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International Journal of Biological... Nov 2020In this study, two synthetic layered doubled hydroxides (LDH), including Mg/Al-CO-LDH (LDH1) and Zn/Al-CO-LDH (LDH2), were prepared using the co-precipitation method and...
In this study, two synthetic layered doubled hydroxides (LDH), including Mg/Al-CO-LDH (LDH1) and Zn/Al-CO-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.
Topics: Enzyme Stability; Enzymes, Immobilized; Hydrogen-Ion Concentration; Hydrolysis; Hydroxides; Kinetics; Lipase; Olive Oil; Saccharomycetales; Surface-Active Agents; Temperature
PubMed: 32562729
DOI: 10.1016/j.ijbiomac.2020.06.145 -
Preparative Biochemistry & Biotechnology 2022Lipid-rich wastewater from the local dairy industry (cheese whey) in the Galilee, Israel was hydrolyzed by using two different sources of lipase as hydrolytic enzymes:...
Lipid-rich wastewater from the local dairy industry (cheese whey) in the Galilee, Israel was hydrolyzed by using two different sources of lipase as hydrolytic enzymes: fungal () and animal porcine pancreatic lipase(PPL). Pretreatment efficiency was verified by comparative biodegradability tests of raw and treated wastewater samples. Simultaneous hydrolysis and anaerobic digestion in the same reactors were also tested. Enzymatic pretreatment of these samples at a concentration of 0.05 w v showed organic matter removal of 90% and methane formation increases of 140% for the fungal source enzyme (i.e., AY), while for the animal source enzyme (i.e., PPL) was 86 and 130%, respectively. Enzymatic pretreatment led to significant methane formation which was obtained only for moderate substrate concentration (initial chemical oxygen demand of 15 gL); While in high concentrated lipid-rich wastewater led to methane yield inhibition. The main finding was that the combination of AY enzyme with (i.e., enzyme mixture) led to a high efficiency in methane production (+152%) and organic materials removal (more than 90%). In summary, the use of fungal hydrolytic lipase mixed with fungus is a promising method for enhancing methane production during the biodegradation of fat and grease-rich wastewaters.
Topics: Animals; Biodegradation, Environmental; Hydrolysis; Industrial Microbiology; Lipase; Lipid Metabolism; Lipids; Methane; Saccharomycetales; Swine; Wastewater
PubMed: 33787468
DOI: 10.1080/10826068.2021.1901233 -
Tropical Animal Health and Production Apr 2021Mycotic mastitis is a neglected problem type of incurable chronic mastitis in sheep flock of many countries which associated with wide economic burden. In the current...
Mycotic mastitis is a neglected problem type of incurable chronic mastitis in sheep flock of many countries which associated with wide economic burden. In the current study, a total of 600 ewes at Menofia governorate, Egypt, were subjected to clinical and molecular examination using PCR-RFLP to estimate the prevalence of chronic mycotic mastitis and identify the causative agent. A structured questionnaire is distributed to shepherds in the study area to identify the risky behavioral practices being followed and lead to increase the prevalence of mycotic mastitis cases. The results showed that out of 600 ewes examined, 150 showed clinical signs of mastitis (25%). A total of 25 ewes with clinical mastitis did not respond to antibiotic treatment for long time and suffered from mycotic mastitis (16.7%, CI 11.1-23.6%). A total of 31 fungal isolates were identified: 14 yeast spp., Candida albicans, Candida parapasilosis, Candida rugosa, and Saccharomyces spp. and 17 mold spp., Alternaria spp., and Fusarium spp. Results showed also the widespread of risky practices among shepherds which could be responsible for the increase the prevalence of mycotic mastitis among ewes in the study area including presenting of decayed food to sheep, uncontrolled usage of antibiotics for mastitis treatment, lack of usage of antiseptics, and keeping of chronic infected animals in flocks for breeding. In conclusions, using of specific ITS1 and ITS4 primer sets with PCR-RFLP technique provided a suitable method for rapid identification and genotyping of Candida spp., Scaccharomyces, Alternaria, and Fusarium vertolliodes isolated from chronic mastitis in sheep. Furthermore, this study is considered up to our knowledge one of scarce estimates available on mycotic mastitis in sheep flocks in Egypt. Mycotic mastitis existed at higher prevalence estimates in the study area and educational campaigns to shepherds are much required to increase their awareness on the threat of risky of behaviors responsible for spread of the disease among their animals.
Topics: Animals; Causality; Egypt; Female; Genotype; Mastitis; Milk; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Prevalence; Saccharomycetales; Sheep; Sheep Diseases
PubMed: 33866417
DOI: 10.1007/s11250-021-02703-5 -
Mikrochimica Acta Nov 2023A sensitive and accurate chemiluminescence (CL) method was developed for one-step determination of diphenyl ether herbicides at trace level with nitrofen...
A sensitive and accurate chemiluminescence (CL) method was developed for one-step determination of diphenyl ether herbicides at trace level with nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether) as a model analyte. Candida rugosa lipase (CRL) was immobilized on a nanocarrier of amine-linked covalent organic framework (named as COF-300-AR) through a self-assembly strategy. The formed nanocomposite of COF-300-AR@CRL owns dual enzymatic catalytic activities. It can directly catalyze luminol-dissolved oxygen reaction to produce an intense CL emission by virtue of oxidase mimic activity of COF-300-AR but also effectively decompose nitrofen to release phenolic compounds by the immobilized CRL. The released phenolic compounds own strong reducing capacity and in turn decrease the CL signal sharply. Under the optimal conditions, the decreased CL intensity presents a good linear response to nitrofen concentration in the 0.02-50.0 μM range. The limit of detection (LOD, 3s/S) is 11 nM and the precision is 2.0% for replicate measurements of 50.0 nM nitrofen solution (n = 11). This method has the advantages of rapid analytical efficiency, good selectivity, satisfactory stability, and recyclability. Recovery experiments were conducted on spiked vegetable and fruit samples with the recoveries falling in the range 90.0-107.0%.
Topics: Vegetables; Fruit; Phenyl Ethers; Phenols; Herbicides; Lipase
PubMed: 38032482
DOI: 10.1007/s00604-023-06077-3 -
Molecules (Basel, Switzerland) May 2024The development of immobilized enzymes with high activity and stability is critical. Metal-organic frameworks (MOFs) have attracted much academic and industrial interest...
The development of immobilized enzymes with high activity and stability is critical. Metal-organic frameworks (MOFs) have attracted much academic and industrial interest in the field of enzyme immobilization due to their unique properties. In this study, the amino-functionalized ionic liquid (NIL)-modified metal-organic framework (UiO-66-NH) was prepared to immobilize lipase (CRL), using dialdehyde starch (DAS) as the cross-linker. The results of the Fourier transform infrared (FT-IR) spectra, X-ray powder diffraction (XRD), and scanning electronic microscopy (SEM) confirmed that the NIL was successfully grafted to UiO-66-NH. The CRL immobilized on NIL-modified UiO-66-NH (UiO-66-NH-NIL-DAS@CRL) exhibited satisfactory activity recovery (79.33%), stability, reusability, and excellent organic solvent tolerance. The research results indicated that ionic liquid-modified UiO-66-NH had practical potential for application in enzyme immobilization.
Topics: Lipase; Ionic Liquids; Enzymes, Immobilized; Metal-Organic Frameworks; Enzyme Stability; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction; Starch; Saccharomycetales; Phthalic Acids
PubMed: 38792242
DOI: 10.3390/molecules29102381 -
Enzyme and Microbial Technology Aug 2021Oil palm leaves (OPL) silica (SiO) can replace the energy-intensive, commercially produced SiO. Moreover, the agronomically sourced biogenic SiO is more biocompatible...
Oil palm leaves (OPL) silica (SiO) can replace the energy-intensive, commercially produced SiO. Moreover, the agronomically sourced biogenic SiO is more biocompatible and cost-effective enzyme support, which properties could be improved by the addition of magnetite (FeO) and graphene oxide (GO) to yield better ternary support to immobilize enzymes, i.e., Candida rugosa lipase (CRL). This study aimed to optimize the Candida rugosa lipase (CRL immobilization onto the ternary OPL-silica-magnetite (FeO)-GO (SiO/FeO/GO) support, for use as biocatalyst for ethyl valerate (EV) production. Notably, this is the first study detailing the CRL/SiO/Fe3O4/GO biocatalyst preparation for rapid and high yield production of ethyl valerate (EV). AFM and FESEM micrographs revealed globules of CRL covalently bound to GL-A-SiO/FeO/GO; similar to Raman and UV-spectroscopy results. FTIR spectra revealed amide bonds at 3478 cm and 1640 cm from covalent interactions between CRL and GL-A-SiO/FeO/GO. Optimum immobilization conditions were 4% (v/v) glutaraldehyde, 8 mg/mL CRL, at 16 h stirring in 150 mM NaCl at 30 °C, offering 24.78 ± 0.26 mg/g protein (specific activity = 65.24 ± 0.88 U/g). The CRL/SiO/FeO/GO yielded 77.43 ± 1.04 % of EV compared to free CRL (48.75 ± 0.70 %), verifying the suitability of SiO/FeO/GO to hyperactivate and stabilize CRL for satisfactory EV production.
Topics: Biocatalysis; Enzyme Stability; Enzymes, Immobilized; Esterification; Ferrosoferric Oxide; Graphite; Lipase; Saccharomycetales; Silicon Dioxide; Valerates
PubMed: 34116744
DOI: 10.1016/j.enzmictec.2021.109807 -
Journal of the Science of Food and... Dec 2023Pine sterol ester is a type of novel food source nutrient with great advantages in lowering blood cholesterol levels, inhibiting tumors, preventing prostate enlargement,...
BACKGROUND
Pine sterol ester is a type of novel food source nutrient with great advantages in lowering blood cholesterol levels, inhibiting tumors, preventing prostate enlargement, and regulating immunity. Macroporous resins with large specific surface area, stable structures, and various functional groups (epoxy, amino, and octadecyl groups) have been selected for immobilization of Candida rugosa lipase (CRL) to improve its stability and efficiency in the synthesis of pine sterol esters. A solvent-free strategy using oleic acid (substrate) as an esterification reaction medium is an important alternative for avoiding the use of organic solvents.
RESULTS
The immobilization conditions of CRL immobilized on several types of commercial macroporous resins were optimized. Fortunately, by adsorption (hydrophobic interaction), a high immobilization efficiency of CRL was obtained using macroporous resins with hydrophobic octadecyl groups with an immobilization efficiency of 86.5%, enzyme loading of 138.5 mg g and enzyme activity of 34.7 U g . The results showed that a 95.1% yield could be obtained with a molar ratio of oleic acid to pine sterol of 5:1, an enzyme amount of 6.0 U g (relative to pine sterol mass) at 50 °C for 48 h.
CONCLUSION
The hydrophobic macroporous resin (ECR8806M) with a large specific surface area and abundant functional groups was used to achieve efficient immobilization of CRL. CRL@ECR8806M is an efficient catalyst for the synthesis of phytosterol esters and has the potential for further large-scale applications. Therefore, this simple, green, and low-cost strategy for lipase immobilization provides new possibilities for the high-efficiency production of pine sterol esters and other food source nutrients. © 2023 Society of Chemical Industry.
Topics: Lipase; Solvents; Enzymes, Immobilized; Oleic Acid; Biocatalysis; Candida; Sterols; Hydrophobic and Hydrophilic Interactions; Enzyme Stability; Esters
PubMed: 37467367
DOI: 10.1002/jsfa.12869 -
International Journal of Biological... Dec 2023Lipase adsorption on solid supports can be mediated by a precise balance of electrostatic and hydrophobic interactions. A suitable fine-tuning could allow the...
Lipase adsorption on solid supports can be mediated by a precise balance of electrostatic and hydrophobic interactions. A suitable fine-tuning could allow the immobilized enzyme to display high catalytic activity. The objective of this work was to investigate how pH and ionic strength fluctuations affected protein-support interactions during immobilization via physical adsorption of a Candida rugosa lipase (CRL) on MgFeO. The highest amount of immobilized protein (IP) was measured at pH 4, and an ionic strength of 90 mM. However, these immobilization conditions did not register the highest hydrolytic activity (HA) in the biocatalyst (CRLa@MgFeO), finding the best values also at acidic pH but with a slight shift towards higher values of ionic strength around 110 mM. These findings were confirmed when the adsorption isotherms were examined under different immobilization conditions so that the maximum measurements of IP did not coincide with that of HA. Furthermore, when the recovered activity was examined, a strong interfacial hyperactivation of the lipase was detected towards acidic pH and highly charged surrounding environments. Spectroscopic studies, as well as in silico molecular docking analyses, revealed a considerable involvement of surface hydrophobic protein-carrier interactions, with aromatic aminoacids, especially phenylalanine residues, playing an important role. In light of these findings, this study significantly contributes to the body of knowledge and a better understanding of the factors that influence the lipase immobilization process on magnetic inorganic oxide nanoparticle surfaces.
Topics: Lipase; Molecular Docking Simulation; Candida; Enzymes, Immobilized; Nanoparticles; Enzyme Stability
PubMed: 37652323
DOI: 10.1016/j.ijbiomac.2023.126615 -
Colloids and Surfaces. B, Biointerfaces Nov 2020A novel integrated lipase nanogel based on functional ionic liquid modification and polymerization immobilization with improved stability was designed. Characterization...
A novel integrated lipase nanogel based on functional ionic liquid modification and polymerization immobilization with improved stability was designed. Characterization before and after modification and polymerization was conducted using infrared spectroscopy, Circular dichroism spectroscopy, fluorescence spectroscopy, and scanning electron microscopy. It was shown that the modification of the ionic liquid influenced the catalytic behavior of lipase significantly due to the changed structure and surface properties of lipase. The enzymatic properties, including acid-base stability, thermal stability, organic solvents stability, and storage stability of CRL nanogel, were investigated in the p-nitrophenyl palmitate hydrolysis reaction (CRL, Lipase from Candida Rugosa). The results indicated that CRL nanogel has a better pH, heat, and organic solvent tolerance after immobilization. After seven weeks of storage, the natural CRL gradually lost its enzymatic activity, and only 17.5±1.7 % of the catalytic activity remained, the residual activity of CRL nanogel was 97.3±1.8 %. It was indicated that the novel CRL nanogel was an excellent biocatalyst.
Topics: Biocatalysis; Enzyme Stability; Enzymes, Immobilized; Ionic Liquids; Lipase; Nanogels; Saccharomycetales
PubMed: 32739774
DOI: 10.1016/j.colsurfb.2020.111275 -
Applied Biochemistry and Biotechnology Mar 2021Fatty acid amides (FAAs) are of great interest due to their broad industrial applications. They can be synthesized enzymatically with many advantages over chemical...
Fatty acid amides (FAAs) are of great interest due to their broad industrial applications. They can be synthesized enzymatically with many advantages over chemical synthesis. In this study, the fatty acid moieties of lipids of Cunninghamella echinulata ATHUM 4411, Umbelopsis isabellina ATHUM 2935, Nannochloropsis gaditana CCAP 849/5, olive oil, and an eicosapentaenoic acid (EPA) concentrate were converted into their fatty acid methyl esters and used in the FAA (i.e., ethylene diamine amides) enzymatic synthesis, using lipases as biocatalysts. The FAA synthesis, monitored using in situ NMR, FT-IR, and thin-layer chromatography, was catalyzed efficiently by the immobilized Candida rugosa lipase. The synthesized FAAs exhibited a significant antimicrobial activity, especially those containing oleic acid in high proportions (i.e., derived from olive oil and U. isabellina oil), against several human pathogenic microorganisms, insecticidal activity against yellow fever mosquito, especially those of C. echinulata containing gamma-linolenic acid, and anticancer properties against SKOV-3 ovarian cancer cell line, especially those containing EPA in their structures (i.e., EPA concentrate and N. gaditana oil). We conclude that FAAs can be efficiently synthesized using microbial oils of different fatty acid composition and used in specific biological applications.
Topics: Amides; Cunninghamella; Eicosapentaenoic Acid; Fungi; Olive Oil; Saccharomycetales
PubMed: 33191449
DOI: 10.1007/s12010-020-03450-3