Bioconversion of Cheese Whey and Food By-Products by Phaeodactylum tricornutum into Fucoxanthin and n-3 Lc-PUFA through a Biorefinery Approach.

This study investigates the potential of utilizing three food wastes: cheese whey (CW), beet molasses (BM), and corn steep liquor (CSL) as alternative nutrient sources for the cultivation of the diatom Phaeodactylum tricornutum, a promising source of polyunsaturated eicosapentaenoic acid (EPA) and the carotenoid fucoxanthin. The CW media tested did not significantly impact the growth rate of P. tricornutum; however, CW hydrolysate significantly enhances cell growth. BM in cultivation medium enhances biomass production and fucoxanthin yield. The optimization of the new food waste medium was conducted through the application of a response surface methodology (RSM) using hydrolyzed CW, BM, and CSL as factors. The results showed a significant positive impact of these factors (p < 0.005), with an optimized biomass yield of 2.35 g L-1 and a fucoxanthin yield of 3.64 mg L-1 using a medium composed of 33 mL L-1 of CW, 2.3 g L-1 of BM, and 2.24 g L-1 of CSL. The experimental results reported in this study showed that some food by-products from a biorefinery perspective could be utilized for the efficient production of fucoxanthin and other high-added-value products such as eicosapentaenoic acid (EPA).

Bio-Based Materials for Packaging.

Plastic pollution is currently one of the most pressing environmental problems, especially in countries with a low recycling rate that is mainly due to the insufficient collection of plastic waste [...].

Properties and Biodegradability of Films Based on Cellulose and Cellulose Nanocrystals from Corn Cob in Mixture with Chitosan.

The increase in consumer demand for more sustainable packaging materials represents an opportunity for biopolymers utilization as an alternative to reduce the environmental impact of plastics. Cellulose (C) and chitosan (CH) are attractive biopolymers for film production due to their high abundance, biodegradability and low toxicity. The objective of this work was to incorporate cellulose nanocrystals (NC) and C extracted from corn cobs in films added with chitosan and to evaluate their properties and biodegradability. The physicochemical (water vapor barrier, moisture content, water solubility and color) and mechanical properties of the films were evaluated. Component interactions using Fourier-transform infrared (FTIR) spectroscopy, surface topography by means of atomic force microscopy (AFM), biodegradability utilizing a fungal mixture and compostability by burying film discs in compost were also determined. The C-NC-CH compared to C-CH films presented a lower moisture content (17.19 ± 1.11% and 20.07 ± 1.01%; w/w, respectively) and water vapor permeability (g m−1 s−1 Pa−1 × 10−12: 1.05 ± 0.15 and 1.57 ± 0.10; w/w, respectively) associated with the NC addition. Significantly high roughness (Rq = 4.90 ± 0.98 nm) was observed in films added to NC, suggesting a decreased homogeneity. The biodegradability test showed larger fungal growth on C-CH films than on CH films (>60% and <10%, respectively) due to the antifungal properties of CH. C extracted from corn cobs resulted in a good option as an alternative packaging material, while the use of NC improved the luminosity and water barrier properties of C-CH films, promoting strong interactions due to hydrogen bonds.

Formulation of New Media from Dairy and Brewery Wastes for a Sustainable Production of DHA-Rich Oil by Aurantiochytrium mangrovei.

Mozzarella stretching water (MSW) is a dairy effluent generated from mozzarella cheese production that does not have a real use and is destined to disposal, causing environmental problems and representing a high disposal cost for dairy producers. Spent brewery yeast (SBY) is another promising food waste produced after brewery manufacturing that could be recycled in new biotechnological processes. Aurantiochytrium mangrovei is an aquatic protist known as producer of bioactive lipids such as omega 3 long chain polyunsaturated fatty acids (ω3 LC-PUFA), in particular docosahexaenoic acid (DHA). In this work MSW and SBY have been used to formulate new sustainable growth media for A. mangrovei cultivation and production of DHA in an attempt to valorize these effluents. MSW required an enzymatic hydrolysis to enhance the biomass production. The new media obtained from hydrolysed MSW was also optimized using response surface methodologies, obtaining 10.14 g L-1 of biomass in optimized medium, with a DHA content of 1.21 g L-1.

Biopolymers as Food Packaging Materials.

Oil-derived plastics are the most commonly used materials for packaging because of their features, low cost, and availability of resources for manufacturing [...].

Black Edible Films from Protein-Containing Defatted Cake of Nigella sativa Seeds.

Black biodegradable/edible protein-based films were prepared from defatted cake waste obtained from Nigella sativa (black cumin) seeds as by-product of oil extraction process. The effects of pH, glycerol concentrations, and transglutaminase-catalyzed protein cross-linking activity on the stability of film-forming solutions were studied to determine the best experimental conditions to produce handleable films. Proteins contained in the analyzed defatted cake were shown to be able to act as transglutaminase acyl donor and acceptor substrates being polymerized when incubated in vitro in the presence of the enzyme. Film-forming solutions containing 20% glycerol and casted at pH 8.0 after treatment with the enzyme gave rise to morphologically more homogeneous films possessing mechanical and barrier properties, as well as antimicrobial activity, compatible with their possible applications as food packaging materials and mulching sheets. These findings confirm the validity of the strategy to consider the seed oil processed cakes as protein-based renewable sources to produce not only fertilizers, animal feed, or culinary food but also further valuable products such as bioplastics.

Tuning the Functional Properties of Bitter Vetch (Vicia ervilia) Protein Films Grafted with Spermidine.

Bitter vetch protein films containing positively charged spermidine, alone or with low amounts of glycerol, showed high tensile strength that progressively decreased by increasing the plasticizer concentration. Accordingly, lower film elongation at break and higher Young's module values were detected in the presence of the polyamine without or with small amounts of glycerol. These data suggest that spermidine not only acts as a plasticizer itself by ionically interacting with proteins, but that it also facilitates glycerol-dependent reduction of the intermolecular forces along the protein chains, consequently improving the film flexibility and extensibility. Thus, spermidine may be considered not only as a primary, but also as a secondary plasticizer because of its ability to enhance glycerol plasticizing performance. Such double behavior of the polyamine was confirmed by the film permeability tests, since spermidine increased the barrier properties to gases and water vapor, while glycerol emphasized this effect at low concentrations but led to its marked reversal at high concentrations. Film microscopic images also substantiated these findings, showing more compact, cohesive, and homogeneous matrices in all spermidine-containing films.

Plasticizing Effects of Polyamines in Protein-Based Films.

Zeta potential and nanoparticle size were determined on film forming solutions of native and heat-denatured proteins of bitter vetch as a function of pH and of different concentrations of the polyamines spermidine and spermine, both in the absence and presence of the plasticizer glycerol. Our results showed that both polyamines decreased the negative zeta potential of all samples under pH 8.0 as a consequence of their ionic interaction with proteins. At the same time, they enhanced the dimension of nanoparticles under pH 8.0 as a result of macromolecular aggregations. By using native protein solutions, handleable films were obtained only from samples containing either a minimum of 33 mM glycerol or 4 mM spermidine, or both compounds together at lower glycerol concentrations. However, 2 mM spermidine was sufficient to obtain handleable film by using heat-treated samples without glycerol. Conversely, brittle materials were obtained by spermine alone, thus indicating that only spermidine was able to act as an ionic plasticizer. Lastly, both polyamines, mainly spermine, were found able to act as "glycerol-like" plasticizers at concentrations higher than 5 mM under experimental conditions at which their amino groups are undissociated. Our findings open new perspectives in obtaining protein-based films by using aliphatic polycations as components.

Physical, Structural, Barrier, and Antifungal Characterization of Chitosan-Zein Edible Films with Added Essential Oils.

Edible films (EFs) have gained great interest due to their ability to keep foods safe, maintaining their physical and organoleptic properties for a longer time. The aim of this work was to develop EFs based on a chitosan-zein mixture with three different essential oils (EOs) added: anise, orange, and cinnamon, and to characterize them to establish the relationship between their structural and physical properties. The addition of an EO into an EF significantly affected (p < 0.05) the a* (redness/greenness) and b* (yellowness/blueness) values of the film surface. The EFs presented a refractive index between 1.35 and 1.55, and thus are classified as transparent. The physical properties of EFs with an added EO were improved, and films that incorporated the anise EO showed significantly lower water vapor permeability (1.2 ± 0.1 g mm h-1 m-2 kPa-1) and high hardness (104.3 ± 3.22 MPa). EFs with an added EO were able to inhibit the growth of Penicillium sp. and Rhizopus sp. to a larger extent than without an EO. Films' structural changes were the result of chemical interactions among amino acid side chains from zein, glucosamine from chitosan, and cinnamaldehyde, anethole, or limonene from the EOs as detected by a Raman analysis. The incorporation of an EO in the EFs' formulation could represent an alternative use as coatings to enhance the shelf life of food products.

Trehalose-containing hydrocolloid edible films prepared in the presence of transglutaminase.

In this article, edible hydrocolloid films were prepared by using Citrus pectins and the protein phaseolin in the presence of microbial transglutaminase, an enzyme able to catalyze isopeptide bonds between endo-protein-reactive glutamine and lysine residues. For the first time, trehalose, a nonreducing homodisaccharide into which two glucose units are linked together by a α-1,1-glycosidic linkage, was used as a component of hydrocolloid films constituted of both proteins and carbohydrates. Our data have demonstrated that these films act as very effective barriers to gases, especially to CO2 . They also present a high antioxidant capability as measured by the 2,2-diphenyl-1-picrylhydrazyl scavenging assay. In addition, the films were characterized using Atomic Force Microscopy, a powerful tool used to evaluate film surface topography and roughness. The results of our experiments clearly indicate that the trehalose-containing films prepared both in the presence and absence of transglutaminase are composed of nanoparticles with a smooth surface, having similar roughness values (Rα). In conclusion, according to barrier and antioxidant properties and to their structure, it is possible to consider the trehalose-containing films as innovative bioplastics potentially able to protect different kinds of foods.

Effect of Amine, Carboxyl, or Thiol Functionalization of Mesoporous Silica Particles on Their Efficiency as a Quercetin Delivery System in Simulated Gastrointestinal Conditions.

Quercetin (Q) dietary supplements exhibit poor oral bioavailability because of degradation throughout gastrointestinal digestion (GD), which may be overcome using mesoporous silica particles (MSPs) as an oral delivery system (ODS). This study aimed to elucidate the effect of the functionalization of MSPs with amine-(A-MSP), carboxyl-(C-MSP), or thiol-(T-MSP) groups on their efficiency as a quercetin ODS (QODS). The type and degree of functionalization (DF) were used as factors in an experimental design. The Q-loaded F-MSP (F-MSP/Q) was characterized by gas physisorption analysis, loading capacity (LC), and dynamic light scattering and kinetics of Q release at gastric and intestinal pHs. Antioxidant capacity and Q concentration of media containing F-MSP/Q were evaluated after simulated GD. A-MSP showed the highest LC (19.79 ± 2.42%). C-MSP showed the lowest particle size at pH 1.5 or 7.4 (≈200 nm). T-MSP exhibited the maximum Q release at pH 7.4 (11.43%). High DF of A-MSP increased Q retention, regardless of pH. A-MSP preserved antioxidant capacity of Q-released gastric media (58.95 ± 3.34%). Nonetheless, MSP and F-MSP did not protect antioxidant properties of Q released in intestinal conditions. C-MSP and T-MSP showed essential features for cellular uptake and Q release within cells that need to be assessed.

Higher susceptibility to amyloid fibril formation of the recombinant ovine prion protein modified by transglutaminase.

Prion proteins are known as the main agents of transmissible spongiform encephalopathies affecting humans as well as animals. A recombinant ovine prion protein was found to be in vitro able to act as an effective substrate for a microbial isoform of transglutaminase, an enzyme catalyzing the formation of isopeptide bonds inside the proteins. We proved that transglutaminase modifies the structure of the prion protein by leading to the formation of three intra-molecular crosslinks and that the crosslinked protein form is more competent in amyloid formation compared to the unmodified one. In addition, the crosslinked prion protein was shown also to be more resistant to proteinase K digestion. Our findings suggest a possible use of transglutaminase in stabilizing the prion protein three-dimensional structure in order to investigate the molecular basis of the conversion of the protein into its pathological form.

Transglutaminase-mediated modification of ovomucoid: effects on its trypsin inhibitory activity and antigenic properties.

Hen egg can cause food hypersensitivity in infants and young children, and ovomucoid is the most allergenic factor among proteins contained in egg white. Since proteinase treatment, a well-recognized strategy in reducing food allergenicity, is ineffective when applied to ovomucoid because of its ability to act as trypsin inhibitor, we investigated the possibility of reducing the ovomucoid antiprotease activity and antigenic properties by covalently modifying its structure. The present paper reports data showing the ability of the Gln115 residue of ovomucoid to act as an acyl donor substrate for the enzyme transglutaminase and, as a consequence, to give rise to a covalent monodansylcadaverine conjugate of the protein in the presence of both enzyme and the diamine dansylated derivative. Moreover, we demonstrated that the obtained structural modification of ovomucoid significantly reduced the capability of the protein to inhibit trypsin activity, also having impact on its anti-ovomucoid serum-binding properties.

Study of Physico-Chemical Properties of Dough and Wood Oven-Baked Pizza Base: The Effect of Leavening Time.

The research objective was to investigate the morpho-rheological, chemical, and structural changes of dough and Neapolitan pizza TSG as the leavening time varies and to evaluate their effects on the digestibility of starch and on the formation of acrylamide during baking. Pizza dough leavening was monitored for 48 h at 22 °C/80% RH, and the analyses were conducted at selected leavening times (0, 4, 8, 16, 24, and 48 h). It was observed that in 30 h the volume tripled and the viscoelastic dough relaxed in the first 4 h, as evidenced by the lower value of the relaxation percentage "a" and the higher rate of decay "b", associated with a high value of the compression work, indicating the presence of a very strong gluten mesh. In the following hours, the dough lost elasticity, and in fact, the G' modulus decreased due to the weakening of the weak interactions between the gluten proteins and the starch. This suggests that a long leavening improved the extensibility of the pizza disc, facilitating the action of the pizza maker. Thermal (TGA and DSC) and morphological (SEM) analyses evidenced the highest water removal rate from the dough, a wider starch gelatinization temperature range, a ∆H of 0.975 ± 0.013 J/g, and a more open and weak gluten structure in dough balls leavened for 16 h. As the leavening time increased, both dough and pizza base samples showed an increase in reducing sugars and free amino groups, while the rapidly digestible starch decreased in the dough following the metabolism of the yeasts and increased in the pizza base due to the starch gelatinization that occurs during baking, which makes it much more susceptible to α-amylase. Finally, the levels of acrylamide remained at the same values despite the higher availability of reducing sugars and its precursors during leavening.

Properties of Active Levan-Bitter Vetch Protein Films for Potential Use in Food Packaging Applications.

ζ-potential and Z-average were determined on film-forming solutions of bitter vetch-levan-based films prepared at different ratios in the absence and presence of glycerol as a plasticizer. The casting method was used to obtain manageable films. The results revealed that levan increases the elongation at break of bitter vetch protein films and reduces the tensile strength. The optimal result was obtained through the film that was prepared with the ratio of 50% bitter vetch proteins and 50% levan, in terms of mechanical properties. The surfaces of the prepared films appeared to be more compact and smooth. On increasing the glycerol concentration in the bitter vetch protein-levan films, the oxygen and water vapor permeability increased compared to the control (P < 0.05). Based on the overall results, the reinforcement of bitter vetch proteins with levan at a ratio of 1:1 represents optimal film properties in the presence of a low concentration of glycerol. The proposed film is suggested as an innovative packaging system for beef meat to preserve its quality over time.

Development of Functional Pizza Base Enriched with Jujube (Ziziphus jujuba) Powder.

Functional and enriched foods are increasingly in demand in the global market due to their benefits for human health and their prevention of several diseases. The aim of this work was to develop a functional pizza base, produced in the Neapolitan style, exploiting the beneficial properties of jujube. The jujube fruit is rich in phenolic compounds with high antioxidant activity and represents a good candidate for functional food development. The doughs were prepared by replacing the wheat flour with 2.5%, 5.0%, and 7.5% (w/w) of Ziziphus jujube powder (ZJP) and were subsequently cooked. Chemical analyses showed that both total phenolic compounds and antioxidant activity grew with the increase of ZJP. The addition of ZJP darkened the pizza base and raised its hardness, gumminess, and chewiness. However, no difference was found in the springiness and cohesiveness of the samples with or without ZJP. These results suggest that jujube powder can be successfully introduced into pizza dough as a functional ingredient.

Basil (Ocimum basilicum L.) Leaves as a Source of Bioactive Compounds.

Basil (Ocimum basilicum L.) is an annual spicy plant generally utilized as a flavouring agent for food. Basil leaves also have pharmaceutical properties due to the presence of polyphenols, phenolic acids, and flavonoids. In this work, carbon dioxide was employed to extract bioactive compounds from basil leaves. Extraction with supercritical CO2 (p = 30 MPa; T = 50 °C) for 2 h using 10% ethanol as a cosolvent was the most efficient method, with a yield similar to that of the control (100% ethanol) and was applied to two basil cultivars: "Italiano Classico" and "Genovese". Antioxidant activity, phenolic acid content, and volatile organic compounds were determined in the extracts obtained by this method. In both cultivars, the supercritical CO2 extracts showed antiradical activity (ABTS●+ assay), caffeic acid (1.69-1.92 mg/g), linalool (35-27%), and bergamotene (11-14%) contents significantly higher than those of the control. The polyphenol content and antiradical activity measured by the three assays were higher in the "Genovese" cultivar than in the "Italiano Classico" cultivar, while the linalool content was higher (35.08%) in the "Italiano Classico" cultivar. Supercritical CO2 not only allowed us to obtain extracts rich in bioactive compounds in an environmentally friendly way but also reduced ethanol consumption.

Exploiting Potential Biotechnological Applications of Poly-γ-glutamic Acid Low Molecular Weight Fractions Obtained by Membrane-Based Ultra-Filtration.

Since the potentialities of applications of low molecular weight poly-γ-glutamic acid (γ-PGA) chains have been so far only partially explored, the separation of diverse molecular families of them, as well as their characterization for potential bioactivity and ability to form films, were investigated. Two different approaches based on organic solvent precipitation or on ultra- and nano-filtration membrane-based purification of inexpensive commercial material were employed to obtain size-specific γ-PGA fractions, further characterized by size exclusion chromatography equipped with a triple detector array and by ultra-high-performance liquid chromatography to assess their average molecular weight and their concentration. The γ-PGA low molecular weight fractions, purified by ultra-filtration, have been shown both to counteract the desiccation and the oxidative stress of keratinocyte monolayers. In addition, they were exploited to prepare novel hydrocolloid films by both solvent casting and thermal compression, in the presence of different concentrations of glycerol used as plasticizer. These biomaterials were characterized for their hydrophilicity, thermal and mechanical properties. The hot compression led to the attainment of less resistant but more extensible films. However, in all cases, an increase in elongation at break as a function of the glycerol content was observed. Besides, the thermal analyses of hot compressed materials demonstrated that thermal stability was increased with higher γ-PGA distribution po-lymer fractions. The obtained biomaterials might be potentially useful for applications in cosmetics and as vehicle of active molecules in the pharmaceutical field.

Transglutaminase crosslinked pectin- and chitosan-based edible films: a review.

The production of biodegradable and edible films with desired mechanical characteristics and gas barrier properties represents one of the most advanced challenges in the field of food wrapping and coating. New edible films can serve not only to provide food with physical protection but also to reduce loss of their moisture, to restrict absorption of oxygen, to lessen migration of lipids, to improve their mechanical handling features, and as materials, to apply in direct contact with internal food to realize a multilayer food packaging. Polymers derived from natural products, like carbohydrates and proteins, offer the greatest opportunities as component of edible films since their biodegradability and environmental compatibility are assured and they can also supplement the nutritional value of specific foods. However, excessive water solubility and poor water vapor barrier properties, and often poor mechanical resistance, have their application limited until the present time. Numerous studies have been carried out to improve their properties by preparing composite and multi-component films or by physically and chemically crosslinking their natural components. In the present review we summarize the main results obtained by crosslinking with the enzyme transglutaminase different proteins contained in multi-component pectin- and chitosan-based edible films, having the aim to create environmentally-friendly "bioplastics" with mechanical and permeability properties similar to the ones exhibited by plastics of petrochemical origin.

Putrescine-polysaccharide conjugates as transglutaminase substrates and their possible use in producing crosslinked films.

Putrescine (1,4-diaminobutane) was covalently linked to alginate and low-methoxyl pectin to synthesize new aminated polysaccharides. Both putrescine-pectin and -alginate conjugates, although the latter at higher concentrations, were found to be able to act as effective acyl acceptor transglutaminase substrates in vitro using both dimethylated casein and soy flour proteins as acyl donors. Monodansylcadaverine, a well known acyl acceptor transglutaminase substrate, dose-dependently counteracted the covalent binding of the aminated polysaccharides to the proteins. Putrescine-pectin conjugate was also tested to prepare, in combination with soy flour proteins, edible films in the presence of purified microbial transglutaminase. Characterization of the enzymatically crosslinked films showed a significant decreased water vapor permeability, with respect to the ones obtained with non-aminated pectin in the presence of transglutaminase, as well as improved mechanical properties, such as high extensibility. Possible biotechnological applications of hydrocolloid films containing putrescine-polysaccharide derivatives enzymatically crosslinked to proteins were suggested.