SciELO - Scientific Electronic Library Online

 
vol.24 issue3Phenolic compounds, antihemolytic, anti-inflammatory and antibacterial activity of propolis from southern SonoraChemical and functional characterization of raw and cooked bean flours from the Pinto Saltillo and Black varieties, from the State of Durango author indexsubject indexsearch form
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

Related links

  • Have no similar articlesSimilars in SciELO

Share


Biotecnia

On-line version ISSN 1665-1456

Biotecnia vol.24 n.3 Hermosillo Sep./Dec. 2022  Epub June 19, 2023

https://doi.org/10.18633/biotecnia.v24i3.1750 

Artículos

Production of extracellular lipase by Enterococcus faecium E68 with olive oil waste as substrate

Producción de lipasa extracelular por Enterococcus faecium E68 en residuos de aceite de oliva como sustrato

Esra Acu Merih-Kivanc1  * 

1Eskisehir Technical University, Science of Faculty, Department of Biology, Eskişehir, Turkey


Abstract

Green technologies eliminate the damages caused by agro-technological wastes to the environment. Our study aimed to both prevent the environmental harm by olive oil waste, and produce lipase enzyme, which is an important biotechnological product. E. faecium E68 obtained from milk and dairy products was used for lipase enzyme production. E. faecium E68 was cultured in lipase production medium containing 10 % olive waste, pH 6.5, at 37 oC with 120 rpm agitation for 48 h. The effect of temperature, pH metal ion, surfactant, and NaCl was also determined. The molecular weight of the partially purified extracellular lipase enzyme was estimated to be around 19-20 kDa by SDS-PAGE. The optimum temperature was 45°C, while the enzyme exhib-ited appreciable thermostability retaining activity at 55°C for 48h. The optimum lipase activity was at pH 10. One mM Ca2+, Mn2+, Cu2+, Ni2+, Zn2+, Mg2+ and K+ ions modulated the enzyme activity, but was inhibited by Hg2+, SDS and Triton X-100. The enzyme is halophilic and 25 % NaCl salt increased the activity.

Keywords: Enterococcus faecium; Lipase activity; Olive oil waste; lipase

Resumen

Con las tecnologías verdes se eliminan los daños que ocasionan los desechos agrotecnológicos al medio ambiente. En nuestro estudio, el objetivo era prevenir el daño de los residuos de aceite de oliva al medio ambiente y producir la enzima lipasa, que es un producto biotecnológico importante. E. faecium E68 obtenido de leche y productos lácteos se utilizó en la producción de la enzima lipasa. E. faecium E68 se desarrolló en medio de producción de lipasa con un 10% de orujo de aceituna, pH 6,5, a 37 oC con agitación a 120 rpm durante 48 h. También se determinó el efecto de la temperatura, el pH del ion metálico, el surfactante y el NaCl. El peso molecular de la enzima lipasa extracelular parcialmente purificada se estimó en alrededor de 19-20 kDa mediante SDS-PAGE. La temperatura óptima fue de 45 °C, mientras que la enzima exhibió una termoestabilidad apreciable reteniendo la actividad a 55°C durante 48 h. La actividad óptima de la lipasa fue a pH10. Los iones Ca2+, Mn2+, Cu2+, Ni2+, Zn2+, Mg2+ y K+ (1 mM) modularon la actividad de la enzima, pero fueron inhibidos por Hg2+, SDS y Triton X-100. La enzima es halófila y la sal de NaCl al 25 % aumentó la actividad.

Palabras clave: Enterococcus faecium; actividad de lipasa; residuos de aceite de oliva; lipasa

Introduction

By-products generated in the agricultural industry cause much environmental pollution and adverse health effects (Hamrouni et al., 2020; Leite et al., 2021). Waste resulting from olive oil extraction creates severe environmental problems with its highly polluting properties (Mantzavinos and Kalogerakis, 2005; Sarika et al., 2005). Olive waste contains biodegradable compounds and phytotoxic phenolic compounds. Phytotoxic wastes correspond to about 80 % of olive oil production. These phenols tend to change into condensed high molecular weight polymers, which are difficult to degrade on storage (Ayed et al., 2005). For these reasons, olive oil wastes may lead to acute odor problems and, more importantly, serious risks for water and soil quality (Mantzavinos and Kalogerakis, 2005; Sarika et al., 2005). Today, with the increasing awareness of environmental protection, the use of biomass has gained importance (Hamrouni et al., 2020). Olive oil waste contains simple and complex sugars, lipids, residual oil, proteins, and mineral elements, besides phytotoxic wastes. These compounds can be directly recovered by chemical extraction followed by purification (Fki et al., 2005; Papadimitriou et al., 2005). Olive oil waste can be used as the basic compound for fermentative production processes (Fenice et al., 2003; Angenent et al., 2004). Various agro-industrial wastes have been used for biotechnological purposes, especially for enzyme production (Mahanta et al., 2008). Lipases are one of those enzymes, which are commercially important since they catalyze the hydrolysis of long chain fatty acids to glycerol and fatty acids.

Most commercial lipases are produced by microorganisms (Babu and Rao, 2007; Treichel et al., 2010; Bharathi and Rajalakshmi, 2019; Adetunji and Olaniran, 2021). Especially since the lipase enzyme produced from lactic acid bacteria is considered safe, it is preferred in the food industry (Meyers et al., 1996; Liu et al., 2001; Lopes et al., 2002; Couto and Sanroman, 2006; Ramakrishnan et al., 2013; 2015; 2016; Sukohidayat et al., 2018; Dellali et al., 2020; Acu et al., 2021).

In our study, it was aimed to produce, partially purify and characterize the lipase enzyme from E. faecium E68 strain in olive oil waste.

Material and methods

Bacteria

In the study, Enterococcus faecium obtained from Eskişehir Technical University microbiology unit was used. E. faecium E68 was inoculated in M17 broth and incubated at 37 °C for 24 h under 10 % CO2 conditions. Growing cultures from M17 broth were inoculated on M17 agar and incubated at 37 °C for 24 h under 10 % CO2 conditions. The morphological features of the colonies formed after incubation were examined. Then, the purity of the cultures was checked by microscopic examination by Gram staining.

Olive oil waste

Olive oil waste with dry matter content of 0.82 g, nitrogen content of 1.593 % and oil content of 0.036 % was obtained from olive oil production facilities and used in the studies.

Lipase production

E. faecium E68 was inoculated onto M17 agar and incubated at 37 °C for 48 h. It was then inoculated as a single colony into M17 broth and incubated at 37 °C for 24 h. Culture was set to an Optical Density (OD) of 1 at 600 nm in the spectrophotometer. Then, 1 % of the culture was inoculated into the lipase determination medium.

The study was carried out in 2 parts. In the first part, without adding olive oil waste to the medium, and in the second part, by adding 10 % olive oil waste to the medium.

For enzyme production, 500 mL of lipase assay medium was prepared, added with 5 % peptone as nitrogen source, 3 % glucose as carbon source and other components (0.1g/L CaSO4, 0.5g/L KH2PO4, 0.1g/L MgSO4 x 7H2O, 1 % tributrine) and the pH adjusted to 6.5-7. It was incubated for 48 h in a 37°C shaking oven (120 rpm) under 10 % CO2 conditions. After incubation, it was centrifuged at 9,798 x g for 30 min at 4 °C. The cooled acetone was mixed with the obtained filtrate at a 1:5 ratio, kept at 4 °C for 24 h, and centrifuged at 9,798xg, 4 °C for 15 min.

In the application where olive oil waste is used, the same processes were applied by adding 10 % olive waste to the lipase medium (0.1 g/L CaSO4, 0.5 g/L KH2PO4, 0.1 g/L MgSO4 x 7H2O, 1 % tributrine).

Then, 3 mL of the enzyme was placed into the dialysis tube (Sigma PURX12015), which in turn was placed in Tris HCl buffer. Tris HCl buffer was changed every 24 h. After 48 h, partially purified enzyme was obtained. Partially purified enzymes were used in the experiments. Experiments were carried out in two replicas each.

Assay of lipase activity

Para-nitrophenyl palmitate (p -NPP) method, which is a spectrophotometric method, was used for lipase activity determination (Arora, 2013). This method determined lipase activity by measuring p-NPP at a wavelength of 405 nm in a spectrophotometer. One unit (U) of lipase activity is the amount of enzyme that releases 1 µmol p-nitrophenol per unit time (min).

Determination of the molecular weight of lipase

The protein amount of the lipase enzyme was determined by the Bradford method (Bradford, 1976) in a spectrophotometer at 590 nm using Coomassie Brilliant Blue G-250 dye. The molecular weight of the enzyme was determined by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) according to the method of Laemmli (1970).

Determination of Factors Affecting Partially Purified Ex-tracellular Enzyme Activity

Effect of temperature and pH on enzyme activity

To determine the effect of temperature on partially purified extracellular enzyme activity, the enzyme was incubated for 1 h at 5 oC, 20 oC, 30 oC, 37 oC, 40 oC, 45 oC, 55 oC and 65 oC in 50 mM sodium phosphate buffer (pH 7).

To determine temperature stability, lipase enzyme was placed in 50 mM sodium phosphate buffer (pH 7) and incubated at 5 oC, 20 oC, 30 oC, 45 oC and 55 oC for 5 min, 1 h, 4 h, 24 h and 48 h. After cooling, the remaining enzyme activity was measured (Esteban-Torres et al., 2015).

The effect of pH on lipase activity was determined by keeping the enzyme in buffers prepared between pH 3-11. Acetic acid-sodium acetate buffer was used for pH 3-5, sodi-um phosphate buffer for pH 6, Tris-HCl buffer for pH 7-8, and glycine NaOH buffer for pH 9 (Esteban-Torres et al., 2015).

In order to determine the enzyme pH resistance, 200 µL of buffers at different pHs were placed in the microtubes. On top of it, 200 µL of enzyme were added and incubated at 45 °C for 2 and 3.5 h. The remaining enzyme activity was determined (Esteban-Torres et al., 2015).

Effect of some surfactants and cations on enzyme activity

To determine the effect of some cations, surfactants, and solutions on the enzyme activity, MnCl2, CuCl2, MgCl2, KCl, NiCl2, CaCl2, HgCl2, and ZnCl2 of 1 mM were added, and the enzyme activity determined at 405 nm in a spectrophotometer.

The effect of urea, EDTA, SDS, tween 20, tween 80, and triton X-100 on enzyme activity was determined by adding 1 µL to the medium (Esteban-Torres et al., 2015, Ramakrishnan, et al., 2016).

Effect of salt on enzyme activity

To determine the effect of sodium chloride (NaCl), it was added to the buffer at concentrations of 0 %, 1 %, 5 %, 10 %, 15 %, 20 %, 25 % (w/v) and the enzyme activity determined in spectrophotometer at 405 nm (Esteban Torres et al., 2015).

Result and discussion

In recent years, producing useful substances from waste materials has been of great importance. Thus, products with economic importance can also be obtained while preventing environmental pollution. In the study, it was determined that olive waste could be used in the nutrient medium and, in this way, a product of high economic importance can be obtained.

The extracellular lipase enzyme obtained without using olive oil waste and using 10 % olive oil was partially purified and used in the tests. The wet and dry weights and protein contents of the obtained lipase enzymes are given in Table 1.

Table 1 Dry weight and protein amounts of extracellular lipase enzymes from E. faecium E68. 

Tabla 1 Peso seco y concentración proteica de las enzimas lipasa extracelulares de E. faecium E68 

Protein (BSA mg/mL) Dry weight (mg/mL) Wet weight (mg/mL)
Medium 0.315 0.396 1.078
Olive oil waste 0.691 0.360 1.133

The protein amounts in the extracellular enzymes were found to be 0.315 mg/mL for the lipase enzyme produced in the medium, while it was 0.691 mg/mL for the partially purified enzyme produced in olive oil waste.

As a result of E. faecium E68 SDS-PAGE analysis, the molecular weight of lipase enzymes was determined at around 19-20 kDa (Figure 1).

Figure 1 SDS-PAGE analysis of lipase enzymes from partially purified E. faecium E68. M. marker. 

Figura 1 Análisis SDS-PAGE de enzimas lipasa de E. faecium E68 parcialmente purificado. Marcador M. 

The effects of temperature, pH, cations, and surfactants on the enzyme activity were determined.

The partially purified extracellular enzyme showed high lipolytic activity between 5 °C and 65 °C. The highest activity of the enzyme produced in the lipase medium was between 5 - 20 oC (Figure 2). Partially purified extracellular enzyme activity obtained with E. faecium E68 in olive waste was highest at 45 oC (Figure 2). Temperatures above 45 oC caused a decrease in enzyme activity (Figure 2). It has been reported that the partially purified extracellular enzyme obtained from E. durans E114 shows maximum activity at 30 - 45 oC (Acu et al., 2021). Maximum activity in the E. faecium lipase enzyme was observed at 40 °C (Ramakrishnan et al., 2016). Dellali et al. (2020) reported that the optimum lipase activity of E. faecium strains was 30-40 oC.

Figure 2 The effect of temperature on the E. faecium E68 extracellular lipase enzyme activity. Olive oil waste, medium. 

Figura 2 Efecto de la temperatura sobre la actividad enzimática de la lipasa extracelular de E. faecium E68.Residuos de aceite de oliva, medio. 

Lipase enzyme production was performed with E. faecium E68 in lipase medium containing 10 % olive oil waste (pH 6.5) after 48 h of incubation at 120 rpm at 37 oC.

Higher enzyme activity was obtained in the extracellular enzyme produced by E. faecium E68 in the fattening medium where olive waste was used. Lipid sources, such as natural oils have been shown to stimulate lipase production. Olive oil is one of the best inducers of lipase production (Zarevúcka, 2012). The presence of a certain amount of olive oil in olive oil waste stimulated lipase production. The activity of intracellular and extracellular lipases increases with increasing lipid concentration (Zarevúcka, 2012). The reason for the higher activity of the lipase enzyme produced using olive oil waste may be related to the increased lipid concentration.

Partially purified enzyme activity produced in olive waste decreased at 20 oC and remained stable at 55 oC, although it was below the optimum activity. The lipase enzyme produced in the lipase medium maintained its activity at 20 oC (Figure 3). The temperature tolerance of the extracellular enzyme remained quite stable after an incubation period of 24-48 h at 30-45 °C. Ramakrishnan et al. (2016) reported that the E. faecium lipase enzyme activity is stable between 30-70 oC. Researchers have reported that the enzyme activity is stable at 80-100 oC, and that enzyme activity does not remain after 100oC. Esteban Torres et al. (2015) observed that the maximum activity of the L. plantarum esterase enzyme is at 40 oC. They reported that the enzyme showed only 40 % of its activity at 5 oC, and, after 10 h of incubation at 55 oC and 65 oC, 40 % of the activity remained. Francisco et al. (2019) reported that the decrease in enzyme activity with temperature is associated with the change in its three-dimensional structure. It has been found that the alpha helix decreases at temperatures above 50 oC. At temperatures above 70 oC, the beta sheet increases while maintaining a low alpha helix. Opening the protein results in permanent inactivation and denaturation (Ismail et al., 2021).

Figure 3 Temperature resistance of E. faecium E68 extracellular lipase enzyme according to different residence times at different temperatures. A; olive oil, B; Medium.5 min,1 h,4 h,24 h, 48 h. 

Figura 3 Estabilidad a la temperatura de la enzima lipasa extracelular de E. faecium E68. A; aceite de oliva, B; Medio. 5 min,1 h, 4 h,24 h,48 h. 

The highest activity of the lipase enzyme, produced in olive waste and partially purified, was obtained at pH 10 (Figure 4). Partially purified enzymes were alkaline in nature. The optimum pH was found to be 10. While the lowest activity is obtained at pH 6, the enzyme has higher activity at pH 3. Acidic pH activity has been observed for a lipase from E. du-rans 27 isolated from fish processing waste. Lipase from ED-27 showed optimal activity at pH 4.6 and at temperature 30 °C (Ramakrishnan et al., 2015). A highly alkaline extracellular lipase that exhibits maximum hydrolytic activity at pH 10.8 has been reported from E. faecium MTCC5695 (Ramakrishnan et al., 2016). Dellali et al. (2020) reported that the optimum activity of the enzyme produced by E. faecium strains is between pH 6 and 9.

Figure 4 Effect of pH on the activity of the E. faecium E68 extracellular lipase enzyme. Olive oil waste (U/mL), Medium (U/mL) 

Figura 4 Efecto del pH sobre la actividad de la enzima lipasa extracelular de E. faecium E68. Residuos de aceite de oliva (U/mL), Medio (U/mL) 

The pH stability of the extracellular enzymes obtained from E. faecium E68 was determined by incubating them at different pH values for 2 h and 3.5 h at 45 oC. The lipase produced in olive oil waste by E. faecium E68 remained significantly stable after 2 h and 3.5 h at pH 3 (respectively % 77,36 and % 70,79). However, the lipase activity produced in the lipase production medium was lower at pH 3. Similar acidic pH activity has been observed for a lipase from E. durans NCIM5427 from fish waste isolated from slaughterhouse waste (Ramakrishnan et al., 2015). For the enzyme produced in the lipase production medium, the activity loss was higher after 2 h and 3.5 h at pH 3. The highest loss of activity was observed at pH 5. Enzymes remained stable at alkaline pH for 2 h and 3.5 h of standing (Figure 5).

Figure 5 pH resistance of E. faecium E68 extracellular lipase enzyme according to different residence times (2 and 3.5 h) at different pH. Olive waste 2 h, Olive waste 3,5h; Medium 2 h, Medium 3,5 h. 

Figura 5 Resistencia al pH de la enzima lipasa extracelular de E. faecium E68 según diferentes tiempos de residencia (2 y 3,5 h) a diferentes pH. Residuos de aceituna 2 h, Residuos de aceituna 3,5 h; Medio 2 h, Medio 3.5 h. 

Lipolytic isoenzymes from a thermophilic Bacillus sp. have also been observed. It showed optimum activity at pH 8.5 and was reported to be very stable at pH 6.0 - 8.0 (Nawani and Kaur, 2007).

The effects of some ions and additives on the enzyme activity are given in Table 2. One mM Ca2+, Mn2+, Cu2+, Ni2+, Zn2+, Mg2+ and K+ ions significantly increased the lipase activities produced in olive oil wastes. Contrary to our findings, Ramakrishnan et al. (2015) reported that it significantly reduced Ca2+ and Mg2+ lipase activity. Mercury (Hg2+) led to a strong decrease in lipase activity. In the lipase enzyme produced in the lipase environment, metals other than Mg decreased the enzyme activity (Table 2). EDTA, which can affect the interface region between substrate and lipase, increased enzyme activity, however, some studies have reported that it reduces activity (Sztajer et al., 1992). The activity of lipase Lp_3562 was strongly inhibited by Hg2+, Cu2+ and SDS (Esteban-Torres et al., 2014a). Urea, Hg+2, Mn+2, Cu+2, Ni+2, Zn+2 and SDS, inhibited the activity of the enzyme produced in the lipase medium. Similarly, Dellali et al. (2020) reported that although the effect of metal and additive ions on esterase activity varies from bacteria to bacteria, they inhibit SDS, NaN3, CuCl2, EDTA, AgNO3 and HgCl2 enzyme activity.

Table 2 Effect of additives on the lipolytic activity of E. faecium E 68. 

Tabla 2 Efecto de aditivos sobre la actividad lipolítica de E. faecium E 68. 

Additives Relative activity (%)
Medium Olive oil waste
Control 100 100
HgCl2 41.7 48.9
CaCl2 104.7 79.3
MnCl2 112.8 88.2
CuCl2 133.9 85.8
NiCl2 110.0 86.1
KCl 130.8 90.9
ZnCl2 105.2 89.2
MgCl2 150.9 123.3
EDTA 101.4 108.4
Urea 128.7 92.8
SDS 87.3 91.2
TritonX-100 81.5 120.5
Tween20 104.6 93.5
Tween80 105.4 128.5

While Triton X-100 and Tween 80 of surfactants increased enzyme activity, Tween 20 and SDS decreased enzyme activity.

This enzyme has high salt resistance and shows halophilic properties, an important feature in the preparation of foods. Even an increase in enzyme activity was observed (Figure 6). The salt resistance of this enzyme is high, and it showed halophilic properties. This feature is important for the preparation of foods.

Figure 6 Effect of sodium chloride on the lipolytic activity of E. faecium E 68. Olive oil waste; medium. 

Figura 6 Efecto del cloruro de sodio sobre la actividad lipolítica de E. aecium E 68. Residuos de aceite de oliva; medio. 

The lipase enzyme produced in olive waste is promising in the food industry due to its resistance to 55 oC for 48 h, not losing its activity at low temperatures, and its halophilic properties. The use of these lipases is important as they can provide some advantages in food production.

The activity of lipase enzyme produced in olive oil waste was higher. The reason for this may be the presence of a small amount of olive oil residue in it. Olive oil has a significant effect for increasing lipase activity. It has been reported that the most suitable inducer in lipase production is olive oil. This has been associated with high levels of unsaturated grade free fatty acids, particularly oleic acid, in oil (Amenaghawon et al., 2022). This has been shown to facilitate cell growth and consequently increase both intracellular and extracellular lipase activity (Suci et al., 2018). A similar observation was reported by Brozzoli ve ark. (2009) and Rajendran and Thangavelu (2012).

Conclusion

The results revealed that natural substrate such as olive oil waste has good inducing properties for lipase synthesis. Therefore, it may be beneficial to use olive oil waste as a cost-effective source for lipase production. The relative stability of E. faecium E68 lipase at high temperatures may make it usable for biotechnological processes, as enzymes that can withstand high temperatures longer, attract the attention of industries. It is important that the lipase enzyme produced by E. faecium E68 has high activity at 45 oC and pH 10, as well as showing activity in acidic conditions such as pH 3. It is promising in the food industry with its resistance to 55 oC for 48 h, its effectiveness at low temperatures and its halophilic feature. Olive oil can be an important substrate for waste lipase production. Thus, environmental pollution can be prevented, and a biotechnological product is also obtained.

References

Acu, E., Kılıç, V., and Kıvanç, M. 2021. Production and characterization of extracellular lipase from Enterococcus durans. The J of Food 46(2):474-487 doi: 10.15237/gida. GD21020. [ Links ]

Adetunji, A.I., and Olaniran, A.O. 2021. Production strategies and biotechnological relevance of microbial lipases: a review. Braz J Microbiol 52(3):1257-1269. doi: 10.1007/s42770-021-00503-5. [ Links ]

Amenaghawon, A.N., Orukpe, P.I., Nwanbi-Victor, J., Okedi, M.O., and Aburime, E.I. 2022. Enhanced lipase production from a ternary substrate mix of agricultural residues: A case of optimization of microbial inducers and global sensitivity analysis. Bioresour Technol Reports 17, 101000. doi. org/10.1016/j.biteb.2022.101000. [ Links ]

Angenent, L.T., Karim, K., Al-Dahhan, M.H., Wrenn, B.A., and Domiguez-Espinosa, R. 2004. Production of bioenergy and bio-chemicals from industrial and agricultural wastewater. Trends Biotechnol 22:477-485. doi: 10.1016/j. tibtech.2004.07.001. [ Links ]

Arora, P.K. 2013. Staphylococcus lipolyticus sp. nov., a new cold-adapted lipase producing marine species. Annals of Microbiology, 63(3), 913-922. doi: 10.1007/s13213-012-0544-2 [ Links ]

Ayed, L., Assas, N., Sayadi, S. and Hamdi, M. 2005. Involvement of lignin peroxidase in the decolourization of black olive millwastewaters by Geotrichum candidum. Lett Appl Microbiol 40:7-11. doi:10.1111/j.1472-765X.2004.01626. x. [ Links ]

Babu, I.S. and Rao, G.H. 2007. Lipase production by Yarrowia lipolytica NCIM 3589 in solid state fermentation using mixed substrate. Research Journal of Microbiology, 2(5), 469-474. doi:10.3923/JM.2007.469.474. [ Links ]

Bharathi, D., and Rajalakshmi, G. 2019. Microbial lipases: an overview of screening, production and purification. Biocatal Agric Biotechnol 22:101368 doi:10.1016/j.bcab.2019.101368 [ Links ]

Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry,72(1-2), 248-254. [ Links ]

Brozzoli, V., Crognale, S., Sampedro, I., Federici, F., D’Annibale, A., and Petruccioli, M., 2009. Assessment of olive-mill wastewater as a growth medium for lipase production by Candida cylindracea in bench-top reactor. Bioresour. Technol. 100, 3395-3402. doi: 10.1016/j.biortech.2009.02.022. [ Links ]

Couto, S.R., Sanromán, M.A. 2006. Application of solid-state fermentation to food industry-a review. J Food Eng. 76: 291-302. doi:10.1016/J.JFOODENG.2005.05.022 [ Links ]

Dellali, A., Karam, Z.H. and Karam, N.E., 2020. Lipase and esterase activities of lactic acid bacteria isolated from different biotopes. African J of Biotechnol 19(4):156-164. doi: 10.5897/ AJB2020.17106. [ Links ]

Esteban-Torres, M., Mancheno, J. M., de las Rivas, B., and Munoz, R. 2015. Characterization of a halotolerant lipase from the lactic acid bacteria Lactobacillus plantarum useful in food fermentations. LWT-Food Sci and Technol, 60(1): 246-252. doi:10.1016/J.LWT.2014.05.063 [ Links ]

Fenice, M., Sermanni, G.G., Federici, F., and D’Annibale, A., 2003. Submerged and solid-state bioprocesses forlaccase and manganese-peroxidase production by Panustigrinuson olive-mill wastewater-based media. J Biotechnol 100:77-85. doi: 10.1016/s0168-1656(02)00241-9. [ Links ]

Filipe, D., Fernandes, H., Castro, C., Peres, H., Oliva-Teles, A., Belo, I., and Salgado, J.M. 2020. Improved lignocellulolytic enzyme production and antioxidant extraction using solid-state fermentation of olive pomace mixed with winery waste. Biofuels Bioprod. Biorefining-Biofpr. 14:78-91. doi: 10.1002/ bbb.2073. [ Links ]

Fki, I., Allouche, N. and Sayadi, S. 2005. The use of polyphenolic extract, purified hydroxytyrosol and 3,4-dihydroxyphenyl acetic acid from olive mill wastewater for the stabilization of refined oils: a potential alternative to synthetic antioxidants. Food Chem 93:197 - 204. doi:10.1016/J.FOODCHEM.2004.09.014 [ Links ]

Francisco, C.C., Luis, C.J., Marina, E.J., Javier, C.F., Alexis, L.A., Del Carmen, S.H., and Alfredo, R.I.. 2019. Effect of temperature and pH on the secondary structure and denaturation process of jumbo squid hepatopancreas cathepsin D. Protein Pept Lett. 26(7):532-541. doi: 10.2174/0929866526666190405124353. [ Links ]

Hamrouni, R., ClaeysBruno, M., Molinet, J., Masmoudi, A., Roussos, S., and Dupuy, N. 2020. Challenges of enzymes, conidia and 6-pentyl-alpha-pyrone production from solid state fermentation of agroindustrial wastes using experimental design and T. asperellum strains. Waste and Biomass Valorization 11:5699-5710. doi:10.1007/s12649-019-00908-2. [ Links ]

Ismail, A.R., Kashtoh, H., and Baek, K.H. 2021. Temperature-resistant and solvent-tolerant lipases as industrial biocatalysts: Biotechnological approaches and applications. Int J Biol Macromol. 187:127-142. doi: 10.1016/j. ijbiomac.2021.07.101. [ Links ]

Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-885. [ Links ]

Leite, P., Salgado, J.M., Venâncio, A., Domínguez, J.M., and Belo, I. 2016. Ultrasounds pretreatment of olive pomace to improve xylanase and cellulase production by solid-state fermentation. Bioresour Technol 214:737-746. doi: 10.1016/j.biortech.2016.05.028 [ Links ]

Leite, P., Sousa, D., Fernandes, H., Marta Ferreira, M., et al. 2021. Recent advances in production of lignocellulolytic enzymes by solid-state fermentation of agro-industrial wastes. Current Opinion in Green and Sustainable Chemistry 27:100407. doi: 10.1016/j.cogsc.2020.100407. [ Links ]

Liu, S.Q., Holland, R., Crow, V.L. 2001. Purification and properties of intracellular esterases from Streptococcus thermophilus. International Dairy Journal 11:27-35. doi:10.1016/S0958-6946(01)00035-8. [ Links ]

Lopes, M.F.S., Leitao, A.L., Regalla, M., Marques, J.J.F., Carrondo, M.J.T., and Crespo, M.T.B. 2002. Characterization of a highly thermostable extracellular lipase from Lactobacillus plantarum. Int J of Food Microbiol 76:107-115. doi: 10.1016/ s0168-1605(02)00013-2. [ Links ]

Mahanta, N., Gupta, A., Khare, S.K. 2008. Production of protease and lipase by solvent tolerant Pseudomonas aeruginosa PseA in solid-state fermentation using Jatropha curcas seed cake as substrate. Bioresour. Technol . 99, 1729-1735. doi: 10.1016/j.biortech.2007.03.046. [ Links ]

Mantzavinos, D. and Kalogerakis, N. 2005. Treatment of olive milleffluents. Part I. Organic matter degradation by chemical and biological processes. An overview. Environ Int 31:289-295. doi:10.1016/J.ENVINT.2004.10.005 [ Links ]

Meyers, S.A., Cuppett, S.L., and Hutkins, R.W. 1996. Lipase production by lactic acid bacteria and activity on butter oil. Food Microbiology 13(5):383-389. DOI:10.1006/ FMIC.1996.0044 [ Links ]

Nawani, N., and Kaur, J. 2007. Studies on lipolytic isoenzymes from a thermophilic bacillus sp: production, purification and biochemical characterization. Enzym. Microb. Technol. 40, 881-887. doi: 10.1016/j.enzmictec.2006.07.006. [ Links ]

Papadimitriou, V., Maridakis, G.A., Sotiroudis, T.G. and Xenakis, A. 2005. Antioxidant activity of polar extracts from olive oil and olive mill wastewaters: an EPR and photometric study. Eur J Lipid Sci Technol 107:513-520. doi:10.1002/ejlt.200501165 [ Links ]

Rajendran, A., Thangavelu, V., 2012. Optimization and modeling of process parameters for lipase production by Bacillus brevis. Food Bioprocess Technol. 5, 310-322. doi:10.1007/ s11947-010-0387-4 [ Links ]

Ramakrishnan, V., Goveas, L.C., Suralikerimath, N., Jampani, C., Halami, P.M., Narayan, B. 2016. Extraction and purification of lipase from Enterococcus faecium MTCC5695 by PEG/ phosphate aqueous-two phase system (ATPS) and its biochemical characterization. Biocatalysis and Agricultural Biotechnology 6:19-27. doi: 10.1016/j.bcab.2016.02.005 [ Links ]

Ramakrishnan, V., Goveas, L.C., Narayan, B., and Halami, P.M. 2013. Comparison of lipase production by Enterococcus faecium MTCC 5695 and Pediococcus acidilactici MTCC 11361 using fish waste as substrate: optimization of culture conditions by response surface methodology. ISRN Biotechnology ID 980562, doi:10.5402/2013/980562 [ Links ]

Ramakrishnan, V., Goveas, L.C., Halami, P.M., Narayan, B. 2015. Kinetic modeling, production and characterization of an acidic lipase produced by Enterococcus durans NCIM5427 from fish waste. J Food Sci Technol 52(3):1328-1338 doi: 10.1007/s13197-013-1141-1145. [ Links ]

Sarika, R., Kalogerakis, N. and Mantzavinos, D. 2005. Treatment of olive mill effluents. Part II. Complete removal of solids by direct flocculation with poly-electrolytes. Environ Int 31:297-304. doi:10.1016/j.envint.2004.10.006. [ Links ]

Suci, M., Arbianti, R., Hermansyah, H. 2018. Lipase production from Bacillus subtilis with submerged fermentation using waste cooking oil. In: IOP Conference Series: Earth and Environmental Science. Institute of Physics Publishing, p. 12126. doi:10.1088/1755-1315/105/1/012126. [ Links ]

Sukohidayat, N.H.E., Zarei, M. , Baharin, B.S., and Manap, M.Y. 2018. Purification and characterization of lipase produced by Leuconostoc mesenteroides Subsp. mesenteroides ATCC 8293 using an aqueous two-phase system (ATPS) composed of triton X-100 and maltitol. Molecules 23, 1800; doi:10.3390/ molecules23071800 [ Links ]

Sztajer, H., Lunsdorf, H., Erdmann, H., Menge, U., Schmid, R. 1992. Purification and properties of lipase from Penicillium simplicissimum. Biochim. Biophys. Acta 1124: 253-261. doi: 10.1016/0005-2760(92)90137-k. [ Links ]

Treichel, H., Oliveira, D., Mazutti, M.A., Luccio, M.D., Oliveira, J.V. 2010. A Review on microbial lipases production. Food Bioprocess Technol 3:182-196 doi: 10.1007/s11947-009-0202-2 [ Links ]

Zarevúcka, M. 2012. Olive Oil as Inductor of Microbial Lipase, Olive Oil- Constituents, Quality, Health Properties and Bioconversions, Dr. Dimitrios Boskou (Ed.), ISBN: 978-953-307-921-9, InTech, Available from: http://www.intechopen.com/books/olive-oil-constituents-quality-health-properties-and-bioconversions/olive-oil-as-inductor-of-microbial-lipase. [ Links ]

Received: May 12, 2022; Accepted: August 08, 2022

*Autor para correspondencia: Merih Kivanc. Correo electrónico: mkivanc@eskisehir.edu.tr

The authors declare no conflict of interest.

Creative Commons License Este es un artículo publicado en acceso abierto bajo una licencia Creative Commons