Esterification of dextran by octenyl succinic anhydride (OSA): Physicochemical characterization and functional properties assessment.

The chemical modification of biopolymers to enhance their functional properties in the food, cosmetic, and pharmaceutical industries is an area of particular interest today. In this study, different molecular weight dextrans were chemically modified for the first time with octenyl succinic anhydride (OSA). This reaction involves an esterification process wherein the hydroxy groups of dextran are partially substituted by a carbonaceous chain, imparting hydrophobic properties to dextran molecules and, consequently, an amphiphilic nature. To assess and quantify the incorporation of OSA into the dextran structure, reaction products were analysed using NMR and FTIR. Additionally, the thermal properties, the Z-potential and the foaming and emulsifying capacity of both native and modified dextrans were examined. The introduction of OSA groups to dextran molecules, with degrees of substitution between 0.028 and 0.058, increased the zeta potential and the thermal stability of the polymer. Furthermore, the chemical modification of dextran backbone with this radical conferred a hydrophobic nature to the biopolymer, which enhance its foaming and emulsifying capacity. Therefore, these results demonstrate that the incorporation of hydrophobic moieties into dextran polymers improves their functional properties and broadens their potential applications in the industry.

Antioxidant Activity of Egg Yolk Protein Hydrolysates Obtained by Enzymatic and Sub-Critical Water Hydrolysis.

Obtaining peptides with antioxidant properties by enzymatic hydrolysis has been widely described; however, the use of non-enzymatic methods to obtain peptides with antioxidant capacities has been poorly investigated. In this study, non-soluble proteins obtained from delipidated egg yolk granules were hydrolyzed with trypsin, and with a non-enzymatic method using sub-critical water hydrolysis under a non-oxidizing (nitrogen) and oxidizing (oxygen) atmosphere. The effect of the sub-critical water hydrolysis on the amino acids' composition of the hydrolysates was assessed. Furthermore, the antioxidant capacities of the hydrolysates were evaluated using the ABTS•+ scavenging assay, the DPPH radical scavenging activity assay, and by measuring the reducing power of the peptides, the peptides' ferrous ion chelating capacities, and the antioxidant effect of the peptides on beef homogenates. The hydrolysate obtained by sub-critical water hydrolysis under a nitrogen stream showed similar or better results in the antioxidant tests than those obtained using trypsin hydrolysis, except in the ferrous chelating capacity, where the trypsin hydrolysate showed the best performance. The oxidizing environment promoted by the oxygen in the other sub-critical water hydrolysis method tested produced the peptides with the lowest antioxidant capacities, due to changes in the primary structure of the peptides. These results suggest that the sub-critical water hydrolysis method under a nitrogen stream, in comparison with the enzymatic hydrolysis, is a reliable method to obtain peptides with good antioxidant capacities.

Kinetic-conceptual model of the hydrolysis of bovine plasma proteins - ALCALASE® 2.9L: The role of inhibition by product.

In the present work, the inhibitory effect of the peptide fractions, obtained through enzymatic hydrolysis of bovine plasma was evaluated, on the enzyme used in the reaction (Alcalase 2.4 L). In this sense, Ultra-filtered peptide fractions of different molecular sizes (A: Fraction>10; B: Fraction 10-3 kDa; and C: Fraction <3 kDa), were used to verify the impact on the total hydrolysis rate. The Fractions between 3 and 10 kDa were refined to fit a conceptual kinetic model which considers inhibition by product and substrate. Additionally, the inactivation of the enzyme through the reaction time was evaluated and its effects incorporated into the model. It was shown that some peptides released in the successive stages of the reaction can in turn inhibit the activity of the hydrolyzing enzyme. The model evaluated suggests a time-varying expression of inhibition parameters as a function of the initial substrate concentration in the reaction. This is based on the kinetic changes of the product profiles for each reaction time in the evaluated operating conditions (S0 variable). A greater inhibitory effect due to the products is evidenced when the reaction occurs with a higher load of the initial substrate (S0 = 20 g/L).

Effect of Birch Sap as Solvent and Source of Bioactive Compounds in Casein and Gelatine Films.

Birch sap consists of a natural water-based solution with valuable compounds such as minerals, sugars, organic acids and phenolic compounds that can be used advantageously in the preparation of edible films. In this study, gelatine- and casein-based films were prepared using birch sap as biopolymer solvent and source of bioactive compounds with the aim of developing new bioactive materials for food packaging. The physical, mechanical, barrier, antioxidant and iron-chelating properties of the obtained films were investigated. Birch sap enhanced the mechanical properties of the films by increasing puncture strength and flexibility, as well as their ultraviolet-visible light barrier properties. In addition, the presence of bioactive compounds endowed the birch sap films with an antioxidant capacity of almost 90% and an iron-chelating capacity of 40-50% with respect to the control films. Finally, to test these films as food packaging material, a photosensitive curcumin solution was packed and exposed to ultraviolet light. Tested films were able to protect curcumin against photodegradation, and the presence of bioactive compounds inside the birch-sap-enriched materials offered an additional 10% photoprotective effect compared to control films. Results showed the potential of birch sap as an environmentally friendly biopolymer solvent and plasticizer that can improve the mechanical and photoprotective properties of the prepared materials.

Bio-production of lactic and lactobionic acids using whey from the production of cow's milk Wagashi cheese in Benin.

Traditional cheese is the main milk derivative in Bénin. This traditional process is not efficient and generate a lot of whey which has no real use until now. It is just disposed without being environmentally treated. Its use as a source for lactobionic and lactic acids production by Pseudomonas taetrolens and Lactobacillus casei is studied in this work, being also a proposal that can greatly boost economically the dairy sector in the country and reduce the end-of-cycle impact of the residue. To our knowledge, no data is available in the metabolization of Bénin's traditional cheese whey and its potential transformation into commercially valuable products such as lactobionic and lactic acids. With bulk filtration, non-controlled pH batch fermentations and without nutrients supplementation, 66 and 22% of lactose in the traditional cheese whey have been converted into lactobionic acid and lactic acid using Pseudomonas taetrolens and Lactobacillus casei, respectively. Those are important results that encourage to enhance the bioprocesses used in a cost-effective way in order to scale up an industrial production.

Downstream Approach Routes for the Purification and Recovery of Lactobionic Acid.

The successful development of a lactobionic acid (LBA) bioconversion process on an industrial scale demands the selection of appropriate downstream methodological approaches to achieve product purification once the bioconversion of LBA is completed. These approaches depend on the nature of the substrate available for LBA production, and their necessary implementation could constitute a drawback when compared to the lesser effort required in downstream approaches in the production of LBA obtained by chemical synthesis from refined lactose. Thus, the aim of this research is to separate LBA from an acid whey substrate after bioconversion with Pseudomonas taetrolens. Freeze drying, crystallization, adsorption with activated carbon, microfiltration, centrifugation, and precipitation with 96% (v/v) ethanol were carried out to separate and purify LBA. The closest product to commercial LBA was obtained using precipitation with ethanol, obtaining a white powder with 95 ± 2% LBA concentration. The procedure described in this paper could help to produce LBA on an industrial scale via microbial bioconversion from acid whey, developing a promising biotechnological approach for lactose conversion.

Preparation of Edible Films with Lactobacillus plantarum and Lactobionic Acid Produced by Sweet Whey Fermentation.

Cheese whey, one of the most abundant by-products of the dairy industry, causes economic losses and pollution problems. In this study, deproteinised sweet whey was fermented by Pseudomonas taetrolens LMG 2336 to produce a prebiotic compound (lactobionic acid, LBA). Endotoxins produced by these microorganisms were successfully removed using microfiltration techniques, allowing the fermented whey permeate to be used in the food industry. The fermented whey permeate was used to develop prebiotic edible films by adding two different concentrations of gelatine (0.45 and 0.9 g gelatine g-1 LBA; LBA45 and LBA90). Furthermore, Lactobacillus plantarum CECT 9567 was added as a probiotic microorganism (LP45 and LP90), creating films containing both a prebiotic and a probiotic. The mechanical properties, water solubility, light transmittance, colour, and microstructure of the films were fully characterised. Additionally, the LBA and probiotic concentration in LP45 and LP90 were monitored under storage conditions. The strength and water solubility of the films were affected by the presence of LBA, and though all these films were homogeneous, they were slightly opaque. In LP45 and LP90, the presence of LBA as a prebiotic improved the viability of L. plantarum during cold storage, compared to the control. Therefore, these films could be used in the food industry to coat different foodstuffs to obtain functional products.

The antimicrobial and bioactive properties of lactobionic acid.

Lactobionic acid (LBA) is a bioactive molecule that has generated keen interest in different industries. However, its future application in the food area is one of the most promising. Chemically, it is a polyhydroxy acid formed by the union of two molecules (galactose and gluconic acid) linked by an ether-bond, showing many interesting and unusual properties due to its structure and composition, although it is traditionally known in the food industry for its chelating, moisturizing, gelling, and antioxidant properties. There has been much research into the production of LBA, either by microbial fermentation or biocatalytic approaches such as enzymatic synthesis, but its use in foodstuffs, to produce new functional products and to evaluate its antimicrobial activity against food-borne pathogens, is a relatively new topic that has attracted the interest of the international research community recently. Furthermore, in spite of the potential of LBA, it has been approved only by the US Food and Drug Administration, and for its use as the salt form, but the publication of new comprehensive studies, able to agglutinate all the new food-related LBA research results, could disseminate knowledge about this compound and have an influence on its current regulation status. The aim of the present review is to describe the most recent advances and research on its antimicrobial potential, as well as summarizing the significant aspects that make LBA a promising bioactive compound for the food sector. © 2022 Society of Chemical Industry.

Peptides recovery from egg yolk lipovitellins by ultrafiltration and their in silico bioactivity analysis.

The lipoproteins that remain after the extraction of phosvitin from the egg yolk granular fraction possess low industrial applicability. In this study, these lipoproteins were hydrolysed using trypsin, and the bioactivity of the resulting peptides was assessed by in silico analysis. In addition, in order to isolate the most valuable previously detected peptides, their transmission through a polyethersulfone (PES) membrane and a stabilised cellulose (SC) based membrane was also evaluated at several pHs. A pH of 4.0 gave the highest observed transmission of peptides through both membranes for every peptide identified in the permeate streams. Regarding the PES membrane, six peptide sequences detected in the permeate were predicted to be antihypertensive, although only one of them showed a bioactivity score higher than 0.5 according to Peptide Ranker. When the SC membrane was assessed, five peptides with a bioactivity score higher than 0.5 were detected in the permeate streams and eight peptides were predicted as antihypertensive. The in silico analysis performed showed that K.VQWGIIPSWIK.K was the most promising antihypertensive peptide found in the permeates.

Novel Bovine Plasma Protein Film Reinforced with Nanofibrillated Cellulose Fiber as Edible Food Packaging Material.

Proteins, such as those in blood from slaughterhouses, are a good option for developing edible films. However, films made exclusively from proteins have low strength and high water solubility, which makes them difficult to use in the food industry. The use of cellulosic material, such as nanofibrillated cellulose (NFC), can improve the properties of these films. In the present work, bovine plasma was acidified and treated with ethanol to precipitate its proteins, and these proteins were used to prepare films reinforced with several concentrations of NFC. In addition, control films prepared with untreated bovine plasma and reinforced with NFC were prepared as well. These new edible films were characterized according to their mechanical properties, water vapor permeability, light transmittance, and microstructure. Furthermore, the film with the best properties was selected to be additivated with nisin to test its antimicrobial properties by wrapping meat previously contaminated with Staphylococcus aureus. In this sense, films prepared with the extracted proteins showed better properties than the films prepared with untreated plasma. In addition, the results showed that the reinforcement of the films with a 10% (w/w) of NFC decreased their water solubility and improved their puncture strength and water vapor barrier properties. Finally, the addition of nisin to the films prepared with extracted protein from bovine plasma and NFC gave them antimicrobial properties against S. aureus.

Egg Yolk Oil as a Plasticizer for Polylactic Acid Films.

Polylactic acid (PLA) is known to be one of the most extensively used biodegradable thermoplastic polyesters, with the potential to replace conventional petroleum-based packaging materials; however, the low flexibility of films prepared using PLA has limited the applications of this biopolymer. In this study, in order to improve the mechanical properties of PLA films and to provide them with antioxidant properties, egg yolk oil was used as a biobased plasticizer. For this purpose, PLA films with increasing concentrations of egg yolk oil were prepared and the effects of this oil on the light transmission, transparency, colour, water vapour permeability, solubility, antioxidant activity and mechanical properties of the films were characterized. In addition, electron microscopy of the structure of the transverse section of the films was also performed. Results showed that the formulations with higher concentrations of egg yolk oil increased the films' elasticity, and their light barrier and antioxidant properties. Finally, in order to test the films as a packaging material for food applications, extra virgin olive oil and resveratrol, both photosensitive compounds, were packed and exposed to ambient light. Overall, the results show the potential of egg yolk oil as an environmentally friendly plasticizer that can improve the flexibility of PLA films and provide them with additional photoprotective properties.

Enhanced Cu(II) adsorption using sodium trimetaphosphate-modified cellulose beads: equilibrium, kinetics, adsorption mechanisms, and reusability.

The current study is focused on the simple synthesis of two novel biosorbent beads: BASB/STMP and CNFB/STMP, derived respectively from bleached almond shell (BAS) and cellulose nanofiber from almond shell (CNF) by means of chemical crosslinking with sodium trimetaphosphate (STMP). These biosorbents were thoroughly characterized in terms of structure (FTIR), texture (N2 adsorption-desorption), thermal behavior (TGA/DTG), morphology (SEM), and surface properties (XPS). The adsorption kinetics of Cu(II) ions onto BASB/STMP and CNFB/STMP materials proved the chemisorption interaction between Cu(II) ions and the STMP functionalized beads. The BASB/STMP equilibrium data were successfully described by the Redlich-Peterson model and the CNFB/STMP data by the Sips model which disclosed maximum adsorption capacities of 141.44 mg g-1 and 147.90 mg g-1, respectively. Furthermore, the BASB/STMP bioadsorbent offers easy regeneration and better reusability with high efficiency (> 83%). This study sheds light on the preparation of low-cost adsorbents for wastewater treatment in order to improve the competitiveness and eco-friendliness of agrowaste-based processes.

Synthesis and characterization of eco-friendly cellulose beads for copper (II) removal from aqueous solutions.

In this study, novel cellulose-bead-based biosorbents (CBBAS) were successfully synthesized from almond shell using a simple three-step process: (i) dissolution of bleached almond shell in ionic liquid (1-butyl-3-methylimidazolium chloride), (ii) coagulation of cellulose-ionic liquid solution in water and (iii) freeze-drying. Their morphological, structural and physicochemical properties were thoroughly characterized. These biomaterials exhibited a 3D-macroporous structure with interconnected pores, which provided a high number of adsorption sites. It should be noted that CBBAS biosorbents were efficiently employed for the removal of copper (II) ions from aqueous solutions, showing high adsorption capacity: 128.24 mg g-1. The biosorption equilibrium data obtained were successfully fitted to the Sips model and the kinetics were suitably described by the pseudo-second-order model. Besides, CBBAS biosorbents can be easily separated from the solution for their subsequent reuse, and thus, they represent a method for the removal of copper (II) from aqueous solutions that is not only eco-friendly but also economical.

Exploring encapsulation strategies as a protective mechanism to avoid amensalism in mixed populations of Pseudomonas taetrolens and Lactobacillus casei.

Pseudomonas taetrolens constitutes an efficient platform for the biosynthesis of lactobionic acid, a potentially prebiotic compound. Unfortunately, an amensalistic interaction has been demonstrated between P. taetrolens and probiotic lactic acid bacteria (LAB), characterized by the competitive exclusion of P. taetrolens, hindering the in situ production of fermented dairy products with synbiotic properties. In the present research, encapsulation was explored as a barrier to the diffusion of the antimicrobial metabolites generated by LAB. Mixed fermentations involving P. taetrolens LMG 2336 and Lactobacillus casei CECT 475 were cultivated, entrapping both microorganisms alternately. Alginate, alginate/starch and carboxymethyl cellulose/k-carrageenan were tested as encapsulating agents. The immobilization of L. casei in 2% alginate/2% starch beads was found to be the best strategy, improving the production of lactobionic acid by 182% with respect to co-cultures with free cells. This study proves the potential of LAB encapsulation for the protection of sensitive strains in mixed food fermentations.

Liquid-phase food fermentations with microbial consortia involving lactic acid bacteria: A review.

Microbial associations are frequent in traditional fermented foods and beverages, conferring upon them their characteristic organoleptic, physical and nutritional properties. Moreover, the search for novel products that satisfy the needs of consumers, especially foods with health-giving properties, means that new combinations of microorganisms are tested on a wide variety of substrates. In these microbial consortia, lactic acid bacteria (LAB) are frequently the major bacterial starters, usually combined with other species such as propionic and acetic acid bacteria and yeasts. The ability of LAB to produce a wide variety of metabolites is a determining factor in the establishment of associations with the interacting microbiota, which can be positive, negative or neutral. In liquid-phase fermentations, the control of these interactions represents a great challenge, due to the rapid rate of bioconversion and the direct liberation of the metabolites into the medium. Therefore, the understanding of the co-culture dynamics is of vital importance. The present review compiles data referring to the microbial consortia involving LAB, traditionally used to obtain artisanal products, as well as the new associations proposed and tested for the development of novel fermented liquid foods and beverages. The different types of interactions that have been found in these microbial consortia are discussed, including some mathematical models that have been proposed to simulate and control the development of fermentations. Some of the strategies, techniques and devices that are being developed and implemented to improve the efficiency of co-cultures are finally presented.

Human plasma gels: Their preparation and rheological characterization for cell culture applications in tissue engineering.

Tissue engineering is one of the fields of clinical medicine that has forged ahead in recent years, especially because of its role as a potential alternative to organ transplantation. The main aim of this study has been the development of biocompatible materials to form extracellular matrix (ECM) structures in order to provide the necessary conditions for the settlement, proliferation and differentiation of dermal cells such as fibroblasts. To this end, human plasma gels were synthesized with the addition of increasing concentrations of transglutaminase (TGase), which catalyses the formation of covalent bonds between Lys and Glu residues. These materials were structurally characterized using rheology and texturometry and were found to have good structural resistance and elasticity for fibroblast culture. A remarkable improvement in the mechanical properties of the human plasma gels was detected when the two highest TGase concentrations were tested, which may be interpreted as an increase in the number of covalent and non-covalent bonds formed between the plasma protein chains. Furthermore, a human fibroblast primary culture was seeded on human plasma scaffolds and satisfactorily proliferated at 37 °C. This was verified in the images obtained by optical microscopy (OM) and by scanning electron microscopy (SEM), which confirmed that the structure of this type of material is suitable for the growth and proliferation of dermal fibroblasts.

The effect of ultrasound on the alkali extraction of proteins from eggshell membranes.

BACKGROUND: Eggshell contains two layers formed by a dense network of fibrous proteins. These proteins are highly insoluble in a broad variety of solvents, but their composition makes them suitable for a broad range of applications. In this study, in order to extract and solubilise these proteins, the eggshell membranes were treated in an alkali solution. A Box-Behnken design was employed to determine the influence of the treatment variables on the amount of protein solubilised. Furthermore, the effect of ultrasound on the protein recovery yield was also evaluated and compared with the unmodified process. RESULTS: A solubilised protein yield close to 100% of the total eggshell membrane protein was obtained. The optimal conditions could be set at 70 °C in a 1.0 mol L-1 NaOH solution for 60 min. However, when ultrasound was applied, it was possible to decrease the time of reaction by half. In the two processes, the temperature was found to be the most important independent variable evaluated. Finally, the antioxidant properties of the proteins obtained in each case were similar. CONCLUSIONS: Ultrasound favours the detachment of big clumps of proteins from the eggshell membrane, facilitating the solubilisation of its compounds. The ultrasound had no effect on the protein properties tested in this study. © 2017 Society of Chemical Industry.

Edible films from residual delipidated egg yolk proteins.

Commercial extraction with organic solvents of valuable lipids from egg yolk produces a highly denatured protein waste that should be valorized. In this work, the delipidated protein waste remaining after ethanol extraction was used to prepare edible films. This material was also treated with transglutaminase, obtaining films that have also been characterized. When compared with gelatin and caseinate edible films, the films made with egg yolk delipidated protein showed poorer mechanical properties, but improved light barrier properties, low water solubility and a high degree of transparency. It is particularly interesting that the presence of phosvitin in the egg yolk gives the films important ferrous chelating properties. When the egg yolk delipidated protein was treated with transglutaminase, the strength of the film was improved in comparison with films made with untreated protein. Finally, addition of thymol and natamycin in the preparation of these films is shown to be an interesting alternative, providing them with antibacterial and antifungal capacities.

Synbiotic Fermentation for the Co-Production of Lactic and Lactobionic Acids from Residual Dairy Whey.

Besides its properties as an antioxidant, stabilizer, or acidifier, lactobionic acid has emerged as a potential prebiotic compound, raising the possibility of being included together with the probiotic microorganism Lactobacillus casei in novel functional fermented foods with synbiotic characteristics. Their manufacturing strategy could benefit from the recently implemented microbial synthesis of lactobionic acid by the strong producer Pseudomonas taetrolens, employing residual dairy whey as raw material. The phenomenon of amensalism established between Pseudomonas and Lactobacillus makes simultaneous fermentation unfeasible. A novel sequential process has been developed in which L. casei is inoculated in a second step. Its ability to utilize lactobionic acid as a carbon and energy source was previously tested. Experimental results showed the capacity of L. casei to work efficiently on the residual substrate fermented by P. taetrolens, producing lactic acid by degrading the remaining lactose, with a lactic acid yield on substrate and productivity of 0.95 g g-1 and 0.20 g L-1 h-1 , respectively. Lactobionic acid was barely consumed in this complex growth medium, thus ensuring its presence in the resulting fermented product. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1250-1256, 2017.

Microbial amensalism in Lactobacillus casei and Pseudomonas taetrolens mixed culture.

Pseudomonas taetrolens has recently been revealed as an effective microbial producer of lactobionic acid from carbohydrates contained in dairy byproducts. In terms of food industrial applications, the implementation of lactobionic acid biosynthesis coupled with the classic bacterial production of lactic acid appears an important goal. This research paper studies the simultaneous fermentation of residual cheese whey by P. taetrolens and Lactobacillus casei to co-produce lactic and lactobionic acids. Experimental data showed the importance of the interactions established between the two microorganisms. Changes in physiology, viability, growth, and productive capacity were tested experimentally. Lactobacillus was not seen to suffer any appreciable stress, but considerable variations were observed in the Pseudomonas behavior presumably owing to inhibitory lactic metabolites, interaction that can be classified as microbial amensalism. As to production, lactic acid remained without significant changes in mixed fermentations, whereas the production of lactobionic acid decreased sharply due to the competitive exclusion of Pseudomonas.