Kinetic modeling and optimization of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on clayey support.

Mono- and diglycerides play a crucial role in the food industry as multifunctional food additives and emulsifiers. Their importance stems from their unique properties, which allow them to improve the quality, texture, and stability of various food products. Here, results of the kinetic modeling of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on the clayey support Spectrogel® type C are reported. The support was characterized by TEM, SEM, and FTIR. Firstly, the influence of pH and lipase load on the immobilization process was analyzed, resulting in an enzymatic activity of 93.2 ± 0.7 U g-1 under optimized conditions (170.9 U g-1 of lipase and pH of 7.1). Afterward, the effects of reaction temperature and concentration of immobilized biocatalyst in the feedstock conversion were evaluated. At optimized parameters, a triglycerides conversion of 97% was obtained at 36.5 °C, 7.9 vol.% of enzyme, a glycerol to feedstock molar ratio of 2:1, and 2 h. The optimized conditions were used to determine the kinetic constants of the elementary reactions involved in the glycerolysis, where a fit superior to 0.99 was achieved between experimental values and predicted data.

Recycled polyvinyl chloride microplastics: investigation of environmentally relevant concentrations on toxicity in adult zebrafish.

Recycled polyvinyl chloride (PVC) microplastics have been detected in the aquatic environment. These recycled microparticles contain chemicals that are released into the environment reaching different organisms. Although the problem of the presence of recycled PVC microparticles in the environment is evident, the toxicological consequences of this contaminant to exposed organisms remains to be better determined. The aim of this study was to investigate the toxicity attributed to exposure to environmentally relevant concentrations of recycled PVC microplastics in adult zebrafish (Danio rerio). The experimental groups were: negative control, vehicle control, positive control, and recycled microplastics (20 ± 5 µm) at 5, 10 or 20 µg/L. Zebrafish (D. rerio) were exposed to respective treatments for 96 hr. Locomotion and oxidative status parameters were measured and mortality recorded. The positive control group presented increased mortality rates and decreased locomotor activity. Animals from the vehicle group did not show marked differences. Finally, no significant disturbances were found in survival rate, locomotion pattern and oxidative status of animals exposed to recycled PVC microparticles at 5, 10 or 20 µg/L. Taken together our results suggest that recycled PVC microplastics in this particle size range do not appear to exert harmful effects on exposed adult D. rerio. However, these results need to be carefully observed due to limitations including size of particle and duration of exposure parameters that might affect ecological consequences. It is suggested that additional studies applying other particles sizes and chronic exposure are needed to more comprehensively verify the toxicity of the contaminant investigated here.

Juçara Fruit (Euterpe Edulis Martius) Valorization Combining Emergent Extraction Technologies and Aqueous Solutions of Alkanediols.

Anthocyanins from juçara fruits were extracted by pressurized liquid extraction (PLE) or ultrasound-assisted extraction (UAE), using aqueous solutions of 1,2-alkanediols and glycerol ethers as biobased solvents. The PLE (100 bar, 13 min, 1 mL/min flow rate) in the optimal extraction conditions originated 23.1 mganthocyanins·gdry biomass-1. On the other hand, the UAE was 10 min long, and the optimal conditions using 1,2-propanediol were 42.6 wt%, 160 W, and pH 7.0, leading to 50 mganthocyanins·gdry biomass-1. Extractions at the UAE optimized conditions, with aqueous solutions of five different 1,2-alkanediols and three glycerol ethers were performed, and compared to water and ethanolic extracts. The biobased solvent solutions presented anthocyanin yields up to 33% higher than water, and were shown to be as efficient as ethanol/water, but generated extracts with higher antioxidant capacity. The anthocyanin-rich extract of juçara, obtained with 1,2-propanediol, was used in the production of a natural soap and incorporated into a cream, showing that the addition of the juçara extract resulted in an antioxidant capacity in both products.

Micronized Curcumin Causes Hyperlocomotion in Zebrafish Larvae.

Zebrafish larvae have been widely used in neuroscience and drug research and development. In the larval stage, zebrafish present a broad behavioral repertoire and physiological responses similar to adults. Curcumin (CUR), a major component of Curcuma longa L. (Zingiberaceae), has demonstrated the ability to modulate several neurobiological processes relevant to mental disorders in animal models. However, the low bioavailability of this compound can compromise its in vivo biological potential. Interestingly, it has been shown that micronization can increase the biological effects of several compounds. Thus, in this study, we compared the effects of acute exposure for 30 min to the following solutions: water (control), 0.1% DMSO (vehicle), 1 µM CUR, or 1 µM micronized curcumin (MC) in zebrafish larvae 7 days post-fertilization (dpf). We analyzed locomotor activity (open tank test), anxiety (light/dark test), and avoidance behavior (aversive stimulus test). Moreover, we evaluated parameters of oxidative status (thiobarbituric acid reactive substances and non-protein thiols levels). MC increased the total distance traveled and absolute turn angle in the open tank test. There were no significant differences in the other behavioral or neurochemical outcomes. The increase in locomotion induced by MC may be associated with a stimulant effect on the central nervous system, which was evidenced by the micronization process.

Evaluation of Resveratrol and Piceatannol Anticonvulsant Potential in Adult Zebrafish (Danio rerio).

Epilepsy is a common neurological disorder which affects 50 million people worldwide. Patients with epilepsy may present cognitive deficits and psychological impairment. Currently, 30% of patients fail to respond to any available antiseizure drug, and a significant number of patients do not well tolerate the offered treatments. Then, it is necessary to find out alternatives for controlling epileptic seizures. Studies have shown that despite its neuroprotective effects, resveratrol shows poor anticonvulsant properties. Resveratrol analog, piceatannol, possesses higher biological activity than resveratrol and could be an alternative to control seizure. Thus, the present study investigated the effects of resveratrol and piceatannol in pentylenetetrazole-induced seizures in adult zebrafish (Danio rerio). Only the experimental positive control (diazepam) showed anticonvulsant effect in this study. In addition, no behavioral changes were observed 24 h after seizure occurrence. Finally, the expression of genes related to neuronal activity (c-fos), neurogenesis (p70S6Ka and p70S6Kb), inflammatory response (interleukin 1ß), and cell apoptosis (caspase-3) did not change by pentylenetetrazole-induced seizures. Therefore, we failed to observe any anticonvulsant and neuroprotective potential of resveratrol and piceatannol in adult zebrafish. However, resveratrol and piceatannol benefits in epilepsy are not discharged, and more studies are necessary.

Curcumin and n-acetylcysteine cocrystal produced with supercritical solvent: characterization, solubility, and preclinical evaluation of antinociceptive and anti-inflammatory activities.

Curcumin presents a promising anti-inflammatory potential, but its low water-solubility and bioavailability hinder its application. In this sense, cocrystallization represents a tool for improving physicochemical properties, solubility, permeability, and bioavailability of new drug candidates. Thus, the aim of this work was to produce curcumin cocrystals (with n-acetylcysteine as coformer, which possesses anti-inflammatory and antioxidant activities), by the anti-solvent gas technique using supercritical carbon dioxide, and to test its antinociceptive and anti-inflammatory potential. The cocrystal was characterized by differential scanning calorimetry, powder X-ray diffraction and scanning electron microscopy. The cocrystal solubility and antichemotaxic activity were also assessed in vitro. Antinociceptive and anti-inflammatory activities were carried out in vivo using the acetic acid-induced abdominal writhing and carrageenan-induced paw oedema assays in mice. The results demonstrated the formation of a new crystalline structure, thereby confirming the successful formation of the cocrystal. The higher solubility of the cocrystal compared to pure curcumin was verified in acidic and neutral pH, and the cocrystal inhibited the chemotaxis of neutrophils in vitro. In vivo assays showed that cocrystal presents increased antinociceptive and anti-inflammatory potency when compared to pure curcumin, which could be related to an improvement in its bioavailability.

Micronized Resveratrol Shows Anticonvulsant Properties in Pentylenetetrazole-Induced Seizure Model in Adult Zebrafish.

Epilepsy affects 50 million people around the world, and the patients experience cognitive, psychological and social consequences. Despite the considerable quantity of antiepileptic drugs available, 30% of patients still suffer in seizure. Therefore, the advance in therapeutic alternatives is mandatory. Resveratrol has been attracting the attention of many researchers because of its pharmacological potential. However, despite its neuroprotective and anti-epileptic effects, clinical resveratrol use is impaired by its low bioavailability. Here, we applied the supercritical fluid micronization technology (SEDS) to overcome this deficit, and investigated the anticonvulsant potential of micronized resveratrol in a PTZ-induced seizure model in adult zebrafish (Danio rerio). SEDS permits obtaining significantly reduced particle size with a fine size distribution in comparison with the starting material. It can improve the pharmacotherapeutic efficacy. Our data showed that micronized resveratrol decreased the occurrence of the tonic-clonic seizure stage and slowed the development of the seizures in a similar manner of diazepam. Non-processed resveratrol was not able to protect the animals. Furthermore, diazepam decreased the locomotion and exploratory behavior. Differently from diazepam, the micronized resveratrol did not induce behavioral adverse events. In addition, our data showed that the PTZ-induced seizures increased the c-fos transcript levels following the neural excitability. However, the increase in c-fos levels was prevented by micronized resveratrol. In conclusion, our results demonstrate that the micronized resveratrol shows anticonvulsant effect, like the classical antiepileptic drug diazepam in a PTZ-induced seizure model. Excitingly, different from diazepam, micronized resveratrol did not provoke behavioral adverse events.

Improvement of tangential microfiltration of gelatin solution using a permanent magnetic field.

Membrane technology is an interesting alternative to conventional gelatin clarification methods, resulting in the elimination of refining chemical agents. In this work, the application of a permanent magnetic field as a pre-treatment of the gelatin solutions was proposed as a strategy to improve the microfiltration (MF) performance. Filtration tests were performed using a 1.5% swine gelatin solution at 40 °C through cellulose acetate membranes in a tangential flow module. Prior to the filtrations, the feed solutions were pretreated by the circulation of the solution through magnetic fields with different flux densities, 0.7 T and 1.34 T, for 2 h. The magnetic induction of the solution significantly increased the permeate flux and the recovery of hydraulic permeance by 63% and 122%, respectively, showing the application of the magnetic field in the solution of gelatin is an attractive alternative to improve the performance of the process.

Co-precipitation of anthocyanin in PHBV by the SEDS technique.

Anthocyanins are pigments of plant origin responsible for most blue, purple and all shades of red found in flowers, fruits and some stems and roots of plants, besides comprising a class of potent antioxidant phenolic compounds. Due to the relevance of anthocyanins this work aims to encapsulate anthocyanin extracted from the wine lees through the Solution Enhanced Dispersion by Supercritical Fluids (SEDS) technique and to evaluate the thermal stability of encapsulated versus non-encapsulated anthocyanin. The highest encapsulation efficiency obtained was approximately 66%. Submicron size particles ranging from 0.22 to 0.30 µm were obtained and they were free of residual organic solvent. In relation to the thermal stability, it was verified that the particles degraded about six times less than the non-encapsulated sample, which allows numerous applications since one of the barriers of anthocyanin use is its sensitivity to high temperatures.

Improving the soluble lipase-catalyzed biodiesel production through a two-step hydroesterification reaction system.

The application of lipases in liquid formulation instead of immobilized forms in the enzymatic biodiesel synthesis can make the process cost-efficient, more competitive, and sustainable. However, despite the benefits, the long reaction times required to achieve satisfactory yields is still a drawback of this biotechnological process. In this sense, employing the novel low-cost soluble NS40116 lipase, this paper proposes an innovative two-step hydroesterification reaction (TSHR) system as a technique of improving the reaction rate of an enzymatic biodiesel production. With the employment of two central composite statistical design to optimize the parameters of each of the reactions involved, the influence of the parameters "water concentration added to the reaction," "methanol-to-oil molar ratio," and "lipase load" on the process yield, besides the acid value of the samples, was investigated. After only 8 h of reaction, the highest fatty acid methyl ester yield reached was 97.1% with an acid value of 4.62 mg KOH g-1 utilizing a total of 8 wt% water, methanol-to-oil molar ratio of 6.3:1, and 0.70 wt% of lipase. Furthermore, the statistical models for both reactions indicated to be significant with 95% of reliability. Considering that the papers published using soluble lipases in a one-step batch process normally reach similar yields to those obtained in this research after 16 h to 24 h of reaction, the proposed system demonstrated to be a promising option of process configuration for the enzymatic production of biodiesel.

Soluble lipase-catalyzed synthesis of methyl esters using a blend of edible and nonedible raw materials.

This work investigates the use of blends of edible and nonedible raw materials as an alternative feedstock to fatty acid methyl esters (FAME) production through enzymatic catalysis. As biocatalyst, liquid lipase from Thermomyces lanuginosus (Callera™ Trans L), was used. Under reaction conditions of 35 °C, methanol to feedstock molar ratio of 4.5:1 and 1.45% of catalyst load, the best process performance was reached using 9% of water concentration in the medium-yield of 79.9% after 480 min of reaction. In terms of use of tallow mixed with soybean oil, the best yield was obtained when 100% of tallow was used in the process-84.6% after 480 min of reaction-behavior that was associated with the degree of unsaturation of the feedstock, something by that time, not addressed in papers of the area. The results show that tallow can be used as an alternative to FAME production, catalyzed by soluble lipase.

Fatty acid profile of pecan nut oils obtained from pressurized n-butane and cold pressing compared with commercial oils.

This short note compares the chemical profile of pecan nut oil samples obtained from pressurized n-butane and cold pressing with two commercial oils. The conventional cold pressing technique yielded 58.9 wt%, while pressurized n-butane yielded from 53 to 65 wt%, being the highest yield at 55 °C, and pressure of 40 bar. The n-butane behaves nearly like a piston fluid within the experimental conditions used. The results showed that the extraction variables had a slight influence on the fatty acid composition of the samples. Extraction with n-butane thus showed to be a promising alternative technique to conventional extraction methods, as very mild operating conditions and eco-friendly solvent can be used to provide good results without any residues in the final product.

Phytochemical profile, antioxidant and antimicrobial activity of extracts obtained from erva-mate (Ilex paraguariensis) fruit using compressed propane and supercritical CO2.

Traditionally, Ilex paraguariensis leaves are consumed in tea form or as typical drinks like mate and terere, while the fruits are discarded processing and has no commercial value. The aim of this work to evaluate phytochemical properties, total phenolic compounds, antioxidant and antimicrobial activity of extracts of Ilex paraguariensis fruits obtained from supercritical CO2 and compressed propane extraction. The extraction with compressed propane yielded 2.72 wt%, whereas with supercritical CO2 1.51 wt% was obtained. The compound extracted in larger amount by the two extraction solvents was caffeine, 163.28 and 54.17 mg/g by supercritical CO2 and pressurized propane, respectively. The antioxidant activity was more pronounced for the supercritical CO2 extract, with no difference found in terms of minimum inhibitory concentration for Staphylococcus aureus for the two extracts and better results observed for Escherichia coli when using supercritical CO2.

Extraction, chemical characterization and antioxidant activity of Litchi chinensis Sonn. and Avena sativa L. seeds extracts obtained from pressurized n-butane.

The extraction of litchi (Litchi chinensis Sonn.) and oat (Avena sativa L.) seeds were investigated using n-butane as pressurized solvent by evaluating the effect of pressure in the range of 7-100 bar and temperature from 25 to 70 °C on the extract yield and chemical composition together with the antioxidant activity of the extracts obtained. It was experimentally observed extraction yields for both seeds up to ~3.5 wt%, with a total phenolic content around 126.4 mg GAE/100 g of extract, and an antioxidant activity up to 78.36%. Oat seeds extract presented higher values of these parameters evaluated compared to litchi extract. Based on the results found, it seems that n-butane may be a promising solvent to conventional extraction methods, as mild operating conditions and eco-friendly solvent can be used to provide good results without any residues in the final product.

Encapsulation of eugenyl acetate in PHBV using SEDS technique and in vitro release evaluation.

Eugenyl acetate obtained via enzymatic esterification using Lipozyme TL IM enzyme was encapsulated in biopolymer poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) through solution-enhanced dispersion by supercritical fluids (SEDS). Produced particles were characterized by SEM and confocal microscopy techniques and in addition in vitro release assays were performed in isopropanol and ethyl acetate. Experimental micronization conditions comprised 8 and 10 MPa, 308 and 313 K and eugenyl acetate concentration ranging from 5 to 20 mg mL-1, keeping PHBV concentration constant (20 mg mL-1 in dichloromethane). The maximum encapsulation efficiency was 58.0 % for 5 mg mL-1of eugenyl acetate at 8 MPa and 308 K. The morphology of the encapsulated particles for most of the trials was spherical, with particle size ranging from 0.061 to 0.276 µm. Regarding the release in ethyl acetate and isopropanol solvents the higher the affinity of the encapsulated ester of these solvents, the faster the release was observed. These results demonstrate the importance of essential clove oil esterification reaction and encapsulation of the ester by SEDS method so that this encapsulated ester can be used in different industrial applications.

Enzyme-catalyzed production of biodiesel by ultrasound-assisted ethanolysis of soybean oil in solvent-free system.

This work reports the transesterification of soybean oil with ethanol using a commercial immobilized lipase, Novozym 435, under the influence of ultrasound irradiation, in a solvent-free s. The experiments were performed in an ultrasonic water bath, following a sequence of experimental designs to evaluate the effects of temperature, enzyme and water concentrations, oil to ethanol molar ratio and output irradiation power on the reaction yield. Besides, a kinetic study varying the substrates molar ratio and enzyme concentration was also carried out. Results show that ultrasound-assisted lipase-catalyzed transesterification of soybean oil with ethanol in solvent-free system might be a potential alternative route to conventional alkali-catalyzed and/or traditional enzymatic methods, as high reaction yields (~78 wt%) were obtained at mild irradiation power supply (~132 W), and temperature (63 °C) in a relatively short reaction time, 1 h. Additionally, a study regarding the enzyme reuse was carried out at the experimental condition that afforded the best reaction yield.

Desulfurization and denitrogenation of heavy gas oil by Rhodococcus erythropolis ATCC 4277.

Some of the noxious atmospheric pollutants such as nitrogen and sulfur dioxides come from the fossil fuel combustion. Biodesulfurization and biodenitrogenation are processes which remove those pollutants through the action of microorganisms. The ability of sulfur and nitrogen removal by the strain Rhodococcus erythropolis ATCC 4277 was tested in a biphasic system containing different heavy gas oil concentrations in a batch reactor. Heavy gas oil is an important fraction of petroleum, because after passing through, the vacuum distillation is incorporated into diesel oil. This strain was able to remove about 40% of the nitrogen and sulfur present in the gas heavy oil. Additionally, no growth inhibition occurred even when in the presence of pure heavy gas oil. Results present in this work are considered relevant for the development of biocatalytic processes for nitrogen and sulfur removal toward building feasible industrial applications.

Continuous lipase-catalyzed esterification of soybean fatty acids under ultrasound irradiation.

This work investigates the continuous production of alkyl esters from soybean fatty acid (FA) charges using immobilized Novozym 435 as catalyst. The experiments were performed in a packed-bed bioreactor evaluating the effects of FA charge to alcohol (methanol and ethanol) molar ratio, from 1:1 to 1:6, substrate flow rate in the range of 0.5-2.5 mL/min and output irradiation power up to 154 W, at fixed temperature of 65 °C, on the reaction conversion. Results showed that almost complete conversions to fatty acids ethyl esters were achieved at mild ultrasonic power (61.6 W), FA to ethanol molar ratio of 1:6, operating temperature (65 °C) and remained nearly constant for long-term reactions without negligible enzyme activity losses.

A review on lipase-catalyzed reactions in ultrasound-assisted systems.

The named "green chemistry" has been receiving increasing prominence due to its environmentally friendly characteristics. The use of enzymes as catalysts in processes of synthesis to replace the traditional use of chemical catalysts present as main advantage the fact of following the principles of the green chemistry. However, processes of enzymatic nature generally provide lower yields when compared to the conventional chemical processes. Therefore, in the last years, the ultrasound has been extensively used in enzymatic processes, such as the production of esters with desirable characteristics for the pharmaceutical, cosmetics, and food industry, for the hydrolysis and glycerolysis of vegetable oils, production of biodiesel, etc. Several works found in the open literature suggest that the energy released by the ultrasound during the cavitation phenomena can be used to enhance mass transfer (substrate/enzyme), hence increasing the rate of products formation, and also contributing to enhance the enzyme catalytic activity. Furthermore, the ultrasound is considered a "green" technology due to its high efficiency, low instrumental requirement and significant reduction of the processing time in comparison to other techniques. The main goal of this review was to summarize studies available to date regarding the application of ultrasound in enzyme-catalyzed esterification, hydrolysis, glycerolysis and transesterification reactions.

Effect of magnetic field on the ultrafiltration of bovine serum albumin.

This work evaluates the effects of a static magnetic field on the permeation of bovine serum albumin (BSA) in a tangential ultrafiltration membrane module. Experimental tests were carried out at different pHs using a poly(sulfone) membrane with molecular weight cut off of 60 kDa under the influence of a 0.4 T neodymium-iron-boron magnetic field. Results showed an increase in the permeate flux of water after the cleaning procedures of the new and reused membranes in the presence of the magnetic field. The elusive mechanism of magnetic memory is also shown to take place for the water fluxes fully recovered after the cleaning procedures when the magnetic field was applied to the system before the permeation. When the magnetic field was applied during permeation, the water fluxes presented lower percent of recuperation after the cleaning procedures, thus suggesting that the BSA solution may have somewhat been influenced by magnetic memory.