Brazilian Journal of Biological Sciences (ISSN 2358-2731)

Home Archive v. 5, no. 9 (2018) Carvalho-Gonçalves


Vol. 5, No. 9, p. 57-68 - Apr. 30, 2018


Lipases and biosurfactants production by the newly isolated Burkholderia sp.

Laís Campos Teixeira de Carvalho-Gonçalves and Krystyna Gorlach-Lira

Lipases and biosurfactants are biocompounds produced by microorganisms involved in the metabolism of oily substrates. In this way, our study aimed to evaluate these molecules production by bacteria isolated from contaminated soil with waste vegetable oil and evaluate the optimal culture conditions for lipase production using the response surface methodology. The lipolytic activity was tested on tributyrin agar and rhodamine B agar with olive or soybean oil. All 66 isolates of bacteria were positive on tributyrin medium, while the percentage of lipolytic bacteria on rhodamine B medium varied from 31 (soybean oil, pH 6.0) to 38 (olive oil, pH 7.0 and 8.0; soybean oil, pH 8.0). The oil-spreading technique revealed that all isolates produced biosurfactants and oil emulsification and hemolytic activity tests detected biosurfactants in 60% and 88% of isolates, respectively. Lipolytic activity and biomass value varied de 8.7 to 12.4 U/mL and 2.5 to 4.04 mg/mL, respectively, in nutrient broth with olive oil medium. Six isolates with higher lipase activity were identified as Burkholderia sp., according to phylogenetic analysis based 16S rRNA sequences. Only Burkholderia sp. O19 strain produced rhamnolipids among bacteria studied. The surface response methodology revealed that the production of lipases by Burkholderia sp. O19 occurs in a wide range of pH and temperature with maximum response achieved at pH 8.5 and 65 oC (18.7 U/mL). The results obtained in this study are relevant as they show the simultaneous production of two biocompounds with broad industrial applications.

Bacteria; Biosurfactants; Lipase; Soil.


Full text

Bhosale, H.; Shaheen, U.; Kadam, T. Characterization of a hyperthermostable alkaline lipase from Bacillus sonorensis 4R. Enzyme Research, v. 2016, p. 1-10, 2016.

Ciafardini, G.; Zullo, B. A.; Iride, A. Lipase production by yeasts from extra virgin olive oil. Food Microbiology, v. 23, p. 60-67, 2006.

Ciccillo, F.; Fiore, A.; Bevivino, A.; Dalmastri, C.; Tabacchioni, S.; Chiarini, L. Effects of two different application methods of Burkholderia ambifaria MCI 7 on plant growth and rhizospheric bacterial diversity. Environmental Microbiology, v. 4, p. 238-245, 2002.

Colla, L. M.; Rizzardi, J.; Pinto, M. H., Reinehr, C. O.; Bertolin, T. E.; Costa, J. A. V. Simultaneous production of lipases and biosurfactants by submerged and solid-state bioprocess. Bioresource Technology, v. 101, p. 8308-8314, 2010.

Das, M.; Das, S. K.; Mukherjee, R. K. Surface active properties of the culture filtrates of a Micrococcus species grown on n-alkenes and sugars. Bioresource Technololy, v. 63, p. 231-235, 1998.

Franzetti, A.; Gandolfi, I.; Raimondi, C.; Bestetti, G.; Banat, I. M.; Smyth, T. J. P.; Papacchini, M.; Cavallo, M.; Fracchia, L. Environmental fate, toxicity, characteristics and potential applications of novel bioemulsifiers produced by Variovorax paradoxus 7bCT5. Bioresource Technology, v. 108, p. 245-251, 2012.

Geys, R.; Soetaert, W.; Bogaert, I. V. Biotechnological opportunities in biosurfactant production. Current Opinion of Biotechnology, v. 30, p. 66-72, 2014.

Gopinath, S. C. B.; Anbu, P.; Lakshmipriya, T.; Hilda, A. Strategies to characterize fungal lipases for applications in medicine and dairy industry. BioMed Research International, v. 2013, p. 1-10, 2013.

Gudiña, E. J.; Teixeira, J. A.; Rodrigues, L. R. Biosurfactants produced by marine microorganisms with therapeutic applications. Marine Drugs, v. 14, No. 2, 38, p. 1-15, 2016.

Gupta, R.; Gupta, N.; Rathi, P. Bacterial lipases: an overview of production, purification and biochemical properties. Applied of Microbiology and Biotechnology, v. 64, p. 763-781, 2004.

Gupta, R.; Kumari, A.; Syal, P.; Singh, Y. Molecular and functional diversity of yeast and fungal lipases: their role in biotechnology and cellular physiology. Progress in Lipid Research, v. 57, p. 40-54, 2015.

Hasan, F.; Shah, A. A.; Hameed, A. Industrial applications of microbial lipases. Enzyme Microbiology and Technology, v. 39, p. 235-251, 2006.

Hörmann, B.; Müller, M. M.; Syldatk, M.; Hausmann, R. Rhamnolipid production by Burkholderia plantarii DSM 9509T. European Journal of Lipid Science and Technology, v. 112, No. 6, p. 674-680, 2010.

Irorere, V. U.; Tripathi, L.; Marchant, R.; McClean, S.; Banat, I. M. Microbial rhamnolipid production: a critical re-evaluation of published data and suggested future publication criteria. Applied Microbiology and Biotechnology, v. 101, p. 3941-3951, 2017.

Kim, E. K.; Jang, W. H.; Ko, J. H.; Kang, J. S.; Noh, M. J.; Yoo, O. J. Lipase and its modulator from Pseudomonas sp. strain KFCC 10818: proline-to-glutamine substitution at position 112 induces formation of enzymatically active lipase in the absence of the modulator. Journal of Bacteriology, v. 183, p. 5937-5941, 2001.

Ko, W. H.; Wang, I. T.; Ann, P. J. A simple method for detection of lipolytic microorganisms in soils. Soil Biology and Biochemistry, v. 37, p. 597-599, 2005.

Kouker, G.; Jaeger, K. E. Specific and sensitive plate assay for bacterial lipases. Applied Environmental and Microbiology, v. 53, p. 211-213, 1987.

Liu, C. H.; Lu, W. B.; Chang, J. S. Optimizing lipase production of Burkholderia sp. by response surface methodology. Process Biochemistry, v. 41, p. 1940-1944, 2006.

Lotfabad, T. B.; Shourian, M.; Roostaazad, R.; Najafabadi, A. R.; Adelzadeh, M. R.; Noghabi, K. A. An efficient biosurfactant-producing bacterium Pseudomonas aeruginosa MR01, isolated from oil excavation areas in south of Iran. Colloids Surfaces B: Biointerfaces, v. 69, No. 2, p. 183-193, 2009.

Lu, Y.; Lu, F.; Wang, X.; Bie, X.; Sun, H.; Wuyundalai; Lu, Z. Identification of bacteria producing a thermophilic lipase with positional non-specificity and characterization of the lipase. Annals of Microbiology, v. 59, p. 565-571, 2009.

Ma, Q. X.; Sun, S.; Gong, S.; Zhang, J. Screening and identification of a highly lipolytic bacterial strain from barbecue sites in Hainan and characterization of its lipase. Annals of Microbiology, v. 60, p. 429-437, 2010.

Morikawa, M.; Hirata, Y.; Imanaka, T. A study on the structure-function relationship of the lipopeptide biosurfactants. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, v. 1488, p. 211-218, 2000.

Moran, A. C.; Martinez, M. A.; Sineriz, F. Quantification of surfactin in culture supernatant by hemolytic activity. Biotechnology Letters, v. 24, p. 177-180, 2002.

Paula, A. V.; Barboza, J. C. S.; Castro, H. F. Study of the influence of solvent, carbohydrate and fatty acid in the enzymatic synthesis of sugar esters by lipases. Quimica Nova, v. 28, p. 792-796, 2005.

Peil, G. H. S.; Kuss, A. V.; Rave, A. F. G.; Villarreal, J. P. V.; Hernandes, Y. M. L.; Nascente, P. S. Bioprospecting of lipolytic microorganisms obtained from industrial effluents. Anais da Academia Brasileira de Ciências, v. 88, p. 1769-1779, 2016.

Ramette, A.; Lipuma, J. J.; Tiedje, J. M. Species abundance and diversity of Burkholderia cepacia complex in the environment. Applied Environmental and Microbiology, v. 71, p. 1193-1201, 2005.

Rehm, S.; Trodler, P.; Pleiss, J. Solvent-induced lid opening in lipases: a molecular dynamics study. Protein Science, v. 19, p. 2122-2130, 2010.

Shaini, V. P.; Jayasree, S. Isolation and characterization of lipase producing bacteria from windrow compost. International Journal of Current Microbiology and Applied Sciences, v. 5, p. 926-933, 2016.

Siegmund, I.; Wagner, F. New method for detecting rhamnolipids excreted by Pseudomonas species grown on mineral agar. Biotechnology Letters, v. 5, p. 265-268, 1991.

Stuer, W.; Jaeger, K. E.; Winkler, U. K. Purification of extracellular lipase from Pseudomonas aeruginosa. Journal of Bacteriology, v. 168, no. 3, p. 1070-1074, 1986.

Yang, J.; Guo, D.; Yan, Y. Cloning, expression and characterization of a novel thermal stable and short-chain alcohol tolerant lipase from Burkholderia cepacia strain G63. Journal of Molecular Catalysis B: Enzymatic, v. 45, p. 91-96, 2007.

Zafar, S.; Shafiq, A.; Nadeem, S. G.; Hakim, S. T. Isolation and preliminary screening of biosurfactant producing bacteria from oil contaminated soil. Brazilian Journal of Biological Sciences, v. 3, p. 285-292, 2016.

Zarinviarsagh, M.; Ebrahimipour, G.; Sadeghi, H. Lipase and biosurfactant from Ochrobactrum intermedium strain MZV101 isolated by washing powder for detergent application. Lipids in Health and Disease, v. 16, p.177-189, 2017.