Preparation of Highly Pure n-3 PUFA-Enriched Triacylglycerols by Two-Step Enzymatic Reactions Combined with Molecular Distillation

Highly pure n-3 polyunsaturated fatty acids (PUFA)-enriched triacylglycerols (TAG) have attracted considerable attention due to their nutritional benefits and pharmacological effects. In this study, an alternative approach to the conventional method for the synthesis of highly pure n-3 PUFA-enriched TAG by using a multi-step process was reported. First, glyceride mixtures were synthesized through Novozym 435 [Novozymes A/S (Bagsvaerd, Denmark)] catalyzed esterification of n-3 PUFA-enriched FA and glycerol. Second, partial glycerides in the resulting glyceride mixtures were hydrolyzed to FA by immobilized partial glycerides-selective lipase from Malassezia globosa. The purity of TAG reached 99.84% under the optimized conditions: buffer solution of pH 6.0, water content of 100% (w/w, with respect to the oil mass), enzyme loading of 120 U/g (U/w, with respect to oil mass) and reaction temperature of 30 °C. During hydrolysis, the immobilized SMG1-F278N exhibited good reusability and TAG purity of over 94% was maintained after being used for six cycles. Subsequently, purification of TAG was accomplished by molecular distillation at low temperature (150 °C) and highly pure (99.85%) TAG with 88.73% n-3 PUFA was obtained. The final glyceride mixtures with low acid, peroxide and anisidine value were promising products for medical and dietetic purposes. Compared with the conventional one-step synthesis of n-3 PUFA-enriched TAG by enzymatic esterification or glycerolysis or the two-step method by combined transesterification and ethanolysis, this improved process allows for higher purity of n-3 PUFA-enriched TAG and significant reduction in reaction time.     

Fungal fucoidanase production by solid-state fermentation in a rotating drum bioreactor using algal biomass as substrate

Fucoidanase enzymes able to degrade fucoidan were produced by solid-state fermentation (SSF). The fermentation assays were initially carried out in a laboratory-scale rotating drum bioreactor, and two fungal strains (Aspergillus niger PSH and Mucor sp. 3P) and three algal substrates (untreated, autohydrolyzed, and microwave processed seaweed Fucus vesiculosus) were evaluated. Additionally, fermentations were carried out under rotational (10 rpm) and static conditions in order to determine the effect of the agitation on the enzyme production. Agitated experiments showed advantages in the induction of the enzyme when compared to the static ones. The conditions that promoted the maximum fucoidanase activity (3.82 U L^?1) consisted in using Mucor sp. 3P as fungal strain, autohydrolyzed alga as substrate, and the rotational system. Such conditions were subsequently used in a 10 times larger scale rotating drum bioreactor. In this step, the effect of controlling the substrate moisture during the enzyme production by SSF was investigated. Moreover, assays combining the algal substrate with an inert support (synthetic fiber) were also carried out. Fermentation of the autohydrolyzed alga with the moisture content maintained at 80% during the fermentation with Mucor sp. 3P gave the highest enzyme activity (9.62 U L^?1).     

Commercial bentonites as detoxifier of broiler feed contaminated with aflatoxin

Some bentonites have been probed to be efficient as sequestering agents for aflatoxins; they decreased the bioavailability of the toxin in the gastrointestinal tracts of birds when they are incorporated in the diets. The binding capacity of these adsorbents varied with the rheological source and even among batches of a given source. Three bentonites from different sources in Argentina, which have very different aflatoxins (AFs) adsorption capacity, were studied. The characterization comprises chemical analysis, scanning electron microscopy (SEM), X-ray power diffraction (XRD), Hg intrusion porosimetry, swelling capacity, etc. The main factors affecting the adsorption of AFs seem to be related to the isomorphic substitution of the montmorillonite and to electrostatic interactions generated by the surface charge of the samples. Neither the mean pore size nor the percentage of quartz has any effect upon the AFs adsorption.     

Antifungal activity mode of Aspergillus ochraceus by bacillomycin D and its inhibition of ochratoxin A (OTA) production in food samples

The inhibition effects of bacillomycin D on the growth of Aspergillus ochraceus and the production of ochratoxin A (OTA) in food samples were investigated. The mycelia growth and sporulation were completely inhibited by 30 μg/mL of bacillomycin D. Microscopic morphological changes such as the distortion of hyphae and the disruption of spores at 20 μg/mL of bacillomycin D were significantly observed. The use of bacillomycin D resulted in cell damage, nucleic acids and proteins divulge, and more production of reactive oxygen species (ROS), and all these factors actively contributed to the promotion of apoptosis of A. ochraceus. In addition, 90 μg/g of bacillomycin D completely inhibited the growth of A. ochraceus and the production of OTA in food samples. Our results suggested that bacillomycin D showed a significant antifungal activity against A. ochraceus that could be used as a potential natural antimicrobial to control food contamination and ensure food safety.     

Intelligent release of cinnamon oil from engineered proteoliposome via stimulation of Bacillus cereus protease

Inspired by the hydrolysis of casein by protease, a new approach to deliver antimicrobials against bacterial infections was developed in this study. As a natural antibacterial agent, cinnamon oil was encapsulated into engineered liposomes inlaid with casein. The average particle size of proteoliposomes was 615.0 nm and their entrapment efficiency (EE) was 40.0%. In this work, Bacillus cereus (B. cereus) was chosen as model bacterium. The controlled release of liposome-encapsulated cinnamon oil was realized via bacterial protease secreted from B. cereus. As a result, 99.99% of the bacteria could be efficiently inhibited in rice and wheat flour.     

Astaxanthin extraction from Haematococcus pluvialis using CO2-expanded ethanol

Microalgae represent diverse branch of microorganism that can produce a wide range of unique functional ingredients that can be used in food, cosmetics, pharmaceuticals, and energy. Among them, Haematococcus pluvialis is known for accumulating the highest levels of a potent natural antioxidant, astaxanthin, which has demonstrated positive health effects. Therefore, the aim of numerous studies has been to develop novel and efficient extraction techniques to produce high-quality (purity and antioxidant activity) extracts, while complying with the Green Chemistry Principles. Supercritical CO₂ (scCO₂) emerges as an alternative to organic solvents because of its high selectivity and bioactivity-preserving qualities. Nevertheless, astaxanthin is a large molecule with low solubility in scCO₂ that usually requires long extractions at high pressures. Ethanol has been used as co-solvent to increase astaxanthin solubility in scCO₂. In this work, a Box–Behnken experimental design was used to study the effects of operating pressure (20–35 MPa), temperature (40–70 °C), and ethanol content in scCO₂ (0–13%, w/w) on the yield, astaxanthin content, and antioxidant activity of H. pluvialis extract. Results showed that ethanol content in CO₂ has a more significant effect on all responses than pressure and temperature. These results lead us to investigate the effect of a further increase in ethanol content, up to the region of gas-expanded liquids. We studied the effects of temperature (30–60 °C) and ethanol content (50–70%, w/w) at a fixed pressure (7 MPa) on the same response variables using CO₂-expanded ethanol (CXE). Results showed that temperature and ethanol content had a significant influence on astaxanthin yield and antioxidant activity. Also, the overall responses of CXE surpassed scCO₂ extractions to match conventional extraction with acetone, maintaining high quality extracts, thus validating the use of this new type of green technology for extraction of high-value compounds.     

Microwave dielectric properties of glass-ceramic composites for low temperature co-firable ceramics

Ba–B–Si glass was added to Ba–Nd–Sm–Bi–Ti–O (BRT₁₁₄) microwave dielectric material for LTCC applications. Conventional one-step processing method for preparing glass-BRT₁₁₄ composite materials yields low dielectric constant, since the glass was easy to react with BRT₁₁₄ and forms a low dielectric constant phase, Ba₃B₆Si₂O₁₆. A large proportion of pores appeared. The nature of glass, whether it is sol-gel derived or fused, shows marked influence on the microstructure and microwave dielectric properties of the composites. A two-step process containing precoating the BRT₁₁₄ powders with a thin layer of glass, followed by conventional samples preparation process, tremendously improved the densification behaviour of the material. The formation of pores and interactions between glass and BRT₁₁₄ was greatly suppressed such that materials with high dielectric constant (ε_r=40) were achieved by sintering 9 wt.% glass-containing composite at 950 °C for 2.5 h.