gel electrophoresis (PFGE) facility and invaluable help and cooperation during this research work.

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  1. Department of Biotechnology, Modern Institute of Technology (MIT), Dhalwala, Rishikesh, 249201, Uttarakhand, India

    Madhuri Kaushish Lily, Ashutosh Bahuguna & Koushalya Dangwal

  2. Department of Biosciences and Biotechnology, Banasthali University, Rajasthan, India

    Veena Garg

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  1. Madhuri Kaushish Lily
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  2. Ashutosh Bahuguna
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Correspondence to Koushalya Dangwal.

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Lily, M.K., Bahuguna, A., Dangwal, K. et al. Optimization of an inducible, chromosomally encoded benzo [a] pyrene (BaP) degradation pathway in Bacillus subtilis BMT4i (MTCC 9447). Ann Microbiol 60, 51–58 (2010). https://doi.org/10.1007/s13213-009-0010-y

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