Antimicrobial activity of whey protein based edible films incorporated with oregano,rosemary and garlic essential oils

The use of edible films to release antimicrobial constituents in food packaging is a form of active packaging. Antimicrobial properties of spice extracts are well known, however their application to edible films is limited. In this study, antimicrobial properties of whey protein isolate (WPI) films containing 1.0–4.0% (wt/vol) ratios of oregano, rosemary and garlic essential oils were tested against Escherichia coli O157:H7 (ATCC 35218), Staphylococcus aureus (ATCC 43300), Salmonella enteritidis (ATCC 13076), Listeria monocytogenes (NCTC 2167) and Lactobacillus plantarum (DSM 20174). Ten millilitres of molten hard agar was inoculated by 200 μl of bacterial cultures (colony count of 1 × 10⁸ CFU/ml) grown overnight in appropriate medium. Circular discs of WPI films containing spice extracts, prepared by casting method, were placed on a bacterial lawn. Zones of inhibition were measured after an incubation period. The film containing oregano essential oil was the most effective against these bacteria at 2% level than those containing garlic and rosemary extracts (P < 0.05). The use of rosemary essential oil incorporated into WPI films did not exhibit any antimicrobial activity whereas inhibitory effect of WPI film containing garlic essential oil was observed only at 3% and 4% level (P < 0.05). The results of this study suggested that the antimicrobial activity of some spice extracts were expressed in a WPI based edible film.     

Spray-dried conjugated linoleic acid encapsulated with Maillard reaction products of whey proteins and maltodextrin

Encapsulation performance of Maillard reaction products (MRPs) produced by whey proteins and maltodextrin were examined for microencapsulation of conjugated linoleic acid (CLA) using spray-drying. CLA was encapsulated using 3 different wall materials containing a single constituent such as whey protein concentrate (WPC), whey protein isolate (WPI), and maltodextrin (MD), and 4 different MRPs. For the development of MRPs, ratios of WPC to MD were 1:2 and 1:3, whereas those of WPI to MD were 1:5 and 1:10. CLA and wall material were mixed and homogenized, and then spraydried. The physical properties of encapsulated CLA powders were characterized by particle size and morphology, ζ-potential, flowability, solubility, and dispersibility. The CLA powders coated with WPI-based MRPs have better encapsulation efficiency, water solubility, and smaller particle size than those coated with WPI, WPC, or MD alone. These encapsulated CLA powders have a number of possible utilities as ingredients in a variety of foods.     

Fish oil encapsulation with chitosan using ultrasonic atomizer

An encapsulation technique was developed using an ultrasonic atomizer and three processing steps: emulsification, ultrasonic atomization, and freeze drying. Emulsion preparation variables such as concentration of wall materials [chitosan (CS), maltodextrin (MD) and whey protein isolate (WPI)] and tuna oil were optimized. The size and stability of the emulsion droplet and the properties of the encapsulated powders after freeze drying were characterized. At 20 g/100 g tuna oil, the optimum ratios of CS to MD and of CS to WPI were 1:10 and 1:1, respectively. There was a significant difference (P<0.05) in the emulsion particle sizes when the preparation conditions were varied. The combination of CS and MD giving the smallest particle size had the highest emulsion stability. The EPA and DHA content (240 mg/g) of the encapsulated powder were slightly higher than commercial specification (100 mg/g) and they had low moisture content and water activity, acceptable appearance and encapsulation efficiency. The ultrasonic technology used in this study could lead to application in the food industry improving the stability of tuna and other oils.     

Sensitivity of Listeria monocytogenes to rosemary (Rosmarinus officinalis L.)

There are limited reports on antilisterial activities of spices and herbs in culture media and fewer still in foods. The present study was undertaken to identify spices that possess antilisterial properties and to evaluate effects in media and food. Of 18 spices screened on BHI agar (48 h, 35°C), only rosemary (≥0·5%, w/v) and cloves (≥1%, w/v) were listericidal. Tests were expanded with rosemary to include also its essential oil and four of the major oil constituents, oleoresin, encapsulated oil, and antioxidant extract of rosemary. Effects on Listeria monocytogenes strain Scott A in BHI broth were examined after 24 and 48 h at 35°C. The inhibitory concentrations of rosemary oil were 10 wl/100 ml broth. Of the major constituents of rosemary oil (cineole, borneole, α-pinene and camphor), only α-pinene (0·1 wl/100 ml) delayed listerial growth. The oleoresin, the encapsulated oil, and the antioxidant extract were also antilisterial (100 mg, ≥ 1 g and 0·02 g per 100 ml, respectively). The antilisterial activity of the aqueous extract of rosemary was considerably lower than that of the ground herb or the ethanolic extract. Addition of 0·5% finely ground rosemary or 1% rosemary oil to ready-to-eat pork liver sausage prior to cooking delayed listerial growth during refrigerated storage. The encapsulated oil (5%) and the antioxidant extract (0·3-0·5%) inhibited growth of listeriae, whereas the oil (1%) was ineffective.     

Stability of flocculated particles in concentrated and high hydrophilic solid layer-by-layer (LBL) emulsions formed using whey proteins and gum Arabic

The objective of the present study was to investigate flocculation in layer-by-layer (LBL) emulsion systems with high total solids content and deflocculation at various pH conditions, and the effects of whey protein isolate (WPI) concentration and total solids content on the stability of LBL emulsions. WPI (1.96% (1WPI) or 10.71% (10WPI), w/w in water) was prepared in water and high-pressure homogenized with sunflower oil (10%, w/w, of total emulsion). Gum Arabic (0.15%, w/w, in total emulsion) was added to assemble electrostatically on WPI at oil particle interfaces at pH 3.5 using aqueous citric acid (10% w/w) forming LBL emulsion. The ζ-potential measurements showed charge reversal upon addition of gum Arabic solution into single layer (SL) emulsion confirming the formation of LBL interface. Trehalose:maltodextrin mixture (1:1, w/w, total emulsion, 28.57% (28) or 57.14% (57), w/w, in water) was used in the continuous phase. The high total solids content of the system results in depletion flocculation of the particles leading to bridging flocculation without coalescence as deflocculation into individual particles occurred with increasing pH from pH 3.5 to pH 6.5 in 10WPI systems. Deflocculation was evident in 10WPI-28 and 10WPI-57 as found from a decreased ζ-average diameter and visually under microscope. Coalescence was observed in 1WPI systems. Viscosity of the systems was significantly (P < 0.05) increased with higher total solids content. Accelerated destabilization test showed that systems at higher WPI and total solids contents exhibited the highest stability against creaming. Deflocculation in LBL systems can be controlled by pH while high solids in the aqueous phase provide stability against creaming.     

Rheology and microstructure of cross-linked waxy maize starch/whey protein suspensions

The objective of the present work was to investigate the effect of the heating process on the structural and rheological properties of whey protein isolate/cross-linked waxy maize starch (WPI/CWMS) blends depending upon the concentration and the starch/whey protein ratio. Starch concentration ranged from 3 to 4% (w/w) and the protein content was of 0.5, 1 and 1.5% (w/w). The blend (pH 7, 100 mM ionic strength) was heated using a jacketed vessel at two pasting temperatures: 90 and 110 °C. The particle size distribution of the WPI suspension (1.5%) displayed three distinct classes of aggregates (0.3, 65 and 220 μm), whereas the size of swollen starch granules varied from 48 to 56 μm according to the pasting temperature. When the two components were mixed together, the peak attributed to swollen starch granules was attenuated and broadened towards higher values (up to 88 μm) due to protein aggregates (260–410 μm). This effect was more pronounced as the protein concentration increased. When compared to starch alone, the rheology of the mixed system was dramatically modified for the flow behaviour as well as for the viscoelastic properties which changed from a solid-like (3–4% starch) to a liquid-like behaviour (3–4% starch/1.5% protein). Microscopic observations showed aggregated proteins located in the continuous phase and swollen starch granules as the dispersed phase. Protein aggregates were of different sizes, part of them appeared adsorbed onto swollen starch granules while another part was unevenly distributed in the continuous phase, yielding discontinuous network which could explain the peculiar viscoelastic behaviour of such suspensions.     

Proposing Novel Encapsulating Matrices for Spray-Dried Ginger Essential Oil from the Whey Protein Isolate-Inulin/Maltodextrin Blends

The aim of this study was to evaluate the effects of the blending of whey protein isolate (WPI) with maltodextrin (MD) and inulin (IN) biopolymers as encapsulating matrices for spray-dried ginger essential oil. Encapsulation was performed by ultrasound-assisted emulsification and using spray drying, and the stability parameters of the emulsion (with or without ultrasound-assisted) were evaluated. The influence of these different wall material systems was investigated based on various functional properties of microparticles such as stability of the emulsion, encapsulation efficiency, reconstitution properties, chemical profile, microparticle stability, morphology, particle size distribution, and crystallinity. Higher viscosity values were obtained for the emulsions prepared with WPI and IN which had the apparent viscosity increased by the ultrasound-assisted emulsification process. Creaming index values indicated that ultrasound-assisted emulsions had higher stability. The composition of the wall materials did not affect the solubility and the moisture content of the particles. The wettability property of the powders was improved by the addition of IN. The lowest level of water adsorption under conditions of high relative humidity was also observed in microparticles containing IN. The partial replacement of WPI by MD significantly affected the efficiency of encapsulation. Moreover, MD led to high thermal microparticle stability. Larger particles were observed in the powders prepared with WPI. The powders obtained from WPI, WPI:IN, and WPI:MD treatments exhibited amorphous structures and did not have any cracks on the surface. The findings of this study indicate that IN and MD together with WPI proved to be good alternative secondary wall materials for spray-dried ginger oil.     

Lipid and water crystallization in protein-stabilised oil-in-water emulsions

Phase and state transitions occurring during freezing and thawing of oil-in-water emulsions with different water phase formulations, interfacial compositions and two lipid types were studied as crucial factors affecting emulsion stability. Emulsions containing 0–40% (w/w) sucrose in the water phase at pH 7, and 10, 20, 30, 40% (w/w) dispersed lipid phase (sunflower oil, SO or hydrogenated palm kernel oil, HPKO) with whey protein isolate, WPI, or sodium caseinate, NaCAS, (protein:lipid = 1:10 and 2:10) as emulsifier were prepared. Phase/state behaviour of the continuous and dispersed phases was determined by differential scanning calorimetry (DSC). Emulsion stability and morphology were derived from DSC data, gravitational separation and particle size analysis during 4 freeze-thaw cycles. Systems were stable when only lipid crystallization occurred. DSC data showed that lipid crystallization prior to water crystallization (i.e. emulsions containing HPKO) caused destabilisation at low sucrose concentrations (0, 2.5 and 5% w/w). Emulsions were stable if the dispersed oil phase crystallized after the dispersing water phase (i.e. emulsions containing SO). A concentration of sucrose ≥10% (w/w) in the aqueous phase gave stable emulsions. At 10:1 lipid to protein ratio, WPI showed better stabilising properties than NaCAS at 2.5 and 5% (w/w) sucrose. Double concentration of WPI (lipid:protein = 10:2) at 0% (w/w) sucrose significantly improved systems stability, whereas no positive effect was observed when the concentration of NaCAS was increased. From morphology study, in addition to lipid destabilisation, thickening and flocculation caused instability of the systems. These were extensive in systems containing WPI and were ascribed to interactions between whey proteins during thermal cycling.     

Tuna oil and Mentha piperita oil emulsions and microcapsules stabilised by whey protein isolate and inulin: characterisation and stability

Whey protein isolate (WPI) and its polysaccharide complexes have been widely used to prepare oil‐in‐water emulsions. The aim of this study was to evaluate the emulsions and spray‐dried microcapsules containing tuna oil and/or mint oil and stabilised by combination of WPI with inulin in terms of physicochemical characteristics and storage stability. Stable emulsions were formed before drying. Tuna oil + Mentha piperita oil emulsions had smaller viscosity, surface tension and size than did tuna oil emulsions. Surface morphology showed that spray‐dried microcapsules were spheres but had many dents and apparent shrinkage. During storage, tuna oil and tuna oil + M. piperita oil microcapsules became larger. In the blend oil microcapsules, menthone was reduced to form menthol, loss of DHA and EPA was slightly less, the degree of oxidation characterised using peroxide value and headspace propanal was less but basically greater than half of that of WTI microcapsules.     

Rheological properties of reduced-fat and low-fat ice cream containing whey protein isolate and inulin

Instrumental analyses were used to evaluate the rheological properties of regular (10%), reduced-fat (6%) and low-fat (3%) ice cream mixes and frozen ice creams stored at −18 °C. The reduced-fat and low-fat ice creams were prepared using 4% whey protein isolate (WPI) or 4% inulin as the fat replacement ingredient. The composition, colour, apparent viscosity, consistency coefficient, flow behaviour index, hardness and melting characteristics were measured. No effect of WPI or inulin was obtained on the colour values. Compared with regular ice cream, WPI changed rheological properties, resulting in significantly higher apparent viscosities, consistency indices and greater deviations from Newtonian flow. In addition, both hardness and melting resistance significantly increased by using WPI in reduced-fat and low-fat ice creams. Inulin also increased the hardness in comparison to regular ice cream, but the products made with inulin melted significantly faster than the other samples.     

Influence of chitosan/clay functional bionanocomposite activated with rosemary essential oil on the shelf life of fresh silver carp

A combination of chitosan biopolymer, nanoclay and rosemary essential oil was prepared as a functional bionanocomposite (FBN). Its ability to improve the shelf life of refrigerated (4 ± 1 °C) silver carp fillets was studied. The fresh fillets were left untreated as a control or coated with chitosan, chitosan/clay bionanocomposite and chitosan/clay/rosemary essential oil (Ch/clay/REO) FBN. Then, they were evaluated for chemical, microbial and sensory properties over 16‐day storage. The samples coated with the FBN had the lowest pH and total volatile basic nitrogen. Ch/clay/REO coating efficiently retarded lipid oxidation by decreasing peroxide, free fatty acid and thiobarbituric acid production in the samples. The coating also reduced total viable and psychrotrophic count of the fillets more than 1.5 log by the end of storage.     

Whey protein nanoparticles prepared with desolvation with ethanol: Characterization,thermal stability and interfacial behavior

Whey protein isolate (WPI) nanoparticles were prepared by diluting an alkaline solution of protein in ethanol at concentrations varying between 50 and 80%. The nanoparticles were then immediately diluted in buffer. While the nanoparticles were not stable at pH 7, they showed no changes in size when diluted at pH 3. When 75–80% ethanol was added during preparation, the size of the WPI nanoparticles ranged between 10 and 100 nm, with no change in size after dilution and storage at pH 3 for 96 h at 22 °C. When heating was applied, particle aggregation occurred, and large aggregates (>1 μm) were observed at temperatures > 60 °C. The particle size of the heat-induced aggregates could be reduced by homogenization. The nanoparticles prepared by desolvation showed interfacial pressure values similar to those of the corresponding protein solutions, indicating similar interfacial properties and the potential to be used to stabilize emulsions but as supramolecular aggregates of WPI.     

Effect of chlorophyll removal and particle size upon the nutritional and technological properties of powdered by‐products from artichoke (Cynara scolymus,L.) industrial canning

In this study, the effect of the particle‐size fractionation (Ø < 0.212 mm and 0.212 mm < Ø < 0.991 mm) and chlorophyll extraction on the nutritional and technological properties of the powdered artichoke ingredient was evaluated. The contents of minerals, protein, fat, carbohydrates and dietary fibre together with the content in bioactive compounds such as inulin and phenolics were determined. Other properties such as water‐ and oil‐holding capacities, water activity and antioxidant capacity were measured. The ingredient with chlorophyll and the lowest particle size presented the highest phenolic content and antioxidant capacity (8.4 mg of vitamin C equivalents per 100 g of dry matter) and water‐ and oil‐holding capacities. The removal of chlorophyll increased the oil‐holding capacity (from 59.7% to 94.6%), which was much higher than in the coarse ingredient (34%), but has a deleterious effect reducing the antioxidant capacity and the inulin content. The ingredients also showed high dietary fibre (22.5–33%), inulin (9–16%) and mineral (7.5–7.8%) contents. Although the ingredient with chlorophyll and smaller particle size had the higher antioxidant capacity, the removal of chlorophyll improved the technological properties to be used as food ingredient without affecting significantly to the nutritional value.     

Physical characteristics of submicron emulsions upon partial displacement of whey protein by a small molecular weight surfactant and pectin addition

O/W emulsions (6 wt.% olive oil) were prepared at pH 3.3 using different WPI:Tween 20 weight ratios (1:0, 3:1, 1:1, 1:3, 0:1) at 1 wt.% total concentration. The emulsion droplet size was found to decrease with an increase in Tween 20. A minimum droplet size of d_3,2 300 nm was found for Tween systems alone, similar to that found (360 nm) for a 1:1 WPI:Tween 20 combination (p < 0.05). This specific composition showed a value for the interfacial tension close to that of Tween 20 alone. However, the emulsions presented low stability regardless of the WPI:Tween 20 ratio. To increase their stability, pectin was added, in various concentrations (0.2, 0.4 and 0.6 wt.%), using the Layer by Layer technique. In the presence of pectin, the ζ-potential of the oil droplets became negative; indicating that negatively charged pectin was absorbed onto the positively-charged droplet surface forming a secondary layer. The additional layer resulted in a wide range of emulsion stability. For all pectin concentrations, the 1:1 ratio of WPI:Tween 20 showed the highest stability. In most emulsions, extensive aggregation of oil droplets was observed, and their viscosity increased. Insufficient amounts of pectin to form the secondary layers led to bridging flocculation phenomena of oppositely charged pectin and proteins, leading to aggregation of the oil droplets. The higher the concentration of pectin, the greater the stability of the emulsion due to higher viscosity. All in all, the addition of a second layer consisting of pectin can be used to increase the stability of an emulsion containing emulsion droplets in the sub-micron range.     

Replacing modified starch by inulin as prebiotic encapsulant matrix of lipophilic bioactive compounds

The purpose of this work was to replace modified starch (SF) by inulin (IN), a prebiotic carbohydrate, during emulsification assisted by ultrasound. Oregano extract was encapsulated using five proportions of IN and SF as wall materials. The effect of such substitution on the microparticle characteristics was evaluated. Attempting to contribute with the increasing demand for prebiotic consumption, mixing one part of SF with three parts of IN (1:3, mass basis) yielded encapsulation efficiency equal to 66 ± 1% and the largest thymol retention: 84 ± 9%. Besides the entrapment of thymol, high amount of other compounds present in oregano extract could be entrapped in the polymeric matrix: 92 ± 1%. Reduction of the microparticles sizes when increasing the proportion of inulin was also observed. Comprising such results and those presented for powder morphology, surface extract, particle size distribution, X-ray diffraction and thermal stability, the proportion 1:3 (SF:3IN) is a favorable prebiotic encapsulant matrix for encapsulating oregano extract and retaining target bioactive compounds.     

Effect of temperature,calcium and protein concentration on aggregation of whey protein isolate: Formation of gel-like micro-particles

Protein aggregation occurs in biological systems and industrial processes, affecting protein solubility and functional properties. In this study, whey protein isolate (WPI) obtained from bovine milk was used as a model to study the dependence of aggregation on pre-heating temperature and on protein and calcium concentrations. WPI solutions (0.1–5.0%, w/v) were heated at 25–85 °C for 30 min prior to cooling and calcium addition. Tryptophan shifted to a more hydrophilic environment as WPI concentrations and pre-heating temperatures increased. Pre-heated WPI solutions yielded soluble particles, which aggregated to form porous gel-like particles by addition of calcium chloride. WPI microgel particles could be prepared by using a cold gelation method and preheated the protein above 65 °C. The particle size was monodisperse with sizes of about 190 nm and 255 nm, respectively in solutions pre-heated to 75 or 85 °C and containing 5 mm calcium.     

Microencapsulated Lactobacillus rhamnosus GG in whey protein and resistant starch matrices: Probiotic survival in fruit juice

This study compared the survival of spray dried microencapsulated Lactobacillus rhamnosus GG (LGG) added into apple juice or citrate buffer (pH 3.5) and stored at 4 or 25 °C over a 5-week period. The LGG was encapsulated in matrices comprising (i) whey protein isolate (WPI) alone, (ii) WPI in combination with a physically-modified resistant starch (RS) at various ratios (4:1, 1:1 and 1:4), or (iii) RS alone. All microencapsulated LGG formulations containing WPI alone or WPI in combination with RS provided better protection to LGG in apple juice or citrate buffer compared to the formulation containing RS alone. We suggest that the protection afforded by formulations containing WPI alone or in combination with RS is due to the ability of WPI to create a buffered microenvironment within the hydrated colloid particle surrounding the embedded LGG, thus isolating the bacteria from the stresses of the low pH external environment.     

Chitosan-whey protein isolate composite films for encapsulation and stabilization of fish oil containing ultra pure omega-3 fatty acids

Chitosan (1.5%, w/v)-whey protein isolate (WPI, 5% w/v) composite films were developed for encapsulating and stabilizing fish oil (FO) containing 93.7% eicosapentaenoic acid (EPA). Chitosan-WPI film-forming solutions (FFS) were incorporated with 1.5% or 2% FO (w/v), 2% (w/v) glycerol, Tween 80 (3 times weight of FO), and 0.5% (w/v) oregano or rosemary essential oil (EO), and cast for films at room conditions. Dried films were stored at 2 °C for 30 d for evaluating encapsulation efficiency (EE), lipid stability, and film functionality. Total oil contents in films from FFS incorporating 1.5% or 2% FO were 28.1% to 32.5% and 33.4% to 37.3%, respectively, and free oil contents were 13.5% to 14.7% and 15.5% to 16.3%, respectively. EE, moisture content, and water activity of the films were 47.8% to 66%, 18.7% to 24.9%, and 0.42% to 0.50%, respectively, without significant difference among differently formulated films. Increasing FO concentration from 1.5% to 2% in FFS decreased tensile strength of the films from 0.57-0.73 to 0.34-0.44 MPa, but not the film elongation. Addition of oregano EO in FFS retarded lipid oxidation of the fish oil encapsulated in the films, in which a 43% to 53% reduction in thiobarbituric acid-reactive substances value and 39% to 51% reduction in peroxide value were achieved. Chitosan-WPI composite films with incorporation of oregano essential oil could be applied as a simple and economic means for encapsulating and stabilizing fish oil for fortifying omega-3 fatty acids in various applications.     

Inulin potential for encapsulation and controlled delivery of Oregano essential oil

The ability of inulin, a prebiotic material, as encapsulation matrix was explored. Microcapsules of Raftiline were produced by spray drying inulin solutions at different solids content (5, 15 and 25%) at 120, 155 and 190 °C, according to a Central Composite Rotatable design. Produced capsules were analysed for morphology and size by SEM and physiochemical characterized by DSC, IR and RAMAN. Oregano essential oil was incorporated in the inulin solutions at 15% solids basis and the emulsions dried at the same conditions. The above mentioned methodologies were applied to evaluate the encapsulation ability and the changes induced by the presence of the EO in capsules morphology and structure. Furthermore the kinetics and amount of release was assessed by a spectrophotometric method. Results showed that it was possible to produce regular spherical inulin microcapsules (3–4.5 μm) for all the tested experimental conditions. According to IR and Raman results mainly drying temperature affected the structure of the capsules, three groups being clearly formed. These groups could be related to the morphology of inulin crystals. The EO was successfully encapsulated in the system as demonstrated by IR and Raman analysis. The differences found in the EO releasing amount, make clear that different degrees of core material retention is achieved, what should be related to structural changes in the matrix wall, denoting in some processing conditions interactions phenomena among inulin and EO. Those different releasing profiles patterns may be quite useful in finding different potential uses for the encapsulates.     

Co-precipitation of whey and pea protein – indication of interactions

The aim was to optimise the yield of co-precipitation of whey protein isolate (WPI) and pea protein isolate (PPI) and compare co-precipitates and protein blends with respect to solubility. The yield of co-precipitates was tested with different protein ratios of WPI and PPI in combination with different temperatures and acid precipitation (pH 4.6). The highest precipitation yield was obtained at protein ratios WPI < PPI, high temperature and alkaline protein solvation. The solubility was measured by an instability index and absorption spectroscopy of re-suspended precipitated proteins at pH 3, 7 and 11.5. Co-precipitates had significantly lower solubility than protein blends. Protein ratios WPI > PPI, low precipitation temperature and high pH showed the highest solubility. Differences in protein composition between co-precipitates and protein blends were observed with SDS-PAGE and matrix-assisted laser desorption ionisation time-of-flight, and indicated different protein–protein interaction in samples, which needs further investigations.