Microencapsulation of Morinda citrifolia L. extract by spray-drying

Microcapsules containing Morinda citrifolia L. microparticles were produced by a spray-drying technique using various proportions of κ-carrageenan and maltodextrin as the binding materials. In this work, the effects of spray-drying on the encapsulation yield, particle size, moisture content, DPPH scavenging activity, total phenolic content and total flavonoid content of the bioactive components of M. citrifolia L. were determined for different volume ratios in the inlet air temperature range of 90–140 °C. The results showed that the percentage of 2,2-diphenyl picrylhydrazyl (DPPH) scavenging activity of the spray-dried powder was the highest for the 1:2 ratio (volume ratio of M. citrifolia L. extract to additive solution) at 90 °C, with maltodextrin at a concentration of 33 mg/ml. The results also showed that the microcapsules had a regular spherical shape. The spray-dried M. citrifolia fruit extract showed high antioxidant activity (28.36% DPPH activity), thus suggesting that it might be useful as a food additive and/or ingredient under the above optimum operating conditions.     

Bioethanol production from corn meal by simultaneous enzymatic saccharification and fermentation with immobilized cells of Saccharomyces cerevisiae var. ellipsoideus

The simultaneous enzymatic saccharification and fermentation (SSF) of corn meal using immobilized cells of Saccharomycescerevisiae var. ellipsoideus yeast in a batch system was studied. The yeast cells were immobilized in Ca-alginate by electrostatic droplet generation method. The process kinetics was assessed and determined and the effect of addition of various yeast activators (mineral salts: ZnSO₄ · 7H₂O and MgSO₄ · 7H₂O, and vitamins: Ca-pantothenate, biotin and myo-inositol) separately or mixed, was investigated. Taking into account high values of process parameters (such as ethanol concentration, ethanol yield, percentage of the theoretical ethanol yield, volumetric productivity and utilized glucose) and significant energy savings the SSF process was found to be superior compared to the SHF process. Further improvement in ethanol production was accomplished with the addition of mineral salts as yeast activators which contributed to the highest increase in ethanol production. In this case, the ethanol concentration of 10.23% (w/w), percentage of the theoretical ethanol yield of 98.08%, the ethanol yield of 0.55 g/g and the volumetric productivity of 2.13 g/l·h were obtained.     

Growth of Escherichia coli and Salmonella typhi inhibited by fractal silver nanoparticles supported on zeolites

Silver nanoparticles were dispersed on clinoptilolite zeolite. These materials were evaluated as biocides for Escherichia coli and Salmonella typhi. Ag-zeolites were shown to be efficient in eliminating both E. coli and S. typhi present in nutritive media. Particularly, E. coli was eliminated at very short times. In the presence of a zeolite free of silver, bacteria use the zeolite to reproduce rapidly.     

Rich bioactive phenolic extract production by microbial biotransformation of Brazilian Citrus residues

Flavanones in Citrus are molecules that play an important role in antioxidant activities in nutraceutical products. Recent studies indicate that molecules of the simplest classes of phenolics have higher biological activity and absorption capacity. However, the molecules that have been shown to be very important bioactive compounds of Citrus, such as hesperetin, naringenin and ellagic acid, are found in trace concentrations in the fruit. An interesting environmentally friendly alternative that deserves attention regarding phenolic compound obtaining is the biotransformation of these molecules. The aim of this study was to develop a process of biotransformation of phenolics from Brazilian Citrus residues by solid-state fermentation with the microorganism Paecilomyces variotii. The optimized fermentation conditions were 10 g of Citrus residues (2.0 mm of substrate particle size), 20 mL distilled water, at 32 °C after 48 h of incubation. The development of this process has generated, simultaneously, an increase of 900, 1400 and 1330% of hesperetin, naringenin and ellagic acid concentration, respectively, and an increase of 73% of the antioxidant capacity. These results give strong evidence that microbial biotransformation does not only produce phenolic compounds but also compounds with high biological activity, such as hesperetin and naringenin.     

Preliminary separation and purification of rutin and quercetin from Euonymus alatus (Thunb.) Siebold extracts by macroporous resins

In this study, the performances of rutin and quercetin from Euonymus alatus (Thunb.) Siebold extracts on five macroporous resins with different physical and chemical properties were investigated. The results of static tests indicated that AB-8 resin was the most appropriate and its adsorption data were well fitted to the Langmuir and Freundlich isotherms. In order to optimize the separation process, different pH values of sample solution, different concentrations and pH values of ethanol solution also have been investigated. Column packed with AB-8 resin was used to perform dynamic adsorption and desorption experiments. After the treatment with AB-8 resin and optimal conditions, the contents of rutin and quercetin in the product were 8.45-fold and 13.14-fold increased with recovery yields of 63.1% and 72.3%, respectively. The results showed that the present method was suitable for large-scale preparation of rutin and quercetin from Euonymus alatus (Thunb.) Siebold or other herbal materials.     

The partitioning of protease from Calotropis procera latex by aqueous two-phase systems and its hydrolytic pattern on muscle proteins

The protease from the latex of Calotropis procera was isolated by an aqueous two-phase system (ATPS). Polyethylene glycol (PEG 1000, 2000 and 3000) at a concentration of 12, 15, and 18% (w/w) with salts ((NH₄)₂SO₄, K₂HPO₄ and MgSO₄) at a concentration of 14, 17, and 20% (w/w) were investigated. The highest protease recovery (74.6%) was found in the PEG-rich phase of the system (p < 0.05), comprising of 18% PEG 1000 and 14% MgSO₄. Protein patterns and activity staining showed that the isolated protease had a molecular weight of ∼31 kDa without the oligosaccharide attached to the molecule. Degradation of muscle proteins in beef, farmed giant catfish, and squid was observed by the electrophoresis of sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE). The degradation of myofibrillar proteins (myosin heavy chain: MHC and actin: AC) of farmed giant catfish was higher than that of beef and squid muscles as indicated by the degradation proteins with lower molecular weight.     

Solubility of Jatropha and Aquilaria oils in supercritical carbon dioxide at elevated pressures

This study investigated experimental equilibrium solubilities of Jatropha curcas and Aquiliaria crassna oils dissolved in supercritical carbon dioxide at temperatures of 318-338 K and pressures of 20, 25, 30, 35 MPa. The highest solubility of J. curcas and A. crassna oil were 29.8 and 28.4 mg L⁻¹, respectively, at 338 K and 35 MPa. The oil solubilities and the concentration of triglycerides both increased with increasing temperature and pressure. Triglyceride molecules surrounded by carbon dioxide molecules may be proposed since solubilities increased with the flux of supercritical carbon dioxide. The solubility of these two oils linearly increased with the density of supercritical carbon dioxide. Experimental data of the oil solubility were successfully correlated by the Chrastil equation.     

The crystal modulus of silk (Bombyx mori)

It is shown that experimentally determined values of the crystal modulus of Bombyx mori silk agree reasonably well with the computationally determined values, if account is taken of the stress relaxation, which occurs during the experimental measurements. The experimental are 16-22 GPa depending on the sample and the computational are 13 or 16 GPa depending on the method of analysis.     

Radical-scavenging activity of extracts from Cordia verbenacea DC obtained by different methods

The possibility of using the leaves of Cordia verbenacea as a new source of natural antioxidant compounds was investigated. In the present work, extracts from C. verbenacea were obtained using different extraction methods: supercritical fluid extraction (SFE), Soxhlet (SE), hydrodistillation and maceration, with the objective to evaluate the methods in terms of yield and antioxidant potential. The high-pressure technique was applied using pure CO₂ and CO₂ with co-solvent at different temperatures and pressures (30, 40 and 50 °C and 100, 200, and 300 bar). Organic solvents with different polarities were used to obtain extracts by low-pressure extraction processes. The extracts were evaluated according to their antioxidant activity using total phenolic content, scavenging abilities on DPPH radical, total antioxidant activities (ABTS^•+), superoxide anion radical-scavenging (O₂^•) and protection against lipid peroxidation in vitro (LPO). Ethyl acetate fraction obtained by maceration and extract isolated by SE using 25% aqueous mixture of ethanol possessed the highest scavenger activity against DPPH radical (IC₅₀ = 9.2 ± 0.4 μg/ml, IC₅₀ = 27.4 ± 0.1 μg/ml, respectively). The SFE with 8% ethanol as a co-solvent produced extracts with distinguished increase in the antioxidant activity. The Soxhlet extract with ethyl acetate exhibited a strong reduction of lipid peroxidation (IC₅₀ = 209 ± 3 μg/ml) value comparable to the standard rutin (IC₅₀ = 203 ± 2 μg/ml). The results indicate that extracts of C. verbenacea have important potential as a source of bioactive compounds with antioxidant activity.     

Effect of culture pH on the antioxidant properties of Antrodia camphorata in submerged culture

Effects of culture pH ranging from pH 3.0 to 6.0 on yields of bioactive compounds, antioxidant activity, and free radical scavenging ability of methanolic extracts from mycelia and filtrates of Antrodia camphorata in a submerged culture were investigated. Culture pH significantly affected antioxidant and scavenging free radical activities of both the methanolic extracts of the mycelia and the filtrates. A low culture pH (3.0) favored cell growth, but higher culture pH (5.0) favored antioxidant properties. Antioxidant activities of the methanolic extracts of the mycelia and the filtrates were found to be correlated with total phenolic content, polysaccharide content, and protein/polysaccharide ratios. Antioxidant production was maximum at pH 5.0 at which the total phenolics and polysaccharide/protein ratio in the methanolic extracts of the mycelia were 152.2 mg/g and 0.335, respectively. The total phenolics might be the compounds responsible for the antioxidant activities of the methanolic extracts of the mycelia and the filtrates on lipid peroxidation and scavenging effects of ferrous ions. The maximum polysaccharide content of the methanolic extracts of the filtrates was 55.3 mg/g at pH 5.0. Higher yields of antioxidants were obtained in the submerged culture as compared with that of fruiting bodies. An alternative approach for producing antioxidants of A. camphorata in a submerged culture instead of solid state cultures was demonstrated in this study.     

Modified electrodes for probing the metal-reductase activity of metalloproteins: the reduction of iron(III) catalyzed by Desulfovibrio vulgaris Hildenborough cytochrome c3

Cytochromes c₃ are polyheme c-type cytochromes characterized by low redox potentials, that have been shown to develop metal-reductase activity. In this paper, different strategies are explored to immobilize one of them, Desulfovibrio vulgaris Hildenborough cytochrome c₃, a highly basic tetraheme cytochrome, including adsorption, covalent bonding, imprisonment in a layer-by-layer assembly, and entrapment within cast films or a dialysis membrane. The performance and efficiency of modified (carbon or gold) electrodes have been evaluated using electrochemical (cyclic and square-wave voltammetry, current-time curves) techniques in the presence of a soluble Fe(III) complex, ammonium Fe(III) citrate acting as the soluble substrate, and chosen as a model system. The advantages and drawbacks of each strategy are discussed with the view of further extension of environmental interest to more toxic metal contaminants.     

Microencapsulation of Radix salvia miltiorrhiza nanoparticles by spray-drying

About 133.5 nm Radix salvia miltiorrhiza nanoparticles were prepared by high speed centrifugal sheering pulverizer and the nanoparticles were characterized by TEM in this study. Microcapsules containing R. salvia miltiorrhiza nanoparticles were produced by spray-drying technique using different proportions of gelatin and sodium salt of carboxymethylcellulose (CMC-Na) as wall materials. The effects of inlet temperature, flow rate, spray-gas flow and the ratio of M_core/M_wall on encapsulation yield (EY) and encapsulation efficiency (EE) were investigated. The EE was determined by reverse high performance liquid chromatography (HPLC); the resulting microcapsules were characterized by FT-IR, SEM, and X-ray diffraction analysis. In addition, in vitro release characters of R. salvia miltiorrhiza raw powder, spray-dried powder and microcapsules were also studied. The results showed that spray-dried microcapsules had a regular spherical shape but the majority presented rough surfaces or invaginations with a diameter of 2-5 μm. R. salvia miltiorrhiza nanoparticles were embedded in the wall system consisting of gelatin and CMC-Na. Higher EE and EY were obtained under the inlet temperature of 80 °C and the ratio of M_core/M_wall of 1/4. In vitro release study showed that R. salvia miltiorrhiza microcapsules could regulate drug release. This study may be helpful to the pharmaceutical application of R. salvia miltiorrhiza.     

Biodegradation of tetracyanonickelate by Klebsiella oxytoca under anaerobic conditions

The aims of this study were to evaluate the biodegradability of tetracyanonickelate (II) [K₂[Ni(CN)₄] (TCN) by Klebsiella oxytoca under anaerobic conditions. Anaerobic serum bottles and fermenter experiments were conducted to evaluate the effects of nitrogen sources and pH on the TCN biodegradation potential by K. oxytoca. Results reveal that TCN can be biotransformed to methane by resting cells of K. oxytoca. Results show that TCN biodegradation was inhibited by the addition of nitrate, nitrite, or ammonia at higher concentrations (5 and 10 mM). Moreover, it was found that the optimum pH for TCN conversion by K. oxytoca was about 7.1. Results from the fermenter experiment show that TCN can be completely degraded within 14 days. K. oxytoca is capable of using TCN as the nitrogen source under anaerobic conditions. TCN could be biotransformed to non-toxic end product (methane) by resting cells of K. oxytoca. Higher TCN biodegradation rate can be obtained under alkaline conditions. This study provides us insight into the characteristics of TCN conversion by K. oxytoca under anaerobic conditions. These findings would be helpful in designing a practical system inoculated with K. oxytoca for the treatment of cyanide-containing wastewater.     

Electrochemical activity of Geobacter sulfurreducens biofilms on stainless steel anodes

Stainless steel was studied as anode for the biocatalysis of acetate oxidation by biofilms of Geobacter sulfurreducens. Electrodes were individually polarized at different potential in the range −0.20 V to +0.20 V vs. Ag/AgCl either in the same reactor or in different reactors containing acetate as electron donor and no electron acceptor except the working electrode. At +0.20 V vs. Ag/AgCl, the current increased after a 2-day lag period up to maximum current densities around 0.7 A m⁻² and 2.4 A m⁻² with 5 mM and 10 mM acetate, respectively. No current was obtained during chronoamperometry (CA) at potential values lower than 0.00 V vs. Ag/AgCl, while the cyclic voltammetries (CV) that were performed periodically always detected a fast electron transfer, with the oxidation starting around −0.25 V vs. Ag/AgCl. Epifluorescent microscopy showed that the current recorded by chronoamperometry was linked to the biofilm growth on the electrode surface, while CVs were more likely linked to the cells initially adsorbed from the inoculum. A model was proposed to explain the electrochemical behaviour of the biofilm, which appeared to be controlled by the pioneering adherent cells playing the role of “electrochemical gate” between the biofilm and the electrode surface.     

The bioelectrocatalytic properties of cytochrome C by direct electrochemistry on DNA film modified electrode

The direct electrochemistry of cytochrome C can be performed in weak acidic and basic aqueous solutions. Cytochrome C can be deposited as a stable and electrochemically active film on a deoxyribonucleic acid (DNA) modified glassy carbon electrode. These films can also be produced on gold, platinum, and transparent semiconducting tin oxide electrodes. Two-layer modified electrodes containing cytochrome C and a DNA film were prepared by the deposition of cytochrome C on a DNA film modified electrode. The cytochrome C/DNA film was electrocatalytically oxidation active for l-cysteine in a pH 8.3 tris(hydroxymethyl)aminomethane (TRIS)-buffered aqueous solution through both Fe^III and Fe^IV species. The electrocatalytic oxidation current developed from the anodic peak of the redox couple. The electrocatalytic oxidation properties of ascorbic acid, NH₂OH, N₂H₄, and SO₃²⁻ by a cytochrome C/DNA film were also determined, and shown to be electrocatalytically active. An electrochemical quartz crystal microbalance, cyclic voltammetry, and direct spectroelectrochemistry were used to study in situ DNA deposition on a gold disc electrode and cytochrome C deposition on DNA/Au and DNA/GC films. The direct electrochemistry of cytochrome C can also be performed, and it can be deposited as a stable and electrochemically active film on polyvinyl sulfonate, polystyrene sulfonate, TiO₂, and polyethylene glycol modified glassy carbon electrodes. The results show that cytochrome C interacts with, and deposits on, a DNA film modified electrode, and that the cytochrome C (Fe^III) oxidized form is more easily deposited on a DNA film than the cytochrome C (Fe^II) reduced form.     

Electrochemical reduction of oxygen catalyzed by Pseudomonas aeruginosa

Pseudomonas aeruginosa has already been shown to catalyze oxidation processes in the anode compartment of a microbial fuel cell. The present study focuses on the reverse capacity of the bacterium, i.e. reduction catalysis. Here we show that P. aeruginosa is able to catalyze the electrochemical reduction of oxygen. The use of cyclic voltammetry showed that, for a given range of potential values, the current generated in the presence of bacteria could reach up to four times the current obtained without bacteria. The adhesion of bacteria to the working electrode was necessary for the catalysis to be observed but was not sufficient. The electron transfer between the working electrode and the bacteria did not involve mediator metabolites like phenazines. The transfer was by direct contact. The catalysis required a certain contact duration between electrodes and live bacteria but after this delay, the metabolic activity of cells was no longer necessary. Membrane-bound proteins, like catalase, may be involved. Various strains of P. aeruginosa, including clinical isolates, were tested and all of them, even catalase-defective mutants, presented the same catalytic property. P. aeruginosa offers a new model for the analysis of reduction catalysis and the protocol designed here may provide a basis for developing an interesting tool in the field of bacterial adhesion.     

Regeneration of Bombyx mori silk by electrospinning. Part 3: characterization of electrospun nonwoven mat

Regenerated Bombyx mori silk fibroin in formic acid was electrospun and the morphological, chemical and mechanical properties of these nanofibers were examined by field emission environmental scanning electron microscopy (FESEM), Raman spectroscopy (RS), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and tensile testing. FESEM indicated that the average fiber diameter was less than 100 nm and circular in cross section. This paper maps the silk fibroin molecular conformations of each step of the sample preparation and the electrospinning process. The secondary structural compositions (random and β-sheet) of the fibroin were determined by FTIR and RS. The crystallinity index of the electrospun fiber, calculated as the intensity ratio of 1624 (β-sheet) and 1663 (random) cm⁻¹ FTIR bands was higher than that of the pristine fiber. Raman spectra of the amide I (1665 cm⁻¹, random) to amide III (1228 cm⁻¹, β-sheet) ratio of the electrospun fiber was less than that of the pristine fiber indicative of higher β-sheet content. The fiber crystallinity, determined by XRD, showed a lower value for the electrospun fiber. The electrospun fiber shows small but significant increases in the β-sheet content in comparison with the pristine fiber. Dissolution of fibroin in formic acid enhances β-sheet crystallization and may facilitate β-sheet formation in electrospun fiber. The electrospun random silk mat had a Young's modulus, ultimate tensile strength and strain of 515 MPa, 7.25 MPa and 3.2%, respectively.     

Regeneration of Bombyx mori silk by electrospinning—part 1: processing parameters and geometric properties

We studied the effect of electrospinning parameters on the morphology and fiber diameter of regenerated silk from Bombyx mori. Effects of electric field and tip-to-collection plate distances of various silk concentrations in formic acid on fiber uniformity, morphology and diameter were measured. Statistical analysis showed that the silk concentration was the most important parameter in producing uniform cylindrical fibers less than 100 nm in diameter.     

Leucaena diversifolia a new raw material for paper production by soda-ethanol pulping process

Pulping and papermaking of Leucaena diversifolia by soda-anthraquinone-ethanol was studied using an experimental design in order to investigate the effects of cooking variables: temperature, time, soda concentration, ethanol concentration and wash-disintegrate temperature on the pulp yield and the physico-chemical characteristics of paper sheets (tensile index, burst index, tear index and brightness). Previously, in order to assess the potential of plants of this raw material grown over short periods, its results were compared with those of other leucaena varieties and the best crop among three grown for 1–3 years was selected. The results were evaluated using the response surface methodology with a view to identifying the most suitable operating conditions. In accordance with biomass production and the features of the raw materials and cellulose pulp obtained, the L. diversifolia grown for 2 years was found to be the most suitable choice for obtaining pulp and paper among the five leucaena varieties examined. Suitable physical characteristics of paper sheets (tensile index, burst and tear index) and acceptable yield pulping and brightness could be obtained by operating at medium temperature, active alkali concentration, pulping time, ethanol concentration and wash-disintegrate temperature.     

Production of biodiesel by ultrasound assisted esterification of Oreochromis niloticus oil

This paper evaluates the production of methyl esters from Oreochromis niloticus (Nile tilapia) oil and methanol. The reaction was carried out applying low-frequency high-intensity ultrasound (40 kHz) under atmospheric pressure and ambient temperature. Response surface methodology (RSM) was used to evaluate the influence of alcohol to oil molar ratio, catalyst concentration (sulfuric acid) and temperature on the yield of O. niloticus oil into methyl esters. Analysis of the operating conditions by RSM showed that the most important operating condition affecting the reaction was the alcohol to FFA molar ratio. The highest yield observed was of 98.2% after 90 min of reaction. The optimal operating condition was obtained applying an alcohol to oil molar ratio of 9.0 and a catalyst concentration of 2.0% w/w and temperature of 30 °C.