The 17^th International
Hydrocolloids Conference
Nov 12-15, 2024, Palmerston North, New Zealand

Food products might have varying qualities depending on how they are prepared. To test this hypothesis, the rheological, microstructural, and in vitro digestible characteristics of the binary mixtures resulting from the interaction of potato starch (PS) and either casein (CA) or whey protein (WP) were ascertained using two different raw material preparations. The experimental treatments were the mixtures obtained from the interaction of gelatinized PS and CA or WP solution (process I, P1) and from the interaction of native PS and CA or WP in a powder form (process II, P2) before mixing with deionized water and heated at 65 or 75°C. The results showed that the mixtures from both processes exhibited as a weak gel with G′ > G′′. Compared to P2, the rheological property of P1 was more solid-like, and in vitro digestion of P1 revealed less starch hydrolysis. The binary mixtures from P1 had a complicated microstructure with milk protein aggregated networks, whereas the mixtures from P2 had greater void areas that caused the structure to loosen. The temperature at 75°C seemed to promote the starch digestion in the mixtures worse than the temperature at 65°C, attributed to a complete milk protein denaturation, found in WP rather than in CA. This could better prevent starch digestion from enzyme hydrolysis in the mixtures containing WP than in those with CA. These findings are useful to manufacturers who use PS and milk protein as the main ingredients in their potato starch-based products with different preparation of raw materials.

Keywords: milk protein, potato starch, in vitro digestion, binary mixture, food interaction

Ganoderma lucidum (G. lucidum) is a type of fungus that has been utilized for 4000 years in traditional Chinese medicine. As one of the main polysaccharides, G. lucidum β-D-glucans possessed various pharmacological activities, and demonstrated the advantages of biocompatibility and safety. Some findings demonstrated that reducing the molecular weight (Mw) of β-glucan can enhance its anti-inflammatory properties. However, the effect of further degradation of G. lucidum β-glucan into oligosaccharides with lower Mws on its anti-inflammatory activity has not been fully elucidated. It would be beneficial for further applications to have a better understanding of the anti-inflammatory properties of G. lucidum β-glucooligosaccharides.

This study systematically investigated the intestinal anti-inflammatory activities and the corresponding mechanisms of β-D-glucans and oligosaccharides from G. lucidum with different Mws. The anti-inflammatory activity studies showed that all fractions could significantly inhibit LPS-induced levels of cytokines including TNF-α and IL-1β in Caco-2 cells at certain concentrations in vitro. In the dextran sulfate sodium (DSS)-induced mice model, it was discovered that supplement with β-D-glucans and oligosaccharides could alleviate typical symptoms of ulcerative colitis (UC), including weight loss, colon shortening, reductions in the colitis DAI scores, and pathological changes in colon. The primary mechanisms of action for β-D-glucans and oligosaccharides in UC were modulation of inflammatory factors, regulation of intestinal immunity, regulation of the intestinal microbiota, and regulation of SCFAs metabolism. Among them, microwave assisted degraded fraction (GLPW) with lower Mw (1.33×104 and 3.51×103 g/mol) and mainly contained β-glucooligosaccharides with degree of polymerization (DP) 2-24 were exhibited better ability to regulate inflammatory factors, intestinal microbiota and SCFAs metabolism than the original high Mw fraction (GLPS, 2.42x106 g/mol). These findings suggested that the further degradation of G. lucidum β-glucan into oligosaccharides could retain or even enhance the biological activity of the components, in addition to further lowering the molecular weight of β-glucan, improving water solubility and solution characteristics. The analysis of anti-inflammatory mechanism also provided a feasible and theoretical basis for potential use of degraded β-D-glucans in the prevention and treatment of UC.

Growing global interest in healthier and cleaner ingredients has prompted a shift in the composition of gluten-free products. This study focused on Bazlama bread, known for its simple ingredient list. The objective was to explore the potential of using clean-label hydrocolloids as alternatives to xanthan gum for producing high-quality gluten-free bread. The functional ingredients studied included psyllium husk gum (P5), xanthan gum (X5), and fenugreek gum (F5), each added at 5.0 g (per 100 g rice/chickpea flour base). Additionally, a combination of xanthan and fenugreek gums in a 50:50 ratio was tested at 3.0 (XF3), 5.0 (XF5), and 6.0 (XF6) g addition levels. The study assessed the pasting properties of hydrocolloid-flour slurries, the rheological behavior of gluten-free batters, and the overall quality of gluten-free Bazlama bread.

Results indicated that adding P5 increased RVA slurry viscosities and bread thickness compared to the X5 recipe. However, there was no significant synergistic effect observed when combining xanthan and fenugreek gum. Increasing the addition level of the xanthan-fenugreek mix from 3.0 to 5.0 g led to a 15.0% to 147.0% increase in slurry viscosities compared to the gum-free recipe. Furthermore, GFBB with the XF3 mix exhibited higher moisture content, greater thickness, and a lower spread ratio than those with F5. Gluten-free Bazlama bread with P5 and X5 were characterized by their elevated thickness and springiness, while gluten-free Bazlama bread with F5 and XF3 demonstrated higher spread ratio, hardness, diameter, and chewiness. Importantly, the inclusion of hydrocolloids did not significantly alter the starch digestibility of the gluten-free Bazlama bread.

Many polyphenolic compounds demonstrate low bioavailability, primarily due to poor solubility and inadequate physicochemical stability. Through encapsulation and delivery, zein-based composite nanoparticles can improve water dispersibility, physicochemical stability, targeted delivery, and gradual release of polyphenolic chemicals at certain regions in vitro under simulated gastrointestinal circumstances. Our team's previous research showed that stabilized zein nanoparticles based on hydroxypropyl-β-cyclodextrin (HP-β-CD) have positive impacts on the improvement of the physicochemical stability of polyphenolic compounds and outstanding polyphenol encapsulation properties. In additionally, we found that the in vitro gastrointestinal and simulated digestive features of the composite system may be impacted by the presence of salt ions. In the present study, we examined the fluorescence properties, bioactivity, and particle microstructure characteristics of polyphenols at various stages of digestion, and the effects of various ionic species and salt ion concentrations on the digestive properties of the polyphenol composite delivery system in an in vitro gastrointestinal simulated digestion solution. In the stomach phase, Ca2+-induced curcumin-loaded zein/HP-β-CD (ZHC) composite nanoparticles showed good enzyme resistance. Ca2+-modulated ZHC composite nanoparticles demonstrated improved digestion and slow-release features by creating a surface reticulated pore structure in the presence of Ca2+ after simulating digestion. The findings of this investigation can be used to create theories for the investigation of the absorption and digestive properties of polyphenol-loaded zein-based nano-delivery systems during in vitro gastrointestinal simulation.

In the Korean market, wild-simulated ginseng (WSG) is traditionally packaged in a polystyrene-board box with moss. Although the moss maintains a high humidity in the headspace at refrigeration temperature and prevents moisture loss from the WSG, the WSG would deteriorate during storage over two months due to micro-organisms, particularly fungi. Antimicrobial hydrogels incorporating oregano oil-loaded nanostructured lipid carriers (oregano-NLC gels) were employed to replace the moss and delay deterioration during storage. The oregano-NLC gels released moisture and achieved a relative humidity of over 95% in the box headspace at a rate comparable to that of moss. Furthermore, the oregano-NLC gels demonstrated remarkable efficacy in inactivating both spore germination and mycelium growth of Fusarium solani, a soil-originated fungal pathogen that can infect animals and plants, including humans and WSGs, respectively. Consequently, the packaging box installed with the compartments containing oregano-NLC gels successfully prevented the microbial deterioration of WSGs coated with oregano-NLC gels during storage at ~0°C for 60 days. Moreover, the WSG demonstrated the ability to maintain its moisture content, water activity, and respiration rate even after the storage period. The findings of this study may provide a novel perspective on the rational design of active packaging systems for the preservation of horticultural products, utilizing hydrogels containing essential oil-loaded NLCs.

Dry fractionation (DF) of pulses is proposed as a more sustainable process compared to wet fractionation for producing protein ingredients. Understanding the relationship between functional properties and processing methods is critical for their applications in foods. This study examines the effects of Calcium Sulphate (CaSO4) salt concentration (0.0%, 0.1% and 0.5%) and protein ingredient concentration (12, 14 and 16%) on the characteristics of gels from untreated and heat-treated faba bean protein ingredients. The ingredients were obtained via milling and air-classification and analysed for their functional properties. Heat-treatment reduced protein solubility by 62% and foaming capacity by 79%, while increasing water-holding capacity 1.47-fold and oil-holding capacity by 9%. The denaturation of proteins in the raw material induced by the heat-treatment, exposing their hydrophilic ends, likely explains the decreased solubility and foaming capacity, and increased water-holding and oil-holding capacity. The hardness of heat-induced gels was significantly increased by the combination of heat-treatment and salt addition. Water-holding capacity increased with salt addition but decreased with prior heat-treatment of the protein ingredient. Heat-treatment impacts functional properties of DF pulse protein, and subsequently affects their gelling behaviour.

Introduction
Menopause is associated with declining estrogen level leading to subjects vulnerable to various diseases like hormone dependent cancers, Osteoporosis, arthritis, CVD, hypertension, diabetes etc. Flaxseed lignans can bind with estrogen receptors and serve as weak estrogen therapies in estogen deficient female. Flaxseed contain highest amount of lignan i.e. Secoisolariciresinol diglucoside (SDG) which is having structural and functional similarities with the human estrogen (17 β-estradiol). Hormone Replacement Therapy (HRT) the single medical support for treating estrogen deficiency and problems associated with several side effects and sometimes not recommended. Studies clearly indicated that flaxseed can be served as an alternative to traditional hormone replacement therapy during estrogen imbalance and deficiency in female. Considering all the aspects, a research project is being proposed for development and validation of milk-flaxseed-based fermented beverage for female health. Milk can be a source of calcium, whereas, flaxseed can provide phytoestrogen and functional Lactobacillus organism can be useful in digestion, nutrient absorption and overall health management of female.

Objective:
The objective of the study was to develop and evaluate the efficacy of milk-flaxseed based fermented beverage in blood estradiol level in ovariectomized female rat model.

Materials & Methods:
Flaxseed is being roasted and used in further study. The functional organism Lactiplantibacillus plantarum (NCDC 296) was used to ferment low fat milk using 3% inoculum for a fermentation time of 7 h. The milk-flaxseed-based fermented beverage was prepared by adding roasted flaxseed flour, stabilizer, and sugar in fermented curd. The stability of functional component of flaxseed (SDG) was evaluated through simulated in vitro gastrointestinal digestion study using HPLC method. The efficacy of the developed product was estimated in ovariectomized female rat of 3 months old for 6 weeks and blood estradiol level was evaluated.

Results & Discussion:
Roasting enhanced sensory and nutritional properties of flaxseed flour by reducing antinutrients level. The optimized product had an overall acceptability score of 8.21±0.5 and a functional Lactobacilli count of 108 CFU/ml. The nutritional composition of product was 2.6% fat, 4.47% protein, and 0.71% ash. The beverage had a shear thinning behavior with the respective L*, a*, and b* values of 69.87±0.30, 2.06±0.67, and 10.83±0.21. The SDG content of product was 32.14 mg per 100 ml of product. The bioactive component SDG minorly degraded during simulated in vitro gastrointestinal study. The optimized beverage showed favorable effect on blood estradiol level in ovariectomized murine model.

Conclusion:
The developed beverage was found suitable in addressing hormonal imbalance in female as established by ovariectomized murine study. Milk-flaxseed-based fermented beverage may be a suitable option for general health management of the female population.

Keywords:
Milk, Flaxseed, Phytoestrogen, Lactobacilli, Estradiol, Beverage, ovariectomized murine model

Funding Support: The work had been conducted under the DST funded project ‘Development of flaxseed-rich probiotic dairy foods to address menopause symptom (DST/SEED/WS/58)’

Introduction:
The concept of sustainable food production is being a major concern of global considerations in current years. The estimated population is to reach 10 billion by 2050 globally which cannot be sustained with current dietary pattern and recent food production system. Ethical, environmental and health concerns around dairy products are driving a fast-growing industry for plant-based dairy alternatives. Although plant-based dairy alternatives are nutritious and useful for health but sensorially less acceptable due to the presence of ’beany off-flavour’ coming naturally from the source material. It is actually difficult to mitigate beany off-flavour and fermentation process is considered one of the best measure. Although fermentation of milk by lactic acid bacteria is well documented, limited work has been carried out in the area of fermentation of plant-based milks. Considering all these aspects, a study was undertaken to identify the flavour profile of a novel plant milk and evaluating the effect of lactic fermentation on flavour profile of novel plant milk.
Materials & methods:
The novel plant-based milk was provided by Saber Foods Limited (Palmerston North, New Zealand) and was primarily made up of specific lentil seeds using a patent pending technology. This substrate was evaluated for flavour profile before and after lactic acid fermentation. The lactic cultures were manufactured by DANISCO, Australia Pvt. Ltd., Australia and supplied by International Flavors & Fragrances (Australia) Pvt. Ltd., Lane Cove NSW 2066, Australia. The novel plant milk was subjected to fermentation by five mix lactic culture at a rate of 1% (w/v) for 6h and samples were collected at every 2h and kept at -20ºC till further analysis. The flavour profile of the samples were analysed in a Shimadzu GC-MS system using SPME fiber (CAR–PDMS-DVB) exposed to the headspace above of the samples for 30 min.
Results & Discussion:
A total of 30 volatile compounds were estimated in unfermented plant milk substrate. The volatile components were composed of alcohols (8), aldehydes (6), ketones (5), aliphatic substances (3), acids (4), esters (2), terpene (1) and furan (1). The beany flavour of substrate can be attributed majorly to hexanal (31.218%) which was found in high proportion. The other components associated with beany flavour were 2-methyl-4-pentenal (0.262%), hexanol (13.433%), 2-heptanol (0.384%), 1-octen-3-ol (0.09%), 1-hepten-4-ol (0.027%), 2-undecanone (1.127%), 2,3-octane dione (0.25%), alfa-pinene (0.19%), 3-hydroxy dodecanoic acid (0.237%). However, these components were present in the plant milk in low individual proportions. Lactic acid fermentation caused maximum reduction in hexanal content when fermented with a particular starter mix VEGE 047 LYO consisting of Streptococcus thermophilus, Lactobacillus bulgaricus, Bifidobacterium lactis and Lactobacillus acidophilus. This culture mix was also responsible for the enhancement of 2-nonanone contents which was favourable. The mechanism of beany off- flavour reduction was probably a result of bioconversion of hexanal into hexanol, increment in proportion of compounds like 2-nonanone and generation of ester and ketone like heptyl formate, 2,3-butanodione, ethyl benzoate, etc. Alcohol dehydrogenase (ADH) enzyme is responsible for bioconversion of hexanal to hexanol. The presence of ADH enzyme in selected microorganisms (L. acidophilus, Lactiplantibacillus plantarum, S. thermophilus) has been reported in bioinformatics data.
Conclusion:
Lactic acid fermentation may be considered an useful strategy for improvement of flavour profile of plant milk
Key words: alcohol dehydrogenase, beany flavour, hexanal, lactic fermentation, plant milk
Acknowledgements. The work had been conducted at Riddet Institute, Massey University New Zealand with the concerned researcher availed fellowship of IDP-NAHEP, New Delhi, India.

Introduction:
The size of starch granules plays an important role in determining their structural and functional properties, which in turn influence their suitability for various applications. The unique properties of small granule starches (< 10 µm in diameter) can be advantageous in certain applications, but was less studied compared to the commercial starches that were generally medium (10 to 25 µm) and large (> 25 µm) sizes. Microwave treatment has emerged as a novel physical modification method with unique advantages, such as rapid heating, enabling continuous operation while preserving appearance and quality. Various studies have suggested that the effect of microwave treatment on starches were largely dependent on the starch type and treatment conditions. Current research on the effect of microwave treatments of starches was majorly done with a short time period at < 100 °C using commercial starches. To better apply the technique, it is necessary to analyze the changes in physicochemical properties and structures from various food materials at high temperature (over 100 °C) and discuss the mechanisms behind the changes. Meanwhile, to increase the utilization of the small granule starches, it is important to comprehensively understand the changes in their properties during various modification processes.

Method:
Quinoa starch and amaranth (1–3 µm) were selected as small granule starches and compared with commercial maize starch (15 µm). Microwave treatments with temperatures ranging from 25 to 150 °C, and durations up to 2 h at 150 °C, were applied. The energy input was from 11.93 kJ/g starch to 472.09 kJ/g starch. The morphology, physicochemical properties (including swelling power, pasting properties, rheological properties and etc.) and thermal properties of treated starch samples were analyzed. The structure of samples was analyzed by FTIR, XRD, and the weight-based chain length distribution was studied.

Results and Discussions:
At treatment temperatures below the gelatinization temperatures, small granule starches exhibited increasing peak viscosity and gelatinization temperature range, suggesting partial annealing, while no significant changes occurred with large granule starch. As treatment temperature increased above gelatinization temperature but below 100 °C, small granule starches showed complete loss of crystallinity compared to a 7% decrease in relative crystallinity for maize starch. A transition from A- to B-type polymorphous structure was found in maize starch. Rheology profile revealed the existence of two types of remnants in the treated samples. At 150 °C, continuous degradation of amylose was observed with increasing treatment time. The proportion of long-chain amylopectin also decreased by 18%. Based on the changes in structure and properties, a three-stage mechanism was proposed. In conclusion, the susceptibility of starches to microwave-induced modifications was strongly influenced by their granule size. Small granule starches, such as amaranth and quinoa, had more pronounced changes in crystallinity, thermal properties, and pasting behavior than large granule maize starch. These findings provide new insights into the mechanisms of microwave-induced modifications in starches with different granule sizes.

This research aimed to optimize the encapsulation conditions for kratom leaf extract using spray drying. The study focused on various encapsulating agents (resistant maltodextrin, gum arabic, and a 1:1 v/v mixture of both) and two inlet temperatures for the spray dryer (150°C and 160°C), keeping the extract to encapsulating agent ratio constant at 1:2 (v/v). The results indicated that the highest yield was achieved with resistant maltodextrin at an inlet temperature of 150°C. Conversely, using gum arabic at 160°C resulted in the highest encapsulation efficiency. All kratom leaf extract microcapsules exhibited over 90% solubility, moisture content below 8%, water activity below 0.6, and a light yellow-green color. Microcapsules made with resistant maltodextrin had a smoother surface compared to other samples. Furthermore, the microcapsules using gum arabic at 160°C displayed the highest levels of bioactive compounds and antioxidant activity, with total phenolic content, total tannin content, total flavonoid content, antioxidant activity (measured by DPPH and FRAP methods), and mitragynine content being 36.78±3.08 mg GAE/g db., 36.42±2.45 mg TAE/g db., 5.22±0.12 mg QE/g db., 293.35±3.28 µM TE/100g db., 522.74±2.47 µM TE/100g db., and 1.96±0.04 mg/g db., respectively.

INTRODUCTION
Carrageenan and pectin are two of the most widely used biopolymers as gelling agents, texturizers, and stabilizers in food products. Carrageenan is considered as an industrially important marine polysaccharide resource, with the kappa-type carrageenan being consumed in large quantities in the food industry. On the other hand, pectin is used in several food applications due to it being considered safe in all countries since it is a natural component of plant materials. The texture of the products of these two polysaccharides can be fine-tuned by changing cations, polysaccharide concentration and by blending with different types of polysaccharides. In this study, we aim to make different mixtures of kappa carrageenan and citrus pectin and investigate their physicochemical properties, specifically viscoelasticity and phase separation structure.

METHODS
Kappa carrageenan (KC) and citrus pectin (CP) mixtures were prepared at different volume fractions and represented as KCxCPy, where x and y would equal to 100. The total polysaccharide concentration for each mixture was fixed at 1.5% w/w. Rheological and differential scanning calorimetry (DSC) measurements were conducted to investigate the macroscopic characteristics of the mixtures while fluorescence video microscopy with particle tracking was used to assess the local properties of the gels on a microscopic length scale.

RESULTS
The rheological and DSC results suggested that citrus pectin governed the viscoelastic properties of the mixtures before gelation and kappa carrageenan was the dominant polysaccharide upon gelation. Mixtures containing a higher ratio of pectin to carrageenan had higher final elastic modulus value than those with low ratio of pectin. Video microscopy with particle tracking probed the microscopic domain of the mixtures and showed results that agree with those obtained from rheological and DSC results.

DISCUSSION
It has been found that although pectin has a thickening effect, it does not form a network structure by itself. However, the results obtained suggest that in mixtures containing both pectin and carrageenan, pectin promotes the formation of the network structure of kappa carrageenan.

Introduction:
Commercial comminuted meat products typically comprise a continuous protein matrix filled with emulsified fat droplets, a composite gel system in which myofibrillar proteins play a principal structural role. Emulsion gels can be used to improve the food structure, develop healthy food products, overcome texture problems caused by lipid particles in food, simulate the effect of fat on the hardness of food, improve the water-holding capacity of meat products, and control the release of intestinal drugs. Emulsion gels have high stability as the gel network fixes the emulsion droplets, thereby avoiding aggregation. In addition, the stability of the oil droplets influences the stability and properties of emulsion gels. Therefore, the research aims to compare the effects of emulsions with various oil types on the rheological, water-binding, and structural properties of MP-emulsion composite gels.

Method:
MP (1 mg/mL) was an emulsifier in canola and olive oil (10% v/v)-based oil-in-water (O/W) emulsions. High-speed homogenization (8,000 rpm) was used to prepare emulsions. Emulsion particle size, ζ-potential, and morphological properties were analyzed. Conformational stability (differential scanning calorimetry) and surface hydrophobicity of MP were measured. MP (5% w/v) gels embedded with emulsified oil were tested for water-holding capacity, dynamic rheological properties, and gel strength. Morphological characteristics were observed using confocal laser scanning microscopy.

Results:
Particle size and ζ-potential studies show a significant effect on the physicochemical and rheological properties of the emulsions, moreover, the results are comparable to the viscosity outcomes, where with the decreasing of particle size, the viscosity values increase. The inclusion of olive oil and coconut oil in a ratio of 4:6 in the emulsion significantly increased the gel strength value, it is also noticeable as the highest indicator of storage modulus. Similarly, the addition of oils in the ratios of 10:0 and 0:10 significantly increased the WHC, but the values of storage modulus and gel strength decreased. Compared to control gels, MP gels with different oil ratios were more effective in moisture retention capacity.

Significance:
MP-stabilized O/W emulsions with various oil types have different effects on the physical properties of MP-emulsion composite gels. Based on the results, the processing of comminuted meat products should incorporate emulsification strategies based on emulsion particles allowing them to achieve different target textural attributes.

Introduction:
Excessive consumption of sodium has been linked to hypertension, promoting many studies on salt substitutes. Potassium chloride (KCl) provides is a major strategy to potentially lower blood pressure. However, the usage of KCl in food products as a sodium alternative remains limited due to unfavorable sensory attributes. Water-in-oil-in-water double emulsion systems can be also used for novel techniques in terms of masking undesirable sensory attributes by encapsulation of negative taste compound in the primary emulsion. Hydrophilic biopolymers such as polysaccharides have been further focused on food nutrition delivery and drug control by masking undesirable sensory attributes. Complex gum systems, such as the mixture of xanthan gum (XG) and locust bean gum (LBG), form a gel that inhibits the dilution of the inner water phase and effectively masks the bitterness of KCl. Therefore, this study aimed to investigate the physicochemical and sensory properties of gel-in-oil-in-water (G/O/W) double emulsion.

Method:
To prepare the gel phase, different concentrations of XG/LBG complex (0, 0.1, 0.5% w/w) and KCl (0.48% w/w) were dissolved in distilled water. Additionally, the same concentration of NaCl (0.28% w/w) was dissolved in the water phase. An 80% (w/w) oil phase with 20% (w/w) gel phase was pre-mixed and homogenized to prepare a gel-in-oil phase (G/O) at 700 rpm for 3 minutes and 15,000 rpm for 5 minutes, respectively. Moreover, 80% (w/w) of the G/O with 20% (w/w) of the water phase was pre-mixed and homogenized at 700 rpm for 3 minutes and 10,000 rpm for 3 minutes, respectively. The oil-in-water (O/W) emulsion was prepared using the same procedure, including 0.48% (w/w) KCl and 0.28% (w/w) NaCl in the water phase to compare sensory attributes. The G/O/W double emulsions were measured for appearance, structural, rheological, thermal, and sensory properties.

Results:
The addition of the XG/LBG complex had no phase separation between G/O and W for 14 days that is highly correlated with ζ-potential. Additionally, the XG/LBG complex as a gel phase is well encapsulated in the oil droplets. The primary emulsions are formed a homogenous shape and highly packed in the outer water phase. Moreover, the particle size of the emulsions is decreased as an increased concentration of the XG/LBG complex with the decreased in encapsulation efficiency and increased in apparent viscosity, respectively. In the strain sweep test, the tan δ is higher in the following order: MCT0>MCT0.1>MCT0.5 and CA0.5>CA0.1>CA0. The loss modulus of the emulsions only had frequency dependence, while storage and loss modulus had temperature dependence with the overshoot tendency. In the sensory test, the double emulsions were assessed that significantly lower saltiness and bitterness than O/W emulsion (p<0.05). Additionally, the saltiness is significantly increased while the bitterness is significantly decreased in the double emulsions with MCT oil (p<0.05).

Significance:
This study investigated the effect of the XG/LBG complex with different concentrations on G/O/W double emulsion. Overall, the double emulsion using the XG/LBG complex can increase the KCl and lower the NaCl improving the sensory attributes.

Chickpea protein isolate (CPI) has good functional properties, such as high solubility and foaming properties, and has wide applications in the food industry. Cellulose nanocrystals (CNC) exhibit excellent properties as stabilizers and thicknesses in aqueous systems. This study aimed to characterize the interactions of CPI with CNC and to understand the impact of CNC concentration on protein digestion. CPI dispersions (3%) with CNC (0, 0.5, 1, and 3%) and their digested counterparts were characterized in terms of charge, size, turbidity, protein solubility, surface hydrophobicity, secondary structure (FT-IR), and molecular weight distribution (SDS-PAGE). These results suggest that the surface properties and molecular ordered structure of CPI were significantly (p<0.05) affected by the presence of CNC. The negative charge of CPI (-35 mV) significantly decreased in the presence of CNC (-32 mV). The z-average and turbidity increased with CNC. Protein solubility did not differ significantly in the presence of CNC. Tryptophan fluorescence was reduced by the CNC. The hydrogen bonds between CPI and CNC were observed using FT-IR spectroscopy. As the CNC content increased, modifications occurred in the β-sheet and β-turn, indicating interactions between the CPI and CNC. CPI-CNC samples showed a significant decrease in the normalized intensity associated with the free–OH groups, indicating that these groups may participate in the formation of hydrogen bonds between the proteins and CNC. With a threshold of 3% CNC, a significant increase in the in vitro digestibility of chickpea proteins was observed, which could be associated with less aggregation of chickpea proteins at the gastric level and changes in their secondary structures. The incorporation of CNC into the beverage system as a stabilizer ingredient could improve the digestibility of CPI at a certain concentration level because they modified the β-sheet and β-turn structures. This research enriches the application of CPI and CNC in plant-based texture-modified beverages.

Plant proteins have recently gained interest as alternatives to animal proteins in food emulsions due to their versatile nature, sustainability, and economical cost. However, they are poorly soluble in water, due to their complex structures. Various methods for increasing protein solubility such as homogenization, or heating are being investigated. The focus of this work is on the impact of exposing pea protein solutions to highly alkaline conditions and then reducing the solution pH to pH 7 (so called pH shifting). This was found to reduce the average size of protein aggregates in solution. The solubility of the protein increased from 14 to 47%. Combining pH shifting with the application of ultrasound increased the protein solubility to 77%. The protein surface hydrophobicity increased indicating that all treatments exposed more hydrophobic groups on the protein surface. SDS-page revealed no significant changes in the electrophoretic pattern; hence the primary structure of protein was not changed by modification approaches. Protein behaviour at interfaces was investigated by measuring the interfacial tension and rheology. The modified protein adsorbed more slowly at the oil-water interface. Future experiments will explore the effect of other processing techniques, including high-pressure homogenization and heating on the physicochemical properties and interfacial behaviour of plant proteins.

A humidity-responsive active packaging technology was designed to address food spoilage by leveraging the increase in internal humidity as spoilage progresses. Lemongrass essential oil (LGEO) was emulsified and incorporated into polyvinyl alcohol (PVA) to create a film. This film exhibited variable release of essential oil depending on humidity levels, demonstrating antimicrobial effects when applied to food. However, the release was continuous rather than targeted at specific humidity levels, leading to rapid depletion of the antimicrobial agent and diminished effectiveness over time. To address this, we modified the emulsifier composition in the film to identify factors influencing the release behavior.

Particle size analysis of LGEO emulsions revealed that larger hydrophilic-lipophilic balance (HLB) values corresponded to smaller particle sizes, with Span 60 (HLB 4.7) producing the largest particles at 120.7 nm and Tween 20 (HLB 16.7) producing the smallest at 72.9 nm. Zeta potential measurements indicated predominantly negative values, with larger HLB values generally leading to higher zeta potentials. Notably, solutions containing Span 60 exhibited higher zeta potentials despite lower HLB values.

Post-drying analysis showed higher LGEO residuals with increased Span emulsifier content, while higher Tween 20 content resulted in lower residuals. A 72-hour release study demonstrated varied release profiles, with faster release from films containing more Tween emulsifiers. Response surface regression analysis confirmed that the Tween ratio had the greatest impact on release behavior, followed by HLB value.

Tween emulsifiers, with relatively high HLB values, are more hydrophilic, possibly weakening their interaction with LGEO or altering emulsion stability under humidity. This study suggests that optimizing emulsifier composition, particularly the Tween to Span ratio, is critical for achieving targeted release of essential oils in active packaging applications.

Introduction:
Barley (Hordeum vulgare) is a versatile and sustainable crop, contains 70% unique bimodal starch and has the potential to replace commercial starches. Typically, maize, potato, cassava, and wheat are the predominant conventional starch sources. However, the high starch content in barley and its 90% utilization for animal feeding and malt production presents an opportunity to shift this paradigm from animal to human feed. This study emphasizes the significance of barley starch by analyzing its unique physicochemical, functional and molecular properties with their correlations. Furthermore, this research highlights the most promising barley starches for industrial applications as sustainable food hydrocolloid sources.

Methods:
Twenty-five barley samples from New Zealand, Australia, Pakistan, and China were collected and analyzed. Starch was extracted using dry milling, steeping, blending, screening, de-proteinization, centrifugation and purification. Physicochemical (moisture content, total starch content, amylose content, swelling power, water solubility index), morphological (particle size distribution, and scanning electron microscopy), molecular (high-performance size exclusion chromatography), thermal (differential scanning calorimetry) and rheological (pasting, gel texture and dynamic oscillation) properties were measured.

Results:
A high diversity has been observed among the results of different barley starch cultivars. The moisture content of different barley starches ranged from 8-12%, with a total starch purity of more than 98%. The amylose contents ranged from 25% to 35%, determined through apparent amylose, true amylose and size exclusion chromatography. The morphological studies of barley starch revealed its unique bimodal characteristics as A and B type (5 out of 25 varieties). The cooking properties of barley starch showed that unimodal starches have more gelatinization enthalpy than the bimodal, irrespective of their overall temperature range, which is due to the non-uniformity in the double helical pattern.

Pasting properties showed a direct relation between breakdown viscosity and adhesiveness and an inverse relation between starch stability and hardness, suitable for the paper industry. The rheological properties of starch particles depict that in bimodal starches, there is less difference between storage modulus and loss modulus due to their small particle size as compared to unimodal starches during different temperatures, frequencies, and amplitude sweeps. Overall, the storage modulus value of barley starch pastes was more than the loss modulus, and their loss tan less than 1, indicating their elastic behaviour than viscous, which is highly desirable for noodles, pasta, bread, dough and chewy candies.

Discussion:
The results of the principal component analysis showed that amylose contents played a significant role in determining the rheology, pasting and cooking behaviour of the starch. Moreover, the ratio of amylose-amylopectin, their interaction, particle size distribution, structure-functionality relationship, and geographical distribution were the primary parameters to decide the characteristics of the starch molecules, which determines their suitability for industrial applications.

Conclusion:
This research provides valuable insights into the physicochemical properties and molecular structure of barley starch and their correlations to enhance its application as a sustainable hydrocolloid in the food industry beyond its traditional uses in animal feed and malt production.

Introduction:
Pickering emulsion is an emulsion stabilized by solid particles instead of conventional surfactants. Proteins are highly preferred in production of stable food-grade Pickering emulsions due to their desirable techno-functional properties and irreplaceable nutritive value. Faba bean protein isolates (FBPIs) are popular because of their high protein content and easy cultivation. Pea protein isolates (PPIs) are also valued for their high digestibility, low environmental impact. However, native plant proteins, particularly from legumes, often lack adequate emulsifying capacity due to their compact globular structures, making it essential to enhance this property for effective use in food applications. One such approach to enhance emulsifying abilities is heat treatment, which can alter the structures of proteins, causing unfolding and denaturation. Furthermore, ultrasound treatment disrupts intermolecular interactions and breaks down protein aggregates through acoustic cavitation. Therefore, this study aims to investigate the fabrication of physically modified particles of FBPIs and PPIs, and to characterize high internal phase Pickering emulsions (HIPPEs) stabilized with these modified protein isolates, focusing on the effects of varying ultrasonication times.

Method:
Plant proteins (4 wt% FBPIs and PPIs) were subjected to heating (90 ˚C for 2h) and ultrasonication (20kHz, 120 W for 5, 10 and 20 min). The properties of physically modified plant protein particles (particle size, ζ-potential, surface hydrophobicity, solubility and turbidity) were measured. High-speed homogenization (10,000 rpm, 5 min) was used to prepare HIPPEs (φ=0.7). The emulsion droplet size, ζ-potential and rheological properties were analyzed. Additionally, morphological characteristics were observed using optical microscopy and confocal laser scanning microscopy.

Results:
Heating and ultrasonication significantly affected the physicochemical properties of the particles. Physically modified particles exhibited a significant decrease in particle size compared to untreated particles, leading to decreased turbidity and increased surface hydrophobicity. Comparison between the two proteins showed that FBPIs exhibited smaller particle size and lower turbidity, while its solubility was consistently higher than PPI across all treatments. These modifications resulted in HIPPEs with smaller droplet sizes. However, both HIPPEs stabilized by FBPI and PPI showed a decrease in droplet size with ultrasonication up to 10 minutes, followed by an increase after 10 minutes. Rheological behaviors suggested that all HIPPEs were non-Newtonian fluids and exhibited a higher G′ than G″. As ultrasonication time increased, viscosity decreased due to the protein particles creating a steric hindrance effect. For morphology, HIPPEs stabilized by physically modified proteins showed smaller droplets and a more densely packed structure. CLSM images indicated the formation of a protein layer at the oil-water interface.

Significance:
This study provides valuable insights into the potential of physically modified FBPIs and PPIs as stabilizers in HIPPEs. Controlled ultrasonication and heat treatments significantly improve the emulsifying properties of plant proteins, enhancing stability and reducing droplet size, offering potential for clean-label, sustainable food products without synthetic surfactants.

Some polysaccharides have prebiotic effects on probiotic strains including Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium bifidum. Their prebiotic effects can be used to modulate gut microbiome thus gut health. However, due to the low solubility and high viscosity of some polysaccharides, their application as prebiotics is restricted. Moreover, there is limited research regarding the relationship between molecular weight, conformation, viscosity of polysaccharides and gut microbiome. Our preliminary data has demonstrated that certain types of polysaccharides, e.g., pectic polysaccharides, can modulate the proliferation performance of some specific probiotic strains.

The objective of the current study was to investigate the effect of molecular weight, conformation, and viscosity of pectic polysaccharides on probiotic stains and gut microbiome. To achieve this, fractions of pectic polysaccharides, namely citrus pectin and water-soluble yellow mustard mucilage (WSM), were obtained using pectinase hydrolysis. Molecular weight and conformation were measured using GPC/SEC, viscosity was determined by a Rheometer. The prebiotic effects were evaluated using probiotic strains including Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium bifidum. The effect on microbiome was investigated using the in vitro method by adding the polysaccharide fractions into human stool samples.

Six fractions were obtained from citrus pectin and WSM, respectively. Their molecular weight varied from 106 to 103 Dalton, with corresponding decreased viscosity. The results have indicated that all the fractions have demonstrated prebiotic effects on the 5 tested prebiotic strains. Fractions with smaller Mw showed a better growth promotion on the probiotic strains, which may attribute to the better mobility of the pectin and WSM molecules, thus leading to a better bioavailable as a carbon source for the tested bacteria. The molecular conformations of WSM showed no effect on the bacterial growth. The viscosity of the WSM fractions were correlated to Mw, indicating that a less viscous solution showed a better growth promoting effect on bacterial strains. The microbiome of the human stool sample treated with different fractions also showed varied bacterial population at phylum level. The fractions with lower molecular weights have increased the population of Firmicutes, Actinobacteria, and Verrucomicrobiota, and decreased Proteobacteria. Surprisingly, no difference was detected between citrus pectin and WSM. The result has revealed that the pectic polysaccharides, usually high in molecular weight and viscosity, can be better utilized as prebiotics by reducing their molecular size, and improving their solubility and bioavailability. This information will help developing synbiotic products using pectin and WSM as carriers to protect the probiotic strains, meanwhile modulating the gut microbiome. Further studies will be carried out in vivo using the developed synbiotic products in animal feeds.

Tissue engineering aims to reconstruct living tissues to maintain, restore, or enhance function, with ideal biological scaffolds integral to this goal. Collagen and glycosaminoglycans (GAGs) are major structural components of the extracellular matrix (ECM), making it a natural biopolymer commonly used in biological scaffolds. Collagen's versatility and wide-ranging characteristics, such as low antigenicity and excellent biocompatibility, make it an impactful biomaterial. Furthermore, its ability to mimic the ECM by promoting the attachment, proliferation, and migration of various cells allows it to play a critical role in repairing damaged tissue. The study aimed to investigate the degradation of different collagen substrates by various enzymatic treatments, monitoring the release of key degradation products over multiple time points. GAGs compositional analysis was then measured through colorimetric assays and liquid chromatography-mass spectrometry (LC-MS). Differential scanning calorimetry (DSC) was used to determine the thermal stability of the collagen. The results revealed significant differences in degradation patterns and GAGs profiles between the collagen substrates. The DSC analysis provided valuable data on their thermal characteristics, highlighting the structural complexity of each collagen substrate. These findings contribute to a broader understanding of collagen biochemistry and GAGs composition, with important implications for developing optimized enzymatic protocols and potential applications in tissue engineering

Gelatin, a widely used biopolymer known for its biocompatibility and biodegradability, shows significant potential for various packaging applications. Blending gelatin with other biopolymers and natural extracts has been extensively explored to modify its mechanical properties and introduce beneficial properties such as antioxidant and antibacterial effects. However, the high vapour permeability and sensitivity to environmental conditions of pure gelatin films introduce challenges, particularly in preserving foods susceptible to degradation from moisture. This study aims to address this issue to improve the applicability of gelatin-based films for food packaging. Through the uncommon approach of using a multilayer film, we aim to develop a material with increased resistance to water vapour permeation and a reduced dependence on environmental conditions. The addition of plasticisers like glycerol, combined with proteins such as casein and chitosan, is investigated to further reduce solubility and improve the mechanical properties of the films. The resulting films were characterised for their tensile strength, solubility, swelling capacity, and thermal properties using techniques such as Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The outcome of this study will expand the future application of gelatin-based film with improved moisture barrier properties without sacrificing mechanical durability, offering a sustainable solution for extending the shelf life of various food products.

Introduction:
Probiotics and their derivatives have garnered significant interest for their potential health benefits, particularly in gastrointestinal health. Among these, the use of postbiotics, which are bioactive compounds produced by probiotics, has been investigated for their therapeutic applications. Colon-targeted postbiotics delivery offers a promising avenue for improving colon health by delivering beneficial postbiotics stably to the colon.

Methods:
In this study, a novel colon-targeted gel system was fabricated using a Lacticaseibacillus casei cell-free supernatant (LCFS), inulin, and gums. Industrially produced LCFS was obtained from Cell Biotech Co. Ltd. (Gimpo, Republic of Korea). Three anionic gums, kappa carrageenan, agar, and gellan gum, were selected based on their gelation properties in the LCFS. To fabricate colon-targeted gel system, anionic gums (1% w/w) were initially mixed with inulin (25% w/w) and then added to the LCFS (74% w/w). The rheological and microstructural properties of the gels were assessed. Indole-3-propionic acid (IPA), known for its effectiveness against colitis, was selected as a model postbiotic substance to monitor the release patterns of postbiotics from the gel systems in the gastrointestinal tract.

Results:
The salt composition of LCFS was suitable for gel formation using anionic gums and inulin. During the simulated oral conditions, the gels released approximately 10.6% (kappa carrageenan-based), 23.4% (gellan gum-based), and 29.4% (agar-based) of encapsulated IPA. Subsequently, the cumulative release of encapsulated IPA from the gels by the end of the simulated gastric phase was approximately 21.6% (kappa carrageenan-based), 32.8% (gellan gum-based), and 44.2% (agar-based). Lastly, the cumulative IPA release from the gels by the end of the simulated small intestinal phase was approximately 37.1% (kappa carrageenan-based), 46.0% (gellan gum-based), and 56.6% (agar-based). The kappa carrageenan-based gels, which had highest storage and loss moduli and provided the best protection among the gels, reliably delivered more than 60% of encapsulated IPA to the colon. This indicates that the novel gel system is a highly effective delivery system compared to control IPA, which is already eliminated in the gastric phase. Furthermore, it is expected that gut microbes could degrade inulin and anionic gums into fermentable monosaccharides, promoting colon health. Importantly, this system can deliver components that promote colon health, such as postbiotics, prebiotics, and LCFS, all without presenting health risks.

Background and purpose: Every year tonnes of macadamia nuts are produced globally, resulting in a large production of agricultural by-products such as macadamia husk. This by-product contains high concentrations of phenolic compounds with health-promoting properties. Utilising these phenolic compounds in food systems is challenging as the stability and biological activities can change when exposed to different types of environmental conditions (e.g., heat, light, oxygen, and pH). Therefore, this study aimed to develop liposomal encapsulation system that can protect the integrity and stability of phenolic compounds from macadamia husk against such environmental conditions towards their delivery in functional foods.

Experimental approach: Different extracts of macadamia husk were prepared by conventional solvent extraction (CSE), accelerated solvent extraction (ASE), and ultrasonic probe-assisted extraction (UPAE). Water and other organic solvents such as ethanol and methanol mixtures of different concentrations of water are used as extraction solvents. The extracts were characterised by their total phenol content (TPC), total flavonoid content (TFC), and antioxidant properties (using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity method). Liposomes composed of soy lecithin were used to encapsulate the phenolic compounds to maintain their stability and biological activity against environmental conditions. Dispersions of liposomes loaded with macadamia husk extract (MHE) were created using a combination of high-shear mixing and ultrasound. The mean particle size, homogeneity, zeta potential, and encapsulation efficiency were used to characterise the liposome properties.

The increasing older population (> 60 years) in the world represents one of the significant challenges for the Food Industry since they have specific nutritional and sensory requirements. Some strategies for designing products focused on nutrition have been proposed, most of which consist of developing functional foods using more efficient nutrient delivery systems, such as nanoemulsions. However, a soft texture is also necessary for easier and safer swallowing of older people due to dysphagia or swallowing disorders, particularly during senescence. This work aimed to evaluate the effect of protein mixtures (whey protein concentrate-WPI and soy protein isolated-SPI) on soft gels' textural and rheological properties based on nanoemulsions. First, a nanoemulsion base (droplet size: 131 nm) was prepared by high-pressure homogenization (700 bar – 4 cycles). Then, different ratios of WPI:SPI (1:3, 1:1, and 3:1) were dispersed into the nanoemulsion (60 min at 200 rpm), followed by thermal treatment at 90ºC for 60 min. In addition, two control samples (WPI and SPI) were prepared. Nanoemulsion-based gels were characterized by Texture Profile Analysis (texturometer) and flow and viscoelastic properties (oscillatory rheometer). The results of TPA showed significant differences (p<0.05) among samples due to the protein-mixture ratio. Hardness increased and cohesiveness decreased as the WPI ratio increased (from 3.1 to 30 N and 0.6 to 0.9, respectively). At the same time, adhesiveness varied slightly with WPI concentration (165 and 170 g·s). Regarding flow properties, all gels showed shear-thinning flow behavior. The gels prepared with higher WPI ratios presented a higher consistency and pseudoplasticity. On the other hand, mechanical spectra indicated a gel-like behavior for all samples since the storage modulus (G’) was higher than the loss modulus (G’’). In addition, G’ and G’’ moduli of mixed-protein gels showed a frequency dependency, indicating a weaker internal structure of these samples. According to the International Dysphagia Diet Standardization Initiative, nanoemulsion-based gel texture can be classified as level 4 (pureed foods), which does not require biting or chewing. In conclusion, it is possible to form soft gels based on nanoemulsions using different protein mixtures, which can help develop high-protein foods for the older population with swallowing problems.

Vitamin E (VitE) is a lipophilic vitamin essential for human health that degrades rapidly due to oxidative instability. Pickering emulsions (PE) can be used as an encapsulation system, where cellulose nanocrystals are proposed as stabilized particles to enhance VitE stability. However, vitamin bioaccessibility depends on various emulsion factors, such as pH and particle interactions with the gastrointestinal fluids. In this context, this study aimed to evaluate the influence of the pH on VitE release from the PE with CNC's through the gastrointestinal tract (GIT)

VitE PEs were stabilized with CNC at a VitE:CNC ratio of 1:2.5. Emulsions were homogenized using an Ultraturrax for 10 min at 10000 rpm, and then with ultrasound for 5 min (ON: 5s – OFF: 5s) at 60% amplitude. The pH of the PEs was adjusted among 3-8. A static in-vitro digestion was carried out following the INFOGEST protocol. Droplet size and zeta potential of initial PEs were characterized by DLS. A digested sample was taken from gastric and intestinal phases at different times (0, 60, 120 min). Then, droplet size and particle morphology changes were analyzed using DLS and confocal microscopy.

Before in-vitro digestion, all the emulsions showed a particle size of 340±16 nm, regardless of pH adjusted. However, significant differences (p<0.05) were observed in the zeta potential, which ranged from -70 mV (pH 8) to -40 mV (pH 3). During gastric digestion, the droplet size of Pickering emulsions increased around 1500 nm at 60 min, indicating the coalescence of oil droplets. Then, droplet size decreased to ~800 nm after 120 min of digestion, mainly observed in emulsions at pH 3, 4, and 5. This variation in the droplet size can be attributed to breaking the Pickering particles under gastric conditions. No differences in particle size were observed in samples at pH 6, 7, and 8, indicating higher stability of Pickering particles during this phase. On the other hand, the reduction of droplet size (~250 nm) of samples at pH 3, 4, and 5 was observed in the intestinal phase, indicating the complete breakdown of Pickering particles and, thus, the VitE release. On the contrary, the samples at pH 6, 7, and 8 showed increased droplet size and greater lipid residues, which was corroborated by images obtained from confocal microscopy. This behavior indicated lower lipid digestion, which could decrease the VitE release. Therefore, the initial pH of the emulsion affects the release of VitE during in-vitro digestion, where an acid pH proves better protection of VitE, increasing their bioaccessibility. These findings have practical implications for the food and pharmaceutical industries, suggesting that adjusting the pH of Pickering emulsions could be a strategy to enhance VitE bioaccessibility in food and drug formulations.

Background: Food security constitutes a worldwide concern closely correlated with the increase in population growth. By 2050, the total global population is expected to reach 9.3 billion. This rise in population is associated with a surge in food demands and an expected rise in metabolic conditions. In this context, looking for alternative and sustainable food and protein sources has become increasingly imperative. Pasture legumes including lucerne (Medicago sativa) and red clover (Trifolium pratense) are becoming popular as they can be used as an alternative protein and functional food sources. As these two crops play an important role in New Zealand’s agriculture, their seeds can be used in human nutrition as alternative food and protein options, however, the presence of anti-nutritional factors (ANF) and their distinct taste make them less favourable for human consumption. Fermentation can be used as a possible strategy to mitigate these limitations.

Methodology: Lactobacillus fermentation was conducted using L. plantarum, L. acidophilus and L. casei. Proximate composition and mineral content were determined following AOAC methods. Total phenols, flavonoids and antioxidant activity (DPPH and ABTS) and ANF including phytic acid, trypsin, and chymotrypsin inhibition were assessed using colourimetric methods. For the enzyme inhibition assays, enzyme-substrate reactions were performed with sample extracts before measurement.

Results: All the legume seeds demonstrated a high nutritional content, with crude protein and crude fibre levels around 40 and 16% respectively. The seeds were also found to contain a rich repertoire of minerals, particularly magnesium, phosphorus, iron and zinc. In addition, fermentation led to an increase in total phenols, flavonoids and antioxidant activity, while significantly reducing the ANF.

Conclusion: Lactobacillus fermentation has proven to be an effective processing technique to enhance the nutritional value of lucerne and red clover seeds. It not only increased the levels of bioactive compounds such as total phenols and flavonoids but also reduced the levels of abundantly present ANF thus improving nutrient digestibility and bioavailability. These findings support the potential of using fermentation to develop novel and sustainable protein sources, contributing to improved dietary quality and nutrition.

Protein bodies are naturally occurring storage organelles in hemp seeds. Major hempseed storage proteins include edestin (11S globulin) and albumin that are primarily located in crystalloids and proteinaceous matrices of hemp protein bodies (HPBs), respectively. The thermal denaturation of hempseed proteins has been studied in isolated systems, but to the best of our knowledge has not yet been explored in HPB systems. Here, we isolated native HPBs using an environmentally sustainable enzymatic method. Then, aqueous suspensions of HPBs (4% protein, w/w) were preheated at selected temperatures (60-100 °C) and pH 7 for 20 min and subsequently hydrolysed by trypsin at pH 7 and 37 °C for 120 min. The thermal aggregation of HPBs was characterised using confocal laser scanning microscopy (CLSM) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The hydrolysis of HPBs by trypsin was monitored by degree of protein hydrolysis (DH) and SDS-PAGE. Insoluble aggregates of HPBs stabilised primarily by disulfide bonds were formed upon preheating, most noticeably at temperatures above 80 °C. Preheating increased DH and altered protein degradation patterns of both acidic and basic subunits of edestin. This could be related to conformational changes in the HPB structure resulting from heat-induced dissociation and association of multiple HPB components. These findings enable a better understanding of structure-hydrolysis relationships of HPBs, which can lead to the use of these protein-rich particles as a new plant-based material for food applications.

The unique properties of quinoa protein, including a well-balanced amino acid profile, high digestibility, and gluten-free nature, make it an ideal ingredient for improving the nutrition of meat analogues. This study aims to develop quinoa-based high moisture extruded meat analogues (HMMA) which will deliver superior nutritional options for the health consciousness. The gastric digestion behaviours of HMMA in comparison to real chicken meat are demonstrated in this study.

HMMAs (approximately 57 % moisture) were produced in a two-screw extruder with mixing high protein quinoa flour (HPQF) (25% protein) and pea protein isolates (80% protein) in the following ratios: 100:0, 95:5, 90:10, 85:15 and 80:20. The maximum barrel temperature of the extruder was 170 °C, with the feed and water rate being 2.8 kg/h and 3.6 kg/h, respectively. The textual properties and microstructure of HMMAs were analysed using Texture Analyser and SEM. The gastric digestion behaviour of samples containing 0% or 20% HPQF and cooked chicken breast was assessed and compared in vitro using a human gastric simulator.

Incorporating HPQF led to reduction in hardness, springiness, chewiness and texturization degree of HMMA. Microscopies revealed a layered structure across all HMMA samples, with samples containing 0% HPQF displaying a dense layer structure and HPQF-containing samples exhibiting a looser and porous structure. HMMA samples and cooked chicken breast showed distinct differences regarding gastric pH profile, digesta size distribution, digesta microstructure changes and protein gastric emptying rates. HMMA bolus initially broke into large particles (100-1000 µm) at the early stage of digestion and these particles gradually reduced to 10-50 µm after 150 min of digestion. However, the disintegration of chicken bolus involved two different debris – large particles (100-1000 µm) and small particles (approximately 10 µm) released from the edge of large particles even at the early stage of digestion, and these two types of debris continued to breaking into smaller particles (approximately 5 µm) over digestion. Moreover, the gastric emptying of proteins for HMMA samples were faster than that of chicken, although the gastric emptying rates of proteins for HMMA containing 0% and 20% HPQF were comparable during the gastric phase. This might be attributed to their structural differences. This study explored the potential of incorporating quinoa protein into HMMA production and provided further understanding on the in vitro digestion behaviour of HMMA compared to chicken breast.

This study investigated the impact of calcium and magnesium chloride at the concentration of 0, 2, 4, 6 and 8mM on physico-chemical properties of milk protein concentrate (MPC)-soy protein isolate (SPI) mixture solutions containing 10% w/w total protein (MPC:SPI=8:2). Increasing divalent ions concentration resulted in a greater increase in viscosity, protein size, sediment formation and greater reduction in pH, hydration, and heat stability on sterilization at 111oC for 15 min. Addition of calcium chloride shows slightly less effective than magnesium chloride for reducing heat stability following the heat sterilization. The results demonstrate that concentration of the calcium and magnesium chloride used strongly impacts heat and physical stability of MPI-SPI mixture based nutritional products.

The growing demand for sustainable, plant-based food ingredients has spurred the search for alternative sources of oil bodies, or oleosomes, which are naturally occurring lipid storage organelles found in seeds, nuts, and other plant tissues. This study investigates safflower seeds as a novel source of oil bodies, with a focus on optimising extraction processes and conducting a thorough structural and physicochemical characterisation to assess their potential applications in various food products. In initial experiments, safflower oil bodies were extracted using an aqueous extraction method at alkaline pH, involving wet grinding, filtration, and centrifugation. Comprehensive compositional analysis, including fat, protein, and moisture content, was conducted using standard analytical methods to determine the composition of the extracted oil bodies. Physicochemical properties, such as zeta potential and particle size distribution, were also measured to evaluate the stability and functionality of the oil bodies in various food formulations. The extracted oil bodies were found to contain 45.22±1.09% fat and 4.44±0.13% protein, indicating a rich lipid content suitable for food applications. The oil bodies exhibited a small particle size with a D3,2 value of approximately 2.163 µm and a zeta potential of –12.53 mV, suggesting good stability in the emulsion system. These preliminary findings highlight the potential of safflower oil bodies as a functional ingredient in plant-based food formulations. Future work will explore enzyme-assisted extraction techniques to enhance oil body recovery while preserving their structural integrity, potentially broadening their applicability in the food industry.

Exosomes are small, 30-150 nm-sized nanovesicles that are generated by both plant and mammalian cells. Depending on where they come from, they include a distinct combination of lipids, proteins, nucleic acids, and metabolites that are all biologically active. All of these are enclosed by a lipid bilayer. An increasing amount of research highlights the critical function of exosomes in various physiological and pathological settings. As exosomes can be absorbed through diet, plant-derived nanovesicles are receiving a lot of attention from researchers. Their bioactive constituents endow them with several therapeutic attributes, including anti-tumorigenic, anti-oxidative, anti-inflammatory, and hepatoprotective. Isolating exosomes from plant sources is a challenging process that needs careful consideration due to a number of factors. Mammalian exosomes have been the subject of several studies, but very little is known about plant-derived exosomes, especially those formed from papaya. This work examined two distinct techniques for separating papaya-derived exosomes (PDEs): Density Gradient Ultracentrifugation and Differential Ultracentrifugation. PDEs were effectively extracted using the differential ultracentrifugation method. They were then characterised using Flow Cytometry (FCM) to determine the numbers of exosomes, FE-SEM for morphological analysis, and Dynamic Light Scattering (DLS) for size and zeta potential analysis. Zeta potential and Z-avg of PDEs have been shown to range from -15 mV to -68 mV and 50 to 150 nm, respectively. PDEs have a spherical vesicle-like form according to FE-SEM imaging. Over 98% of all events below 100 nm are present, according to results from FCM. This work successfully identified and described papaya exosomes, suggesting that further research into the biological functions and applications of these exosomes may be possible. The unique properties of exosomes derived from papayas could provide new insights into plant-derived extracellular vesicles and their potential use in the development of drug delivery systems and functional foods.
Keywords: Exosomes, Plant-derived nanovesicles, Extracellular vesicles, Differential Ultracentrifugation, Density Gradient Ultracentrifugation

This study investigated the effects of calcium sequestering salts (CSS) addition (5 mM trisodium citrate (TSC) or disodium hydrogen phosphate (DSHP) and heat treatment (85 or 95°C for 5 min) on the physicochemical and protein structural properties and the in vitro gastrointestinal digestibility of proteins in blends (0:100, 25:75, 50:50, 75:25, 100:0) of skimmed buffalo and bovine milk. Significant changes (P < 0.05) in pH and calcium activity as a result of TSC or DSHP addition were linked to higher particle size, milk viscosity, and non-sedimentable proteins levels. These changes were mostly observed during heat treatment at 95°C and increased proportionally to the concentration of buffalo skim milk in the milk mixture. Increasing the proportion of buffalo skim in the blend resulted in decreased protein digestibility, irrespective of CSS addition and heat treatment. Digestibility of proteins in milk was not markedly changed by heat treatment alone, but the combined effect of CSS addition and heat treatment greatly impacted the digestibility of κ-casein but not whey protein digestibility. While addition of TSC and DSHP induced comparable κ-casein digestibility in bovine skim milk and milk blends, the addition of TSC and heat treatment effected maximum κ-casein gastrointestinal digestibility in skimmed buffalo milk. α-lactalbumin and κ-casein were the most resistant proteins against degradation by digestive enzymes. This study elucidated the feasibility of adding either TSC or DSHP to buffalo:bovine milk mixture and subsequently provided information to better understand protein digestibility in buffalo and bovine milk blends. These results further shed light on the suitability of processes that can be applied to maximise the gastrointestinal digestion of proteins in these milk systems.

The global demand for protein is projected to rise steadily in the coming years, driven by the growing world population and shifts in food production and consumer diets. To address the current overconsumption of animal protein and meet this increasing sustainability demand, it is essential to boost the production of high-quality, functional, and sustainable plant protein ingredients. Pea, Pisum sativum, is one of the main legume crops utilized to provide protein-rich ingredients in the plant-based food market. Pea protein concentrates are usually obtained through dry fractionation procedures, which involves preservation of the native protein structure at relatively high purity (~50% protein). However, dry fractionation also preserves high amount of non-protein components and antinutritional factors, which can impair protein functionality and nutritional value. In this work, pea protein concentrate (PPC) was hydrothermally treated by subcritical water to obtain enrichened pea protein fractions with improved technological and nutritional characteristics. To achieve this, PPC was solubilized at 10% (w/w) water with and without 0.4 M NaCl addition prior subcritical treatment. Afterwards, subcritical hydrolysis was conducted at 170 bar for 4 s at two different temperatures, 200 ºC (T1) and 340 ºC (T2). After the treatment, the treated PPCs was also fractionated to evaluate the ratio and composition of soluble and insoluble fractions and the whole, and soluble/insoluble fractions obtained were further dried for analyses. In the obtained protein concentrates, protein composition, structure and functionality were evaluated in terms of globulin/albumin composition (reducing and non-reducing SDS-PAGE), molecular weight and aggregation (HPLC-MALS), secondary structure (FTIR), degree of denaturation and stability (DSC, TGA), solubility, size and charge (Z-potential), foaming and heat-induced gelation ability. In addition, the presence of phytic acid, an antinutritional factor with high thermal resistance, was also evaluated. Results denoted that partial denaturation of protein occur, with modification of some of the protein subunits, probably due to slight hydrolysis during the short-time treatment. Futhermore, treatments at higher temperatures of the subcritical water led to higher degree of denaturation, especially if performed in the absence of salt. Meanwhile, water binding capacity and gelation of the ingredients was generally improved, at low temperature conditions (T1>T2) and salt-free treatments. Conversely, protein solubility improved under high-temperature conditions (T2>T1), with the greatest improvement observed in the presence of salt, altough the increase was also observed without salt addition.This would suggest that the partial hydrolysis of the fibre fraction also promoted the solubility of the protein within the matrix. Interestingly, subcritical water also increased the foaming ability of the treated PPC, although the long-term stability of the foams was reduced. Regarding phytic acid, the amount of free phosphorus increased in the soluble fractions, indicating phytic acid (inositol hexaphosphate) was hydrolyzed during subcritical water treatments. Therefore, results suggest that treatment of pea protein concentrate with subcritical water allows to improve the technological and nutritional value of pea protein, increasing the offer of available plant-protein ingredients in the market for oriented food applications and functionalities.

Purpose: Gymnema lactiferum (G. lactiferum) is a medicinal plant containing bioactive phytochemicals deeply rooted in traditional usage for treating various diseases. External adverse factors during processing and digestion can affect the quality and bioavailability of their bioactive compounds. Encapsulation is a viable option for targeted, controlled, and effective delivery of G. lactiferum extract with minimal loss of bioavailability. This technique also helps to mask undesirable odours and flavours while minimising exposure to non-targeted tissues. This is the first attempt of its kind to target systematic extraction and encapsulation of G. lactiferum bioactive compounds.

Methods: Ultrasonication was carried out using 50% ethanol:water (ethanol extract) and 100% water as extracting solvents at temperatures below 50°C. The extracts were analysed for total phenolic content (TPC), total flavonoid content (TFC), and total antioxidant capacity (TAC) to determine the optimal extraction method for encapsulation. To synthesise liposomes, the extract was dissolved in a mixture of soy lecithin and cholesterol (5:1) with Tween 80. The solvent mixture was evaporated under low pressure in a vacuum concentrator until a thin coating formed, then dissolved in a pH 7 phosphate buffer solution and homogenised using a high-shear mixer. The resulting liposome suspension was either filtered through a 0.22 µm membrane or filtered using a gel filtration column (Sephadex G 50). The physicochemical characteristics of the diluted liposome suspensions were analysed at 25°C. Statistical analysis was conducted using one-way analysis of variance (ANOVA), followed by Tukey’s test for post hoc comparison analyses.

Results: The highest TFC (3.53 ± 0.06a mg quercetin equivalents/g), TAC (555 ± 7a mg ascorbic acid equivalents/g) and a TPC value of 8.75 ± 0.15b mg gallic acid equivalents were obtained from the ethanol extract, which was the extraction method selected for encapsulation. The Z-average size, zeta potential and polydispersity index (PDI) values of the liposomes were 224.0 ± 0.2a nm, -26.17 ± 0.32a mV and 0.47 ± 0.05a for the gel column-filtered liposomes, and 486 ± 4b nm, -31 ± 7a mV and 0.490 ± 0.028a for the membrane filtered liposomes, respectively. The Z-average values were significantly different (p < 0.05) between methods whereas zeta potential and PDI values were not significantly different from each other.

Conclusion: Ultrasonication using ethanol as the extraction method is more effective in extracting flavonoids and antioxidative compounds from G. lactiferum compared to water extraction. Gel column-filtered liposomes showed more uniform and monodisperse size distribution compared to membrane-filtered liposomes. This suggests controlled particle size and better stability in gel-filtered liposomes, with membrane-filtered liposomes showing a more heterogeneous distribution

Keywords: G. lactiferum extraction, Encapsulation, Liposomes, Bioactive compounds

Introduction: Cinnamomum zeylanicum is a plant rich in bioactives such as cinnamaldehyde and has been associated with several health benefits, including anti-diabetic, anti-inflammatory, antioxidant, and anti-carcinogenic properties. However, bioactive compounds like cinnamaldehyde are very sensitive to environmental factors (e.g., temperature, light, and pH) and can degrade under such conditions. Encapsulation of these bioactives can be a simple solution to the degradation issues during storage and processing, while also ensuring controlled release at the target site and improving bioavailability.

Methods: In this study, cinnamon was extracted in different ethanolic concentrations (25,50,75,100%) using two different extraction methods; ultrasonic and accelerated solvent extraction. The obtained extracts were then analysed for total phenols, flavonoids, and antioxidant activity using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. The extracts were also analysed for the presence of cinnamaldehyde, cinnamic acid, and eugenol by high-performance liquid chromatography to obtain the best extract for encapsulation. The optimal cinnamon bark extract was then chosen for creating liposomes using cholesterol and soy lecithin. The liposomes were also characterised for z-average mean diameter, zeta potential, and polydispersity index using a Zetasizer Nano (Malvern Instruments Ltd).

Results: The total phenols and flavonoids of the accelerated solvent extracts were significantly higher than those of the ultrasonicated extracts of the same ethanolic concentration (p < 0.05). However, there was no significant difference between the DPPH scavenging activity of the two types of extraction (p > 0.05). The highest total phenols (6.83 ± 0.31 mg/g gallic acid equivalent) and total flavonoids (50.08 ± 0.26 mg/100g quercetin equivalent) were observed with the cinnamon barks extracted with 50% ethanol. The highest cinnamaldehyde (19.33 mg/g dry cinnamon), cinnamic acid (0.18 mg/g dry cinnamon), and eugenol (10.57 mg/g dry cinnamon) content were also seen with 50% ethanolic cinnamon extracted using an accelerated solvent extractor. The particle size of the liposome significantly increased (p < 0.05) with the addition of the cinnamon extract (from 213.1 to 219.8 nm). The addition of cinnamon extract also slightly reduced the negative zeta potential (from -27.56 to -23.26 mV). This change could be due to the interaction of the bioactive components in the extract, such as cinnamaldehyde, with the liposomal surface or the lipid bilayer, altering the surface charge. The values indicate a moderately stable liposome formulation.

Conclusions: Overall, the primary findings of this research demonstrated that using 50% ethanol in accelerated solvent extraction yielded the highest levels of phenols, flavonoids, and key bioactives. Also, liposomes formed with soy lecithin produced a moderately stable encapsulation system, offering a promising approach to enhance the bioavailability of extracts. Characterises such as encapsulation efficiency, stability, and release behaviour of key bioactives are yet to be understood. Further research is also underway to study the structure, distribution and morphology of the formed liposomes under the transmission and scanning electron microscope. Following this, the encapsulated extract will be incorporated into a low-fat yoghurt, to study the effect on the physicochemical and rheological properties, as well as the digestion behaviour (in vitro) of this yoghurt and the release mechanisms of the bioactive compounds.

Keywords: Cinnamaldehyde; Encapsulation efficiency; Cinnamon; Bioactive compounds

Despite ongoing research, gluten-free ingredients to produce wheat-free bakery products capable of mimicking the texture and quality of wheat-based products have yet to be developed. The challenge to mimic wheat-based product quality arises from the characteristics of gluten-free batters, which impede the proper expansion of air bubbles essential for increasing bread volume. While some modern gluten-free alternatives increase batter viscosity, they remain short in providing the elasticity necessary for optimal performance. Balancing viscosity and elasticity is crucial for developing successful gluten-free alternatives. This study conducted a rheological investigation to explore the feasibility of employing a dry conjugation technique for generating conjugates of soy protein isolate, pea protein isolate, chickpea protein concentrate, or zein protein, on the one hand, and psyllium husk gum, on the other hand. The primary objective was to use those conjugates to enhance the quality attributes of gluten-free bread. All four psyllium husk gum-protein conjugates (PPCs) were synthesized at a 3:1 ratio, at 60°C and 75% relative humidity for 3, 7, 11, and 15 days. Their effect on the rheological properties of gluten-free batter was subsequently tested. Confirmation of PPC formation was achieved through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier Transform Infrared Spectroscopy (FTIR). The findings indicated that 11 days at 60°C exhibited the most significant effect on the molecular weight of PPCs as assessed by SDS-PAGE. Furthermore, rheology results showed that chickpea protein-psyllium husk gum conjugate is a promising ingredient for gluten-free bread formulations.

Plant-based proteins continue to gain popularity as a sustainable alternative to animal proteins, due to their nutritional, functional, and health benefits. Meanwhile, dry and wet fractionation methods are increasingly being used for production of value-added pulse protein ingredients.

This study aims to compare nutritional properties, protein quality, secondary structure, and physico-chemical properties of Australian faba bean and yellow pea protein concentrates and protein isolates obtained by dry and wet fractionation. Amino acid scores highlighted that faba bean and yellow pea protein isolates and concentrates were deficient in the sulphur-containing amino acids. Faba bean (63.7%) and yellow pea protein (63.0%) isolates had higher in-vitro protein digestibility-corrected amino acid scores compared to the protein concentrates. Fourier-transform infrared spectroscopy revealed different secondary structures between protein concentrates and isolates, especially for amide I region. Faba bean and yellow pea protein concentrates had higher protein solubility (46.2 and 50.1%, respectively), and higher foaming capacity (65 and 59%, respectively), compared to the protein isolates. Water-holding capacity was higher for faba bean and yellow pea protein isolates, being 4.3 and 4.0 g/g, respectively.

These findings demonstrate that Australian faba bean and yellow pea protein concentrates and isolates have unique functional properties that can be exploited for use in a diverse range of new and existing food applications. This also provides food manufacturers with ingredient options to help with their food formulations for new and existing foods.

Gum Arabic (GA), a natural polysaccharide derived from the exudates of Acacia tree species in the Sehal belt, is widely utilised in food, pharmaceuticals, and cosmetics due to its emulsifying, stabilising, and encapsulating properties, which allows it to uniformly incorporate sensitive bioactive compounds. This makes GA a valuable component in developing innovative delivery systems aimed at optimising the efficacy of functional ingredients across various applications.

When dried slowly in nature, GA self-organises into a glassy state. However, for industrial applications, it is typically spray-dried from non-entangled, low viscosity solution, resulting in a powdery precipitate. The drying kinetics plays a crucial role in determining the structural characteristics of GA assembly. The glassy state, with its dense molecular packing, contrasts with the powdery state, which is characterised by structural defects across multiple length scales, from molecular to micro-structural. These structural differences between both amorphous states significantly influence dissolution kinetics, with the glassy state expected to dissolve more slowly and uniformly than the spray-dried powder. The dissolution kinetics of GA in its glassy state could provide controlled delivery of bioactive compounds. Aiming to reproduce this glassy state via spray drying provides unique challenges and opportunities. Understanding fundamental droplet drying behaviour of GA solutions however is crucial in this endeavour and remains unknown. By simulating GA droplets at different stages of the drying journey, this project aims to understand the dynamic process of spray drying. This is the essential first step before physiochemically-altering methods such as enzymatic hydrolysis can be implemented to attempt to achieve glassy states via spray drying.

Drying kinetics are influenced by processes such as evaporation and diffusion, which contribute to molecular jamming and entanglement and thus determine the amorphous solid state. Therefore, this research aims to first understand GA’s relationship to water in solution and how the bio polymer reacts to evaporation. Insights on the transition from solution to dried particulate occurring during spray drying were gained using a concentration-dependent approach to analyse drying processes of GA solutions. Techniques such as Thermogravimetric and Moisture Balance analysis were used to provide key insights to understand this evaporation behaviour. Because the GA droplet increases in concentration over the course of spray drying as evaporation occurs, this study analysed a range of GA solutions from low to previously unstudied high concentrations (10 – 80% w/w). It was revealed that highly concentrated GA solutions, notably around 65% (w/w) exhibit complex evaporation behaviour indicative of water entrapment. This suggests that close molecular packing, which is needed to produce a dense glassy matrix, could be hindered as delayed evaporation across the droplet causes heterogeneity in drying and pockets of free space. This research underscores an important starting place in understanding drying kinetics and structure-property relationships in GA for enhancing its application in delivery systems.