Poly(ethylene oxide) (PEO) is known for facilitating the electrospinning of biopolymer solutions, that are otherwise not electrospinnable. The objective of this study was to investigate the mechanism by which PEO enables the formation of whey protein isolate (WPI) electrospun fibers under different pH conditions. This investigation revealed that the addition of PEO increased the viscosity of WPI/PEO (10% w/w WPI; 0.4% w/w PEO) solutions. Difference in pH levels of the polymer solutions affected electrospinnability and fiber morphology. Acidic solutions resulted in smooth fibers (700 ± 105 nm) while neutral solutions produced spheres (2.0 ± 1.0 um) linked with ultrafine fibers (138 ± 32 nm). In comparison, alkaline solutions produced fibers (191 ± 38 nm) that were embedded with spindle-like beads (1.0 ± 0.5 um). Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analyses revealed that the native globular configuration of WPI was not altered under neutral conditions. By contrast, the electrophoresis and spectrometry data indicated that WPI was denatured and hydrolyzed under acidic conditions, which facilitated the formation of smooth fibers. C13 nuclear magnetic resonance (NMR) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopies showed that the increase random coil and a-helix secondary structures in WPI contributed to the formation of bead-less electrospun fibers. Also, C13 NMR analysis showed no evidence of chemical interaction between WPI and PEO. Scanning transmission electron microscopy coupled with energy dispersive X-rays (STEM-EDAX) revealed that WPI was uniformly distributed within WPI/PEO electrospun fibers. Observations by scanning electron microscopy (SEM) and field emission scanning electron microscopy (FESEM) indicated that fibers possessed a solid core. All these findings suggested that PEO enables the formation of WPI/PEO electrospun fibers by entanglement/entrapment/deposition. Preliminary studies were conducted on hydroxypropyl methyl cellulose (HPMC). In the absence of PEO, HPMC enabled the formation of WPI electrospun fibers under acidic conditions (124 ± 46 nm). FTIR analyses indicated that there was no interaction between HPMC and WPI, suggesting that HPMC aided in the electrospinning of WPI fibers, also by entanglement/entrapment/deposition. Hence, HPMC and PEO aid in the electrospinning of WPI fibers by entanglement/entrapment/deposition, which can be manipulated by alterations in the protein configuration and solution properties. / Natural Sciences and Engineering Research Council (NSERC) of Canada and the Dairy Farmers of Ontario (DFO)

Master of Science / Department of Food Science / Karen A. Schmidt / Milk proteins are excellent sources of sulfur-containing amino acids methionine and cysteine, in particular whey proteins. Cysteine is synthesized from methionine by γ-cystathionase. However, cysteine has to be included in the diets of certain subpopulations due to diminished γ-cystathionase activity. Cysteine, a heat- liable amino acid, may lose bioavailability during thermal processing. The objective of this research was to enhance cysteine content in yogurt while maintaining its quality. First, yogurt mixes were formulated to a total solids content of 12.5% with nonfat dry milk (NDM) (N) or a combination of NDM (10%) and whey protein isolate (WPI) (2.5%) (W), and processed at 70°C (20 min) (70) or 90°C (7 min) (90). Yogurt was prepared and maintained at 4oC for 60 days. Three replications were performed and data were analyzed using SAS®. The W mixes had 65%, 32% and 190% more cysteine, true protein and whey protein contents respectively, compared to N mixes prior to processing. However in day 1 yogurt, the highest cysteine content (398.3 mg/L) was found in the W70 yogurt and its gel quality was comparable to the N90 yogurt except for firmness. During a 60 day storage period the W70 and N90 were similar in gel quality except for firmness. Secondly, a hedonic test was done on the W70 (HC) and N90 (LC) yogurts which had been reformulated to contain sugar and vanillin. One replication was performed and data were analyzed using SAS®. The LC and HC yogurts did not vary in liking of flavor (6.1), aftertaste (6.1) and overall acceptability (6.3) corresponding to the words of “like slightly” when compared. However, the appearance of the LC yogurt was liked more than the HC yogurt (6.7 vs. 6.1) whereas the thickness of HC yogurt was liked more than the LC yogurt (6.4 vs. 5.8). These results suggest that addition of WPI along with lower process treatment resulted in yogurt with enhanced cysteine; however, further studies may be needed to optimize the WPI addition to improve the visual characteristics of the yogurt for consumer acceptance.

Whey proteins are mainly a group of small globular proteins. Their structures can be modified by physical, chemical and other means to improve their functionality. The objectives of this study were to investigate the effect of radiation on protein-protein interaction, microstructure, and microbiological properties of whey protein-water solutions. Whey protein isolate (WPI) solutions (27-36% protein) were treated with different dosages (10-35 KGy) of gamma radiation. The protein solutions were analyzed for viscosity, turbidity, soluble nitrogen, total plate count, and yeast and mold counts. The interactions between whey proteins were also analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and scanning electron microscopy (SEM). The viscosity of protein solution (27%, w/w) was increased from 2.19 for the control to 4.78 mPa*s for the sample treated at 25 KGy, respectively, and viscosity also increased during storage at 23°C. The soluble nitrogen (10%, w/w) was decreased from 100% to 54.7% for control and the sample treated at 35 KGy. The effects of gamma radiation and storage time on viscosity of whey protein solutions were significant (p

The purpose of this study was to evaluate the effectiveness of a whey protein isolate (WPI), a reported glutathione (GSH) booster, on exercise performance, immune function, and antioxidant status during weight maintenance and energy restriction in humans. Twenty well-trained, college age, male cyclists performed a cycling exercise test for 45 min, the first 7 min at 70% of VO2peak and the remaining 38 min at 55% VO₂peak immediately followed by a performance test set at 90% VO2peak until exhaustion. Blood samples were collected prior to the exercise test, after 45 min of exercise, within 5 min of exhaustion, and 1 h after exercise. Blood samples were analyzed for GSH, GSH/GSSG ratio, glutathione peroxidase (GPx), lipid hydroperoxides (LPO), phagocytosis, oxidative burst, peripheral blood mononuclear cell (PBMC) proliferation, and PBMC phenotyping. Subjects consumed 40g/day of WPI or casein placebo (P) along with their normal diet for 2 wk, repeated the exercise test, and then began a low energy period continuing the same supplementation for 4 d before the final exercise test. WPI was not associated with superior exercise performance or antioxidant status following exercise or weight loss. WPI supplementation did result in 33% greater lymphocyte proliferation capacity following exercise. Following exhaustive exercise for all trials, tGSH and GPx increased 7% and 11%, respectively, while WBCGSH decreased 13%. For WPI, GPx activity was 10% lower than P following exhaustive exercise for all trials combined. Weight loss (2.67 ± 0.26 kg) resulted in increases in phagocytosis (65%), white blood cell (WBC) GSH (40%), and GPx (35%) while decreasing the GSH/GSSG ratio (55%) and LPO (16%). Exhaustive exercise caused a 28% increase in CD8+ PBMCs and decreased CD4+ (34%), CD3+ (15%), the CD4+/8+ ratio (45%), and phagocytosis (8%) with all values returning to baseline after 1 h recovery. Supplementation with WPI did not enhance GSH status or exercise performance in trained cyclists, during weight maintenance or energy restriction. Following exercise, WPI is associated with greater lymphocyte proliferation of PBMCs which may help maintain an athlete's health during heavy training or competition. / Ph. D.

Having access to clean water is not a certainty for every human being. Today, there are major problems with polluted water that not just affect us humans, but also the ecosystem around us. In recent years, research into making aerogels from protein nanofibrils (PNF) has increased. What is interesting about these gels is their properties of adsorbing contaminants in the water, such as organic molecules and metal ions. In this report, hydrogels and non-dry aerogels (called “non-dry” due to there being one additional heat-treatment step that can be performed to “dry” the aerogel to make it sturdier and more water-proof), have been produced by whey protein isolate (WPI), which is extracted from the dairy industry. This is a sustainable, cheap, and renewable raw material. The goals are thus: (1) to examine if there is a difference in adsorption capacity between hydrogels and non-dry aerogels in a static setup and (2) to understand and examine which product parameters give the gels the best adsorption efficiency. To achieve these goals, eight different gels were synthesized with eight different contents. The parameters have varied from the following; dialyzed or non-dialyzed WPI solution, straight or curly seeds and whether or not salt had been added to the gels. The samples’ adsorption efficiency was analyzed by micro equilibrium dialysis (MED) and UV-VIS spectroscopy. For hydrogels, the dialyzed solution with salt and curly seeds was the variant with the highest binding capacity for ThT at 34% of the initial ThT concentration. For non-dry aerogels the dialyzed solution with curly seeds and no added salt was the best variant for binding ThT, at 84% of initial ThT content bound. It was also seen that the gels that have non-dialyzed WPI solution or that the solutions contained salt increased the adsorption capacity. The conclusions drawn were that salt has a positive effect on the adsorption capacity of the gels that have formed a matrix, and a negative effect on the adsorption of loose fibrils as well as that the solutions may not need to be dialyzed. The type of seeds or fibrils effect on adsorption capacity was inconclusive. The adsorption capacity for non-dry aerogels was three times higher compared to hydrogels. / Att ha tillgång till rent vatten är inte en självklarhet för varje människa. Idag är det stora problem med förorenat vatten som inte bara påverkar oss människor utan också ekosystemet. På senare år har forskning inom att tillverka aerogeler av protein nanofibriller (PNF) ökat. Det som är intressant med dessa geler är deras egenskaper att adsorbera föroreningar i vattnet, som till exempel organiska molekyler och metalljoner. I detta arbete har icke-torra aerogels (kallas "icke-torra" på grund av att det finns ytterligare ett värmebehandlingssteg som kan utföras för att "torka" aerogelen för att göra dem robustare och mer vattentålig) och hydrogels producerats av vassleproteinisolat WPI, som utvinns från mjölkindustrin. Detta protein är ett hållbart, billigt och förnybart råmaterial. Målet med denna rapport är att (1) undersöka om det finns skillnader i adsorptionskapaciteterna mellan hydrogels och icke-torra aerogels i ett statiskt medium (2) förstå och undersöka vilka produktionsparametrar som ger den bästa adsorptionskapaciteten. För att uppnå dessa mål har åtta olika geler syntetiserats med åtta olika innehåll. De varierande parametrarna var följande; dialyserad eller icke dialyserad WPI-lösning, raka eller krulliga frön och om salt har tillsatts eller inte. Proverna analyserades med mikro-jämvikts-dialys och UV-VIS spektroskopi. För hydrogeler var den dialyserade lösningen med salt och krulliga seeds den variant som hade högst bindningskapacitet för ThT, vilket låg på 34 % av den orginellaThT-koncentrationen. För icke-tora aerogels var lösningen som var dialyserad med krulliga frön och inget tillsatt salt den variant som var bäst på att binda ThT, med en kapacitet på 84% av den orginella koncentrationen. Vi såg även att gelerna som hade odialyserad WPI-lösning alternativt att lösningarna som innehöll salt ökade adsorptionskapaciteten. Slutsatserna som dragits är att salt har en positiv påverkan på gelernas adsorptionsförmåga när de format en fibermatris och en negativ påverkan när de bara består av fria fibriller, samt att lösningarna kanske inte behöver dialyseras. Typ av frön eller fibrillers påverkan på adsorptionsförmåga kunde inte påvisas. Icke-torra aerogels hade en adsorptionsförmåga på ThT som är tre gånger högre jämfört med hydrogels.

Crispness is one of the most desirable textural characteristics of breaded fried foods. Consumers often judge the quality of breaded fried foods based on the perceived crispness of the product. Furthermore, today's consumers are showing increasing concern over fat intake. As a result, there is great interest in being able to enhance the crispness and reduce the fat uptake in breaded fried foods without sacrificing other quality attributes. To achieve these goals, modifications to both frying equipment and product formulation have been explored in this study. In this study, two edible film coatings, methylcellulose (MC) and whey protein isolate (WPI) were incorporated into the batter and pre-dust to determine their effect on the crispness of breaded fried chicken nuggets held under a heat lamp for varying time intervals. Crispness was evaluated by both objective (ultrasonic non-destructive evaluation system) and subjective methods. An untrained sensory panel was used to obtain subjective measurements of product crispness. Panelists rated product attributes such as crispness, juiciness, oiliness and flavor on a simple intensity scale. Additionally, panelists rated the liking of the products on a nine-point hedonic scale (1=dislike extremely, 9=like extremely). Two pressure sources (nitrogen gas and steam naturally released from the food material) were used to determine their effects on product crispness, texture, pressed juice, moisture content, fat content and color. Products fried with nitrogen gas as the pressurizing medium produced samples that were comparable to or exceeding the quality of products generated by frying with steam, as it relates to product crispness, texture, pressed juice, moisture content, fat content and color. As related to objective crispness, chicken nuggets fried with nitrogen were significantly crispier (p<0.05) than those fried with steam. Coating type and application also had a significant effect on product crispness. Samples coated with MC in the pre-dust were crispier than samples coated with WPI. However, no significant differences were found in product crispness, juiciness, oiliness or flavor, and overall liking among samples tested by the sensory panel. The results of this study demonstrated that applying an edible film coating to the pre-dust and using nitrogen gas as the pressurizing medium can enhance and maintain the crispness of breaded fried foods. / Master of Science

Under flera månaders tid har ett kandidatexamensarbete utförts med syftet att optimera produktionen av protein-nanofibrer av vassleproteinisolat. Vassleproteinisolat består till stor del av proteinet β-laktoglobulin. Detta protein kan under upphettning bilda nanofibrer i sur miljö. Det var därför med avseende på parametrarna värme, koncentration och inkubationstid som processen optimerades eftersom det redan existerar ett pH-optimum vid pH-värdet 2. Lösningar av vassleproteinisolat med olika koncentrationer inkuberades under 24 timmar vid fyra olika temperaturer. Samtliga lösningar hade pH-värdet 2. För varje temperatur och inkubering togs proverna ut en åt gången för att sedan analyseras. De olika proverna analyserades sedan med Thioflavin T fluorescens för att se indikationer på fibrillering. De erhållna ThT spektrumen visade på fibrillbildning och resultaten för detta experiment visar på att utbytet av fibrilleringsreaktionen blir högre i takt med att hydrolysens hastighetskonstant blir lägre samt att lägre temperaturer kan gynna fibrillbildning . Ytterligare försök, tid och resurser bör läggas ner på detta område för att med säkerhet kunna optimera produktionen av nanofibrer av vassleproteinisolat. / This bachelor’s degree project’s aim was to optimize the production of protein nanofibrils originating from whey protein isolate. Whey protein isolate largely consists of the protein β-lactoglobulin, which can form nanofibrils while immersed in an acidic environment when heated. Therefore, the process was attempted to be optimized with regards to the yield of the final product of protein nanofibrils by varying parameters such as incubation time, initial concentration and temperature, with a constant pH-value of 2. Solutions of the whey protein isolate at different concentrations were incubated during a time interval of 24 hours and at different temperatures. For every temperature and time period of incubation, one sample at a time was taken out to be measured and analyzed, a total of four samples per initial concentration. The samples were analyzed with Thioflavin T fluorescence to see indications of the existence of fibrillation. The obtained ThT spectra showed intensity diagrams that can be related to the amount of formed nanofibrils, and this experiment shows that the yield of fibrils increases while the rate constant of the hydrolysis decreases, and that the fibrillation is favoured by lower temperatures. To optimize the production of nanofibrils of whey protein with certainty, further experiments, time and resources should be invested in this area.

Naturally occurring food antimicrobials such as plant essential oils are receiving tremendous interest as intervention systems to enhance microbiological safety and quality. Poor water solubility of essential oils makes it difficult to incorporate them in foods, impacting visual appearance, antimicrobial effectiveness, and possibly organoleptic properties. Engineered nanoscale delivery systems can principally solve these challenges, but those based on low-cost food ingredients and inexpensive and scalable processes are currently scarce. This dissertation presents a simple and scalable two-step technology to prepare nano-delivery systems. The first encapsulation step, based on emulsion-evaporation, involves preparing emulsions composed of an oil phase with thymol or eugenol, major compounds in extracts from thyme and clove respectively, in hexane and an aqueous phase with conjugates of whey protein isolate and maltodextrin, followed by evaporation of hexane by spray drying. The second step is to hydrate spray dried capsules to enable the formation of nanoscale particles. The encapsulation performance and dispersion characteristics were affected by amounts and types of conjugates (ratio of protein: maltodextrin and maltodextrin chain length), volume fraction and composition of the oil phase. The optimal conditions corresponded to 55.8 % encapsulation efficiency and 12.6 % loading for thymol and 47.9 % encapsulation efficiency and 7.9 % loading for eugenol. Dispersions prepared from the identified capsules contained particles smaller than 100 nm and were transparent at pH 3.0-7.0 and 0-50 mM before and after heating at 80°C for 15 min. Nano-dispersions and free oil were tested for antimicrobial activity against Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, and Salmonella typhimurium. Nano-dispersed and free antimicrobials had similar effectiveness at various pH and temperatures in tryptic soy broth and apple cider, while in 2 % reduced fat milk, nano-dispersed antimicrobials were consistently more effective than unencapsulated ones. Therefore, the commercially viable nanoscale technology presented in this study enables the delivery of lipophilic antimicrobials for enhanced microbial safety and quality, without compromising visual appearance of foods, especially clear beverages.

Lors de la production des poudres protéiques laitières, des précautions sont prises pour assurer des fonctions technologiques optimales selon leurs utilisations. Cependant, des évolutions structurales et fonctionnelles surviennent lors du stockage. Ce projet vise ainsi à comprendre les mécanismes de modification qui interviennent lors du stockage des poudres d’isolat de protéines solubles du lait (WPI). Pour ce faire, des poudres de WPI ont été stockées en conditions contrôlées (15 mois à 20°C, 40°C et 60°C), et leurs évolutions structurales et fonctionnelles après réhydratation ont été suivies expérimentalement à intervalles réguliers.Les résultats montrent que le vieillissement suit une trajectoire définie impliquant d’abord la lactosylation d’une partie des protéines, puis leur agrégation à l’état sec. De plus, cette trajectoire est caractérisée par un phénomène de rattrapage des modifications observées aux basses températures vers les plus hautes. L’impact des évolutions structurales sur les propriétés fonctionnelles est contrasté : les résultats montrent que les propriétés moussantes et interfaciales sont peu affectées, alors que les propriétés d’agrégation thermo-induites sont grandement modifiées. Pour minimiser ces modifications, deux pistes d’amélioration ont été suivies : 1. la granulométrie des poudres et 2. la teneur en lactose résiduel. La différence de granulométrie n’a pas eu d’influence sur le vieillissement alors que la diminution de la teneur en lactose a permis de limiter significativement l’étendue des modifications induites lors du stockage. / During dairy powder manufacture, precautionary measures are taken to ensure optimal technological functionalities regarding their use requirements. However, changes in structural and functional properties appear during storage. This project aimed understand the mechanisms of changes that occur during storage of whey protein isolate (WPI) powders. To do this, WPI powders were stored under controlled conditions (20°C, 40°C and 60°C for 15 months), and their structural and functional changes after rehydration were experimentally monitored at regular intervals. The results showed that ageing follows a specific path involving first protein lactosylation and then their aggregation in the dry state. In addition, this path was characterized by a catching up behaviour from the changes obtained at lower temperatures to those at highest temperatures. The impact of these structural changes on the functional properties was mixed:the results showed that the foaming and interfacial properties were only slightly affected, while the heat-induced aggregation properties were greatly modified. To minimize these storage-induced changes, several areas for improvement were followed, playing on either the powder particle size, or on lactose content whose role appeared to be crucial for the development of the Maillard reaction. The study showed that the difference in powder particle size had no influence on their ageing path while lowering the lactose concentration allowed to significantly reduce the extent of the storage-induced changes.

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