Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti

Temitayo Omotunde Olowomofe; Olaoluwa Jacob Oluyege; Paul Ikechukwu Orjiakor; Ayodele Oluwayemisi Ogunlade; Solomon Temitayo Olaoye

doi:doi:10.11648/j.ijmb.20200503.14

Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti

Temitayo Omotunde Olowomofe, Olaoluwa Jacob Oluyege, Paul Ikechukwu Orjiakor, Ayodele Oluwayemisi Ogunlade, Solomon Temitayo Olaoye

Published in International Journal of Microbiology and Biotechnology (Volume 5, Issue 3)

Received: 21 April 2020 Accepted: 13 May 2020 Published: 20 June 2020

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Abstract

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.

Published in	International Journal of Microbiology and Biotechnology (Volume 5, Issue 3)
DOI	10.11648/j.ijmb.20200503.14
Page(s)	97-102
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2020. Published by Science Publishing Group

Keywords

Bioremediation, Laccase, PCR Amplification, Hydrocarbon-degrading Bacteria

References

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[13]	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.
[14]	Barathi, S. and Vasudevan, N. (2001). Utilization of petroleum hydrocarbons by pseudomonas flourescens isolated from petroleum contaminated soil. Environmental International. 26: 413-416.
[15]	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.
[16]	Kiiskinen, L. L., Rättö, M., Kruus, K. (2004). Screening for novel laccase-producing microbes. Journal of applied microbiology, 97 (3): 640-646.
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Temitayo Omotunde Olowomofe, Olaoluwa Jacob Oluyege, Paul Ikechukwu Orjiakor, Ayodele Oluwayemisi Ogunlade, Solomon Temitayo Olaoye. (2020). Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti. International Journal of Microbiology and Biotechnology, 5(3), 97-102. https://doi.org/10.11648/j.ijmb.20200503.14

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ACS Style

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. Int. J. Microbiol. Biotechnol. 2020, 5(3), 97-102. doi: 10.11648/j.ijmb.20200503.14

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AMA Style

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. Int J Microbiol Biotechnol. 2020;5(3):97-102. doi: 10.11648/j.ijmb.20200503.14

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@article{10.11648/j.ijmb.20200503.14,
  author = {Temitayo Omotunde Olowomofe and Olaoluwa Jacob Oluyege and Paul Ikechukwu Orjiakor and Ayodele Oluwayemisi Ogunlade and Solomon Temitayo Olaoye},
  title = {Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {5},
  number = {3},
  pages = {97-102},
  doi = {10.11648/j.ijmb.20200503.14},
  url = {https://doi.org/10.11648/j.ijmb.20200503.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200503.14},
  abstract = {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.},
 year = {2020}
}

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TY  - JOUR
T1  - Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti
AU  - Temitayo Omotunde Olowomofe
AU  - Olaoluwa Jacob Oluyege
AU  - Paul Ikechukwu Orjiakor
AU  - Ayodele Oluwayemisi Ogunlade
AU  - Solomon Temitayo Olaoye
Y1  - 2020/06/20
PY  - 2020
N1  - https://doi.org/10.11648/j.ijmb.20200503.14
DO  - 10.11648/j.ijmb.20200503.14
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 97
EP  - 102
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20200503.14
AB  - 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.
VL  - 5
IS  - 3
ER  -

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Author Information

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

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APA Style

Temitayo Omotunde Olowomofe, Olaoluwa Jacob Oluyege, Paul Ikechukwu Orjiakor, Ayodele Oluwayemisi Ogunlade, Solomon Temitayo Olaoye. (2020). Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti. International Journal of Microbiology and Biotechnology, 5(3), 97-102. https://doi.org/10.11648/j.ijmb.20200503.14

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ACS Style

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. Int. J. Microbiol. Biotechnol. 2020, 5(3), 97-102. doi: 10.11648/j.ijmb.20200503.14

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AMA Style

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. Int J Microbiol Biotechnol. 2020;5(3):97-102. doi: 10.11648/j.ijmb.20200503.14

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@article{10.11648/j.ijmb.20200503.14,
  author = {Temitayo Omotunde Olowomofe and Olaoluwa Jacob Oluyege and Paul Ikechukwu Orjiakor and Ayodele Oluwayemisi Ogunlade and Solomon Temitayo Olaoye},
  title = {Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {5},
  number = {3},
  pages = {97-102},
  doi = {10.11648/j.ijmb.20200503.14},
  url = {https://doi.org/10.11648/j.ijmb.20200503.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200503.14},
  abstract = {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.},
 year = {2020}
}

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TY  - JOUR
T1  - Phenotypic and Molecular Screening of Laccase-producing Bacteria Isolated from Automobile Workshop Soil Samples in Ado-Ekiti
AU  - Temitayo Omotunde Olowomofe
AU  - Olaoluwa Jacob Oluyege
AU  - Paul Ikechukwu Orjiakor
AU  - Ayodele Oluwayemisi Ogunlade
AU  - Solomon Temitayo Olaoye
Y1  - 2020/06/20
PY  - 2020
N1  - https://doi.org/10.11648/j.ijmb.20200503.14
DO  - 10.11648/j.ijmb.20200503.14
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 97
EP  - 102
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20200503.14
AB  - 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.
VL  - 5
IS  - 3
ER  -

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