Lipase/acyltransferase‐catalysed interesterification of fat blends containing n‐3 polyunsaturated fatty acids

The lipase/acyltransferase from Candida parapsilosis is an original biocatalyst that preferentially catalyses alcoholysis over hydrolysis in biphasic aqueous/organic media. In this study, the performance of the immobilised biocatalyst in the interesterification in solvent‐free media of fat blends rich in n‐3 polyunsaturated fatty acids (n‐3 PUFA) was investigated. The interesterification activity of this biocatalyst at a water activity (a_w) of 0.97 was similar to that of commercial immobilised lipases at a_w values lower than 0.5. Thus, the biocatalyst was further used at an a_w of 0.97. Response surface modelling of interesterification was carried out as a function of medium formulation, reaction temperature (55–75 °C) and time (30–120 min). Reaction media were blends of palm stearin (PS), palm kernel oil and triacylglycerols (TAG) rich in n‐3 PUFA (“EPAX 4510TG”; EPAX AS, Norway). The best results in terms of decrease in solid fat content were observed for longer reaction time (>80 min), lower temperature (55–65 °C), higher “EPAX 4510TG” content and lower PS concentration. Reactions at higher temperature led to final interesterified fat blends with lower free fatty acid contents. TAG with high equivalent carbon number (ECN) were consumed while acylglycerols of lower ECN were produced.     

Production of Human Milk Fat Substitutes by Interesterification of Tripalmitin with Ethyl Oleate Catalyzed by Candida parapsilosis Lipase/Acyltransferase

In human milk fat, the saturated fatty acids, namely palmitic acid, are located at the sn-2 position of triacylglycerols (TAG) while unsaturated fatty acids (e.g. oleic acid) are esterified at position sn-1,3. Thus, sn-1,3-dioleoyl-2-palmitoylglycerol (OPO) is the target TAG to be used as human milk fat substitutes (HMFS) in infant formulas. In this study, the noncommercial recombinant lipase/acyltransferase from Candida parapsilosis (CpLIP2) was immobilized in Accurel MP1000, and used as a biocatalyst for the interesterification of tripalmitin with ethyl oleate in a solvent-free medium, to obtain structured lipids used as HMFS. Different molar ratios (MR) of ethyl oleate to tripalmitin (2:1–8:1) were used. After 4 h reaction at 60°C, about 30 mol% of oleic acid incorporation was already observed for all tested MR. An apparent equilibrium was reached after 8–24 h, with 32–51 mol% final incorporation, increasing with the MR. The incorporation of oleic acid into TAG was compared with the maximum predicted values when a random or a sn-1,3-regioselective biocatalyst was used. The obtained values are consistent with the maximum incorporation expected for a sn-1,3-regioselective enzyme. In fact, the amount of oleic acid at position sn-2 was approximately 15% for all the MR tested, which is explained by the acyl migration phenomenon. CpLIP2 exhibited higher activity than most commercial immobilized lipases (e.g. faster reaction in solvent-free media, low enzyme load, and low MR needed), and showed a recognized sn-1,3 regioselective behavior.     

Production of Human Milk Fat Substitutes Catalyzed by a Heterologous Rhizopus oryzae Lipase and Commercial Lipases

This study aims to produce human milk fat substitutes by an acidolysis reaction between lard and the free fatty acids (FFA) from a fish oil concentrate rich in docosahexaenoic acid, in solvent-free media. The immobilized commercial lipases from (1) Rhizomucor miehei (Lipozyme RM IM), (2) Thermomyces lanuginosa (Lipozyme TL IM) and (3) Candida antarctica (Novozym 435) were tested as biocatalyst. Also, the heterologous Rhizopus oryzae lipase (rROL), immobilized in Accurel^® MP 1000, was tested as a feasible alternative to the commercial lipases. After 24 h of reaction at 50 °C, similar incorporations of polyunsaturated fatty acids (c.a. 17 mol%) were attained with Novozym 435, Lipozyme RM IM and rROL. The lowest incorporation was achieved with Lipozyme TL IM (7.2 mol%). Modeling acidolysis catalyzed by rROL and optimization of reaction conditions were performed by response surface methodology, as a function of the molar ratio FFA/lard and the temperature. The highest acidolysis activity was achieved at 40 °C at a molar ratio of 3:1, decreasing with both temperature and molar ratio. Operational stability studies for rROL in seven consecutive 24-h batches were carried out. After the fourth batch, the biocatalyst retained about 55 % of the original activity (half-life of 112 h).     

Optimized Production of MLM Triacylglycerols Catalyzed by Immobilized Heterologous <Emphasis Type="Italic">Rhizopus oryzae</Emphasis> Lipase

Response surface methodology was used to model and optimize the acidolysis of virgin olive oil with caprylic (C8:0) or capric (C10:0) acids, aimed at the production of low caloric triacylglycerols (TAG) of MLM type, in solvent free media, catalyzed by the heterologous Rhizopus oryzae lipase (r-ROL) immobilized in Eupergit^® C. This lipase was produced in the methylotrophic yeast Pichia pastoris Mut^s phenotype (experiments with C10:0) or a Mut⁺ phenotype (experiments with C8:0), under different operational conditions. The r-ROL used in experiments with C10:0 presented a hydrolytic activity about 5 times of that presented by r-ROL used in acidolysis with C8:0. The experiments were carried out following a central composite rotatable design, as a function of the molar ratio (MR) medium chain fatty acid/TAG (1.6–4.4) and temperature (25–55 °C). Convex surfaces described by second order polynomials as a function of MR and temperature were well fitted to fatty acid incorporation values. After 24-h reaction, the predicted maximum incorporation of caprylic (15.5 mol%) or capric (33.3 mol%) acids in olive oil occurs at 37 and 35 °C, respectively, and at C8:0/TAG of 2.8:1 or C10:0/TAG of 3:1. These predicted optima were experimentally validated. Fermentation conditions used in r-ROL production highly affected hydrolytic activity and to a lesser extent interesterification activity.     

Adsorption of acid orange 7 dye in aqueous solutions by spent brewery grains

Spent brewery grains (SBG), a by-product of the brewing process, were tested as an adsorbent of acid orange 7 dye (AO7), a monoazo acid dye currently used in paper and textile industries. The presence of AO7 in these effluents causes obvious environmental problems.

Kinetics studies of adsorption of AO₇ to SBG (3.75%, m/v) were carried out at 20 °C, using aqueous solutions with different AO7 concentrations (30–834 mg/L). For every situations tested, no significant variation in residual AO7 concentration in solution was detected after 1 h contact between the dye and the adsorbent. The adsorption process followed a pseudo-first order model.

The equilibrium process showed to be well described by both Freundlich and Langmuir isotherm models, at 20 and 30 °C. The maximum adsorption capacity was estimated to be 30.5 mg AO7/g SBG, at 30 °C.

Free energy of adsorption (ΔG°), enthalpy (ΔH°), and entropy (ΔS°) changes were calculated to predict the nature of adsorption. The estimated values for ΔG° were −22.78 and −24.53 kJ/mol, respectively, at 293.3 K (20 °C) and 303.3 K (30 °C), which are rather low indicating that a spontaneous process occurred. The enthalpy changes and entropy of adsorption were 28.66 and 175.36 J/mol K, respectively. The positive value for ΔH° indicates that the adsorption of AO7 dye to SBG is an endothermic process. The positive value of entropy reflects the affinity of the adsorbent for AO7 dye.

The obtained results are very promising since: (i) high levels of colour removal (>90%) were achieved with low contact times adsorbent/dye (less than 1 h contact); and (ii) the whole SBG can be successfully used as adsorbent of AO7 dye in aqueous solution without needing any previous treatments such as milling and/or sieving. Spent grains, being a cheap, and easily available material, can be an alternative for more costly adsorbents used for dye removal in wastewater treatment processes.     

The waterborne polyurethane dispersions based on polycarbonate diol: Effect of ionic content

Three water-based polyurethane dispersions (PUD) were synthesized by modified dispersing procedure using polycarbonate diol (PCD), isophorone diisocyanate (IPDI), dimethylolpropionic acid (DMPA), triethylamine (TEA) and ethylenediamine (EDA). The ionic group content in the polyurethane-ionomer structure was varied by changing the amount of the internal emulsifier, DMPA (4.5, 7.5 and 10 wt.% to the prepolymer weight). The expected structures of obtained materials were confirmed by FTIR spectroscopy. The effect of the DMPA content on the thermal properties of polyurethane films was measured by TGA, DTA, DSC and DMTA methods. Increased DMPA amounts result in the higher hard segment contents and in the increase of the weight loss corresponding to the degradation of the hard segments. The reduction of hard segment content led to the elevated temperature of decomposition and to the decrease of the glass transition temperature and thermoplasticity. The atomic force microscopy (AFM), results indicated that phase separation between hard and soft segment of PUD with higher DMPA content is more significant than of PUD with lower DMPA content. The physico-mechanical properties, such as hardness, adhesion test and gloss of the dried films were also determined considering the effect of DMPA content on coating properties.     

Combustion performance of rubber seed methyl ester using aromatic and phenolic antioxidants in a multi-cylinder diesel engine

This paper analyzes the effect of antioxidants on engine combustion performance of a multi-cylinder diesel engine fueled with rubber seed biodiesel blends. Four antioxidants, namely 2-tert-butylbenzene-1,4-diol, N,N′-diphenyl-1,4-phenylenediamine, 2(3)-tert-Butyl-4-methoxyphenol and N-phenyl-1,4-phenylenediamine, were added at concentrations of 1000 and 2000 ppm to rubber seed biodiesel blends. Antioxidants blends increased the brake power by 4.21% and decreased the brake-specific fuel consumption by 6.82% compared to base biodiesel blends. The NO emissions reduction percentage for antioxidants fuels was 9.78% compared to base line biodiesel. However, the treated biodiesel blends increased carbon monoxide, hydrocarbon and smoke opacity up to 32.20, 42.15 and 43.92%, respectively, compared to non-treated blends. Compared to diesel fuel, antioxidants fuels decreased the brake power and increased the brake-specific fuel consumption, cylinder pressure and heat release rate. But compared to biodiesel blends, the cylinder pressure and heat release rate with antioxidants were reduced by 4.17 and 6.87%, respectively. It can be concluded that antioxidants addition is effective in increasing the oxidation stability and controlling the NO emissions of rubber seed biodiesel fueled diesel engines.     

Ultrathin 2D‐Layered Cyclodextrin Membranes for High‐ Performance Organic Solvent Nanofiltration

Synthetic membranes with a high selectivity for demanding molecular separations and high permeance have a large potential for the reduction of energy consumption in separation processes. Herein, for the first time, the fabrication of an ultrathin layered macrocycle membrane for molecular separation in organic solvent nanofiltration using per‐6‐amino‐β‐cyclodextrin as a monomer for membrane manufacturing by interfacial polymerization is reported. Compared to a regular nonfunctionalized cyclodextrin, a higher reactivity is observed, enabling a very fast membrane formation under mild conditions. The formed membrane is composed of a layered structure of polymerized cyclodextrin, which shows high stability in different organic solvents. The membrane exhibits excellent separation performance for organic solvent nanofiltration, both with nonpolar and polar solvents. Most importantly, this new membrane type can discriminate between molecules with nearly identical molecular weights but different shapes. The unmatched high permeance and shape selectivity of the membranes can be attributed to the ultralow thickness, controlled microporosity, as well as the layered macrocycle structure, which makes the membranes promising for high‐performance molecular separation in the chemical and biochemistry industry.     

Electrospun poly(ε-caprolactone) matrices containing silver sulfadiazine complexed with β-cyclodextrin as a new pharmaceutical dosage form to wound healing: preliminary physicochemical and biological evaluation

Cooperation between researchers in the areas of medical, pharmaceutical and materials science has facilitated the development of pharmaceutical dosage forms that elicit therapeutic effects and protective action with a single product. In addition to optimizing pharmacologic action, such dosage forms provide greater patient comfort and increase success and treatment compliance. In the present work, we prepared semipermeable bioactive electrospun fibers for use as wound dressings containing silver sulfadiazine complexed with β-cyclodextrin in a poly(?-caprolactone) nanofiber matrix aiming to reduce the direct contact between silver and skin and to modulate the drug release. Wound dressings were prepared by electrospinning, and were subjected to ATR-FT-IR and TG/DTG assays to evaluate drug stability. The hydrophilicity of the fibrous nanostructure in water and PBS buffer was studied by goniometry. Electrospun fibers permeability and swelling capacity were assessed, and a dissolution test was performed. In vitro biological tests were realized to investigate the biological compatibility and antimicrobial activity. We obtained flexible matrices that were each approximately 1.0?g in weight. The electrospun fibers were shown to be semipermeable, with water vapor transmission and swelling indexes compatible with the proposed objective. The hydrophilicity was moderate. Matrices containing pure drug modulated drug release adequately during 24?h but presented a high hemolytic index. Complexation promoted a decrease in the hemolytic index and in the drug release but did not negatively impact antimicrobial activity. The drug was released predominantly by diffusion. These results indicate that electrospun PCL matrices containing β-cyclodextrin/silver sulfadiazine inclusion complexes are a promising pharmaceutical dosage form for wound healing.     

Synthesis and characterization of poly(2-hydroxyethyl methacrylate/itaconic acid) copolymeric hydrogels

In the present study poly(2-hydroxyethyl methacrylate-co-itaconic acid) (P(HEMA/IA)) hydrogels were synthesized by free-radical copolymerization of 2-hydroxyethyl methacrylate and itaconic acid in order to evaluate as controlled release drug delivery system. All polymerizations were performed in a mixture of water/ethanol with ethylene glycol dimethacrylate, as crosslinking agent, potassium persulfate, as initiator, and N, N, N′, N′-tetramethylethylene diamine, as activator. The samples were characterized by FTIR and SEM. Swelling kinetics and antibiotic release studies were performed using gravimetry and UV spectrophotometry, respectively. The antibacterial activity studies were performed bearing in mind that infections are the most common cause of biomaterial implant failure and represent a constant menace to the application of medical implants. The hemocompatibility testing was performed as imperative for medical devices intended for direct or indirect blood exposure. In accordance with the results of antibacterial assessment on gels loaded with antibiotics and hemolytic activity testing these gels exerted good bacterial growth inhibition and favorable hemolytic activity. The release profiles of antibiotics, evaluated in vitro conditions, were correspondent for antibacterial therapeutics. Therefore, it was concluded that (P(HEMA/IA)) gels could be propound for the potential application as drug delivery systems for the controlled release of antibiotics.