Volume 5, Issue 3, September 2020, Page: 97-102
Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti
Temitayo Omotunde Olowomofe, Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Nigeria
Olaoluwa Jacob Oluyege, Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Nigeria
Paul Ikechukwu Orjiakor, Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Nigeria
Ayodele Oluwayemisi Ogunlade, Department of Food Technology, Federal Polytechnic, Ado-Ekiti, Nigeria
Solomon Temitayo Olaoye, Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Nigeria
Received: Apr. 21, 2020;       Accepted: May 13, 2020;       Published: Jun. 20, 2020
DOI: 10.11648/j.ijmb.20200503.14      View  38      Downloads  21
This work was carried out to isolate hydrocarbon-degrading bacteria from oil contaminated soil sample in Ado-Ekiti and screen them for laccase production. Soil samples were collected and analyzed using standard microbiological techniques. The isolates were initially screened for hydrocarbon degrading ability on minimal salt medium supplemented with 1% crude oil and incubated for 14days. The isolates were further screened for their ability to produce laccase enzyme using plate screening and molecular techniques. Four of the isolates that gave the best results on tannic-agar plates were selected for PCR amplification of laccase gene using specific primers. The isolates were identified as Lactobacillus sakei, Pseudomonas aeruginosa, Bacillus cereus and Gracilibacter thermotolerans based on 16SrRNA sequencing. The DNA of these bacteria amplified the primer specific for laccase gene with 1300bp, 1400bp, 1600bp and 350bp respectively. For bioremediation to be effective, microorganisms must enzymatically attack the pollutants and convert them to harmless products. Therefore, laccase production potentials in these bacteria make them useful in bioremediation as laccase is known to break heavy phenol containing hydrocarbons. Further work can be done to determine the activity of this enzyme during the degradation of crude oil.
Bioremediation, Laccase, PCR Amplification, Hydrocarbon-degrading Bacteria
To cite this article
Temitayo Omotunde Olowomofe, Olaoluwa Jacob Oluyege, Paul Ikechukwu Orjiakor, Ayodele Oluwayemisi Ogunlade, Solomon Temitayo Olaoye, Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti, International Journal of Microbiology and Biotechnology. Vol. 5, No. 3, 2020, pp. 97-102. doi: 10.11648/j.ijmb.20200503.14
Copyright © 2020 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cerniglia, N. and Sutherland, M. (2001). Application Environmental Microbiology. 63: 4120-4122.
Clemente, A., Dimcheva, N., Ferapontova, E. E., Gorton, L., Ruzgas, T., Stoica, L., Shleev, S., Yaropolov, A. I., Haltrich, D., Thorneley, R. N. and Austin, S. D. (2001). Application of Microbiology and Biotechnology. 16: 1074-1092.
Bird, C. E. Cavanaugh, E. G. Dickerman, J. C. and Fernandes, S. R. (2008). A Study of Vapor Control Methods for Gasoline Marketing Operations. U.S. Environmental Protection Agency. 171: 31-36.
Zouari-Mechichi, H., Mechichi, T., Dhouib, A., Sayadi, S., Martinez, A. T, mMartinez, M. J., (2005). Laccase purification and characterization from Trametes trogii isolated in Tunisia: decolorization of textile dyes by the purified enzyme. Enzyme Microbial Tech., 39: 141-148.
Desai, S., Nityanand, C. (2011). Microbial laccases and their applications: a review. Asian J Biotechnol, 3 (2): 98-124.
Bains J, Capalash N, Sharma P (2003). Laccase from a non-melanogenic, alkalotolerantc-proteobacterium JB isolated from industrial wastewater drained soil. Biotechnol Lett 25 (1): 155-1159.
Michael, M. T., M. G. Georg and R. Astrid (2005). Degradation of Azo dyes by lacase and ultrasound treatment. Apl. Environ. Microbiol. 71: 260-2607.
Elsayed, M. A., Hassan, M. M., Elshafei, A. M., Haroun, B. M. Ot man, A. M. (2012). Optimization of cultural and nutritional parameters for the production of laccase by Pleurotus ostreatus ARC280. British Biotechnology Journal, 2 (3), 115-132.
Baldrian, P. (2006). “Fungal laccases-occurrence and properties,”FEMS Microbiology Reviews, vol. 30, no. 2, pp. 215-242.
Sivakumar R, Rajendran R, Balakumar C, Tamilvendan M (2010). Isolation, screening and optimization of production medium for thermostable laccase production from Ganoderma sp. Int. J. Eng. Sci. Technol. 2 (12): 7133-7141.
Pointing, S. B. (1999). Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi. Fungal Diversity 2, 17-33.
Bourbonnais, R., Leech, D. and Paice, M. G. (1998). “Electrochemical analysis of the interactions of laccase mediators with lignin model compounds,” Biochimica et Biophysica Acta, vol. 1379, no. 3, pp. 381-390.
Majcherczyk, A., Johannes, C. and A. Huttermann, A. (1998). “Oxidation ¨of polycyclic aromatic hydrocarbons (PAH) by laccase of Trametes versicolor,” Enzyme and Microbial Technology, vol. 22, no. 5, pp. 335-341.
Barathi, S. and Vasudevan, N. (2001). Utilization of petroleum hydrocarbons by pseudomonas flourescens isolated from petroleum contaminated soil. Environmental International. 26: 413-416.
Olowomofe T. O, Oluyege J. O, Sowole D. O (2017). Isolation, screening and characterization of hydrocarbon-utilizing bacteria isolated from bitumen-contaminated surface water in Agbabu, Ondo State. J Adv Biol Biotechnol 15: 1-9.
Kiiskinen, L. L., Rättö, M., Kruus, K. (2004). Screening for novel laccase-producing microbes. Journal of applied microbiology, 97 (3): 640-646.
Rahman KSM, Rahman TJ, Kourkoutas Y, Petsas I, Marchant R, Banat I. M. (2003). Enhanced bioremediation of N-alkane in petroleum sludge using bacterial consortium amended with rhamnolipid and micronutrients. Biores. Technol., 90: 159-168.
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