- Volume 34 Issue 3
BACKGROUND: Recent studies have described the importance of bacteria that can degrade polycyclic aromatic hydrocarbons (PAHs). Here we screened bacterial isolates from commercial gasoline for PAH degraders and characterized their ability to degrade PAHs, lipids and proteins as well as their enantioselective epoxide hydrolase activity, salt tolerance, and seawater survival. METHODS AND RESULTS: One hundred two bacteria isolates from commercial gasoline were screened for PAH degraders by adding selected PAHs on to the surface of agar plates by the sublimation method. A clear zone was found only around the colonies of PAH degraders, which accounted for 13 isolates. These were identified as belonging to Bacillus sp., Brevibacterium sp., Micrococcus sp., Corynebacterium sp., Arthrobacter sp., and Gordonia sp. based on 16S rRNA sequences. Six isolates belonging to Corynebacterium sp., 3 of Micrococcus sp., Arthrobacter sp. S49, and Gordonia sp. H37 were lipid degraders. Arthrobacter sp. S49 was the only isolate showing high proteolytic activity. Among the PAH-degrading bacteria, Arthrobacter sp. S49, Brevibacterium sp. S47, Corynebacterium sp. SK20, and Gordonia sp. H37 showed enantioselective epoxide hydrolase activity with biocatalytic resolution of racemic styrene oxide. Among these, highest enantioselective hydrolysis activity was seen in Gordonia sp. H37. An intrinsic resistance to kanamycin was observed in most of the isolates and Corynebacterium sp. SK20 showed resistance to additional antibiotics such as tetracycline, ampicillin, and penicillin. CONCLUSION: Of the 13 PAH-degraders isolated from commercial gasoline, Arthrobacter sp. S49 showed the highest lipid and protein degrading activity along with high active epoxide hydrolase activity, which was the highest in Gordonia sp. H37. Our results suggest that bacteria from commercial gasoline may have the potential to degrade PAHs, lipids, and proteins, and may possess enantioselective epoxide hydrolase activity, high salt tolerance, and growth potential in seawater.
Biodegradation;Commercial gasoline;Polycyclic Aromatic Hydrocarbon (PAH);PAH-degrading bacteria
- Alley, J. F., & Brown, L. R. (2000). Use of sublimation to prepare solid microbial media with water insoluble substrates. Applied and Environmental Microbiology. 66(1), 439-442. https://doi.org/10.1128/AEM.66.1.439-442.2000
- Bezalel, L., Hadar, Y., & Cerniglia, C. E. (1997). Enzymatic mechanisms involved in phenanthrene degradationby the white rot fungus Pleurotus ostreatus. Applied and Environmental Microbiology. 63(7), 2495-2501.
- Hwang, S. S., & Song, H. G. (1999). Biodegradation of pyrene in marine environment. Korean Journal of Microbiology. 35(1), 53-60.
- Hilyard, E. J., Jones-Meehan, J. M., Spargo, B. J., & Hill, R. T. (2008). Enrichment, isolaton, and phylogenetic identification of polycyclic aromatic hydrocarbon-degrading bacteria form Elizabeth river sediments. Applied and Environmental Microbiology. 74(4), 1176-1182. https://doi.org/10.1128/AEM.01518-07
- Janda-Ulfig, K., Ulfig, K., Cano, J., & Guarro, J. (2008). A study of the growth of Pseudaallescheria boydii isolates from sewage sludge and clinical sources of tributyrin, rapeseed oil, biodiesel oil and diesel oil. Annals of Agricultural and Environmental Medicine. 15(1), 45-49.
- Kallimanis, A., Frillingos, S., Drainas, C., & Koukkou, A. I. (2007). Taxonomic identification, phenantrene uptake activity, and membrane lipid alterations of the PAH degrading Arthrobacter sp. strain Sphe3. Applied and Environmental Microbiology. 76(3), 709-717.
- Kanaly, R. A., & Harayama, S. (2000). Biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons by bacteria. Journal of Bacteriology. 182(8), 2059-2067. https://doi.org/10.1128/JB.182.8.2059-2067.2000
- Kazunga, C., & Aitken, M. D. (2000). Products from the incomplete metabolism of pyrene by polycyclic aromatic hydrocarbon-degrading bacteria. Applied and Environmental Microbiology. 66(5), 1917-1922. https://doi.org/10.1128/AEM.66.5.1917-1922.2000
- Kim, G. E. (2014). Isolation of protease producing microorganisms. Journal of Korean Society of Environmental Engineering. 36(4), 265-270. https://doi.org/10.4491/KSEE.2014.36.4.265
- Kim, J. Y., & Lee., S. S. (2008). Biodegradation of kerosene by Pseudomonas aeruginosa K14. Korean Journal of Microbiology. 44(2), 156-163.
- Kim, J. Y., Hu, C. G., Lee, M. G., & Kam, S. G. (2003a). Photodegradation of pyrene, chrysene and benzo[a]pyrene in water. Journal of Environmental Sciences. 12(3), 337-344. https://doi.org/10.5322/JES.2003.12.3.337
- Kim, S. J., Kwon, O., Jones, R.C., Freeman, J.P., Edmondson, R.D., & Cerniflia, C.E. (2007). Complete and integrated pyrene degradation pathway in Mycobacterium vanbaalenii PYR-1 based on systems biology. Journal of Bacteriology. 189(2), 464-472. https://doi.org/10.1128/JB.01310-06
- Kim, S. H., Kang, S. M., Oh, K. H., Kim, S. I., Yoon, B. J., & Khang, H. Y. (2005) Characterization of PAH-Degrading bacteria from soils of reed rhizosphere in Suncheon bay using PAH consortia. Korean Journal of Microbiology. 41(3), 208-215.
- Kim, T. J., Jo, G. S., & Ryu, H. U. (2003b). Degradation of phenanthren and pyrene by Burkholderia sp. D5. Korean Journal of Microbiology. 39(4), 267-271.
- Kim, Y. H., & Freeman., J. P. (2005). Effects of pH on the degradation of phenanthrene and pyrene by Mycobacterium vanbaalenii PYR-1. Applied Microbiology and Biotechnology. 67(2), 275-285. https://doi.org/10.1007/s00253-004-1796-y
- Kwon, T. H., Kim, J. T., & Kim, J. S. (2010). Application of a modified sublimation method to screen for PAH-degrading microorganisms. Korean Journal of Microbiology. 46(1), 109-111.
- Lee, E. J., Lee, S. M., Lee, G. T., Kim, I. S., & Kim, Y. H. (2008). Application of effective microorganisms for bioremediation of crude oil spill in Taean. Journal of Environmental Sciences. 17(7), 795-799. https://doi.org/10.5322/JES.2008.17.7.795
- Maccormack, W.P., & Frail, E.R. (1997). Characterization of a hydrocarbon degrading psychrotrophic Antarctic bacterium. Antarctic Science. 9(2), 150-155.
- Meguro, N., Kodama, Y., Gallegos, M. T., & Watanabe, K. (2005). Molecular characterization of resistance-nodulation-division transporters from solvent-and drug-resistant bacteria in petroleum-contaminated soil. Applied and Environmental Microbiology. 71(1), 580-586. https://doi.org/10.1128/AEM.71.1.580-586.2005
- Moody, J. D., Freeman, J. P., Doerge, D. R., & Cerniglia, C. E. (2001). Degradation of phenanthrene and anthracene by cell suspensions of Mycobacterium sp. Strain PYR-1. Applied and Environmental Microbiology. 67(4), 1476-1483. https://doi.org/10.1128/AEM.67.4.1476-1483.2001
- Moody, J. D., Freeman, J. P., Fu, P. P., & Cerniglia, C. E. (2004). Degradation of benzo[a]pyrene by Mycobacterium vanbaalenii PYR-1. Applied and Environmental Microbiology. 70(1), 340-345. https://doi.org/10.1128/AEM.70.1.340-345.2004
- Nichadhain, S. M., Norman, R. S., Pesce, K. V., Kukor, J. J., & Zylstra, G. J. (2006). Mirobial dioxygenase gene population shifts during polycyclic aromatic hydrocarbon biodegradation. Applied and Environmental Microbiology. 72(6), 4078-4087. https://doi.org/10.1128/AEM.02969-05
- Samanta, S. K., Chakraborti, A. K., & Jain, R. K. (1999). Degradation of phenanthrene by different bacteria: evidence for novel transformation sequences involving the formation of 1-naphthol. Applied Microbiology and Biotechnology. 53(1), 98-107. https://doi.org/10.1007/s002530051621
- Schneider. J., Grosser, R., Jayasimhulu, K., Xue, W., & Warshawsky, D. (1996). Degradation of pyrene, benz[a]anthracene, and benzo[a]pyrene by Mycobacterium sp strain RJGII-135, isolated from a former coal gasification site. Applied and Environmental Microbiology. 62(1), 13-19.
- Seo, J. S., Keum, Y. S., Hu, Y., Lee, S. E., & Li, Q. X. (2006). Phenanthrene degradation in Arthrobacter sp. P1-1: initial 1,2- and 3,4- and 9,10-dioxygenation, and meta- and ortho-cleavages of naphthalene-1,2-diol after its formation from naphthalene-1,2-dicarboxylic acid and hydroxyl naphthoic acids. Chemosphere. 65(11), 2388-2394. https://doi.org/10.1016/j.chemosphere.2006.04.067
- Tiehm, A. (1994). Degradation of polycyclic aromatic-hydrocarbons in the presence of synthetic surfactants. Applied and Environmental Microbiology. 60(1), 258-263.
- Uyttebroek, M., Vermeir, S., Wattiau, P., Ryngaert, A., & Springael, D. (2007). Characterization of cultures enriched from acidic polycyclicaromatic hydrocarbon-contaminated soil for growth on pyrene at low pH. Applied and Environmental Microbiology. 73(10), 3159-3164. https://doi.org/10.1128/AEM.02837-06
- Wang, R. F., Wennerstrom, D., Cao, W., Khan, A. A., & Cerniglia, C. E. (2000). Cloning, expression, and characterization of the katG gene, encoding catalase-peroxidase, from the polycyclic aromatic hydrocarbon-degrading bacterium Mycobacterium sp. strain PYR-1. Applied and Environmental Microbiology. 66(10), 4300-4304. https://doi.org/10.1128/AEM.66.10.4300-4304.2000
- Woo, J. H., & Lee, E. Y. (2014). Enantioselective hydrolysis of racemic styrene oxide and its substituted derivatives using newly-isolated Sphingopyxis sp. exhibiting a novel epoxide hydrolase activity. Biotechnology Letters. 36(2), 357-362. https://doi.org/10.1007/s10529-013-1373-5
- Woo, J. H., Kwon, T. H., Kim, J. T., Kim, C. G., & Lee, E. Y. (2013). Identification and characterization of epoxide hydrolase activity of polycyclic aromatic hydrocarbon-degrading bacteria for biocatalytic resolution of racemic styrene oxide and styrene oxide derivatives. Biotechnology Letters. 35(4), 599-606. https://doi.org/10.1007/s10529-012-1114-1
연구 과제 주관 기관 : National Research Foundation of Korea (NRF)