The Effect of Precipitation pH on Protein Recovery Yield and Emulsifying Properties in the Extraction of Protein from Cold-Pressed Rapeseed Press Cake.

Rapeseed is the second most cultivated oilseed after soybean and is mainly used to produce vegetable oil. The by-product rapeseed press cake is rich in high-quality proteins, thus having the possibility of becoming a new plant protein food source. This study aimed to investigate how the precipitation pH affects the protein yield, protein content, and emulsifying properties when industrially cold-pressed rapeseed press cake is used as the starting material. Proteins were extracted under alkaline conditions (pH 10.5) with an extraction coefficient of 52 ± 2% followed by precipitation at various pH (3.0-6.5). The most preferred condition in terms of process efficiency was pH 4.0, which is reflected in the zeta potential results, where the proteins' net charge was 0 at pH 4.2. pH 4.0 also exhibited the highest protein recovery yield (33 ± 0%) and the highest protein concentration (64 ± 1%, dry basis). Proteins precipitated at pH 6.0-6.5 stabilized emulsions with the smallest initial droplet size, although emulsions stabilized by rapeseed protein precipitated at pH 5.0-6.0 showed the highest emulsion stability at 37 °C for 21 days, with a limited layer of free oil. Overall, emulsion stabilized by protein precipitated at pH 5.0 was the most stable formulation, with no layer of free oil after 21 days of incubation.

Starch granule stabilized Pickering emulsions: an 8-year stability study.

BACKGROUND: Pickering emulsions are known to have advantages over conventional emulsions, in particular, improved and long-term stability against coalescence. This research is an eight-year stability investigation of oil-in-water Pickering emulsions stabilized by quinoa starch granules modified by octenyl succinic anhydride (OSA). Two different concentrations of starch (i.e. 200 and 600 mg mL-1 based on oil) were used at oil fraction (ϕ) of 0.1. The emulsions were prepared using a high-pressure homogenizer. The emulsions were stored in a refrigerator (at 6 °C) and evaluated using particle size analyzer over the storage period and light microscopy at the end of the storage period. RESULTS: Starch granule stabilized Pickering emulsions produced by a high-pressure homogenizer displayed remarkable storage stability over the eight years with no indication of coalescence. In addition, the results showed that increasing the concentration of starch granules resulted in a decrease in droplet sizes. The sizes measured by the particle size analyzer showed a decline over the storage period which was due to dissociation of some networks of aggregated droplets that, in addition to representing smaller droplet sizes, resulted in the release of free and unbound starch entrapped in these networks. CONCLUSIONS: This study showed that Pickering emulsions produced by OSA modified starch granules from quinoa can be used in practical applications for the development of highly stable formulations when prolonged storage is required. © 2020 Society of Chemical Industry.

Chemical methods and techniques to monitor early Maillard reaction in milk products; A review.

Maillard reaction is an extensively studied, yet unresolved chemical reaction that occurs as a result of application of the heat and during the storage of foods. The formation of advanced glycation end products (AGEs) has been the focus of several investigations recently. These molecules which are formed at the advanced stage of the Maillard reaction, are suspected to be involved in autoimmune diseases in humans. Therefore, understanding to which extent this reaction occurs in foods, is of vital significance. Because of their composition, milk products are ideal media for this reaction, especially when application of heat and prolonged storage are considered. Thus, in this work several chemical approaches to monitor this reaction in an early stage are reviewed. This is mostly done regarding available lysine blockage which takes place in the very beginning of the reaction. The most popular methods and their applications to various products are reviewed. The methods including their modifications are described in detail and their findings are discussed. The present paper provides an insight into the history of the most frequently-used methods and provides an overview on the indicators of the Maillard reaction in the early stage with its focus on milk products and especially milk powders.

Characteristics and functionality of appetite-reducing thylakoid powders produced by three different drying processes.

BACKGROUND: Thylakoids, a chloroplast membrane extracted from green leaves, are a promising functional ingredient with appetite-reducing properties via their lipase-inhibiting effect. Thylakoids in powder form have been evaluated in animal and human models, but no comprehensive study has been conducted on powder characteristics. The aim was to investigate the effects of different isolation methods and drying techniques (drum-drying, spray-drying, freeze-drying) on thylakoids' physicochemical and functional properties. RESULTS: Freeze-drying yielded thylakoid powders with the highest lipase-inhibiting capacity. We hypothesize that the specific macromolecular structures involved in lipase inhibition were degraded to different degrees by exposure to heat during spray-drying and drum-drying. We identified lightness (Hunter's L-value), greenness (Hunter's a-value), chlorophyll content and emulsifying capacity to be correlated to lipase-inhibiting capacity. Thus, to optimize the thylakoids functional properties, the internal membrane structure indicated by retained green colour should be preserved. This opens possibilities to use chlorophyll content as a marker for thylakoid functionality in screening processes during process optimization. CONCLUSION: Thylakoids are heat sensitive, and a mild drying technique should be used in industrial production. Strong links between physicochemical parameters and lipase inhibition capacity were found that can be used to predict functionality. The approach from this study can be applied towards production of standardized high-quality functional food ingredients. © 2017 Society of Chemical Industry.

High-Pressure Pasteurization of Oat Okara.

The issue of the short microbiological shelf life of residues from the plant-based beverage industry creates a large food waste problem. Today, the oat beverage residue, in this study referred to as oat okara, is generally converted to energy or used as animal feed. High-pressure pasteurization (200 MPa, 400 MPa, and 600 MPa) was applied to oat okara to investigate the effect on shelf life and microbiological activity. A 4-week microbiological storage study was performed and thermal properties, viscosity, and water and oil holding capacities were analyzed. The total aerobic count, including yeast and mold, was significantly reduced (p < 0.05) by 600 MPa after four weeks of storage at 4 °C. The content of lactic acid bacteria after four weeks of storage was low for untreated oat okara (3.2 log CFU/g) but, for 600 MPa, the content remained at the detection limit (2.3 log CFU/g). Conversely, the treatments of 200 MPa and 400 MPa increased the microbial content of the total aerobic count significantly (p < 0.05) after two weeks in comparison to untreated oat okara. The thermal properties of untreated and high-pressure-treated oat okara demonstrated an increase in protein denaturation of the 12S globulin, avenalin, when higher pressure was applied (400-600 MPa). This was also confirmed in the viscosity measurements where a viscosity peak for avenalin was only present for untreated and 200 MPa treated oat okara. The water holding capacity did not change as a function of high-pressure treatment (3.5-3.8 mL/g) except for the treatment at 200 MPa, which was reduced (2.7 mL/g). The oil holding capacity was constant (1.2-1.3 mL/g) after all treatments. High-pressure pasteurization of 600 MPa reduced the microbial content in oat okara resulting in a shelf life of 2-4 weeks. However, more research is required to identify the microorganisms in oat okara to achieve a microbiologically safe product that can be used for food applications.

High-Pressure Pasteurization of Soy Okara.

Okara is a by-product from the production of soy beverages, which has a high content of protein and fiber. Even though it has a high nutritional value, it is generally discarded or used as animal feed or compost. The problem is its short shelf life due to its high water content and high water activity. The aim of this study was to investigate the effect of high-pressure pasteurization at 200 MPa, 400 MPa, and 600 MPa on the shelf life of soy okara. Microbiological growth, as well as thermal properties, viscosity, water holding capacity, and oil holding capacity, was evaluated after the pressure treatments. Treatment at 600 MPa significantly reduced (p < 0.05) the growth of total aerobic count, yeast and mold, and lactic acid bacteria for up to four weeks of storage at 4 °C. The pasting properties were increased while the water and oil holding capacities of the soy okara did not significantly change (p > 0.05) after high-pressure pasteurization at 400 MPa and 600 MPa. High-pressure pasteurization is therefore a potential application technique for soy okara to produce a microbiologically safe product with maintained functional properties. However, more research is needed to optimize the process and to further investigate the microbiological species present in untreated soy okara to exclude any potential food safety risks.

Synthesis of controlled-size starch nanoparticles and superparamagnetic starch nanocomposites by microemulsion method.

In this study, a synthesis process based on the microemulsion method (ME) was developed with the aim to produce controlled-size starch nanoparticles (SNPs). Several formulations were tested for the preparation of the W/O microemulsions varying the organic/aqueous phase ratios and co-stabilizers concentrations. SNPs were characterized in terms of size, morphology, monodispersity and crystallinity. Spherical shape particles with mean sizes 30-40 nm were prepared. The method was then used to simultaneously synthesize SNPs and iron oxide nanoparticles with superparamagnetic properties. Starch-based nanocomposites with superparamagnetic properties and controlled size were obtained. Therefore, the microemulsion method developed could be considered an innovative technology for the design and development of novel functional nanomaterials. The starch-based nanocomposites were evaluated in terms of morphology and magnetic properties, and they are being considered as promising sustainable nanomaterials for different biomedical applications.

Quinoa starch granules: a candidate for stabilising food-grade Pickering emulsions.

BACKGROUND: Particle-stabilised emulsions, so-called Pickering emulsions, are known to possess many beneficial properties, including being extremely stable. Starch granules isolated from quinoa have been used as emulsion stabilising particles. The granules were intact, 1-3 µm in diameter and modified with octenyl succinic anhydride to increase their hydrophobicity. Starch granules, as opposed to most other particles used to generate Pickering emulsions, are edible, abundant and derived from natural sources. RESULTS: Emulsions produced by high shear homogenisation had droplet sizes of 9-70 µm depending on the starch-to-oil ratio. Droplet size decreased with increasing starch-to-oil ratio, but was unaffected by the oil phase volume over a range of 5-33% oil (v/v). Although the drops were large and subject to creaming, their size remained unchanged over a period of 7 days. By adjusting the starch-to-oil ratio drops could be made to be buoyancy neutral to prevent creaming. Rheological characterisation indicated a gel structure with an elastic modulus in the range 200-2000 Pa depending on droplet size. CONCLUSION: This work has demonstrated the successful use of starch granules to stabilise emulsions which may find applications beyond that of food, for example in cosmetics and pharmaceutical formulations.

Plant-Based Meat Analogues from Alternative Protein: A Systematic Literature Review.

This study aimed to conduct a systematic literature review (SLR) of the research performed in the plant-based meat analogues area. Historical, current, and future tendencies are discussed. The paper offers a comprehensive SLR coupled with a bibliometric analysis of the publication from 1972 to January 2022. The articles were obtained using a research string and precise inclusion and exclusion criteria from two prominent databases, Scopus and Web of Science (WoS). The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) flow technique was used to describe the data screening and selection. In total, 84 publications were selected for further analysis after a thorough literature assessment. From this study, six main themes were identified: (1) objectives of the study; (2) type of plant protein; (3) product type; (4) added ingredients; (5) texturization technique; and (6) quality assessment considered in the studies. Recent trends in publication imply that meat analogue technology is gaining prominence. This review revealed significant research on improving meat analogues via texturization. Even though extrusion is used industrially, the technique is still in its infancy and needs improvement. Future studies should focus more on fiber and protein-protein interactions, macromolecule conformation and mechanisms, diversifying or improving current methods, sensory attributes, and gastrointestinal absorption rate of each novel protein ingredient.

Development and characterization of medium and high internal phase novel multiple Pickering emulsions stabilized by hordein nanoparticles.

Medium and high internal phase W1/O/W2 multiple Pickering emulsions (MPEs) were fabricated by physically-modified hordein nanoparticles. A triphasic system was developed at dispersed phase volume fraction (Φ) of 0.5 with an overrun value of ∼40%. No overrun was detected in high internal phase MPEs (Φ 0.8). Optical and confocal laser scanning microscopy confirmed the formation of MPEs. Monomodal droplet size distribution with a mean diameter of 32.90 and 21.48 µm was observed for MPEs at Φ 0.5 and Φ 0.8, respectively. Static multiple light scattering confirmed that creaming was the main mechanism behind the instability of MPEs. Both MPEs revealed pseudo-plastic behavior and predominant storage modulus (G') over the applied frequency range. The encapsulation efficiency of vitamin B12 in MPEs was 98.3% and remained relatively constant during 28 d. These results suggested the excellent potential of hordein nanoparticles as appropriate candidate for designing multi-structural colloidal systems using plant proteins.

Protein extraction from cold-pressed hempseed press cake: From laboratory to pilot scale.

During the production of industrial hempseed oil, a press cake is formed as a byproduct, which is often used as animal feed although it contains a high amount of protein that could be used for human consumption. Extracting this valuable protein would reduce food waste and increase the availability of plant-based protein. A protein extraction process based on the pH-shift method was adapted to improve the protein extraction yield from industrial hempseed press cake (HPC). Parameters such as alkali extraction pH, time, and temperature, as well as isoelectric precipitation pH, were investigated in laboratory scale and were thereafter carried out in a pilot trial to explore the suitability for future scale up. The phytic acid content of the extracted protein isolate was also analyzed to investigate any potential inhibitory effect on mineral absorption. A final protein yield of 60.6%, with a precipitated protein content of 90.3% (dw), was obtained using a constant alkali extraction pH of 10.5 for 1 h at room temperature, followed by precipitation at pH 5.5. The pilot trial showed promising results for the future production of industrial hemp protein precipitate on a larger scale, showing a protein yield of 57.0% and protein content of 90.8% (dw). The amount of phytic acid in the protein isolate produced in the optimal laboratory experiment and in the pilot trial was 0.595 and 0.557 g phytic acid/100 g dw, respectively, which is 83%-88% less than in the HPC. This is in the range of other plant-based protein sources (tofu, kidney beans, peas, etc.). PRACTICAL APPLICATION: Industrial hempseed press cake is a byproduct in the production of industrial hempseed oil, which is mostly used as animal feed, but has the potential to become an additional source of plant-based protein for human consumption with a suitable protein extraction method. The extracted hemp protein could be used to develop new plant-based dairy or meat analog products.

Chloroplast thylakoids reduce glucose uptake and decrease intestinal macromolecular permeability.

Thylakoid membranes, derived from chloroplasts, have previously been shown to retard fat digestion and lower blood glucose levels after oral intake. The purpose of the present study was to investigate the effect of thylakoid membranes on the passage of methyl-glucose, dextran and ovalbumin over rat intestine in vitro using Ussing chambers. The results show that thylakoids retard the passage of each of the test molecules in a dose-dependent way. The thylakoids appear to be attached on the mucosal surface and a mechanism is suggested that the thylakoids delay the passage of the test molecules by sterical hindrance. The present results indicate that thylakoid membranes may be useful both to control intestinal absorption of glucose and to enhance the barrier function of the intestine.

Chloroplast thylakoid membrane-stabilised emulsions.

BACKGROUND: Thylakoid-stabilised emulsions have been reported to possess satiety-promoting effects and inhibit pancreatic lipase-colipase activity in vitro, which prompted the investigation of their interfacial properties. RESULTS: Thylakoid membranes isolated from spinach were used as an emulsifier/stabiliser in oil (triglyceride)-in-water emulsions. Emulsions were characterised with respect to droplet size, interfacial tension, creaming, surface load and electron microscopy. The effects of pH and thylakoid concentration were also considered. Droplet size decreased with increasing thylakoid concentration, reaching a plateau around 15 microm beyond concentrations of 2 mg protein mL(-1) oil. The resulting emulsions were stable against coalescence but were subject to creaming. The surface pressure (air/water interface) of the thylakoid isolate was 44 mN m(-1) and the surface load 13 mg m(-2) at 10 mg protein mL(-1) oil. Electron micrographs showed thylakoids adsorbed as bunched vesicles on the drop surfaces. The stabilisation mechanism can be described as a combined effect of surface-active molecules, mainly membrane proteins but also membrane lipids, exposed on surfaces of thylakoid membrane vesicles adsorbed as particles. CONCLUSION: Thylakoid membranes effectively stabilise oil-in-water emulsions, which should facilitate their incorporation in food with satiety-promoting effects. To the authors' knowledge, this is the first study on the emulsifying properties of an isolated biological membrane as a functional ingredient.

Development and Characterization of Extrudates Based on Rapeseed and Pea Protein Blends Using High-Moisture Extrusion Cooking.

Rapeseed protein is not currently utilized for food applications, although it has excellent physicochemical, functional, and nutritional properties similar to soy protein. Thus, the goal of this study was to create new plant-based extrudates for application as high-moisture meat analogs from a 50:50 blend of rapeseed protein concentrate (RPC) and yellow pea isolate (YPI) using high-moisture-extrusion (HME) cooking with a twin-screw extruder to gain a better understanding of the properties of the protein powders and resulting extrudates. The effects of extrusion processing parameters such as moisture content (60%, 63%, 65%, 70%), screw speed (500, 700, and 900 rpm), and a barrel temperature profile of 40-80-130-150 °C on the extrudates' characteristics were studied. When compared to the effect of varying screw speeds, targeted moisture content had a larger impact on textural characteristics. The extrudates had a greater hardness at the same moisture content when the screw speed was reduced. The specific mechanical energy (SME) increased as the screw speed increased, while increased moisture content resulted in a small reduction in SME. The lightness (L*) of most samples was found to increase as the target moisture content increased from 60% to 70%. The RPC:YPI blend was equivalent to proteins produced from other sources and comparable to the FAO/WHO standard requirements.

Emulsifying and Anti-Oxidative Properties of Proteins Extracted from Industrially Cold-Pressed Rapeseed Press-Cake.

One of the functional proteins in rapeseed-the amphiphilic protein oleosin-could be used to stabilize emulsions. The objectives of this study were to extract oleosins from cold-pressed rapeseed press-cake, optimize the extraction process, and investigate their emulsifying and anti-oxidative capacity. The proteins were recovered from industrially cold-pressed rapeseed press-cake at different alkali pHs. Emulsifying properties and oxidation rates were assessed. Oleosin extracted at pH 9 stabilized smaller emulsion droplets than oleosin extracted at pH 12, although the protein yield was higher at pH 12. Emulsions were formulated from flaxseed oil and corn oil and were stabilized by oleosin, bovine serum albumin, de-oiled lecithin and Tween 20,h and the emulsions were stored in accelerated conditions (30 °C) for 12 days. Oleosin stabilized emulsions to the same extent as commercial food-grade emulsifiers. Flaxseed oil emulsions stabilized by oleosin had a significantly lower concentration of malondialdehyde (MDA) which indicates a lower oxidation rate compared to BSA, de-oiled lecithin and Tween 20. For corn oil emulsions, oleosin and BSA had a similar capacity to delay oxidation and were significantly more efficient compared to de-oiled lecithin and Tween 20. Rapeseed oleosin recovered from cold-pressed rapeseed press-cake could be a suitable natural emulsifier with anti-oxidation properties.

Effects of Storage Conditions on Degradation of Chlorophyll and Emulsifying Capacity of Thylakoid Powders Produced by Different Drying Methods.

Thylakoid membranes isolated from spinach have previously been shown to inhibit lipase/co-lipase and prolong satiety in vivo. There is a need to develop thylakoid products that not only have the desired characteristics and functionality after processing, but also are stable and provide equivalent effect on appetite over the promised shelf life. The aim of the present study was therefore to evaluate how the thylakoid powders' characteristics and functionality were affected by moisture during storage. Thylakoids produced by drum-drying, spray-drying, and freeze-drying were incubated in controlled atmosphere with different relative humidity (10 RH%, 32 RH%, 48 RH% and 61 RH%) for 8 months. The water content in all powders was increased during storage. The water absorption was moisture-dependent, and the powders were considered hygroscopic. Relative humidity showed a definite influence on the rate of chlorophyll degradation and loss of green color in thylakoid powders after storage which correlated with impaired emulsifying capacity. Spray-dried powder had the overall highest chlorophyll content and emulsifying capacity at all RH-levels investigated. Spray drying was therefore considered the most suitable drying method yielding a powder with best-maintained functionality after storage. The results can be applied towards quality control of high-quality functional foods with appetite suppressing abilities.

Comparison of Three Methods to Determine the Degree of Substitution of Quinoa and Rice Starch Acetates, Propionates, and Butyrates: Direct Stoichiometry, FTIR, and 1H-NMR.

Rice and quinoa starch esters were prepared by acylation using short-chain fatty acid anhydrides with different chain lengths (acetic, propionic, and butyric anhydride). A direct stoichiometric method based on the acylation reaction was used to determine the degree of substitution (DS) and acyl content (AC). In addition, Fourier-transform infrared spectroscopy (FTIR) was used to validate the conformational changes of acylated starch and 1H-NMR was used as a DS reference method. DS by stoichiometric calculation was shown to be in agreement with FTIR and was comparable with DS obtained from Proton nuclear magnetic resonance (1H-NMR). Based on this study, stoichiometric calculation allows rapid and direct determination of substitution levels and acyl content without the loss of samples, which provides efficiency and optimization of manufacturing procedures in producing the desired level of esterified starches.

Development of High-Moisture Meat Analogues with Hemp and Soy Protein Using Extrusion Cooking.

The interest in plant-based products is growing in Western countries, mostly due to health and environmental issues that arise from the consumption and production of animal-based food products. Many vegan products today are made from soy, but drawbacks include the challenges of cultivating soy in colder climates such as northern Europe. Therefore, the present study investigates whether industrial hemp (Cannabis sativa) could substitute soy in the production of high moisture meat analogues (HMMA). A twin screw co-rotating extruder was used to investigate to what extent hemp protein concentrate (HPC) could replace soy protein isolate (SPI) in HMMAs. The substitution levels of HPC were 20 wt%, 40 wt% and 60 wt%. Pasting properties and melting temperature of the protein powders were characterized by Rapid Visco Analyzer (RVA) and Differential Scanning Calorimeter (DSC), respectively and the produced HMMA was analysed by determining the texture and colour attributes. The results showed that it is possible to extrude a mixture with up to 60% HPC. HPC absorbed less water and needed a higher denaturing temperature compared to SPI. Increasing the moisture content by 5% would have resulted in a reduction of hardness and chewiness. The lightness (L* value) was found to be significantly higher in SPI product and decreased in the mixture with higher HPC (p < 0.05).

Thylakoids promote release of the satiety hormone cholecystokinin while reducing insulin in healthy humans.

OBJECTIVE: The effects of a promising new appetite suppressor named "thylakoids" (membrane proteins derived from spinach leaves) were examined in a single meal in man. Thylakoids inhibit the lipase/colipase hydrolysis of triacylglycerols in vitro and suppress food intake, decrease body-weight gain and raise the satiety hormone cholecystokinin (CCK) in rats, but their effects in man remain unclear. The aim of this study was to investigate whether thylakoids, when added to a test meal, affect appetite regulation and blood parameters in healthy individuals. MATERIAL AND METHODS: In an intervention crossover study, healthy individuals of normal weight (n=11) were offered a high-fat meal with and without the addition of thylakoids. Blood samples were taken 0 (prior to meal), 30, 60, 120, 180, 240, 300 and 360 min after the start of the meal. Blood samples were analysed for satiety and hunger hormones (CCK, leptin and ghrelin), insulin and blood metabolites (glucose and free fatty acids). RESULTS: The CCK level increased, in particular between the 120 min time-point and onwards, the ghrelin level was reduced at 120 min and leptin level increased at 360 min after intake of the thylakoid-enriched meal. The insulin level was reduced, whereas glucose concentrations were unchanged. Free fatty acids were reduced between time-point 120 min and onwards after the thylakoid meal. CONCLUSIONS: The addition of thylakoids to energy-dense food promotes satiety signals and reduces insulin response during a single meal in man.

The Effects of Oil Extraction Methods on Recovery Yield and Emulsifying Properties of Proteins from Rapeseed Meal and Press Cake.

The agricultural sector is thought to be responsible for around 30% of the anthropogenic climate change and it is well established that high meat consumption has a tremendous impact on the environment. Rapeseed is mainly used for production of vegetable oil, but press cake has high protein content with the potential for incorporation into new plant protein-based foods. Protein was recovered from press cakes generated from different oil pressing processes. Industrially cold-pressed, hot-pressed, and solvent-extracted rapeseed press cake and the effect of heat treatment in the recovery process was assessed. Protein recovery yield, protein concentration and emulsifying properties were analyzed. Cold-pressed rapeseed press cake (RPC) recovered in the absence of heat, yielded the highest protein recovery (45%) followed by hot-pressed rapeseed meal (RM) (26%) and solvent-extracted RM (5%). Exposure to heat during recovery significantly reduced the yield for cold-pressed RPC but no difference was found for hot-pressed RM. The protein recovery yield was improved for solvent-extracted RM when heat was applied in the recovery process. The ability to stabilize emulsions was highest for protein recovered from cold-pressed RPC, followed by hot-pressed RM and solvent-extracted RM, and was in the same range as commercial emulsifying agents. Heat treatment during recovery significantly reduced the emulsifying properties for all pressing methods examined. This study suggests that cold-pressed rapeseed press cake without heat in the recovery process could be a successful strategy for an efficient recovery of rapeseed protein with good emulsifying properties.