Hydrolysis Of Olive Oil Research Articles

Magnetic polydopamine nanoparticles as stabilisers for enzyme-Pickering emulsions: Application in the interfacial catalytic reaction of olive oil

Solid particles are essential for stabilising Pickering emulsions and improving interfacial catalytic reactions. We constructed magnetic polydopamine nanoparticles to stabilise lipase-Pickering emulsions for olive oil deacidification. The results showed that the nanoparticles had a core-shell structure with an average particle size of 605.8 nm, a zeta potential of −39.3 mV and a contact angle of 55.9°, which effectively stabilised the emulsion. The particles were added to the lipase solution and sonicated to construct the emulsion system. The emulsion droplets were the smallest and most uniformly distributed under 400 W ultrasonic irradiation for 10 min. The lipase adsorbed on the oil-water interface and promoted the hydrolysis of olive oil. The released fatty acid content increased 1.7-fold compared with the non-emulsion. This study not only provides a new immobilisation method for the interfacial catalysis of lipase but also provides ideas for the high-value utilisation of high acid-value oil resources.

Kinetic Model for the Hydrolysis of Olive Oil by Lipase in Absence of Emulsifier

Kinetic Model for the Hydrolysis of Olive Oil by Lipase in Absence of Emulsifier

Mycelium-bound lipase as skillful biocatalysts: Production of fatty acid concentrates from waste oils for the food industry

This study aimed to produce Rhizopus oryzae whole-cells having high lipolytic activity. The hydrolytic activity of 558.99 U.g−1 and total lipase activity of 7440.17 U was obtained in 72 h of cultivation using 30 g.L−1 of canola oil. Optimal activity was observed at pH 7.5 and 30 °C. Kinetic parameters were determined through olive oil hydrolysis (Km = 59.96 mM, Vmax = 344.09 μmol.g−1.min−1). Thermal inactivation constant and half-life were 0.15 h−1 and 4.62 h at 50 °C. Hydrolysis of vegetable oils achieved 80% yield in 24 h. These results suggest that the biocatalyst shows the potential to produce free fatty acids of great industrial interest.

Efficiency of an alkaline, thermostable, detergent compatible, and organic solvent tolerant lipase with hydrolytic potential in biotreatment of wastewater

Discharging the tannery wastewater into the environment is a serious challenge worldwide due to the release of severe recalcitrant pollutants such as oil compounds and organic materials. The biological treatment through enzymatic hydrolysis is a cheap and eco-friendly method for eliminating fatty substances from wastewater. In this context, lipases can be utilized for bio-treatment of wastewater in multifaceted industrial applications. To overcome the limitations in removing pollutants in the effluent, we aimed to identify a novel robust stable lipase (PersiLipase1) from metagenomic data of tannery wastewater for effective bio-degradation of the oily wastewater pollution. The lipase displayed remarkable thermostability and maintained over 81 % of its activity at 60 °C.After prolonged incubation for 35 days at 60°C, the PersiLipase1 still maintained 53.9 % of its activity. The enzyme also retained over 67 % of its activity in a wide range of pH (4.0 to 9.0). In addition, PersiLipase1 demonstrated considerable tolerance toward metal ions and organic solvents (e.g., retaining >70% activity after the addition of 100 mM of chemicals). Hydrolysis of olive oil and sheep fat by this enzyme showed 100 % efficiency. Furthermore, the PersiLipase1 proved to be efficient for biotreatment of oil and grease from tannery wastewater with the hydrolysis efficiency of 90.76 % ± 0.88. These results demonstrated that the metagenome-derived PersiLipase1 from tannery wastewater has a promising potential for the biodegradation and management of oily wastewater pollution.

Optimization of catalytic properties of Mucor racemosus lipase through immobilization in a biocompatible alginate gelatin hydrogel matrix for free fatty acid production: a sustainable robust biocatalyst for ultrasound-assisted olive oil hydrolysis

Immobilization is a key technology that improves the operational stability of enzymes. In this study, alginate-gelatin (Alg-Gel) hydrogel matrix was synthesized and used as immobilization support for Mucor racemosus lipase (Lip). Enzyme catalyzed ultrasound-assisted hydrolysis of olive oil was also investigated. Alg-Gel matrix exhibited high entrapment efficiency (94.5%) with a degradation rate of 42% after 30 days. The hydrolysis of olive oil using Alg-Gel-Lip increased significantly (P < 0.05) as compared to free Lip. Optimum pH and temperature were determined as pH 5.0 and 40 °C, respectively. The Vmax values for free and immobilized Lip were determined to be 5.5 mM and 5.8 mM oleic acid/min/ml, respectively, and the Km values were 2.2 and 2.58 mM/ml respectively. Thermal stability was highly improved for Alg-Gel-Lip (t1/2 650 min and Ed 87.96 kJ/mol) over free Lip (t1/2 150 min and Ed 23.36 kJ/mol). The enzymatic activity of Alg-Gel-Lip was preserved at 96% after four consecutive cycles and 90% of the initial activity after storage for 60 days at 4 °C. Alg-Gel-Lip catalyzed olive oil hydrolysis using ultrasound showed a significant (P < 0.05) increase in hydrolysis rate compared to free Lip (from 0.0 to 58.2%, within the first 2 h). In contrast to traditional methodology, using ultrasonic improved temperature-dependent enzymatic catalyzed reactions and delivered greater reaction yields. Results suggest that Alg-Gel-Lip biocatalyst has great industrial application potential, particularly for free fatty acid production. In addition, the combined use of enzyme and ultrasound has the potential of eco-friendly technology.

Dynamic Modelling of Enzymatic Hydrolysis of Oil Using Lipase Immobilized on Zeolite

Immobilization has been proposed as a way to simplify the separation and repeated reuse of enzymes, which is essential for their feasible application at industrial scales. However, in their immobilized form, enzyme activity is fully utilized, due primarily to the additional diffusion limitations. Here, the immobilization of lipase on zeolite and its use in catalyzing oil hydrolysis is studied. Adsorption isotherms were investigated, and the data identified the model that best describes the process, which is the Sips model. The adsorption capacity of zeolite was determined as 62.6 mg/g, which is relatively high due to the high porosity of the support. The rate of enzymatic hydrolysis of olive oil, using the immobilized lipase, was determined at a pH of 7 and a temperature of 40 °C and was compared to that when using free enzymes. The results determined the parameters for a diffusion-reaction model. The effects of both the surface reaction and diffusion were found to be significant, with a slightly higher effect from surface reactions.

Cetyl trimethyl ammonium bromide-activated lipase from Aspergillus oryzae immobilized with Cu3(PO4)2⋅3H2O via biomineralization for hydrolysis of olive oil

Cetyl trimethyl ammonium bromide (CTAB)-activated lipase-mineral hybrid microflowers (CTAB-Lip-MHFs) were fabricated via biomineralization using Cu3(PO4)2⋅3H2O as the mineral for olive oil hydrolysis. FT-IR spectra indicated that immobilized lipase molecules in CTAB-Lip-MHFs has good structural integrity, and XPS spectra revealed that immobilized lipase molecules were present at the surface of CTAB-Lip-MHFs. Kinetic studies suggested that CTAB-Lip-MHFs had good substrate affinity ascribed to interfacial effect caused by CTAB, meanwhile, the open conformation of lipase activated by CTAB contributed to increased reaction rate with substrate. The generated fatty acids via hydrolysis of olive oil by CTAB-Lip-MHFs were 146% of that by free lipase, moreover, CTAB-Lip-MHFs could retain 76.5% and 93.2% of initial activities after storage at 50 °C for 10 h and 4 °C for 30 days, respectively, indicating the good stability of CTAB-Lip-MHFs. In addition, after 5 recycles, CTAB-Lip-MHFs preserved 72.7% of initial activity, demonstrating the favorable reusability of CTAB-Lip-MHFs for fatty acid production.

The Activation Energies and Optimum Temperatures of Olive Oil Hydrolysis By Lipase Porcine Pancreas

AbstractLipase activity is a perfect indicator for the monitoring of processes of bioremediation of degraded soils. Lipase is also used in the processes of oil hydrolysis in wastewater treatment. To be able to predict and model processes with used lipase in environmental operations, knowledge of the kinetic parameters of the process are required. The paper presents the determined values of activation energies and optimum temperatures for porcine pancreas lipase. The parameters were estimated based on the literature of the activity curvesvs. temperature for hydrolysis of olive oil by lipase. It was noticed that concentration of gum arabic added as an emulsifier during lipase activity measurements influences on the obtained values of determined parameters. A mathematical model describing the effect of temperature on porcine pancreas lipase activity was used. Based on the comparison analysis, the optimum temperatureToptwere obtained in the range from 313.30 ±0.56 to 319.62 ±0.96 K, activation energiesEawere from 51 ±10 to 82.6 ±9.9 kJ/mol, and values of deactivation energiesEdwere in the range from 122.7 ±4.0 to 150.9 ±5.8 kJ/mol.

Construction of lipid layer and monitoring its digestion by optical interferometry

A method for real-time monitoring of lipid digestion based on photonic crystals formed from silica was developed. As an effective “net”, the highly ordered silica colloidal crystal (SCC) film provides structural support for lipid payload. This method based on optical interferometric film kinetics was used to record the whole kinetics progress of olive oil hydrolysis by lipase in real time and calculate the kinetic Michaelis constant. The kinetic parameters were compared with the results determined by the titration method. The effects of bile salt content on lipase and olive oil layer were studied. This method provides a potential evaluation system for real-time digestion and degradation of edible oil in the food field. It also provides a basis for further real-time evaluation of lipid bioavailability in food systems by real-time recording the release and degradation of lipids in the food nano-matrix.

Covalent immobilization of lipase from Candida rugosa on epoxy-activated cloisite 30B as a new heterofunctional carrier and its application in the synthesis of banana flavor and production of biodiesel

In this paper, an epoxy-activated cloisite (ECL) was prepared as a new heterofunctional carrier via a reaction between cloisite 30B (CL) and epichlorohydrin and utilized for covalent immobilization of lipase from Candida rugosa. The lipase immobilized on the ECL (LECL) was successfully used in the olive oil hydrolysis, synthesis of isoamyl acetate (banana flavor), and biodiesel production. The TGA, FT-IR, SEM, and XRD were used to characterize CL, ECL, and LECL. The influences of temperature, pH, thermal stability, and storage capacity were examined in the olive oil hydrolysis. The effects of solvent, temperature, time, water content, and substrates molar ratio on the yields of ester and biodiesel were also investigated. In the optimized conditions, the hydrolytic activity of LECL was 1.85 ± 0.05 U/ mg, and the maximum yield of ester and biodiesel was 91.6% and 95.4%, respectively. The LECL showed good thermal stability and storage capacity compared to the free lipase. Additionally, LECL was reusable for both esterification and transesterification after being used for nine cycles.

The effect of pH on determination of activation energies and the optimum temperatures of hydrolysis of olive oil by lipase from porcine pancreas

The present paper reports the determination of the activation energies and the optimum temperatures of olive oil hydrolysis by porcine pancreas lipase with simultaneous effect of pH. The parameters were estimated based on the literature data on the activity curves versus temperature for olive oil hydrolysis by lipase obtained from porcine pancreas. It was assumed that both the hydrolysis reaction process and the deactivation process of lipase were first-order reactions by the enzyme concentration. A mathematical model describing the effect of temperature on porcine pancreas lipase activity was used. The determinate activation energies Ea were from 31.37 ± 5.38 kJ/mol to 61.60 ± 11.46 kJ/mol, the optimum temperatures Topt were obtained in the range from 305.46 ± 1.26 K to 313.23 ± 1.18 K and the values of deactivation energies Ed were in the range from 65.18 ± 3.19 kJ/mol to 109.27 ± 6.79 kJ/mol. The obtained results (Ea, Ed, Topt) might find application in research on the prognosis of pancreatic cancer.

Retracted: Partial purification and characterization of lipase from locally produced edible oil-seeds and its relevance in industries.

Lipase was extracted from germinating seeds of Helianthus annus (Sunflower), Zea mays (Maize), and Brassica compastris (Mustard). The lipolytic activity was assessed using olive oil as substrate at different germination-time and the maximum-activity was obtained after 120 hr. Partial-purification was executed by precipitating the seed-homogenate with varying concentration of ammonium sulfate solution. 80% ammonium sulfate solution showed maximum lipase activity of 5320IUml-1 , 3500IUml-1 , 3080IUml-1 with 9.6, 6.9, and 4.8-fold purification and total protein content of 162, 84, and 60 mg for partially purified enzyme extracts namely SN5, BN5, and MN5, respectively. The optimum temperature and pH observed for hydrolysis of olive oil were 37°C, and 8.0 respectively. Enzyme was found to be stable upto 6 days at 4°C and its activity was stimulated by Ca+2 ions. Oil-stains removal from cotton fabric was observed to be superior in the presence of lipase and detergent. Moreover, the SN5, BN5, and MN5 lipase increased free fatty acid release upto 4.2, 4.3, and 3.8 mg, respectively than wastewater without treatment of lipase (0.21 mg) and promoted fat hydrolysis to approximately 40, 42, and 48% mass reduction after 6 hr incubation of fat particle at a concentration of 20 mg/ml. Biodiesel produced by catalyzing transesterification of vegetable oil with SN5, BN5, and MN5 lipase provided an acid value of 0.8, 1.08, and 0.5 mg/g, viscosity 5.50, 5.7, and 5.53 mm2 /s and density 0.87, 0.88, and 0.79 g/ml, respectively. To the best of our knowledge, no such study has been conducted prior on lipase from the seeds mentioned above in Azad Kashmir region.

Design for preparation of more active cross-linked enzyme aggregates of Burkholderia cepacia lipase using palm fiber residue.

A new design of cross-linked enzyme aggregates (CLEAs) of Burkholderia cepacia lipase (BCL) based mainly on the use of lignocellulosic residue of palm fiber as an additive was proposed. Different parameters for the preparation of active CLEAs in the hydrolysis of olive oil, such as precipitation agents, crosslinking agent concentration, additives, and coating agents were investigated. The highest activity yield (121.1 ± 0.1%) and volumetric activity (1578.1 ± 2.5 U/mL) were achieved for CLEAs prepared using the combination of a coating step with Triton® X-100 and polyethyleneimine plus the use of palm fiber as an additive. The variations of the secondary structures of BCL-CLEAs were analyzed by second-derivative infrared spectra, mainly indicating a reduction of the α-helix structure, which was responsible for the lipase activation in the supramolecular structure of the CLEAs. Thus, these results provided evidence of an innovative design of BCL-CLEAs as a sustainable and biocompatible opportunity for biotechnology applications.

Isolation, Purification and Analysis of Pancreatic Lipase from ‘Gallus gallus domesticus’

This article discusses the isolation of pancreatic lipase enzyme from the pancreas of Gallus gallus domesticus. Whereas lipase catalyses the hydrolysis and the synthesis of esters formed from glycerol and long-chain fatty acids. Lipases occur widely in nature, it involves applications like organic syntheses, hydrolysis of fats, oils, modification of fats, flavor enhancement in food processing, detergent industries, pharmaceutical industries, chemical analyses, and biodiesel production. Pancreatic lipase was purified to the homogeneity by 70% saturated Ammonium sulphate further, it was dialysate using the dialysis membrane and then gel filtration chromatography was carried out by Sephadex G-75 and DEAE cellulose. The molecular weight of purified lipase sample was determined by SDSPAGE, it was found to be 98KDa. The lipase was active in the pH range of 5-10 with an optimum pH of 6.0. The optimum temperature for the hydrolysis of olive oil was 37ºC in the range of 25ºC - 50ºC.

Make proper surfaces for immobilization of enzymes: Immobilization of lipase and α-amylase on modified Na-sepiolite

This paper has focused on making suitable carriers for the immobilization of α-amylase from Bacillus subtilis and lipase from Candida rugosa via adsorption on modified Na-sepiolite (SEP). Two modified carriers were prepared by changing the nature of SEP with cetrimonium bromide surfactant at concentrations below the CMC of the surfactant, to produce SEP with monolayer surfactant (MSEP) with hydrophobic properties and above the CMC of the surfactant to produce SEP with bilayer surfactant (BSEP) with hydrophilic properties. The enzymatic activity of immobilized lipase on MSEP (MSEPL) and immobilized α-amylase on BSEP (BSEPA) was successfully tested in the hydrolysis of olive oil and starch. The support modification and immobilization process were characterized by BET, XRD, and SEM techniques. The results demonstrated that the specific activity of MSEPL and BSEPA was 1.74 and 2.28 U/mg, respectively. The remained activity of MSEPL (56.7%) and BSEPA (40.4%) after their incubation at 60 °C for 4 h was much higher than that of free enzymes. The residual activity of MSEPL and BSEPA was 77.4% and 66.7%, after 30 days of storage at 4 °C. The MSEPL and BSEPA also showed good reusability, and their relative activities were 54.0% and 44.2% after ten cycles.

Utilization of two modified layered doubled hydroxides as supports for immobilization of Candida rugosa lipase

In this study, two synthetic layered doubled hydroxides (LDH), including Mg/Al-CO3-LDH (LDH1) and Zn/Al-CO3-LDH (LDH2), were prepared using the co-precipitation method and modified with sodium dodecyl sulfate to be utilized as carriers for immobilization of Candida rugosa lipase via the adsorption. The activity of prepared biocatalysts was measured in the olive oil hydrolysis. The effects of lipase concentration, pH, storage stability and thermal resistance of the samples were also studied. The maximum activity was obtained at pH 6.0 for immobilized lipase on modified LDHs with monolayer surfactants, including MLDH1 (0.922 U/mg) and MLDH2 (0.744 U/mg), respectively. The remained activities for immobilized lipase on MLDH1 and MLDH2 after 24 h incubation at 60 °C were 85% and 81%, respectively. During the 25days of storage at 4 °C, immobilized lipases on MLDH1, MLDH2, and free lipase kept 87%, 86%, and 70% of their initial activities. The residual activities for immobilized lipase on MLDH1 and MLDH2 after reusing for ten cycles were 72% and 67% of their initial activities. Adsorption parameters for sorption of lipase on all supports were fitted to the Freundlich and Langmuir isotherms. Kinetic parameters obtained from the Michaelis-Menten equation on MLDH1 and MLDH2 were comparable to free enzyme.

Application of lipase immobilized on a hydrophobic support for the synthesis of aromatic esters

The present study aimed at preparing three biocatalysts via physical adsorption of lipases from Candida rugosa (CRL), Mucor javanicus, and Candida sp. on a hydrophobic and mesoporous support (Diaion HP-20). These biocatalysts were later applied to the synthesis of aromatic esters of apple peel and citrus (hexyl butyrate), apple and rose (geranyl butyrate), and apricot and pineapple (propyl butyrate). Scanning electron microscopy and gel electrophoresis confirmed a selective adsorption of lipases on Diaion, thus endorsing simultaneous immobilization and purification. Gibbs free energy (∆G) evinced the spontaneity of the process (-17.9kJ/mol≤∆G≤-5.1kJ/mol). Maximum immobilized protein concentration of 30mg/g support by CRL. This biocatalyst was the most active in olive oil hydrolysis (hydrolytic activity of 126.0±2.0U/g) and in the synthesis of aromatic esters. Maximum conversion yield of 89.1% was attained after 150Min for the synthesis of hexyl butyrate, followed by the synthesis of geranyl butyrate (87.3% after 240Min) and propyl butyrate (80.0% after 150Min). CRL immobilized on Diaion retained around 93% of its original activity after six consecutive cycles of 150Min for the synthesis of hexyl butyrate.

Study on Lipase-Catalyzed Hydrolysis of Olive Oil at Oil-Water Interface

Abstract Olive oil was selected as the oil substrate and hydrolyzed by Candida sp. 99–125 lipase. The hydrolysis rate of olive oil was used as an indicator. Based on the single factor experiment, the effects of dosage of Candida sp. 99–125 lipase, reacting temperature, pH value and water-oil ratio were investigated. Box-Behnken center combination and response surface methodology were utilized to optimize the hydrolysis rate. The results showed that the significant differences of each single factor on lipase hydrolysis of olive oil on the oil-water interface were different. pH value is the first significance factor, and the significance of water oil ratio on lipase hydrolysis of olive oil is second only to pH value. Finally, the mechanism of Candida sp. 99–125 lipase hydrolyzing olive oil at the oil-water interface was discussed.

Thermostable acidic lipase of Bacillus glycinifermentans-MK840989 isolated from contaminated environment; its optimization, purification and exploring potential applications

This study is the first report about isolation, purification and optimization of lipase from Bacillus glycinifermentans. In this study, Bacillus glycinifermentansMK-840989 was isolated from a local petrol pump. The bacterium showed lipolytic zones of 0.19cm, 0.044cm, and 0.28cm on peptone yeast agar, olive oil hydrolysis agar and chromogenic plate agar, respectively. B. glycinifermentans also produced an extracellular lipase (55.1µmol/ml). This bacterium preferred acidic environment (pH 5) for growing optimally at 80˚C when the medium was supplemented with 1% olive oil. The olive oil induced its growth up to 9h. The protein content of the purified lipase was estimated about 75mg/ml as compared to its crude form, i.e. 350mg/ml. The purified lipase was found to be thermostable acidic in nature as its optimum activity was observed at 90˚C (0.08U/ml) and pH 5 (0.02U/ml). Other optimization factors included 1% olive oil (0.065U/ml), 0.1mM maltose (0.023U/ml), 0.1mM Ca (0.025U/ml), 1% yeast extract (16.8U/ml), 1% wheat waste (0.019U/ml), 1% commercial detergent (0.016U/ml) and 1% tween-20 (0.015 U/ml). The purified lipase showed a polypeptide of 26.7kDa on SDS-PAGE. These features such as thermostability, acidic nature, ability to show activity in wheat waste and tolerance to detergents render the lipase of B.glycinifermentans MK-840989 as an attractive choice for biotechnologists to employ it at industrial level. The purified lipase of B.glycinifermentans MK-840989 can be a potential candidate for detergent and oil-remediation industry. It can help to replace conventional synthetic detergent as it is cost-effective and eco-friendly.

Optimization of Aspergillus niger lipase production by solid state fermentation of agro-industrial waste

Lipase is one of the most used industrial enzymes, which can be produced by Aspergillus niger. Large scale production of Aspergillus niger lipase is more profitable by using solid state fermentation method and utilizing agro-industrial waste as the substrate. Optimization of Aspergillus niger lipase production has been performed in this study. Optimization was performed on solid state fermentation of rice bran and Jathropa seed cake for 5 days with variations of inducer and extractant. Fermentation cake produced was extracted, filtered using muslin cloth, and centrifuged. The supernatant was spray-dried and assayed using olive oil hydrolysis titrimetry. Inducer optimization results showed that 1% of olive oil was the best inducer, yielding dry lipase extract with highest activity unit (176 U/ml enzyme). Extractant optimization results showed that 1% of NaCl – 0.5% of Tween 80 was the best extractant, yielding dry lipase extract with highest activity unit (282 U/ml enzyme).