ISSN (Online): 2321-3418
server-injected
Plant Science and Biotechnology
Open Access

Isolation, Screening, Identification, Molecular Characterization and Optimizatiom of Lipase producing Bacteria from Edible Oil Contaminated Soil in Maiduguri, Borno State, Nigeria

, , , , , , ,
DOI: 10.18535/ijsrm/v13i09.pb04· Pages: 08-32· Vol. 13, No. 09, (2025)· Published: September 16, 2025
PDF
Views: 487 PDF downloads: 445

Abstract

Lipase is an important hydrolytic enzyme used for numerous industrial applications due to their qualities such as versatility, specificity, increased sustainability and environmental friendliness. This work focused on the isolation, screening, identification and molecular characterization of lipase producing bacteria isolated from edible oil contaminated soil in Maiduguri, Borno state as well as their optimization for higher lipase production. Bacteria were isolated using standard microbiological techniques and screened for lipase production using tributyrine, rhodamine B and Tween 80 agar media. Secondary screening for high lipase producing bacteria was done in a mineral salt medium. The highest lipase producing bacteria was suspected to be Pseudomonas sp which was identified as Stutzerimonas stutzeri by sequencing using 16s rRNA gene when subjected to homology search using Basic Local Alignment Search Tool (BLAST). Lipase producing parameters were subjected to varying temperatures and pH for optimal lipase production. The bacteria Stutzerimonas stutzeri were identified as good candidates for lipase production in this research. having the highest lipase activity (10.65 U/mL) with coconut oil as the substrate, glycerol as the carbon source, yeast extract as the nitrogen source, pH 9.0, incubation at 40 °C for 48 h, inoculum size of 4000 µL, and agitation at 120 rpm, resulting in a substantial yield increase.

Keywords

isolationscreeningidentificationMolecular characterizationoptimizationlipase producing bacteriaedible oil contaminated soil

References

  1. A’yuni H, IImi M (2021). Lipase production from Aspergillus aculeatus MS.11 in broth medium with variation of agitation speed. AIP. Conf. Proc. 2353, 030081. https://doi.org/10.1063/5.0052540DOI ↗Google Scholar ↗
  2. Abubakar A, Abioye OP, Aransiola SA, Maddela NR, Prasad R (2024). Crude oil Biodegradation Potential of lipase produced by Bacillus subtilis and Pseudomonas aeruginosa isolated from hydrocarbon contaminated soil. Environmental chemistry and Ecotoxicology. 6: 26-32. https://doi.org/10.1016/j.enceco.2023.12.001DOI ↗Google Scholar ↗
  3. Ali S, Khan SA, Hamayun M and Lee I (2023). The Recent Advances in the Utility of Microbial Lipases: A Review. Microorganisms 11(2) 510; https://doi.org/10.3390/microorganisms11020510.DOI ↗Google Scholar ↗
  4. Babatunde, F. O., and Olayemi, A. B. (2023). Isolation and characterization of lipase-producing bacteria from vegetable oil-contaminated soil in Nigeria. Journal of Applied Microbiology, 135(1), 155-168.Google Scholar ↗
  5. Baron, S., Jorgensen, J. H., and Landry, M. L. (2019). Baron's medical microbiology (5th ed.). Galveston (TX): University of Texas Medical Branch at Galveston.Google Scholar ↗
  6. Baruwa AS, Fadipe TO, Isaac TO, Okonji RE, Oboutor EM (2021). Optimization of lipase production by Bacillus subtilis BF001 isolated from vegetable oil processing site soil. https://www.research gate.net/publication/358547843 DOI:10.13140/RG.2.2.12244.68487.ASM 2021 international conference.DOI ↗Google Scholar ↗
  7. Burcu Bakir Z, Metin K (2017). Production and Characterisation of an Alkaine Lipase from Thermophilic Anoxybacillus sp. HBB16. Chem. Biochem. Eng. Q 31(3) 303-312. https://doi.org/10.15255/CABEQ.2016.990DOI ↗Google Scholar ↗
  8. Chouhan R, Lapaka S, Alpula N, Podeti S (2020). Optimization and Molecular Characterization of Lipase Producing Bacillus subtilis strain Rcps3 and Bacillus fumarioli strain Rcps4 from oil-contaminated soils in Warangal. J pure Appl Microbiol. 14(3) 2063-2074. https://doi.org/10.22207/JPAM.14.3.47DOI ↗Google Scholar ↗
  9. Eprahimpour A, Rahman RNZRA, Ean Ch’ng DH, Basri M, Salleh AB (2008). A Modelling Study by Response Surface Methodology and Artificial Neural Network on Culture Parameters Optimization for Thermostable Lipase Production from a Newly Isolated thermophilic Geobacillus sp. Strain ARM. BMC Biotechnol8, 96. https://doi.org/10.1186/1472-6750-8-96DOI ↗Google Scholar ↗
  10. Ezenwelu CO, Afeez OA, Anthony OU, Promise OA, Mmesoma UC, Henry OE (2022). Studies on Properties of Lipase Produced from Aspergillus sp. Isolated from Compost Soil. Advances in Enzyme Research 10 (2) 46-60. https://doi.org/10.4236/aer.2022.102003DOI ↗Google Scholar ↗
  11. Fakorede CN, Itakorede BO, Olayiwola AT, Anifowose TE (2019). Lipase producing potential of different bacteria species isolated from cooking oil contaminated soils. International journal of of Environment, Agriculture and Biotechnology (IJEAB). 4(2). https://dx.doi.org/10.22161/ijeab/4.2.8DOI ↗Google Scholar ↗
  12. Funke G, Tortora PM, Case CL, and Johnson, T. R. (2019). Microorganisms, an introduction to microbes. 13th ed. Pearson, New York.Google Scholar ↗
  13. Gupta R, Gupta N, Rathi P (2004). Bacterial lipases: an overview of production, purification and biochemical properties. Applied Microbiology and Biotechnology. 64(6): 763–781. https://doi.org/10.1007/s00253-004-1568-8DOI ↗Google Scholar ↗
  14. Han, D., Gao, P., Li, R., Tan, P., Xie, J., Zhang, R., and Li, J. (2020). Multicenter assessment of microbial community profiling using 16S rRNA gene sequencing and shotgun metagenomic sequencing. Journal of Advanced Research, 26, 111-121.Google Scholar ↗
  15. Ibrahim, A. M., and Adamu, K. M. (2023). Isolation and characterization of lipase-producing bacteria from vegetable oil-contaminated soil in Maiduguri, Borno State, Nigeria. Journal of Environmental Science and Health, Part A, 58(1), 127-139.Google Scholar ↗
  16. Ikhwani AZN, Idris I, Elfirta RR, Ferdian PR (2024). Production and activity characterization of lipase from Bacillus flexus B486. JOTCSA, 11:2, 394-404. https://doi.org/10.18596/jotcsa.1333916DOI ↗Google Scholar ↗
  17. Jayaraman M, Narayanan SA, Jeya, KR, Alagendran S (2020). Isolation and Identification of a Novel Lipase Producing Bacteria from Oil Spilled Soil. International Journal of Innovative Research in Science, Engineering and Technology, 3.Google Scholar ↗
  18. Joyruth P, Growther L (2020). Molecular Identification, Production and Optimization of Lipase from Oil Contaminated Soil using Submerged Fermentation. J. Pure Appl. Microbiol. 14(1). 341-349. https://doi.org/10.22207/JPAM.14.1.35.DOI ↗Google Scholar ↗
  19. Kattatheyil H, Sajeela V, Kabeer SS (2024). Screening, Optimization and Molecular Characteriation of cold –active Lipase Producing Bacillus cereus I13 from Arctic Sediment. Biologi.79. Pp 1041-1055. https://doi.org/10.1007/s11756-024-01610-yDOI ↗Google Scholar ↗
  20. Kim YO, Khosasih V, Nam BH, Lee SJ, Suwanto A, Kim HK (2011). Gene cloning and catalytic charectirization of cold adapted lipase of Photobacterium sp MAI-3 isolated from blood clam. J. Biosci.Bioeng. 114: 589-595. https://doi.org/10.1016/j.jbiosc.2012.06.013DOI ↗Google Scholar ↗
  21. Koneman, E. W., Allen, S. D., Janda, W. M., Schreckenberger, P. C., and Winn, W. C. (2019). Koneman's color atlas and textbook of diagnostic microbiology (7th ed.). Philadelphia, PA: Lippincott Williams and Wilkins.Google Scholar ↗
  22. Maytham, A. A. and Hanaa, J. J. A. (2016).Isolation and identification of lipase producing bacteria from oil-contaminant soil. Journal of Biology, Agriculture and Healthcare, 6(20), 1-7.Google Scholar ↗
  23. Messaoudi A, Belguith H, Hamida JB (2011). Three-Dimensional Structure of Aradposis Thaliana Lipase Predicted by Homology Modelling Method. Evolutionary Bioinformatics, 7, 99-105. https://doi.org/10.4137/EBO.S7122.DOI ↗Google Scholar ↗
  24. Murtius WS, Hari PD, Putri IIN (2022). The Effect of Incubation Time to the activity of Lipase Preduced by Bacillus thuringiensis on Coconut (Cocos nucifera L.) Dreg. Lop conf. ser: Earth Environ. Sci. 1059. https://doi.org/10.1088/1755-1315/1059/1/012076DOI ↗Google Scholar ↗
  25. Ojo, AA, Ogunbanwo ST (2022). Optimization of lipase production from Bacillus cereus isolated from soil contaminated with palm oil. Journal of Applied Microbiology, 134(2), 481-493.Google Scholar ↗
  26. Oliveira BH, Coradi GV, Attili‐Angelis D, Scauri C, Luques A H P, Barbosa AM, Lima VM (2013). Comparison of lipase production on crambe oil and meal by Fusarium sp. (Gibberella fujikuroi complex). European Journal of Lipid Science and Technology, 115(12), 1413-1425.Google Scholar ↗
  27. Oluwasaye II, Abiodun EA, Japhel AO, Efemena MO, Adebayo LO (2020). Isolation, optimization and molecular characterization of lipase producing bacteria from contaminated soil. Scientific African. 8:e00297. https://doi.org/10.1016/j.sciaf.2020.e00279DOI ↗Google Scholar ↗
  28. Oyetunde OO, Adebayo, AA, Babatunde, FO (2022). Molecular characterization and enhanced production of a halophilic lipase from Bacillus cereus OOUC1 isolated from a palm oil-contaminated soil. Journal of Biotechnology, 353, 144-154.Google Scholar ↗
  29. Pham VH, Kim J, Chang S, Chung W (2021). Investigating of lipolytic – secreting bacteria from an artificially polluted soilusinf a modified culture method and optimization of their lipase production. Microorganisms. 9:12. 2590. https://doi.org/10.3390/microorganisms9122590DOI ↗Google Scholar ↗
  30. Popoola BM, Olanbiwoninu AA (2021). Characeristics of some Selected Bacterial Isolates from Vegetable Oil Contaminated Soil. Microbiology Research Journal International. 31(2):22-37. https://doi.org/10.9734/mrji/2021/v31i230294.DOI ↗Google Scholar ↗
  31. Rajeshkumar P, Mahendran VS, Balakrishnan V (2013) Isolation and identification of lipase producing organisms from diverse soil samples of Kolli hills. International Journal of Current Microbiology and Applied Sciences, 2(5), 205-210.Google Scholar ↗
  32. Robinson PK (2015). Enzymes: Principles and Biotechnological Application. Essays biochem. 15:59. 1-41. https://doi.org/10.1042/bse0590001DOI ↗Google Scholar ↗
Author details
Sa’ad A. J.
National Biotechnology Research and Development Agency, Abuja
✉ Corresponding Author
👤 View Profile →🔗 Is this you? Claim this publication
Daja A.
Department of Biochemistry, Faculty of Life Science, University of Maiduguri, Borno State
ORCID ↗
👤 View Profile →
Bislava S. B.
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →🔗 Is this you? Claim this publication
Buba F.
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →🔗 Is this you? Claim this publication
Ali H. K
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →
Isa A.
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →🔗 Is this you? Claim this publication
Madu H. M.
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →🔗 Is this you? Claim this publication
Milala A.M.
Department of Biotechnology, Faculty of life sciences, University of Maiduguri, Borno State
👤 View Profile →🔗 Is this you? Claim this publication