Yeast protein-containing compositions
A stable liquid composition with high yeast protein content is achieved by combining a fat component and non-proteic thickening agent, addressing stability issues in yeast protein emulsions, ensuring phase stability and sensory quality for food and beverage applications.
Patent Information
- Application Number
- PCT/EP2025/066360
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2024-06-13
- Filing Date
- 2025-06-12
- Publication Date
- 2025-12-18
AI Technical Summary
Existing technologies face challenges in preparing stable, high-protein liquid compositions, particularly emulsions, using yeast proteins, due to issues such as phase separation, sedimentation, gelation, and fouling during processing and storage, which are exacerbated by high protein content and the presence of fat.
A liquid composition comprising a fat component, non-proteic thickening agent, and a yeast protein source with at least 40wt.% yeast proteins, combined with optional additional protein sources, is prepared through dispersion, addition of a fat component, and homogenization, followed by heat-treatment to stabilize the emulsion.
The composition remains stable during processing and storage, preventing phase separation, sedimentation, and gelation, while maintaining good sensory properties and nutritional quality, suitable for use in various food and beverage products.
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Abstract
Description
[0001] YEAST PROTEIN-CONTAINING COMPOSITIONS
[0002] TECHNICAL FIELD
[0003] The present invention relates generally to the field of yeast protein-based compositions. In particular, the present invention also relates to liquid composition comprising yeast protein source; a powder composition obtained by drying such liquid emulsions; methods and uses involving said liquid compositions or powder compositions; processes for preparing such liquid compositions or powder compositions; and products comprising said liquid compositions or powder compositions.
[0004] BACKGROUND OF THE INVENTION
[0005] In recent years, the need to address pressing global challenges such as food security and sustainability, due to the world population being estimated to increase from the current 8 to almost 10 billion by 2050, has pushed food companies and academic groups towards the identification of alternative protein sources with the potential to replace animal-based ones in food products. Most of the efforts in this direction have so far focused on plant proteins, with a variety of liquid (e.g., ready-to-drink beverages, coffee creamers), semi-solid (e.g., yogurts, cooking creams) and solid foods (e.g., meat and fish analogues) based on these ingredients having been launched on the market (D. J. McClements & Grossmann, 2021).
[0006] While increasing the use of plant proteins in human nutrition will be of paramount importance in the near future in order to ensure protein supply in a sustainable manner, processing of plant-based foods is often challenging, even more so when it comes to liquid products such as ready-to-drink (RTD) beverages (Qamar, Manrique, Parekh, & Falconer, 2020). This is due to the fact that most plant protein ingredients are characterized by highly ordered tertiary and quaternary structures, and thus low solubility in water, which often results in poor overall functionality, unless these are subjected to specific treatments, such as physical, chemical, enzymatic ones, or combinations thereof (Amagliani, Silva, Saffon, & Dombrowski, 2021). Furthermore, heat treatments of plant proteins above their denaturation temperature (e.g., UHT) may trigger the occurrence of phenomena such as sedimentation, fouling and / or gelation inside the heat exchangers, major hurdles towards the development of shelf-stable beverages, particularly when their target protein concentration is higher than those typical of milk analogues (i.e., ~l-3 wt%) (David Julian McClements, Newman, & McClements, 2019). It should also be taken into consideration that, with regard to plant-based beverages, meeting consumer expectations in terms of nutritional and sensory properties is a complex exercise, with only proteins derived from soy or potato displaying a biological value similar to that of animal proteins such as milk- and egg-based ones (Day, 2013), and with many commercial products having low levels of acceptance due to their poor flavour and mouthfeel (Moss et al., 2022).
[0007] In addition to plant proteins, single cell proteins (SCPs) represent a promising option for the development of sustainable, nutrient-rich food products which could enable to cater for the growing world population. The term SCPs refers to proteins derived from microorganisms such as bacteria, fungi, yeasts or algae. These are obtained via fermentation and offer several advantages, including (i) rapid growth rates (typically 1 to 4 days), thus ensuring fast and efficient protein production; (ii) reduced land and water requirements compared to livestock farming, since fermentation is typically performed in bioreactors, as well as lower greenhouse gas emissions; (iii) ability to grow on diverse feedstocks, including a variety of agri-food by-products, which enables the valorization and efficient utilization of resources that might otherwise go to waste, thus reducing environmental impact and promoting sustainability; (iv) high protein concentration, with values which in most cases range from about 40 to >70 wt% on a dry weight basis, depending on microorganism, species and fermentation conditions used (Ritala, Hakkinen, Toivari, & Wiebe, 2017).
[0008] Concerning human nutrition, yeasts are by far the most interesting among SCPs, due to (i) their established history of food use (e.g., spreads, dietary supplements, as processing aids in alcoholic fermentation and baking, and as flavouring agents), which facilitates consumer acceptance; (ii) regulatory aspects, with two species (i.e., Saccharomyces cerevisiae and Candida utilis or Cyberlindnera jadinii, commonly known as Torula) being approved for use in human food in both Europe and the United States; (iii) their good commercial availability (Jach, Serefko, Ziaja, & Kieliszek, 2022; Ma et al., 2023).
[0009] The preparation of high protein liquid compositions, in particular liquid emulsions with yeast protein ingredients have not been reported in the literature. However, there is a significant interest in developing such liquid compositions, in particular emulsions while addressing the challenges associated with their stability during processing and storage. These challenges arise due to the high protein content, the presence of fat, and the liquid state of such compositions.
[0010] In view of this, it would be desirable to prepare liquid compositions comprising a high content of proteins and wherein the proteins include yeast proteins while said compositions remain stable. In particular, it would be desirable that said liquid compositions remain liquid and do not exhibit phase separation, in particular creaming and protein sedimentation during the manufacturing process, including high temperature treatment and / or during storage under chilled or ambient conditions. In addition, it would be desirable that said liquid compositions do not experience excessive gelation and result in undesirable fouling of the manufacturing line during the manufacturing process.
[0011] It would also be desirable to prepare liquid compositions comprising a high content of proteins and wherein the proteins consist only of yeast proteins while said compositions remain stable. In particular, it would be desirable that said liquid compositions remain liquid and do not exhibit do not exhibit phase separation, in particular creaming and protein sedimentation during the manufacturing process, including high temperature treatment and / or during storage under chilled or ambient conditions. In addition, it would be desirable that said liquid compositions do not experience excessive gelation and result in undesirable fouling of the manufacturing line during the manufacturing process.
[0012] It would also be desirable that the above liquid compositions remain shelf-stable, i.e. remain liquid and do not exhibit phase separation, in particular creaming and protein sedimentation, when stored under ambient conditions, over time, even for several months.
[0013] It would also be desirable that the above liquid compositions have a good protein quality, in particular PDCAAS and / or amino acid composition. It would also be desirable that the above liquid compositions are a significant source of protein and energy. It would also be desirable that the proteins represent a significant portion of the energy delivered by the liquid emulsions.
[0014] It would also be desirable that the above liquid compositions have good sensory properties, despite the presence of high content of proteins, in particular yeast proteins. In particular, it would be desirable that the above liquid emulsions have enhanced sensory properties compared to a liquid composition prepared with plant proteins only.
[0015] It would also be desirable that mineral fortification has limited impact on the properties of the yeast protein and the liquid compositions.
[0016] It would also be desirable to prepare liquid compositions that are liquid emulsions having the above-mentioned desirable features and advantages and wherein the emulsions and their fat droplets remain stable during the manufacturing process and storage.
[0017] It would also be desirable to prepare liquid compositions, preferably liquid emulsions in the form of beverages, preferably ready-to-drink beverage, more preferably ambient storage ready-to-drink beverage and that said liquid compositions, preferably liquid emulsions remain stable in this form, i.e. remain liquid and do not exhibit phase separation, in particular creaming and protein sedimentation during manufacturing process, including during high temperature treatment and / or during storage under chilled or ambient conditions. In addition, it would be desirable that said liquid compositions, preferably liquid emulsions in the form of beverages, preferably ready-to-drink beverage, more preferably ambient storage ready-to-drink beverage do not experience excessive gelation and do not result in undesirable fouling of the manufacturing line during the manufacturing process.
[0018] It would also be desirable to provide powder composition, preferably powdered emulsions starting from the above liquid compost, preferably liquid emulsions, said powder compositions, preferably powdered emulsions showing part or all of the desirable advantages and features of the above liquid emulsions, preferably liquid emulsions upon reconstitution in an aqueous liquid.
[0019] It would also be desirable to provide products comprising the above liquid compositions, preferably liquid emulsions or the above powder compositions, preferably powdered emulsions, said products benefiting from part or all of the desirable advantages and features of the above liquid compositions, preferably liquid emulsions and / or the above powder compositions, preferably liquid emulsions.
[0020] Any reference to prior art documents in this specification is not to be considered an admission that such prior art is widely known or forms part of the common general knowledge in the field.
[0021] SUMMARY OF THE INVENTION
[0022] The object of the present invention is to improve the state of the art, and in particular to provide liquid compositions, powder compositions, products, uses, methods, processes that overcome the problems of the prior art and addresses the needs described above, or at least to provide a useful alternative.
[0023] The inventors were surprised to see that the object of the present invention could be achieved by the subject matter of the independent claims. The dependent claims further develop the idea of the present invention. Accordingly, a first aspect of the invention proposes a liquid composition which comprises a fat component, at least one non-proteic thickening agent and a yeast protein source, wherein the yeast protein source comprises at least 40wt.% yeast proteins, and wherein the total protein content of the liquid composition is at least 5.01wt.%.
[0024] A second aspect of the invention proposes a powder composition which is obtained by drying the liquid composition of the first aspect of the invention.
[0025] A third aspect of the invention proposes the use of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention in the preparation and / or formulation of a product selected from the list consisting of: beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake, powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement (ONS), tube feeding and combinations thereof.
[0026] A fourth aspect of the invention proposes a product comprising the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention, wherein the product is selected from the list consisting of beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake, powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement
[0027] (ONS), tube feeding and combinations thereof.
[0028] A fifth aspect of the invention proposes a process for preparing a liquid composition comprising the steps of:
[0029] (a) Dispersing a yeast protein source comprising at least 40wt.% yeast proteins in an aqueous liquid to form a yeast protein dispersion, wherein the yeast protein dispersion comprises at least 5.01wt.% protein,
[0030] (b) Adding a fat component to the yeast protein dispersion to form a liquid pre-emulsion,
[0031] (c) Adding a non-proteic thickening agent to the liquid pre-emulsion,
[0032] (d) Homogenizing and heat-treating the liquid pre-emulsion of step (c) to form a liquid composition, wherein the heat treatment is before and / or after the homogenization.
[0033] A sixth aspect of the invention proposes a process for preparing a powder composition which comprises the step of drying the liquid composition obtained in step (d) of the fifth aspect of the invention or the liquid composition of the first aspect of the invention.
[0034] It has been observed that a combination of yeast protein sources with non-proteic thickening agent, optionally further combined with other protein sources (e.g. milk protein sources, plant protein sources etc...), allows to provide compositions, in particular emulsions comprising high protein content. Despite their high protein content, the compositions, in particular emulsions one or more of the following benefits: remain easy to process, retain good stability during manufacturing and storage (remain liquid (if liquid), do not exhibit phase separation, limited fat coalescences, no protein gelation, no fouling, no line blockage etc...) and exhibit good sensory properties. In addition, in view of its high protein content and / or amino acid profile, the compositions have good nutritional properties (e.g. good PDCAAS, significant source of proteins and energy), n particular, the inventors observed that yeast proteins are capable of both emulsifying and stabilizing the fat droplets in the emulsion, even in the presence of a minor soluble protein fraction, which is somewhat counterintuitive. This observation contrasts with the established understanding in the field that an important soluble protein fraction is essential for the emulsification and stabilization of emulsions. These and other aspects, features and advantages of the invention will become more apparent to those skilled in the art from the detailed description of embodiments of the invention, in connection with the attached drawings.
[0035] BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1: Protein profile (ten most abundant proteins) of yeast protein concentrate 1 (YPC 1; a, b) and 2 (YPC 2; c, d), as determined by UPLC-MS / MS.
[0037] Figure 2: Confocal laser scanning microscopy images of yeast protein concentrate 1 (YPC 1) and 2 (YPC 2) dispersions (3 wt% protein) before and after homogenization. Proteins (green) were fluorescently labelled with Fast Green FCF.
[0038] Figure 3: Confocal laser scanning microscopy images of yeast protein concentrate 2 (YPC 2) dispersion (3 wt% protein). Proteins (green) (e.g. cf. arrow labelled "prot."), fat (red) (e.g. cf. arrow labelled "fat") and fibers (chitin and cellulose, blue) (e.g. cf. arrow labelled fib.) were fluorescently labelled with respectively Fast Green FCF, Nile Red and Calcofluor White, respectively. Images of the dispersion with the filters for the three stains combined (a) and the filter for Calcofluor White only (b) are shown.
[0039] Figure 4: Solubility of yeast protein concentrate 1 (YPC 1) and 2 (YPC 2) dispersions in the pH range 2-9 (1 wt% protein).
[0040] Figure 5: Particle size distribution of ready-to-drink beverage prototypes based on yeast protein concentrate 1 (YPC 1; a) and 2 (YPC 2; b), as well as the milk and soy proteinbased reference, and volume-weighted mean particle diameter of the prototypes (c), as determined with and without SDS. Processing of the yeast protein-based prototypes involved upstream (U), downstream (D) or a combination of upstream and downstream (U+D) homogenization, whereas only the combination of upstream and downstream homogenization was used for the milk and soy protein-based reference.
[0041] Figure 6: Confocal laser scanning microscopy images of ready-to-drink beverage prototypes based on yeast protein concentrate 1 (YPC 1) and 2 (YPC 2), as well as the milk and soy protein-based reference, obtained by means of a combination of upstream and downstream homogenization. Proteins (green) (e.g. cf. arrow labelled "prot.") and fat (red) (e.g. cf. arrow labelled "fat") were fluorescently labelled with respectively Fast Green FCF and Nile Red.
[0042] Figure 7: Viscosity of ready-to-drink beverage prototypes, based on yeast protein concentrate 1 (YPC 1) and 2 (YPC 2), as well as the milk and soy protein-based reference, as determined by controlled shear stress rheology at 10 s-1. Processing of the yeast proteinbased prototypes involved upstream (U), downstream (D) or a combination of upstream and downstream (U+D) homogenization, whereas only the combination of upstream and downstream homogenization was used for the milk and soy protein-based reference.
[0043] Figure 8: Backscattering profiles throughout the height of ready-to-drink beverage prototypes based on yeast protein concentrate 1 (YPC 1) and 2 (YPC 2), as well as the milk and soy protein-based reference, as determined using the Turbiscan Lab Expert upon production (DO) and after 30 days of refrigerated storage (D30). Processing of the yeast protein-based prototypes involved upstream (U), downstream (D) or a combination of upstream and downstream (U+D) homogenization, whereas only the combination of upstream and downstream homogenization was used for the milk and soy protein-based reference.
[0044] Figure 9: Pictures of ready-to-drink beverage prototypes, including those based on yeast protein concentrate 1 (YPC 1) upon production (DO) (a) and after 30 days of refrigerated storage (D30) (b), yeast protein concentrate 2 (YPC 2) on DO (c) and D30 (d), and the milk and soy protein-based reference on DO (e) and D30 (f). Processing of the yeast protein-based prototypes involved upstream (U), downstream (D) or a combination of upstream and downstream (U+D) homogenization, whereas only the combination of upstream and downstream homogenization was used for the milk and soy protein-based reference.
[0045] Figure 10: Overall intensity scale in odour and flavour (a) and monadic RATA scale (b) used for the sensory evaluation of the yeast protein-based ready-to-drink beverage prototypes, and overall intensity scale in odour and flavour (c) and comparative RATA scale (d) used to benchmark the preferred yeast protein-based prototype against the reference.
[0046] Figure 11: Outcomes of the sensory evaluation of ready-to-drink beverage prototypes based on yeast protein concentrate 1 (YPC 1; a) and 2 (YPC 2; b), as obtained by upstream (U), downstream (D) or a combination of upstream and downstream (U+D) homogenization. Only attributes with average perceived intensity >1 are reported in the graphs. Prototypes obtained by a combination of upstream and downstream homogenization were used for the comparison of the sensory properties of YPC 1 vs YPC 2 (c) and the milk and soy protein-based reference vs YPC 2 (d). White bars in (d) indicate significant differences.
[0047] Figure 12: Volume-weighted mean particle diameter (D[4,3]) of yeast protein concentrate 1 (YPC 1) and 2 (YPC 2) dispersions (8.5 wt% protein) in the presence of salts, including calcium chloride (CaCI), tricalcium phosphate (CaP) and tripotassium citrate (KCI) (a, b), added at concentrations required to reach the target calcium (20 or 40 mM) and potassium (35 and 70 mM) concentrations. D[4,3] of YPC 1 and YPC 2 dispersions (8.5 wt% protein) in the presence of salt blends, including CaCI + KCit and CaP + KCit (c, d), added at concentrations required to reach the target calcium (40 mM) and potassium (70 mM) concentrations.
[0048] Figure 13: Protein solubility of yeast protein concentrate 1 (YPC 1) and 2 (YPC 2) dispersions (8.5 wt% protein) in the presence of salts, including calcium chloride (CaCI), tricalcium phosphate (CaP) and tripotassium citrate (KCI) (a, b), added at concentrations required to reach the target calcium (20 or 40 mM) and potassium (35 and 70 mM) concentrations. Protein solubility of YPC 1 and YPC 2 dispersions (8.5 wt% protein) in the presence of salt blends, including CaCI + KCit and CaP + KCit (c, d), added at concentrations required to reach the target calcium (40 mM) and potassium (70 mM) concentrations.
[0049] Figure 14: Influence of mineral fortification of ready-to-drink beverage prototypes based on yeast protein concentrate 2 (YPC 2) on particle size distribution (a), viscosity (b), as determined by controlled shear stress rheology at 10 s-1, and colloidal stability (c), as determined using the Turbiscan Lab Expert upon production (DO) and after 30 days of refrigerated storage (D30). The salts included blends of calcium chloride (CaCI) and tripotassium citrate (KCit) or tricalcium phosphate (CaP) and KCit, added at concentrations required to reach the target calcium (40 mM) and potassium (70 mM) concentrations. The prototypes were obtained by a combination of upstream and downstream homogenization.
[0050] Figure 15: Outcomes of the evaluation of the sensory properties of ready-to-drink beverage prototypes based on yeast protein concentrate 2 (YPC 2) fortified with blends of calcium chloride (CaCI) and tripotassium citrate (KCit) or tricalcium phosphate (CaP) and KCit, added at concentrations required to reach the target calcium (40 mM) and potassium (70 mM) concentrations, benchmarked against the prototype without added salts.
[0051] DETAILED DESCRIPTION OF THE INVENTION
[0052] As used herein, the words "comprise", "comprising" and the like are to be construed in an inclusive sense, that is to say, in the sense of "including, but not limited to", as opposed to an exclusive or exhaustive sense. Likewise, the terms "include," "including" and "or" should all be construed to be inclusive, unless such a construction is clearly prohibited from the context. Nevertheless, the products and compositions, including emulsions disclosed herein may lack any element that is not specifically disclosed herein. Thus, a disclosure of an embodiment using the term "comprising" includes a disclosure of embodiments "consisting essentially of" and "consisting of" the components identified. All numerical ranges should be understood to include each integer, whole or fractions, within the range. Moreover, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.
[0053] All percentages expressed herein are by weight of the total weight of the composition unless expressed otherwise.
[0054] As used herein, "about," "approximately" and "substantially" are understood to refer to numbers in a range of numerals, for example the range of -10% to +10% of the referenced number, preferably -5% to +5% of the referenced number, more preferably -1% to +1% of the referenced number, most preferably -0.1% to +0.1% of the referenced number.
[0055] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a fat component" or "the fat component" includes one fat component but also two or more fat components.
[0056] Unless defined otherwise, all technical and scientific terms have and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
[0057] As used herein, the term "and / or" used in the context of "X and / or Y" should be interpreted as "X," or "Y," or "X and Y.". Similarly, "at least one ofX or Y" should be interpreted as "X," or "Y," or "both X and Y.". For example, "pulses and / or cereals" means "pulses" or "cereals" or "both pulses and cereals".
[0058] As used herein, the terms "non-limiting example", "example", "e.g." and "such as," particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be deemed to be exclusive or comprehensive. But, a disclosure of an embodiment using the term "example" and "such as" includes a disclosure of embodiments where the terms are exclusive and / or comprehensive.
[0059] As used herein, the terms "food," "food product" and "food composition" mean a product or composition that is intended for ingestion by an individual such as a human and provides at least one nutrient to the individual.
[0060] The compositions and products, including emulsions, of the present disclosure, including the many embodiments described herein, can comprise, consist of, or consist essentially of the elements disclosed herein, as well as any additional or optional ingredients, components, or elements described herein or otherwise useful in a diet. As used herein, the terms "beverage" and "drink" refers to a "food", "food product" or "food composition" which is generally consumed by drinking. In particular, the terms "beverage" and "drink" are used interchangeably.
[0061] As used herein, the term "animal" includes, but is not limited to, mammals, which includes but is not limited to rodents; aquatic mammals; domestic animals such as dogs, cats and other pets; farm animals such as sheep, pigs, cows and horses; and humans. Where "animal," "mammal" or a plural thereof is used, these terms also apply to any animal that is capable of the effect exhibited or intended to be exhibited by the context of the passage, e.g., an animal benefiting from improved mitochondrial calcium import. While the term "individual" or "subject" is often used herein to refer to a human, the present disclosure is not so limited. Accordingly, the term "individual" or "subject" refers to any animal, mammal or human that can benefit from the methods and compositions disclosed herein.
[0062] As used herein, a "subject" or "individual" is a mammal, preferably a human. In particular, the terms "subject" and "individual" are used interchangeably.
[0063] As used herein, the term "aged" or "elderly" in the context of a human means an age from birth of at least 50 years, preferably above 55 years, more preferably above 60 years, even more preferably above 63 years, more preferably above 65 years, and most preferably above 70 years.
[0064] As used herein, the term "infant" means a child under the age of 12 months. The expression "young child" or "toddler" means a child aged between one and less than three years. The expression "child" means a between three and seven years of age.
[0065] As used herein, a "preterm" or "premature" means an infant or young child who was not born at term. Generally, it refers to an infant or young child born prior 37 weeks of gestation.
[0066] As used herein, the terms "serving" or "unit dosage form," are interchangeable and refer to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of the liquid compositions, powder compositions or products as disclosed herein, in an amount sufficient to produce the desired effect, preferably in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for the unit dosage form depend on the particular compounds employed, the effect to be achieved, and the pharmacodynamics associated with each compound in the host. In an embodiment, the unit dosage form can be a predetermined amount of liquid housed within a container such as a bottle. As used herein, an "oral nutrition supplement" or "ONS" is a composition comprising at least one macronutrient and / or at least one micronutrient, for example in a form of sterile liquids, semi-solids or powders, and intended to supplement other nutritional intake such as that from food. Non-limiting examples of commercially available ONS products include MERITENE®, BOOST®, NUTREN® and SUSTAGEN®. In some embodiments, an ONS can be a beverage in liquid form that can be consumed without further addition of liquid, for example an amount of the liquid that is one serving of the composition.
[0067] As used herein, the term "food for special medical purpose (FSMP)" refers to formula foods specially processed and prepared in order to meet special needs for nutrient or diet of those suffering from food intake restriction, disorder of digestive absorption, disorder of metabolic or certain diseases. Such foods shall be used alone or together with other foods under the guidance of a doctor or clinical nutritionist. FSMP is special dietary food, not medicine, but not ordinarily eaten by normal people. It is specially developed by clinicians and nutritionists based on scientific facts after extensive medical research.
[0068] As used herein, the expression "infant formula" as used herein refers to a foodstuff intended for particular nutritional use by infants during the first months of life and satisfying by itself the nutritional requirements of this category of person (Article 2(c) of the European Commission Directive 91 / 321 / EEC 2006 / 141 / EC of 22 December 2006 on infant formulae and follow-on formulae). It also refers to a nutritional composition intended for infants and as defined in Codex Alimentarius (Codex STAN 72-1981) and Infant Specialities (incl. Food for Special Medical Purpose). The expression "infant formula" encompasses both "starter infant formula" and "follow-up formula" or "follow-on formula".
[0069] As used herein, a "follow-up formula" or "follow-on formula" is given from the 6th month onwards. It constitutes the principal liquid element in the progressively diversified diet of this category of person.
[0070] As used herein, the expression "baby food" means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.
[0071] As used herein, the expression "infant cereal composition" means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.
[0072] As used herein, the expression "growing-up milk" (orGUM) refers to a milk-based drink generally with added vitamins and minerals, that is intended for young children or children.
[0073] As used herein, the term "fortifier" refers to liquid or solid nutritional compositions suitable for fortifying or mixing with human milk, infant formula, growing-up milk or human breast milk fortified with other nutrients. Accordingly, the fortifier of the present invention can be administered after dissolution in human breast milk, in infant formula, in growing-up milk or in human breast milk fortified with other nutrients or otherwise it can be administered as a stand-alone composition. When administered as a stand-alone composition, the milk fortifier of the present invention can be also identified as being a "supplement". In one embodiment, the milk fortifier of the present invention is a supplement.
[0074] As used herein, the term "paediatric supplement" refers to a product which is intended to supplement the general diet of an infant, a young child or a child.
[0075] As used herein, the expression "weaning period" means the period during which the mother's milk is substituted by other food in the diet of an infant or young child.
[0076] As used herein, the term "mobility" is the ability to move independently and safely from one place to another.
[0077] As used herein, the term "combination", or terms "in combination", "used in combination with" or "combined preparation" as used herein may refer to the combined administration of two or more agents simultaneously, sequentially or separately.
[0078] As used herein, the term "simultaneous" as used herein means that the agents are administered concurrently, i.e. at the same time.
[0079] As used herein, the term "sequential" as used herein means that the agents are administered one after the other.
[0080] As used herein, the term "separate" as used herein means that the agents are administered independently of each other but within a time interval that allows the agents to show a combined, preferably synergistic, effect. Thus, administration "separately" may permit one agent to be administered, for example, within 1 minute, 5 minutes or 10 minutes after the other.
[0081] As used herein, the term "amino acid" as used herein includes free form amino acids, or bound form of amino acids in molecules between 2 and 20 amino acids (referenced herein as "peptides"), and also in longer chains of amino acids (i.e. proteins). Small peptides, i.e., chains of 2 to 10 amino acids, are suitable for the composition alone or in combination with other proteins. The "free form" of amino acid to the monomeric form of the amino acid. When the term "free amino acid" is used, it refers exclusively to "free form", i.e. the monomeric form of the amino acid.
[0082] Each amino acid disclosed herein can be present in the composition as only one type of the amino acid or as a mixture of one or more types of the amino acid, for example one or more (i) peptides containing the amino acid, (ii) longer chains of amino acids (i.e proteins) including the amino acid, or (iii) free form of the amino acid. For example, a disclosure of "composition comprising an aromatic amino acid" or "product comprising an aromatic amino acid" or "composition comprising an aromatic amino acid" constitutes a disclosure of aromatic amino acids only in free form, a disclosure of aromatic amino acids only bound to other amino acids, and a mixture of aromatic amino acids in free form and aromatic amino acids bound to other amino acids. Similarly, in embodiments where the referenced amino acid is in peptides or proteins, optionally the composition can have substantially no free form of the referenced amino acid.
[0083] As used herein, the term "an essential amino acid (EAA)" or an indispensable amino acid as used means an amino acid that cannot be synthesized de novo by the organism at a rate commensurate with its demand, and thus must be supplied in its diet. Of the twenty one amino acids common to all life forms the following nine amino acids are considered essential amino acids in the human diet which include phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine and histidine. Six other amino acids which are considered conditionally essential in the human diet are arginine, cysteine, glycine, glutamine, proline and tyrosine. There are six amino acids are non-essential (dispensable) in human diet, and these six non-essential amino acids are alanine, aspartic acid, asparagine, glutamic acid, serine and selenocysteine.
[0084] As used herein, the term "an aromatic amino acid (AAA)" refers to an amino acid that includes an aromatic ring. Examples of aromatic amino acids include: Phenylalanine (symbol Phe or F); Tryptophan (symbol Trp or W); Tyrosine (symbol Tyr or Y); and Histidine (symbol His or H).
[0085] As used herein, the term "a branched chain amino acid (BCAA)" means an amino acid having an aliphatic side-chain with a branch (a central carbon atom bound to three or more carbon atoms). Among the proteinogenic amino acids, there are three BCAAs: leucine (Leu or L), isoleucine (He or I), and valine (Vai or V). Non-proteinogenic BCAAs include 2- aminoisobutyric acid.
[0086] As used herein, the term "protein" as used herein includes molecules between 2 and 20 amino acids (referenced herein as "peptides"), and also includes longer chains of amino acids (i.e. molecules having more than 20 amino acids). Small peptides, i.e., chains of 2 to 10 amino acids, are suitable for the liquid compositions, powder compositions and products alone or in combination with other proteinsln a preferred embodiment, the term "protein" refers to molecules having more than 20 amino acids only.
[0087] The "free form" of an amino acid or "free amino acid" is the monomeric form of the amino acid. Suitable amino acids include both natural and non-natural amino acids.
[0088] As used herein, the term "vegan" refers to an edible composition which is entirely devoid of animal products, or animal derived products.
[0089] As used herein, the term "vegetarian" refers to an edible composition which is devoid of meat, including fish.
[0090] As used herein, the term "yeast protein concentrate" refers to an ingredient comprising whole yeast cells in a non-living state and / or components / fragments derived from said yeast cells and comprising from 70.0 to 90.0wt.% yeast proteins.
[0091] As used herein, the term "yeast protein isolate" refers to an ingredient comprising whole yeast cells in a non-living state and / or components / fragments derived from said yeast cells and comprising from 90.1 to 99.9wt.% yeast proteins.
[0092] As used herein, the term "yeast biomass" refers to an ingredient comprising whole yeast cells in a non-living state and / or components / fragments derived from said yeast cells and comprising from 40.0 to 69.9wt.% yeast proteins.
[0093] As used herein, the term "non-proteic thickening agent" refers to ingredients other than proteins that increase the viscosity. In a preferred embodiment, non-proteic thickening agent is a polysaccharide.
[0094] In a first aspect, the invention relates to a liquid composition. In a preferred embodiment, the liquid composition is a liquid emulsion. In a further preferred embodiment, the liquid emulsion is an oil-in-water emulsion.
[0095] A liquid emulsion is defined as a stable mixture of two immiscible liquids, in which one liquid is dispersed as droplets within the other. The liquid that is dispersed as droplets is referred to as the dispersed phase (in the present invention, the fat component), while the other liquid is referred to as the continuous phase (in the present invention, the aqueous liquid).
[0096] When the liquid composition is a liquid emulsion, the fat component is in the form of fat droplets and therefore, the liquid emulsion comprises fat droplets formed by the fat component. The fat droplets formed by the fat component are surrounded (i.e. stabilized) at their surface by yeast proteins. In particular, the yeast proteins are adsorbed to the surface of the fat droplets. This results in the stabilization of the fat droplets in the emulsion. Indeed, it has been observed by the inventors that the yeast proteins interact with the fat droplets and not only emulsify but also stabilize the fat droplets, thereby contributing to the overall stability of the liquid emulsion.
[0097] In a preferred embodiment, the liquid composition, preferably the liquid emulsion is a beverage, preferably a ready-to-drink beverage, more preferably ambient storage ready-to- drink beverage. The definition of "ambient storage" and related features are further provided below.
[0098] In particular, it has been observed that the liquid composition, preferably the liquid emulsion of the invention containing yeast proteins remains stable during processing (i.e. remains liquid, no or limited protein gelation, no or limited fouling, no line blocking) and over the shelf-life (i.e. no phase separation such as creaming, sedimentation etc...) while providing significant content of proteins, even when formulated as a beverage, preferably a ready-to- drink beverage, more preferably an ambient storage ready-to-drink beverage.
[0099] In some further embodiment, the liquid composition is in the form of an incomplete nutritional product. As used herein, the "incomplete nutritional product" refers to preferably nutritional products that do not contain sufficient levels of macronutrients (protein, fats and carbohydrates) or micronutrients to be sufficient to be a sole source of nutrition for the subject, in particular human or non-human animal, which consume the nutritional product or the subject, in particular human or non-human animal to which the nutritional product is being administered. Preferably, the non-human animal is a pet.
[0100] In some preferred embodiment, the liquid composition is in the form of a complete nutritional product. As used herein, the "complete nutritional product" refers to nutritional products that contains sufficient levels of macronutrients (protein, fats and carbohydrates) or micronutrients to be sufficient to be a sole source of nutrition for the subject, in particular human or non-human animal, which consume the nutritional product or the subject, in particular human or non-human animal to which the nutritional product is being administered. Preferably, the non-human animal is a pet.
[0101] In some embodiment, the liquid composition is not a dressing. In some embodiment, The liquid composition is not an animal feed. In some embodiment, The liquid composition is suitable for human consumption.
[0102] In some embodiment, the liquid composition is vegetarian. In some embodiment, the liquid composition is vegan. In some embodiment, the liquid composition has a pH of 2 to 9, preferably of 4 to 8, more preferably 5 to 7.
[0103] In some embodiment, the liquid composition has a viscosity between 1 and 100 mPa.s, preferably between 1 and 60 mPa.s, more preferably between 1 and 40 mPa.s, most preferably between 10 and 40 mPa.s.
[0104] The viscosity of the present liquid composition can be suitably determined with Physica MCR 501 controlled shear stress rheometer (Anton Paar GmbH, Graz, Austria) equipped with a concentric cylinder geometry (CC27 / S, Anton Paar GmbH, Graz, Austria) characterized by a rough (sandblasted) surface to prevent wall slip and a gap with the outer cup (C-CC27 / T200 / SS / S, Anton Paar GmbH, Graz, Austria) of 1.13 mmat shear rate of 10 s-1 at 20°C.
[0105] In particular, the liquid composition provides significant source of protein while still retaining liquid consistency, facilitating its consumption by drinking. In particular, its liquid consistency may be suitable for individuals suffering from dysphagia.
[0106] In some embodiment, the liquid composition may be chilled storage liquid composition or ambient storage liquid composition.
[0107] By "chilled storage", it is understood a liquid composition which has a shelf-life of several days when stored under chilled conditions. The term "chilled conditions" refers to temperatures ranging from 2°C to 14°C, preferably from 2°C to 10°C, more preferably from 4°C to 8°C. In particular, a chilled storage liquid composition has a shelf-life of at least 10 day, preferably at least 25 days, more preferably of at least 30 days when stored under chilled conditions. These storage temperatures relate to the storage of the composition before being commercially obtained by an end consumer. Generally, the end consumer is advised to store the composition under the same chilled conditions until consumption, for example in a refrigerator.
[0108] By "ambient storage", it is understood a liquid composition which has a shelf-life of several months when stored under ambient conditions. This also generally applies when it is stored under chilled conditions. The term "ambient conditions" refers to temperatures ranging from 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C. Especially, an ambient storage liquid composition has a shelf-life of at least 1 month, preferably at least 3 months, more preferably at least 6 months, even more preferably 8 months, even more preferably at least 12 months, even more preferably of at least 24 months, even more preferably of at least 30 months when stored under ambient conditions. In some further embodiment, an ambient storage liquid composition has a shelf-life of at most 30 months, preferably at most 18 months, more preferably at most 12 months when stored under ambient conditions. These storage temperatures relate to the storage of the composition before being commercially obtained by an end consumer. Generally, the end consumer is advised to store the composition under the same ambient conditions until consumption, for example in a shelf at room temperature.
[0109] In a preferred embodiment, the liquid composition is ambient storage liquid composition. The definition and features of "ambient storage" liquid composition(s) is provided above.
[0110] The liquid composition, preferably liquid emulsion is advantageous on two aspects.
[0111] First, it provides significant amount of protein while remaining stable upon processing for preparing ambient storage product that include high temperature treatment, e.g. UHT. In particular, it remains liquid and does not exhibit excessive gelation during the manufacturing process, including important temperature treatment. In addition, it does not lead to fouling and blocking of manufacturing lines during the manufacturing process, including important temperature treatment.
[0112] Second, it provides significant amount of protein while remaining stable during storage for several months under ambient conditions. In particular, it remains liquid and does not exhibit phase separation phenomenon, including creaming (i.e. fat separation) and protein sedimentation during storage for several months under chilled and ambient conditions.
[0113] Liquid composition with yeast proteins were observed to be particularly stable in the above conditions, i.e. upon processing including high temperature treatment and during storage under chilled and ambient conditions.
[0114] In particular, when the liquid composition is a liquid emulsion, it shows good emulsion stability properties during the manufacturing process and storage.
[0115] Accordingly, in an advantageous embodiment, the ambient storage liquid composition is shelf-stable for at least 1 month, preferably at least 3 months, more preferably at least 6 months, even more preferably at least 8 months, more preferably at least 12 months, even more preferably of at least 24 months, even more preferably of at least 30 months at 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C. In some further embodiment, the ambient storage liquid composition is shelf-stable for at most 30 months, preferably at most 18 months, more preferably at most 12 months at 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C.
[0116] By "shelf stable", it is understood that the liquid composition remains liquid and does not exhibit any phase separation, in particular creaming and protein sedimentation under the abovementioned storage time and temperature conditions. This can be assessed by visual inspection. In particular, the liquid properties of the composition may be assessed by pouring or agitating the composition and observing if the composition has a liquid flowing consistency. This also can be assessed by measuring the viscosity as disclosed in the example over the shelf life. The separation of phase in the composition may be observed by appearance of several phase in the liquid composition (heterogenous composition), e.g. appearance of fat phase (creaming), appearance of solid particles, in particular generally at the bottom of the liquid composition (sedimentation). The separation of phase may also be assessed by measuring the backscattering of the composition with a Turbiscan Lab Expert (Formulaction, Toulouse, France) according to the method of example 3 over the shelf life.
[0117] In addition, the term "shelf stable" may also include that the liquid composition does not spoil under the abovementioned storage time and temperature conditions.
[0118] In some embodiment, the term "shelf stable" includes that the emulsion remains stable over the shelf life.
[0119] The liquid composition comprises a fat component. The fat component may be any fat suitable for human oral consumption. The fat may be liquid or solid fat. A liquid fat refers to a fat that is generally liquid at room temperature (i.e. 25°C) or below. A solid fat refers to a fat that is generally solid at room temperature (i.e. 25°C) or below.
[0120] The fat component may be selected from the list consisting of vegetable fat, animal fat, microbial fat, and combination thereof.
[0121] Examples of vegetable fat include hydrogenated vegetable oil, vegetable oil, vegetable oil fractions, algal oil, algal oil fractions, lecithin, cocoa butter, cocoa butter fractions, shea butter, shea butter fractions, coconut fat, coconut fat fractions and mixture thereof. Examples of vegetable oil include sunflower seed oil, rapeseed oil, cottonseed oil, walnut oil, peanut oil, olive oil, grapeseed oil, soybean oil, palm oil, high oleic sunflower seed oil, high stearic high oleic sunflower seed oil, high oleic safflower oil, high oleic soybean oil, high oleic rapeseed oil such as high oleic canola oil, high oleic palm oil, high oleic peanut oil, macadamia nut oil, moringa oleifera seed oil, papaya seed oil, hazelnut oil, avocado oil, and mixtures thereof. Examples of animal fat include milk fat, milk fat fractions, fish oil, fish oil fractions and mixtures thereof.
[0122] The microbial fat refers to fat derived from microorganisms such as yeast, bacteria and / or fungi. Examples of microbial fat include microbial oil (e.g. yeast oil), microbial oil fraction (e.g. yeast oil fraction) and mixtures thereof.
[0123] In some embodiment, the fat component may comprise or consist of omega-3 fatty acids. Examples of omega-3 fatty acids include alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), hexadecatrienoic acid (HTA), stearidonic acid (SDA), eicosatrienoic acid (ETE), eicosatetraenoic acid (ETA), heneicosapentaenoic acid (HPA), docosapentaenoic acid (DPA), tetracosapentaenoic acid, tetracosahexaenoic acid and mixtures thereof.
[0124] In some embodiments, when the liquid composition is a liquid emulsion, the fat component comprises at least 80wt.%, preferably at least 90wt%, more preferably 100% fat. In addition, the fat of the fat component is not absorbed to any solids. These conditions, i.e. high fraction of fat in the fat component and no absorption to solids, contribute to facilitation of the formation of fat droplets, and consequently, ensure that an emulsion is not achieved.
[0125] In some embodiments, when the liquid composition is a liquid emulsion, at least 50%, at least 60%, at least 70%, at least 80% or at least 90% of the fat component is in liquid form. Preferably, all the fat component is in liquid form. In particular, the fat component in liquid form is liquid fat, i.e. fat that is liquid at room temperature (i.e. 25°C) or below. Examples of liquid fat include liquid vegetable oil, liquid vegetable oil fractions, liquid algal oil, liquid algal oil fractions, lecithin, and mixture thereof. Examples of liquid vegetable oil include sunflower seed oil, rapeseed oil, cottonseed oil, walnut oil, peanut oil, olive oil, grapeseed oil, soybean oil, palm oil, high oleic sunflower seed oil, high stearic high oleic sunflower seed oil, high oleic safflower oil, high oleic soybean oil, high oleic rapeseed oil such as high oleic canola oil, high oleic palm oil, high oleic peanut oil, macadamia nut oil, moringa oleifera seed oil, papaya seed oil, hazelnut oil, avocado oil, and mixtures thereof. The liquid fat may comprise or consist of omega-3 fatty acids. Examples of omega-3 fatty acids include alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), hexadecatrienoic acid (HTA), stearidonic acid (SDA), eicosatrienoic acid (ETE), eicosatetraenoic acid (ETA), heneicosapentaenoic acid (HPA), docosapentaenoic acid (DPA), tetracosapentaenoic acid, tetracosahexaenoic acid and mixtures thereof. When the liquid composition is a liquid emulsion, the fat component comprises less than 10wt.%, less than 5wt.% or less lwt.% of crystallized fat. Preferably, the fat component is free from crystallized fat.
[0126] When the liquid composition is a liquid emulsion, the fat component is not provided as cocoa or cocoa fat. Indeed, in cocoa, cocoa fat is absorbed to cocoa solids and cocoa fat is crystallised at ambient temperature. As a result, this does not lead to the formation of fat droplets, and consequently, an emulsion is not achieved.
[0127] In some embodiment, the liquid composition comprises at least 0.5wt.%, preferably 0.5 to 20wt.%, more preferably 0.5 to 15wt.%, even more preferably 1 to 10wt%, even more preferably 1 to 5wt.% fat component.
[0128] In some embodiment, the liquid composition comprises significant amount of fat, i.e. comprises 8 to 15wt% fat component.
[0129] It has been observed that the liquid compositions, preferably liquid emulsions of the invention remain stable during processing and over shelf life even in presence of significant amount of fat component as provided above while still remaining liquid.
[0130] Liquid compositions, in particular emulsions with a fat content exceeding 20%, also known as concentrated emulsions, typically exhibit high viscosity due to their high fat content. This high viscosity aids in stabilizing such emulsions. However, liquid compositions, in particular emulsions with a fat content below 20%, particularly below 10%, are generally in a liquid state, and the stabilizing effect of viscosity is often limited or absent, making fat stabilization challenging.
[0131] It has been observed that the liquid compositions, preferably liquid emulsions of the invention remain stable during the manufacturing process and over shelf life, even under nonconcentrated emulsion conditions, i.e. below 20% fat, preferably below 10% fat.
[0132] The liquid composition comprises at least one non-proteic thickening agent. The non- proteic thickening agent may be selected from the list consisting of alginate, xanthan gum, pectin, locust bean gum, gellan gum, carrageenan, guar gum, cellulose, carboxymethylcellulose, agar, gum arabic, starch, konjac gum, tara gum, chitosan, tragacanth gum, psyllium husk and mixture thereof.
[0133] While the yeast protein source stabilizes the fat component and limits its coalescence, the non-proteic thickening agent ensures colloidal stability and avoids protein sedimentation by increasing the viscosity of the continuous phase.
[0134] In a preferred embodiment, the non-proteic thickening agent is gellan gum. It has been observed that gellan gum was particularly effective for colloidal stabilization. The quantity of non-proteic thickening agent in the liquid composition may be from 0.01wt.% to 2wt.%, more preferably 0.05wt.% to 1.5wt.%, even more preferably 0.1 to 1.5wt.%. In particular, it has been observed that good colloidal stabilization may be achieved at low thickening agent concentration.
[0135] The liquid composition comprises a yeast protein source. The yeast protein source comprises at least 40wt.%, preferably at least 50wt.%, more preferably at least 60wt.%, even more preferably at least 70wt.%, even more preferably at least 80% yeast proteins.
[0136] In some embodiment, the liquid composition comprises 40wt.% to 99wt.%, preferably 40wt.% to 90wt.%, more preferably 50wt.% to 90wt.%, even more preferably 60 wt.% to 90wt.%, even more preferably 60wt.% to 80wt.% yeast proteins.
[0137] In some embodiment, the yeast protein source is selected from the list consisting of yeast biomass, yeast protein concentrate, yeast protein isolate, and mixture thereof. In a preferred embodiment, the yeast protein source is yeast protein concentrate. In some embodiment, the yeast protein concentrate comprises 70 wt.% to 90wt.% yeast proteins. Compared to biomass, yeast protein concentrates are advantageous as their protein content is higher, meaning that less ingredient is required for stabilization minimizing undesirable viscosity increase and as their flavour is more neutral. Compared to yeast protein isolates, yeast protein concentrates are advantageous due their higher availability.
[0138] In some preferred embodiment, the yeast protein source is derived from yeast from the genus Saccharomyces, Candida and combination thereof. In some further preferred embodiment, the yeast protein source is derived from Saccharomyces cerevisae, Candida utilis and combination thereof.
[0139] Likewise, the yeast proteins of the yeast protein source and / or the liquid composition are derived from yeast from the genus Saccharomyces, Candida and combination thereof. In some further preferred embodiment, the yeast proteins of the yeast protein source and / or the liquid composition are derived from Saccharomyces cerevisae, Candida utilis and combination thereof.
[0140] It has been observed that yeast protein rich ingredients may be used to prepare protein-rich liquid compositions, preferably emulsions which are stable. The obtained compositions are not only a good nutritional source of protein but also are outstandingly stable despite the presence of significant amount of fat and proteins. This is even observed in presence of limited amount of non-proteic thickening agents. In particular, it has been observed that yeast protein rich ingredients, in particular yeast protein concentrates, effectively stabilize fat droplets in liquid compositions, preferably liquid emulsions. These liquid compositions, preferably liquid emulsions remain stable over time during storage and do not experience significant destabilization phenomena, in particular phase separation such as creaming.
[0141] Furthermore, the yeast proteins from yeast protein rich ingredients exhibit exceptional stability in liquid compositions, preferably liquid emulsions even at high levels, and do not do not undergo sedimentation over time during storage and upon processing, including upon high heat treatment. Additionally, they do not significantly increase the viscosity of the liquid compositions, preferably liquid emulsions, even when subjected to high heat treatment. This advantageous low viscosity-building property of yeast proteins enables the creation of liquid compositions, preferably liquid emulsions that remain in a liquid state upon processing, including high heat treatment and during storage while still providing a substantial level of proteins. This low viscosity-building property also avoids or at least limits fouling and blockages of the manufacturing lines by the composition during the process.
[0142] In addition, yeast protein rich ingredients are also advantageous as they provide beneficial nutrients: minerals, amino acids and so on. In addition, they can impart limited sensory defects in the liquid composition and therefore, the obtained liquid composition can have acceptable sensory properties, including limited yeast, fermented and umami notes.
[0143] In some embodiment, the yeast protein source has a very low or is free from mannoprotein content. The yeast protein source comprises less than 20wt.%, preferably less than 15wt.%, more preferably less than 10wt.%, even more preferably less than 5wt.% mannoproteins. In some embodiment, the yeast protein source may comprise at least 0.05wt.% preferably at least lwt.% mannoproteins.
[0144] Likewise, the liquid composition comprises less than 5wt.%, preferably less than 4wt.%, more preferably less than 3wt.%, even more preferably less than 2wt.% mannoproteins. In some embodiment, the liquid composition may comprise 0.4wt.% to 5wt.%, preferably 0.4wt.% to 4wt.%, more preferably 0.4wt.% to 3wt.%, even more 0.4wt.% to 2wt.% mannoproteins. In some other embodiment, the liquid composition may comprise lwt.% to 5wt.%, preferably lwt.% to 4wt.%, more preferably lwt.% to 3wt.%, even more lwt.% to 2wt.% mannoproteins.
[0145] Less than 20%, less than 10%, preferably less than 5%, more preferably less than 4%of the proteins of the yeast protein source and / or the liquid composition are mannoproteins. It has been observed that it was possible to stabilize liquid compositions, preferably liquid emulsions, in particular stabilize their fat component even when using yeast protein source with limited or no mannoproteins.
[0146] In some embodiment, the yeast protein source comprises an insoluble yeast proteins to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20. Likewise, the liquid composition comprises an insoluble yeast proteins to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20. The insoluble yeast proteins to soluble yeast proteins ratio is a concentration ratio. In particular, the insoluble yeast proteins to soluble yeast proteins ratio corresponds to the ratio between the insoluble yeast protein concentration to soluble yeast protein concentration of a composition (e.g. yeast protein source, liquid composition etc.).
[0147] The soluble protein concentration, such as soluble yeast protein concentration or protein solubility of a composition, such as yeast protein source or liquid composition may be measured as follows. The protein-containing composition (e.g. yeast protein source, liquid composition etc.) is reconstituted in ultrapure water at a concentration of lwt.% protein under low-speed magnetic stirring for 1 hour at room temperature and the pH is adjusted to 6.8 using 0.1-1 M HCI and / or NaOH, as required. Samples is centrifuged at 1000 rpm for 15 min using a Sorvall evolution RC centrifuge (Thermo Fischer, Waltham, MA) equipped with a fixed angle rotor SS-34. The protein concentration of the supernatant is determined by the Kjeldahl method according to the AOAC Official Method 930.29 (AOAC, 2005) using the nitrogen-protein conversion of 6.25. Solubility or soluble protein concentration is calculated as the protein concentration of the supernatant expressed as a percentage of the protein concentration of the initial dispersion.
[0148] The insoluble protein concentration, such as insoluble yeast protein concentration corresponds to the total protein content minus the soluble protein content, such as soluble yeast protein content of a given composition (e.g. yeast protein source, liquid composition etc.).
[0149] It is observed that the use of yeast protein source(s) comprising a higher fractions of insoluble proteins compared to soluble proteins are particularly effective to stabilize liquid compositions, preferably liquid emulsions, in particular their fat component.
[0150] In some embodiment, the yeast protein source comprises at most 30, preferably at most 18wt.% of soluble yeast proteins. Likewise, the liquid composition comprises at most 15wt.% of soluble yeast proteins. The low solubility may be explained that part of the proteins is within yeast cells structure, either within the yeast cells cytoplasm and / or within their cell wall. Without wishing to be bound by theory, it is believed that the yeast cells structure and the proteins they contain can participate in fat droplet stabilization.
[0151] Accordingly, in some embodiment, part of the yeast proteins, in particular insoluble yeast proteins of the yeast protein source are contained within yeast cells, in particular yeast cells cytoplasm and / or entrapped in the cell wall of yeast cells, in particular in the inner cell of yeast cells. Accordingly, in some embodiment, part of the yeast proteins, in particular insoluble yeast proteins of the liquid composition are contained within yeast cells, in particular yeast cells cytoplasm and / or entrapped in the cell wall of yeast cells, in particular in the inner cell of yeast cells.
[0152] In some embodiment, in view of the above, the yeast protein source may have low solubility as the proteins are embedded / contained within insoluble yeast cell structure. In particular, the yeast protein source has a solubility of 5 to 30%, preferably of 5 to 25%, more preferably of 5 to 20% at a pH of 2-9, preferably at a pH of 6.8.
[0153] The liquid composition is a significant source of proteins, in particular yeast proteins. Accordingly, the total protein content of the liquid composition is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0154] At least 10%, at least 20%, at least 30 %, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% of the proteins of the liquid composition are yeast proteins. In some embodiment, the proteins of the liquid composition consist only of yeast proteins.
[0155] In some embodiment, the proteins of the liquid composition comprise or are intact proteins. In some embodiment, the yeast proteins of the liquid composition comprise or are intact yeast proteins.
[0156] In some preferred embodiment, the liquid composition is a significant source of yeast proteins. In some embodiment, the total yeast protein content of the liquid composition is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0157] In some embodiment, the yeast protein source has a good protein quality. In particular the yeast protein source has a minimum PDCAAS of 0.80, preferably of 0.90, more preferably of 0.95, most preferably of 1. Likewise, the liquid composition has a minimum PDCAAS of 0.80, preferably of 0.90, more preferably of 0.95, most preferably of 1.
[0158] The protein digestibility-corrected amino acid score (PDCAAS) is a method of evaluating the quality of a protein based on both the amino acid requirements of humans and their ability to digest it. PDCAAS compares the amount of the essential amino acids in a food to a reference (scoring) pattern based on the essential amino acid requirements of a preschool-age child to determine its most limiting amino acid (amino acid score). This approach is recommended by the Food and Drug Administration (FDA) and is described in the 1991 FAO / WHO Protein Quality Report.
[0159] In an embodiment, the yeast protein source may comprise one or more conditionally essential amino acids (e.g., amino acids conditionally essential in illness or stress) selected from the group consisting of arginine, cysteine, glutamine, glycine, proline, ornithine, serine and tyrosine. In particular, the yeast protein source may comprise 15 to 30g of said conditionally essential amino acids per 100 g yeast protein in the yeast protein source.
[0160] In an embodiment, the yeast protein source may comprise one or more essential amino acids selected from the group consisting of histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. In particular, the yeast protein source may comprise 35 to 50g, preferably 40 to 50g of said essential amino acids per 100 g yeast protein in the yeast protein source.
[0161] In an embodiment, the yeast protein source may comprise one or more branched chain amino acids selected from the group consisting of leucine, Isoleucine and valine. In particular, the yeast protein source may comprise 15 to 50g, preferably 15 to 30g, more preferably 15 to 25g of said branched chain amino acids per 100 g yeast protein in the yeast protein source.
[0162] In an embodiment, the yeast protein source may comprise one or more autophagyinducing amino acids selected from the group consisting of Glycine, Cysteine, Proline, Glutamate, Valine, Tyrosine and any precursors thereof. In particular, the precursors may be selected from the Serine (as a precursor to Glycine), N-Acetyl Cysteine, Methionine (as a precursor to Cysteine). In particular, the yeast protein source may comprise 25 to 50g, preferably 30 to 40g of said one or more autophagy-inducing amino acids per 100 g yeast protein in the yeast protein source.
[0163] In an embodiment, the yeast protein source may comprise one or more anabolic amino acids selected from the group consisting of Leucine, Isoleucine and arginine. In particular, the yeast protein source may comprise 10 to 30g, preferably 15 to 25g of said one or more anabolic amino acids per 100 g yeast protein in the yeast protein source.
[0164] In an embodiment, the yeast protein source may comprise one or more acidic amino acids selected from the group consisting of aspartic acid and glutamic acid. In particular, the yeast protein source may comprise 15 to 30g, preferably 20 to 30g of said one or more acidic amino acids per 100 g yeast protein in the yeast protein source.
[0165] In an embodiment, the yeast protein source may comprise one or more aromatic amino acids selected from the group consisting of Tryptophan, Tyrosine, Phenylalanine, and Histidine. In particular, the yeast protein source may comprise 5 to 30g, preferably 10 to 18g of said one or more aromatic amino acids per 100 g yeast protein in the yeast protein source.
[0166] In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6g of Alanine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6.5g of Arginine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 5 to 15g, preferably 8 to 12g, more preferably 10 to 11.5g of Aspartic acid per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 0.5 to 3g, preferably 0.5 to 2g, more preferably 0.5 to lg of Cysteine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 5 to 15g, preferably 8 to 12g, more preferably 10 to 11.5g of Glutamic acid per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 4 to 5g of Glycine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 3 to 4g of Proline per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6g of Serine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 4 to 5g of Tyrosine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 1 to 10g, preferably 1 to 6g, more preferably 2 to 3g of Histidine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6g of Isoleucine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 5 to 15g, preferably 6 to 10g, more preferably 8 to 9g of Leucine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 5 to 15g, preferably 7 to 12g, more preferably 9 to 10g of Lysine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 0.5 to 5g, preferably 0.5 to 3g, more preferably 1 to 2g of Methionine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6g of Threonine per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 3 to 8g, more preferably 5 to 6g of Tryptophan per 100 g yeast protein in the yeast protein source. In an embodiment, the yeast protein source may comprise 2 to 10g, preferably 5 to 8g, more preferably 6 to 7g of Valine per 100 g yeast protein in the yeast protein source.
[0167] In some embodiment, the liquid composition may further comprise at least one milk protein source and / or at least one plant protein source and / or at least one collagen source and / or at least one collagen peptide source and / or at least one gelatin source and / or at least one fungal protein source and / or at least one bacterial protein source.
[0168] Examples of milk protein source include milk powder, milk protein concentrate, milk protein isolate, whey protein isolate, whey protein concentrate, whey protein hydrolysate, milk protein, hydrolysate, microparticulated whey, caseinate, micellar casein, acid whey, sweet whey, modified sweet whey, fractions of whey protein, beta-lactoglobulin concentrate, beta-lactoglobulin isolate, and mixture thereof. "Modified sweet whey" refers to sweet whey from which the caseino-glycomacropeptide has been removed. For example, the milk powder may be skimmed milk powder.
[0169] In some embodiment, the plant protein source may be selected from the list consisting of plant flour, plant protein concentrate, plant protein isolate and mixture thereof.
[0170] In some embodiment, the protein of said plant protein source may comprise or consist of protein coming from any one of pulses, nuts, oilseeds, cereals, coconut and mixture thereof.
[0171] Examples of pulses include bean, chickpea, faba, lentil, lupine, pea, soy, peanut and mixture thereof. For example, pea may be selected from split pea, cow pea, yellow pea, green pea and mixture thereof. For example, the bean may be selected from the list consisting of navy bean, black bean, butter bean, red bean, green bran, kidney bean, pinto bean, lima bean, cannellini bean, adzuki bean, mung bean, cranberry bean, Great Northern bean, yellow eye bean, black turtle bean, calypso bean, Jacob's cattle bean, tongue of fire bean, and mixture thereof. Examples of nuts include almond, cashew nut, hazelnut, macadamia nut, pecan nut, pine nut, pistachio, tiger nut, walnut and mixture thereof.
[0172] Examples of oilseeds include chia seed, Curcubitaceae seed, cotton seed, flaxseed, linseed, grape seed, hemp seed, rapeseed, sesame seed, sunflower seed, and mixture thereof. For example, the Cucurbitaceae seed may be selected from egusi seed, pumpkin seed, squash seed, watermelon seed, winter melon seed, cucumber seed, calabash seed and mixture thereof.
[0173] Examples of cereal include barley, buckwheat, maize, millet, oat, rice, rye, spelt, teff, quinoa, wheat and mixture thereof.
[0174] In some preferred embodiment, the liquid composition further comprises at least one plant protein source, wherein the proteins of said plant protein source come from pulses and / or cereals and / or oilseeds.
[0175] In particular, the pulses may be selected from the list consisting of bean, chickpea, faba, lentil, lupine, pea, soy and mixture thereof. For example, the pea may be selected from split pea, cow pea, yellow pea, green pea and mixture thereof. For example, the bean may be selected from the list consisting of navy bean, black bean, butter bean, red bean, green bran, kidney bean, pinto bean, lima bean, cannellini bean, adzuki bean, mung bean, cranberry bean, Great Northern bean, yellow eye bean, black turtle bean, calypso bean, Jacob's cattle bean, tongue of fire bean, and mixture thereof.
[0176] In particular, the oilseeds are selected from the list consisting of chia seed, Curcubitaceae seed, cotton seed, flaxseed, linseed, grape seed, hemp seed, rapeseed, sesame seed, sunflower seed, and mixture thereof. The Cucurbitaceae seed may be selected from the list consisting of egusi seed, pumpkin seed, squash seed, watermelon seed, winter melon seed, cucumber seed, calabash seed and mixture thereof.
[0177] In particular, the cereals are selected from the list consisting of barley, buckwheat, maize, millet, oat, rice, rye, spelt, teff, quinoa, wheat and mixture thereof.
[0178] In a further preferred embodiment, the cereals are oat.
[0179] In a further preferred embodiment, the oilseeds are pumpkin seeds.
[0180] In some preferred embodiment, the liquid emulsion comprises a plant protein source coming from oat.
[0181] In some preferred embodiment, the liquid emulsion comprises a plant protein source coming from soy. In some preferred embodiment, the liquid emulsion comprises a plant protein source coming from pea.
[0182] In some preferred embodiment, the liquid emulsion comprises a plant protein source coming from chickpea.
[0183] In some preferred embodiment, the liquid emulsion comprises a plant protein source coming from pumpkin seed.
[0184] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from pea, pumpkin seed and chickpea.
[0185] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from oat.
[0186] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from soy.
[0187] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from pea.
[0188] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from chickpea.
[0189] In some preferred embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from pumpkin seed.
[0190] In some embodiment, the liquid emulsion comprises a plant protein source, wherein the proteins of said plant protein source come from pea, pumpkin seed and chickpea.
[0191] The use of yeast protein source only or in combination with other protein source such as plant protein source is advantageous because it provides compositions, preferably emulsions with good protein quality. In particular, the yeast protein source has a good PDCAAS. It can also improve the protein quality of other protein sources, such as plant protein sources by bringing amino acids that are absent in such other protein sources, in particular plant protein sources. For example, the yeast proteins may bring for example lysine that is missing in plant proteins coming from some cereal or for example methionine and cysteine that is missing in plant proteins coming from some pulse.
[0192] In some embodiment, the liquid composition has a minimum PDCAAS of 0.80, preferably of 0.90, more preferably of 0.95. In some preferred embodiment, the liquid composition has PDCAAS of 1.
[0193] The protein digestibility-corrected amino acid score (PDCAAS) is a method of evaluating the quality of a protein based on both the amino acid requirements of humans and their ability to digest it. PDCAAS compares the amount of the essential amino acids in a food to a reference (scoring) pattern based on the essential amino acid requirements of a preschool-age child to determine its most limiting amino acid (amino acid score). This approach is recommended by the Food and Drug Administration (FDA) and is described in the 1991 FAO / WHO Protein Quality Report.
[0194] When using the PDCAAS method, the highest possible score is a 1.0 meaning, that after digestion, a protein having a PDCAAS of 1.0 provides (per unit of protein) 100% or more of the indispensable amino acids required.
[0195] In some embodiment, the collagen source may be any composition comprising more than 50wt.% collagen, preferably more than 80% collagen, more preferably consisting of collagen. The collagen of the collagen source may be derived from any animal, for example, from mammals such as cows, pigs, chickens, and the like, or fish.
[0196] In some embodiment, the collagen peptides source may be any composition comprising more than 50wt.% collagen peptides, preferably more than 80% collagen peptides, more preferably consisting of collagen peptides. The collagen peptides of the collagen peptides source may be derived from any animal, for example, from mammals such as cows, pigs, chickens, and the like, or fish. The type of collagen peptide is not particularly limited, and may be, for example, I type, II type, or the like.
[0197] In some embodiment, the gelatin source may be any composition comprising more than 50wt.% gelatin, preferably more than 80% gelatin, more preferably consisting of gelatin. The gelatin of the gelatin source may be derived from any animal, for example, from mammals such as cows, pigs, chickens, and the like, or fish.
[0198] The fungal protein source is an ingredient comprising proteins derived from fungi. In some embodiment, the fungal protein source may be selected from fungi biomass, fungal protein concentrate, fungal protein isolate and mixture thereof.
[0199] The bacterial protein source is an ingredient comprising proteins derived from bacteria. In some embodiment, the fungal protein source may be selected from bacteria biomass, bacterial protein concentrate, bacterial protein isolate and mixture thereof.
[0200] The proteins of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the gelatin source and / or at and / or fungal protein source and / or bacterial protein source may be unhydrolyzed, partially hydrolyzed (i.e., peptides of molecular weight 3 kDa to 10 kDa with an average molecular weight less than 5 kDa) or extensively hydrolyzed (i.e., peptides of which 90% have a molecular weight less than 3 kDa), for example in a range of 5% to 95% hydrolyzed. In some embodiments, the peptide profile of hydrolyzed protein of the of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the gelatin source and / or fungal protein source and / or bacterial protein source can be within a range of distinct molecular weights. For example, the majority of peptides (>50 molar percent or >50 wt.%) can have a molecular weight within 1-5 kDa, or 5-10 kDa, or 10-20 kDa.
[0201] The proteins and / or peptides of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the collagen peptides source and / or the gelatin source and / or fungal protein source and / or bacterial protein source can comprise essential amino acids and / or conditionally essential amino acids, e.g., such amino acids that may be insufficiently delivered in a caloric restriction regimen. For example, the proteins and / or peptides of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the collagen peptides source and / or the gelatin source and / or fungal protein source and / or bacterial protein source can comprise one or more essential amino acids selected from the group consisting of histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine; and each of these amino acids (if present) may be administered in the liquid composition in a daily dose from about 0.0476 to about 47.6 mg essential amino acid / kg bw. Notably, lower intake of methionine leads to lower levels of protein translation and ultimately muscle synthesis, the proteins and / or peptides of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the collagen peptides source and / or the gelatin source and / or the fungal protein source and / or the bacterial protein source can comprise one or more conditionally essential amino acids (e.g., amino acids conditionally essential in illness or stress) selected from the group consisting of arginine, cysteine, glutamine, glycine, proline, ornithine, serine and tyrosine; and each of these amino acids (if present) may be administered in the liquid composition in a daily dose from about 0.0476 to about 47.6 mg conditionally essential amino acid / kg bw.
[0202] The liquid composition can comprise essential free amino acids and / or conditionally essential free amino acids. For example, the liquid composition can comprise one or more essential free amino acids selected from the group consisting of histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine; and each of these amino acids (if present) may be administered in the liquid composition in a daily dose from about 0.0476 to about 47.6 mg essential free amino acid / kg bw. Notably, lower intake of methionine leads to lower levels of protein translation and ultimately muscle synthesis. The protein of the liquid composition can comprise one or more conditionally essential free amino acids (e.g., amino acids conditionally essential in illness or stress) selected from the group consisting of arginine, cysteine, glutamine, glycine, proline, ornithine, serine and tyrosine; and each of these amino acids (if present) may be administered in the liquid composition in a daily dose from about 0.0476 to about 47.6 mg conditionally essential free amino acid / kg bw.
[0203] The proteins and / or peptides of the yeast protein source and / or the milk protein source and / or the plant protein source and / or the collagen source and / or the collagen peptides source and / or the gelatin source and / or the fungal protein source and / or the bacterial protein source can comprise one or more branched chain free amino acids. A daily dose of the branched chain free amino acids can include one or more of 0.35- 142.85 mg leucine / kg bw, preferably 0.175-71.425 mg leucine / kg bw; 0.175-71.425 mg isoleucine / kg bw; and 0.175-71.425mg valine / kg bw. The daily dose of the one or more branched chain free amino acids can be provided by one or more servings of the liquid composition per day.
[0204] The liquid composition can comprise one or more branched chain free amino acids. A daily dose of the branched chain free amino acids can include one or more of 0.35- 142.85 mg leucine / kg bw, preferably 0.175-71.425 mg leucine / kg bw; 0.175-71.425 mg isoleucine / kg bw; and 0.175-71.425mg valine / kg bw. The daily dose of the one or more branched chain free amino acids can be provided by one or more servings of the liquid composition per day.
[0205] The liquid composition may be high in protein, including yeast proteins and said proteins may represent a significant portion of the energy delivered by the liquid composition.
[0206] Accordingly, in some embodiment, the liquid composition may have a protein / energy ratio greater than 6 g protein / 100 kcal, preferably greater than 9 g protein / 100 kcal. In an embodiment, the proteins represent at least 24% of the total energy of the liquid composition, more preferably at least 36% of the total energy of the liquid composition.
[0207] In some embodiment, the liquid composition may have a yeast protein / energy ratio greater than 6 g yeast protein / 100 kcal, preferably greater than 9 g yeast protein / 100 kcal. In an embodiment, the yeast proteins represent at least 24% of the total energy of the liquid composition, more preferably at least 36% of the total energy of the liquid composition.
[0208] As non-limiting examples, the liquid composition can be administered in a daily dose that provides an amount of protein greater than 1.0 g protein / kg body weight / day, preferably greater than 1.2 g protein / kg body weight / day; for example up to 2.5 g protein / kg body weight / day (e.g., 1.0-2.5 g protein / kg body weight / day; 1.2-2.5 g protein / kg body weight / day; or 1.5-2.5 g protein / kg body weight / day), preferably up to 2.0 g protein / kg body weight / day (e.g., 1.0-2.0 g protein / kg body weight / day; 1.2- 2.0 g protein / kg body weight / day; or 1.5-2.0 g protein / kg body weight / day), and more preferably up to 1.5 g protein / kg body weight / day (e.g., 1.0-1.5 g protein / kg body weight / day or 1.2- 1.5 g protein / kg body weight / day). The daily dose of the protein can be provided by one or more servings of the liquid composition per day. In some embodiment, the liquid composition can be administered enterally or parenterally, preferably enterally.
[0209] As non-limiting examples, the liquid composition can be administered in a daily dose that provides an amount of yeast protein greater than 1.0 g yeast protein / kg body weight / day, preferably greater than 1.2 g yeast protein / kg body weight / day; for example up to 2.5 g yeast protein / kg body weight / day (e.g., 1.0-2.5 g yeast protein / kg body weight / day; 1.2-2.5 g yeast protein / kg body weight / day; or 1.5-2.5 g yeast protein / kg body weight / day), preferably up to 2.0 g yeast protein / kg body weight / day (e.g., 1.0-2.0 g yeast protein / kg body weight / day; 1.2- 2.0 g yeast protein / kg body weight / day; or 1.5-2.0 g yeast protein / kg body weight / day), and more preferably up to 1.5 g yeast protein / kg body weight / day (e.g., 1.0-1.5 g yeast protein / kg body weight / day or 1.2- 1.5 g yeast protein / kg body weight / day). The daily dose of the yeast protein can be provided by one or more servings of the liquid composition per day. In some embodiment, the liquid composition can be administered orally, enterally or parenterally, preferably orally or enterally.
[0210] Non-limiting examples of suitable high protein concentrations for the liquid composition include 6-20 g protein / 100 ml, e.g., 6-11 g protein / 100 ml; 7- 14 g protein / 100 ml; 7-12 g protein / 100 ml; 8-11 g protein / 100 ml, 8-20 g protein / 100 ml; 9-20 g protein / 100 ml; and 11-20 g protein / 100 ml of liquid composition.
[0211] Non-limiting examples of suitable high yeast protein concentrations for the liquid composition include 6-20 g yeast protein / 100 ml of liquid composition, e.g., 6-11 g yeast protein / 100 ml of liquid composition; 7-14 g yeast protein / 100 ml of liquid composition; 7-12 g yeast protein / 100 ml of liquid composition; 8-11 g yeast protein / 100 ml of liquid composition, 8-20 g yeast protein / 100 ml of liquid composition; 9-20 g yeast protein / 100 ml of liquid composition; and 11-20 g yeast protein / 100 ml of liquid composition.
[0212] In some embodiment, the proteins of the liquid composition consist only of yeast proteins. In particular, it has been observed that it was possible to prepare protein-rich liquid compositions, preferably protein-rich liquid emulsions that are stable in the presence of thickening agent and yeast proteins as sole source of proteins.
[0213] In some embodiment, the proteins of the liquid composition comprise or are intact proteins. In some embodiment, the yeast proteins of the liquid composition comprise or are intact yeast proteins.
[0214] In some embodiment, the liquid composition can be administered orally, enterally or parenterally, preferably orally or enterally.
[0215] In a preferred embodiment, the liquid composition is administered to the individual orally or enterally (e.g. tube feeding).
[0216] In some embodiment, the liquid composition may comprise an aqueous liquid. The aqueous liquid forms the major liquid matrix of the liquid composition in which the other ingredients are dispersed and / or dissolved. The aqueous liquid is preferably water. The liquid composition may comprise from 40wt.% to 90wt.%, preferably 50wt.% to 90wt.%, more preferably 50wt.% to 85wt.%, even more preferably 60 wt.% to 80wt.% aqueous liquid.
[0217] In some embodiment, the liquid composition may further comprise at least one ingredient selected from the list consisting of vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, herbs and combination thereof.
[0218] In some embodiment, the liquid composition may comprise one or more vitamin(s). Non-limiting examples of vitamins include Vitamin A, Vitamin E, Vitamin C, Vitamin Bl, Vitamin B2, Pantothenic Acid, Vitamin B6, Vitamin B12, Niacin, Folic Acid, Biotin and Choline or any combination thereof. In some further embodiment, the vitamins may comprise or consist of added vitamins, i.e. vitamins that do not come from the yeast protein source.
[0219] In some embodiment, the liquid composition may comprise one or more mineral(s). Non-limiting examples of minerals include sodium, potassium, calcium, phosphorus, magnesium, chloride, iron, zinc, copper, manganese, fluoride, chromium, molybdenum, selenium, iodine or any combination thereof. The minerals may be provided in the form of salts. In some further embodiment, the minerals may comprise or consist of added minerals, i.e. minerals that do not come from the yeast protein source. In some embodiment, the minerals may comprise or be any one or more of calcium, potassium and salts thereof. The minerals may be positively or negatively charged, in particular for use as electrolytes.
[0220] In some embodiment, the liquid composition may comprise one ore more free amino acid(s). Non-limiting examples of free amino acids include Alanine, Arginine, Asparagine, Aspartate, Citrulline, Cysteine, Glutamate, Glutamine, Glycine, Histidine, Hydroxyproline, Hydroxyserine, Hydroxytyrosine, Hydroxylysine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Proline, Serine, Taurine, Threonine, Tryptophan, Tyrosine, Valine, HICA (Alpha- Hydroxyisocaproic Acid), HIVA (Alpha- Hydroxyisovaleric Acid), HIMVA (alphahydroxymethylvaleric acid) or any combination thereof . In some further embodiment, the free amino acids may comprise or consist of added free amino acids, i.e. free amino acids that do not come from the yeast protein source. The free amino acid(s) may be added to the liquid composition to achieve a desired amino acid profile / content.
[0221] In some embodiment, the liquid composition may comprise one or more carbohydrate(s). The carbohydrates are different than the non-proteic thickening agent. Nonlimiting examples of carbohydrates include glucose, fructose, maltose, lactose, sucrose, maltodextrin, corn syrup solids, glucose syrup, honey, xylitol, sorbitol or any combination thereof.
[0222] In some further embodiment, the carbohydrates preferably does not represent greater than 50% of the total energy of the liquid composition, more preferably not greater than 36 % of the total energy of the liquid composition, and most preferably not greater than 30% of the total energy of the liquid composition.
[0223] In some additional further embodiment, the liquid composition can have a high protein: carbohydrate energy ratio, for example greaterthan 0.66, preferably greaterthan 0.9 and more preferably greaterthan 1.2. Preferably, the liquid composition can have a high yeast protein: carbohydrate energy ratio, for example greaterthan 0.66, preferably greaterthan 0.9 and more preferably greater than 1.2.
[0224] In some embodiment, the liquid composition may comprise another particular type of carbohydrates: prebiotics.
[0225] In particular, the liquid composition may comprise one or more prebiotics. The prebiotics that may be used in accordance with the present invention are not particularly limited and include all food substances that promote the growth of probiotics or health beneficial micro-organisms in the intestines. They may be selected from the group consisting of oligosaccharides, optionally containing fructose, galactose, and mannose; dietary fibers, in particular soluble fibers, soy fibers; inulin; or mixtures thereof. Non-limiting examples of prebiotics are alpha glucan, beta glucan, fructo-oligosaccharides (FOS), galactooligosaccharides (GOS), isomalto-oligosaccharides (IMO), xylo-oligosaccharides (XOS), arabino-xylo oligosaccharides (AXOS), mannan-oligosaccharides (MOS), soyoligosaccharides, gentiooligosaccharides, glucooligosaccharides, inulin, polydextrose, glycosylsucrose (GS), lactosucrose (LS), lactulose (LA), palatinose-oligosaccharides (PAO), pecticoligosaccharides, malto-oligosaccharides, sugar alcohols, gums and / or hydrolysates thereof, pectins and / or hydrolysates thereof, or any combination thereof. In a particular embodiment, the prebiotics may be fructooligosaccharides and / or inulin. Suitable commercial products that can be used include combinations of FOS with inulin such as the product sold by BENEO under the trademark Orafti, or polydextrose sold by Tate & Lyle under the trademark STA-LITE®.
[0226] The prebiotics can also be a BMO (bovine's milk oligosaccharide) and / or a HMO (human milk oligosaccharide) such as N-acetylated oligosaccharides, sialylated oligosaccharides, fucosylated oligosaccharides and any mixtures thereof.
[0227] A particular example of prebiotic is a mixture of galacto-oligosaccharide(s), N- acetylated oligosaccharide(s) and sialylated oligosaccharide(s) in which the N-acetylated oligosaccharide(s) represent 0.5 to 4.0 wt% of the oligosaccharide mixture, the galacto- oligosaccharide(s) represent 92.0 to 98.5 wt% of the oligosaccharide mixture and the sialylated oligosaccharide(s) represent 1.0 to 4.0 wt% of the oligosaccharide mixture. For example, a composition for use according to the invention can contain from 2.5 to 15.0 wt% CMOS-GOS on a dry matter basis with the proviso that the composition comprises at least 0.02 wt% of an N-acetylated oligosaccharide, at least 2.0 wt% of a galacto-oligosaccharide and at least 0.04 wt% of a sialylated oligosaccharide. W02006087391 and W02012160080 provide some examples of production of such an oligosaccharide mixture.
[0228] In some embodiment, the liquid composition may comprise one or more probiotic(s). As probiotics are preferably microorganisms (alive, including semi-viable or weakened) that could confer health benefits on the host when administered in adequate amounts, more specifically that beneficially affect a host by improving its intestinal microbial balance, leading to effects on the health or well-being of the host. In general, it is believed that these probiotics inhibit and / or influence the growth and / or metabolism of pathogenic bacteria in the intestinal tract. The probiotics may also activate the immune function of the host. Non-limiting examples of probiotics include Aspergillus, Rhizopus, Mucor, Penicillium, Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus, Lactobacillus or a combination thereof.
[0229] In some embodiment, the liquid composition may comprise one or more postbiotic(s). Postbiotics include all substances, in particular metabolites or fragments derived from microorganisms that could confer health benefits on the host. Examples of postbiotics include short-chain fatty acids, microbial lysates, cellular wall fragments of microbial origin, supernatant of microbial original or any combination thereof.
[0230] In some embodiment, the liquid composition may comprise one or more synbiotic(s). The synbiotic is a supplement that contains both prebiotic(s) and probiotic(s). The prebiotic(s) and the probiotic (s) work together to improve the micro flora of the intestine. The synbiotic comprises any combination of the prebiotic(s) and the probiotic (s) referred to above.
[0231] In some embodiment, the liquid composition may comprise one or more low molecular weight surfactant(s). Non-limiting examples of low molecular weight surfactant include lecithin, mono- and diglycerides, polysorbate 80, sorbitan monostearate, sodium stearoyl lactylate, glycerol monostearate, polyglycerol esters of fatty acids, propylene glycol monostearate, sodium lauryl sulfate (SLS), sodium oleate or any combination thereof.
[0232] In some embodiment, the liquid composition may comprise one or more pharmaceutically acceptable carrier(s).
[0233] In some embodiment, the liquid composition may comprise one or more bioactive agent(s).
[0234] In some embodiment, the liquid composition may comprise one or more flavour agent(s). The flavour agent may provide a flavour and / or enhance a flavor and / or decrease a flavor and / or mask a flavor.
[0235] In some embodiment, the liquid composition may comprise one or more colorant(s).
[0236] In some embodiment, the liquid composition may comprise cocoa.
[0237] In some embodiment, the liquid composition may comprise coffee.
[0238] In some embodiment, the liquid composition may comprise malt extract. In some embodiment, the liquid composition may comprise one or more spices. Nonlimiting examples of spices include cinnamon, vanilla, curry, cumin, pepper, paprika, tonka, cardamom, saffron, ginger, nutmeg, chili pepper, allspice, cloves and mixtures thereof.
[0239] In some embodiment, the liquid composition may comprise herbs. Non-limiting examples of herbs include mint, thyme, coriander, basil, verbena, fennel, chervil, rosemary, lemon balm, sage, oregano, and mixtures thereof.
[0240] In some specific embodiment, the liquid composition comprises:
[0241] - a fat component, and,
[0242] - a yeast protein source, and,
[0243] - a carbohydrate, and,
[0244] - at least one non-proteic thickening agent, and,
[0245] -aqueous liquid, wherein the yeast protein source comprises at least 40wt.% yeast proteins, and wherein the total protein content of the liquid composition is of at least 5.01wt.%. In some further embodiment, the yeast protein has less than 20wt.% mannoproteins.
[0246] For sake of clarity, the carbohydrate is different than non-proteic thickening agent.
[0247] In this specific embodiment the fat component, the yeast protein source (incl. mannoprotein content and yeast protein content), the carbohydrate, the non-proteic thickening agent and the total protein content may be as described hereinabove. Preferably, the fat component is high oleic sunflower oil. Preferably, the yeast protein source is yeast protein concentrate. Preferably, the carbohydrate, in particular carbohydrate different than non-proteic thickening agent is sucrose and / or glucose syrup. Preferably, the non-proteic thickening agent is gellan gum or guar gum or a combination of gellan gum and guar gum.
[0248] Also, in this specific embodiment, the quantity of fat component, yeast protein source, non-proteic thickening agent and aqueous liquid in the liquid composition may be as described herein above.
[0249] The quantity of the carbohydrate, in particular carbohydrate different from non- proteic thickening agent in the liquid composition may be from 0.3wt.% to 25wt.%, more preferably 0.5wt.% to 15wt.%, even more preferably 2 to 12wt.%, even more preferably 5 to 12wt.%, even more preferably 8 to 12wt.%. In a second aspect, the invention relates to a powder composition which is obtained by drying the liquid composition according to the first aspect of the invention.
[0250] In a preferred embodiment, the powder composition is a powdered emulsion, preferably powdered oil-in-water emulsion.
[0251] The powder composition may be stored in a sachet and then suspended in an aqueous liquid such as water for use.
[0252] The drying of the liquid composition may be performed by any drying method suitable for food, nutritional and / or pharmaceutical product. For example, the drying of the liquid composition may be performed by freeze drying, roller drying, spray drying, vacuum drying or drum drying.
[0253] The powder composition may have the same features as the liquid composition of the first aspect of the invention, except it is not liquid except it is not liquid and does not comprise liquid components. Or, if any liquid components are used in the preparation of the powder composition, they are no longer in liquid state in the powder composition due to drying.
[0254] In particular, the powder composition, preferably powdered emulsion of the invention provides a liquid composition, preferably liquid emulsion high in protein, in particular yeast protein, upon reconstitution in an aqueous liquid, preferably water. The obtained liquid composition, preferably liquid emulsion has the advantage of the liquid composition, preferably liquid emulsion of the first aspect. In particular, upon reconstitution, the obtained liquid composition, preferably liquid emulsion is stable, in particular, for example it remains liquid, and exhibits limited or no phase separation, such as protein sedimentation and creaming.
[0255] In a third aspect, the invention relates to the use of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention in the preparation and / or formulation of a product selected from the list consisting of beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake, powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement (ONS), tube feeding and combinations thereof.
[0256] As used herein, the "incomplete nutritional composition" refers to preferably nutritional compositions that do not contain sufficient levels of macronutrients (protein, fats and carbohydrates) or micronutrients to be sufficient to be a sole source of nutrition for the subject, in particular human or non-human animal, which consume the nutritional composition or the subject, in particular human or non-human animal to which the composition product is being administered. Preferably, the non-human animal is a pet.
[0257] As used herein, the "complete nutritional composition" refers to nutritional compositions that contain sufficient levels of macronutrients (protein, fats and carbohydrates) or micronutrients to be sufficient to be a sole source of nutrition for the subject, in particular human or non-human animal, which consume the nutritional product or the subject, in particular human or non-human animal to which the nutritional composition is being administered. Preferably, the non-human animal is a pet.
[0258] In some embodiment, the product is vegetarian. In some embodiment, the product is vegan.
[0259] In some embodiment, the product comprises 0.5wt.% to 99wt.%, preferably lwt.% to 99wt.%, more preferably 10wt.% to 99wt.%, even more preferably 30wt.% to 99wt.%, even more preferably 50wt.% to 99wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0260] In some preferred embodiment, the product further comprises at least one plant protein source, wherein the proteins of said plant protein source come from pulses and / or cereals. The pulses and / or cereals may be as provided in the first aspect of the invention.
[0261] The product is a significant source of proteins. In particular, the total protein content of the product is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0262] At least 10%, at least 20%, at least 30 %, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% of the proteins of the product are yeast proteins. In some embodiment, the proteins of the product consist only of yeast proteins.
[0263] In some embodiment, the proteins of the product comprise or are intact proteins. In some embodiment, the yeast proteins of the product comprise or are intact yeast proteins. In some preferred embodiment, the product is a significant source of yeast proteins. In some embodiment, the total yeast protein content of the product is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0264] In some advantageous embodiment, the product comprises an insoluble yeast proteins to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20. The advantage of this ratio is provided in the first aspect of the invention. The insoluble yeast proteins to soluble yeast proteins ratio is a concentration ratio. In particular, the insoluble yeast proteins to soluble yeast proteins ratio corresponds to the ratio between the insoluble yeast protein concentration to soluble yeast protein concentration of a composition (here, of the product).
[0265] In some embodiment, the product comprises at least one non-proteic thickening agent. The non-proteic thickening agent may be selected from the list consisting of alginate, xanthan gum, pectin, locust bean gum, gellan gum, carrageenan, guar gum, cellulose, carboxymethylcellulose, agar, gum arabic, starch, konjac gum, tara gum, chitosan, tragacanth gum, psyllium husk and mixture thereof.
[0266] In a preferred embodiment, the non-proteic thickening agent is gellan gum or guar gum or a combination of gellan gum and guar gum.
[0267] The quantity of non-proteic thickening agent in the product may be of at least 0.01wt.%, preferably from 0.01wt.% to 2wt.%, more preferably 0.05wt.% to 1.5wt.%, even more preferably 0.1 to 1.5wt.%. The advantage of the non-proteic thickening agent is provided in the first aspect of the invention.
[0268] In some embodiment, the product comprises at most 30wt.%, preferably at most 18wt.% of soluble yeast proteins.
[0269] In some embodiment, the product comprises less than 5wt.%, preferably less than 4wt.%, more preferably less than 3wt.%, even more preferably less than 2wt.% mannoproteins. In some embodiment, the product may comprise 0.4wt.% to 5wt.%, preferably 0.4wt.% to 4wt.%, more preferably 0.4wt.% to 3wt.%, even more 0.4wt.% to 2wt.% mannoproteins. In some other embodiment, the product may comprise lwt.% to 5wt.%, preferably lwt.% to 4wt.%, more preferably lwt.% to 3wt.%, even more lwt.% to 2wt.% mannoproteins. The advantage of low mannoprotein content is provided in the first aspect of the invention. Less than 20%, less than 10%, preferably less than 5%, more preferably less than 4% of the proteins of the product are mannoproteins.
[0270] In some embodiment, the product may further comprise at least one milk protein source and / or at least one plant protein source and / or at least one collagen source and / or at least one collagen peptide source and / or at least one gelatin source and / or at least one fungal protein source and / or at least one bacterial protein source. The milk protein source, the plant protein source, the collagen source, the collagen peptide source, the gelatin source, the fungal protein source and the bacterial protein source may be as provided in the first aspect of the invention.
[0271] In some embodiment, the product has a minimum PDCAAS of 0.80, preferably of 0.90, more preferably of 0.95. In some preferred embodiment, the product has PDCAAS of 1.
[0272] In some embodiment, the product may comprise a fat component. In particular, the fat component may be as provided in the first aspect of the invention.
[0273] In some embodiment, the product has a 2 to 9, preferably of 4 to 8, more preferably 5 to 7.
[0274] In some embodiment, the product may be chilled storage product or ambient storage product.
[0275] By "chilled storage", it is understood a product which has a shelf-life of several days when stored under chilled conditions. The term "chilled conditions" refers to temperatures ranging from 2°C to 14°C, preferably from 2°C to 10°C, more preferably from 4°C to 8°C. In particular, a chilled storage product has a shelf-life of at least 25 days, preferably of at least 30 days when stored under chilled conditions. These storage temperatures relate to the storage of the product before being commercially obtained by an end consumer. Generally, the end consumer is advised to store the product under the same chilled conditions until consumption, for example in a refrigerator.
[0276] By "ambient storage", it is understood a product which has a shelf-life of several months when stored under ambient conditions. This also generally applies when it is stored under chilled conditions. The term "ambient conditions" refers to temperatures ranging from 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C. Especially, an ambient storage product has a shelf-life of at least 1 month, preferably at least 3 months, more preferably at least 6 months, even more preferably 8 months, even more preferably at least 12 months, even more preferably of at least 24 months, even more preferably of at least 30 months when stored under ambient conditions. In some further embodiment, an ambient storage product has a shelf-life of at most 30 months, preferably at most 18 months, more preferably at most 12 months when stored under ambient conditions. These storage temperatures relate to the storage of the product before being commercially obtained by an end consumer. Generally, the end consumer is advised to store the product under the same ambient conditions until consumption, for example in a shelf at room temperature.
[0277] In a preferred embodiment, the product is ambient storage product. The definition and features of "ambient storage" product(s) is provided above.
[0278] The product is advantageous on two aspects.
[0279] First, it provides significant amount of protein while remaining stable upon processing for preparing ambient storage product that include high temperature treatment, e.g. UHT. In particular, it does not exhibit excessive gelation during the manufacturing process, including important temperature treatment. For products in liquid format, they remain liquid during the manufacturing process, including important temperature treatment. In addition, it does not lead to fouling and blocking of manufacturing lines during the manufacturing process, including important temperature treatment.
[0280] Second, it provides significant amount of protein while remaining stable during storage for several months under ambient conditions. In particular, it does not exhibit phase separation phenomenon, including creaming (i.e. fat separation) and protein sedimentation during storage for several months under chilled and ambient conditions. In addition, for products in liquid format, they remain liquid during storage for several months under chilled and ambient condition.
[0281] Products with yeast proteins were observed to be particularly stable in the above conditions, i.e. upon processing including high temperature treatment and during storage under chilled and ambient conditions.
[0282] Accordingly, in an advantageous embodiment, the ambient storage product is shelfstable for at least 1 month, preferably at least 3 months, more preferably at least 6 months, even more preferably at least 8 months, more preferably at least 12 months, even more preferably of at least 24 months, even more preferably of at least 30 months at 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C. In some further embodiment, the ambient storage product is shelf-stable for at most 30 months, preferably at most 18 months, more preferably at most 12 months at 15°C to 37° C, preferably 18°C to 37°C, more preferably at 18°C to 30°C, even more preferably at 25°C to 30°C, most preferably at 25°C and / or 30°C.
[0283] By "shelf stable", it is understood that the product keeps its consistency unchanged and does not exhibit protein sedimentation and fat separation, in particular creaming under the abovementioned ambient storage time and ambient temperature conditions. This can be assessed by visual inspection. In particular, the consistency properties of the product may be assessed by pouring or agitating the product and observing if the product has the same consistency. This also can be assessed by measuring the viscosity of the product as disclosed in the example over the shelf life. The separation of phase in the product may be observed by appearance of several phase in the product (heterogenous product appearance), e.g. appearance of fat phase (creaming), appearance of solid particles, in particular generally at the bottom of the product (sedimentation). The separation of phase may also be assessed by measuring the backscattering of the product with a Turbiscan Lab Expert (Formulaction, Toulouse, France) according to the method of example 3 over the shelf life.
[0284] In addition, the term "shelf stable" may also include that the liquid composition does not spoil under the abovementioned storage time and temperature conditions.
[0285] In some embodiment, the term "shelf stable" includes that the emulsion remains stable over the shelf life.
[0286] The product can comprise essential free amino acids and / or conditionally essential free amino acids. For example, the product can comprise one or more essential free amino acids selected from the group consisting of histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine; and each of these amino acids (if present) may be administered in the product in a daily dose from about 0.0476 to about 47.6 mg essential free amino acid / kg bw. Notably, lower intake of methionine leads to lower levels of protein translation and ultimately muscle synthesis. The protein of the product can comprise one or more conditionally essential free amino acids (e.g., amino acids conditionally essential in illness or stress) selected from the group consisting of arginine, cysteine, glutamine, glycine, proline, ornithine, serine and tyrosine; and each of these amino acids (if present) may be administered in the product in a daily dose from about 0.0476 to about 47.6 mg conditionally essential free amino acid / kg bw.
[0287] The product can comprise one or more branched chain free amino acids. A daily dose of the branched chain free amino acids can include one or more of 0.35- 142.85 mg leucine / kg bw, preferably 0.175-71.425 mg leucine / kg bw; 0.175-71.425 mg isoleucine / kg bw; and 0.175- 71.425mg valine / kg bw. The daily dose of the one or more branched chain free amino acids can be provided by one or more servings of the product per day.
[0288] The product may be high in protein, including yeast proteins and said proteins may represent a significant portion of the energy delivered by the product.
[0289] Accordingly, in some embodiment, the product may have a protein / energy ratio greater than 6 g protein / 100 kcal, preferably greater than 9 g protein / 100 kcal. In an embodiment, the proteins represent at least 24% of the total energy of the product, more preferably at least 36% of the total energy of the product.
[0290] In some embodiment, the product may have a yeast protein / energy ratio greater than 6 g yeast protein / 100 kcal, preferably greater than 9 g yeast protein / 100 kcal. In an embodiment, the yeast proteins represent at least 24% of the total energy of the product, more preferably at least 36% of the total energy of the product.
[0291] As non-limiting examples, the product can be administered in a daily dose that provides an amount of protein greater than 1.0 g protein / kg body weight / day, preferably greater than 1.2 g protein / kg body weight / day; for example up to 2.5 g protein / kg body weight / day (e.g., 1.0-2.5 g protein / kg body weight / day; 1.2-2.5 g protein / kg body weight / day; or 1.5-2.5 g protein / kg body weight / day), preferably up to 2.0 g protein / kg body weight / day (e.g., 1.0-2.0 g protein / kg body weight / day; 1.2- 2.0 g protein / kg body weight / day; or 1.5-2.0 g protein / kg body weight / day), and more preferably up to 1.5 g protein / kg body weight / day (e.g., 1.0-1.5 g protein / kg body weight / day or 1.2- 1.5 g protein / kg body weight / day). The daily dose of the protein can be provided by one or more servings of the product per day. In some embodiment, the product can be administered orally, enterally or parenterally, preferably orally or enterally.
[0292] As non-limiting examples, the product can be administered in a daily dose that provides an amount of yeast protein greater than 1.0 g yeast protein / kg body weight / day, preferably greater than 1.2 g yeast protein / kg body weight / day; for example up to 2.5 g yeast protein / kg body weight / day (e.g., 1.0-2.5 g yeast protein / kg body weight / day; 1.2-2.5 g yeast protein / kg body weight / day; or 1.5-2.5 g yeast protein / kg body weight / day), preferably up to 2.0 g yeast protein / kg body weight / day (e.g., 1.0-2.0 g yeast protein / kg body weight / day; 1.2- 2.0 g yeast protein / kg body weight / day; or 1.5-2.0 g yeast protein / kg body weight / day), and more preferably up to 1.5 g yeast protein / kg body weight / day (e.g., 1.0-1.5 g yeast protein / kg body weight / day or 1.2- 1.5 g yeast protein / kg body weight / day). The daily dose of the yeast protein can be provided by one or more servings of the product per day. In some embodiment, the product can be administered enterally or parenterally, preferably enterally.
[0293] Non-limiting examples of suitable high protein concentrations for the product include
[0294] 6-20 g protein / 100 ml, e.g., 6-11 g protein / 100 ml; 7- 14 g protein / 100 ml; 7-12 g protein / 100 ml; 8-11 g protein / 100 ml, 8-20 g protein / 100 ml; 9-20 g protein / 100 ml; and 11-20 g protein / 100 ml of product.
[0295] Non-limiting examples of suitable high yeast protein concentrations for the product include 6-20 g yeast protein / 100 ml of product, e.g., 6-11 g yeast protein / 100 ml of product;
[0296] 7-14 g yeast protein / 100 ml of product; 7-12 g yeast protein / 100 ml of product; 8-11 g yeast protein / 100 ml of product, 8-20 g yeast protein / 100 ml of product; 9-20 g yeast protein / 100 ml of product; and 11-20 g yeast protein / 100 ml of product.
[0297] The product can induce autophagy in muscle, for example a skeletal muscle. Nonlimiting examples of such muscle include one or more of the following: vastus lateralis, gastrocnemius, tibialis, soleus, extensor, digitorum longus (EDL), biceps femoris, semitendinosus, semimembranosus, gluteus maximus, extra-ocular muscles, face muscles or diaphragm.
[0298] In some embodiment, the proteins of the product consist only of yeast proteins.
[0299] In some embodiment, the proteins of the product comprise or are intact proteins. In some embodiment, the yeast proteins of the product comprise or are intact yeast proteins.
[0300] In some embodiment, the product can be administered orally, enterally or parenterally, preferably orally or enterally.
[0301] In some embodiment, the product, in particular comprising high protein content, can be administered to an individual such as a human, e.g., an ageing individual or a critically ill individual, in a therapeutically effective dose. The therapeutically effective dose can be determined by the person skilled in the art and will depend on a number of factors known to those of skill in the art, such as the severity of the condition and the weight and general state of the individual.
[0302] The product is preferably administered to the individual at least two days per week, more preferably at least three days per week, most preferably all seven days of the week; for at least one week, at least one month, at least two months, at least three months, at least six months, or even longer. In some embodiments, the product is administered to the individual consecutively for a number of days, for example at least until a therapeutic effect is achieved. In an embodiment, the product can be administered to the individual daily for at least 30, 60 or 90 consecutive days.
[0303] The above examples of administration do not require continuous daily administration with no interruptions. Instead, there may be some short breaks in the administration, such as a break of two to four days during the period of administration. The ideal duration of the administration of the composition can be determined by those of skill in the art.
[0304] In a preferred embodiment, the product is administered to the individual orally or enterally (e.g. tube feeding).
[0305] In some embodiments, the product is administered to the individual in a single dosage form, i.e. all compounds are present in one product to be given to an individual in combination with a meal. In other embodiments, the product is co-administered in separate dosage forms, for example at least one component separately from one or more of the other components of the composition.
[0306] The product may be liquid, semi-liquid, solid or powder.
[0307] In some embodiment, the product may comprise an aqueous liquid. In particular, if the product is liquid or semi-liquid or solid, the aqueous liquid forms the major liquid matrix of the product in which the other ingredients are dispersed and / or dissolved. The product is preferably water. The product may comprise from 40wt.% to 99wt.%, preferably 50wt.% to 95wt.%, more preferably 50wt.% to 90wt.%, even more preferably 60 wt.% to 90wt.% aqueous liquid.
[0308] In some embodiment, the product may comprise a powdered carrier matrix. In particular, if the product is powder or solid, the powdered carrier matrix forms the major matrix of the product in which the other ingredients are dispersed into or mixed with. Examples of carrier matrix include carbohydrates such as maltodextrin. The product may comprise from 40wt.% to 99wt.%, preferably 50wt.% to 95wt.%, more preferably 50wt.% to 90wt.%, even more preferably 60 wt.% to 90wt.% powdered carrier matrix.
[0309] In some embodiment, the product may further comprise at least one ingredient selected from the list consisting of vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, herbs, and combination thereof. The vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices and herbs may be as provided in the first aspect of the invention.
[0310] In some embodiment, the product is not a dressing. In some embodiment, the product is not an animal feed. In some embodiment, the product is suitable for human consumption.
[0311] In a fourth aspect, the invention relates to a product comprising the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention, wherein the product is selected from the list consisting of beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake, powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement (ONS), tube feeding and combinations thereof.
[0312] The product comprises at least lwt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0313] The product comprises lwt.% to 99wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0314] The product comprises 5wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0315] The product comprises 10wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0316] The product comprises 20wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0317] The product comprises 30wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0318] The product comprises 40wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention. The product comprises 50wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0319] The product comprises 60wt.% to 90wt.% of the liquid composition of the first aspect of the invention or the powder composition of the second aspect of the invention.
[0320] In some embodiment, the product is not a dressing. In some embodiment, the product is not an animal feed. In some embodiment, the product is suitable for human consumption.
[0321] The product may have the same features as the product disclosed in the third aspect of the invention.
[0322] In a fifth aspect, the invention relates to a process for preparing a liquid composition. In some embodiment, the liquid emulsion may be a liquid composition as described in the first aspect of the invention.
[0323] The process comprises a step (a) of dispersing a yeast protein source in an aqueous liquid to form a yeast protein dispersion. The yeast protein source comprises at least 40wt.% yeast proteins. The yeast protein source may be a yeast protein source as provided in the first aspect of the invention. In particular, for example, the yeast protein source may have a mannoprotein content as provided in the first aspect of the invention.
[0324] In some embodiment, the total protein content of the yeast protein dispersion is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0325] At least 10%, at least 20%, at least 30 %, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% of the proteins of the product are yeast proteins. In some embodiment, the proteins of the yeast protein dispersion consist only of yeast proteins.
[0326] In some preferred embodiment, the yeast protein dispersion is a significant source of yeast proteins. In some embodiment, the total yeast protein content of the yeast protein dispersion is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0327] In some embodiment, the proteins of the yeast protein dispersion comprise or are intact proteins. In some embodiment, the yeast proteins of the yeast protein dispersion comprise or are intact yeast proteins.
[0328] In some advantageous embodiment, the yeast protein dispersion comprises an insoluble yeast proteins to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20. The advantage of this ratio is provided in the first aspect of the invention. The insoluble yeast proteins to soluble yeast proteins ratio is a concentration ratio. In particular, the insoluble yeast proteins to soluble yeast proteins ratio corresponds to the ratio between the insoluble yeast protein concentration to soluble yeast protein concentration of a composition (here, of the yeast protein dispersion). The insoluble and soluble yeast protein concentration may be measured as described in the first aspect of the invention. In some embodiment, the yeast protein dispersion comprises at most 30wt.%, preferably at most 18wt.% of soluble yeast proteins.
[0329] In some embodiment, the yeast protein dispersion comprises less than 5wt.%, preferably less than 4wt.%, more preferably less than 3wt.%, even more preferably less than 2wt.% mannoproteins.
[0330] In some embodiment, the yeast protein dispersion may comprise 0.4wt.% to 5wt.%, preferably 0.4wt.% to 4wt.%, more preferably 0.4wt.% to 3wt.%, even more 0.4wt.% to 2wt.% mannoproteins. In some other embodiment, the yeast protein dispersion may comprise lwt.% to 5wt.%, preferably lwt.% to 4wt.%, more preferably lwt.% to 3wt.%, even more lwt.% to 2wt.% mannoproteins. The advantage of low mannoprotein content is provided in the first aspect of the invention.
[0331] Less than 20%, less than 10%, preferably less than 5%, more preferably less than 4% of the proteins of the yeast protein dispersion are mannoproteins.
[0332] In certain embodiments, step a) involves dispersing the yeast protein source in the aqueous liquid together with at least one milk protein source, and / or at least one plant protein source, and / or at least one collagen source, and / or at least one collagen peptide source, and / or at least one gelatin source and / or at least one fungal protein source and / or at least one bacterial protein source. The milk protein source, the plant protein source, the collagen source, the collagen peptide source, the gelatin source, the fungal protein source and the bacterial source may be as provided in the first aspect of the invention.
[0333] In some embodiment, the yeast protein dispersion obtained in step (a) may be mixed with at least one milk protein source and / or at least one plant protein source and / or at least one collagen source and / or at least one collagen peptide source and / or at least one gelatin source and / or at least one fungal protein source and / or at least one bacterial protein source.
[0334] The milk protein source, the plant protein source, the collagen source, the collagen peptide source, the gelatin source, the fungal protein source, the bacterial protein source may be as provided in the first aspect of the invention. In some embodiment, before step (b), the yeast protein dispersion obtained in step (a) may be mixed with an ingredient selected from the list consisting of vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, herbs and combination thereof. The vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, symbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, and herbs may be as provided in the first aspect of the invention.
[0335] The process further comprises a step (b) of adding a fat component to the yeast protein dispersion to form a liquid pre-emulsion. The fat component may be a fat component as provided in the first aspect of the invention. In some embodiment, the fat component may be added to the yeast protein dispersion in an amount of at least 0.5wt.%, preferably 0.5 to 20wt.%, more preferably 0.5 to 10wt.%, even more preferably 1 to 10wt%, even more preferably 1 to 5wt. In some embodiment, the fat component may be added to the yeast protein dispersion in an amount of 8 to 15wt% fat component.
[0336] Hence, in some embodiment, the liquid pre-emulsion may comprise at least 0.5wt.%, preferably 0.5 to 20wt.%, more preferably 0.5 to 10wt.%, even more preferably 1 to 10wt%, even more preferably 1 to 5wt.% fat component, in particular fat. In some other embodiment, the liquid pre-emulsion may comprise 8 to 15wt% fat component, in particular fat.
[0337] In some embodiment, the liquid pre-emulsion is a significant source of protein. In particular, the total protein content of the liquid pre-emulsion is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0338] At least 10%, at least 20%, at least 30 %, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% of the proteins of the liquid pre-emulsion are yeast proteins. In some embodiment, the proteins of the liquid pre-emulsion consist only of yeast proteins.
[0339] In some embodiments, the proteins of the liquid pre-emulsion comprise or are intact proteins. In some embodiments, the yeast proteins of the liquid pre-emulsion comprise or are intact yeast proteins.
[0340] In some preferred embodiment, the liquid pre-emulsion is a significant source of yeast proteins. In some embodiment, the total yeast protein content of the liquid pre-emulsion is of at least 5.01wt.%, preferably of 5.01wt.% to 20wt.%, more preferably of 5.01wt.% to 15wt.%, even more preferably of 5.01wt.% to 10wt.%.
[0341] In some advantageous embodiment, the liquid pre-emulsion comprises an insoluble yeast proteins to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20. The advantage of this ratio is provided in the first aspect of the invention. The insoluble yeast proteins to soluble yeast proteins ratio is a concentration ratio. In particular, the insoluble yeast proteins to soluble yeast proteins ratio corresponds to the ratio between the insoluble yeast protein concentration to soluble yeast protein concentration of a composition (here, of the liquid pre-emulsion). The insoluble and soluble yeast protein concentration may be measured as described in the first aspect of the invention. In some embodiment, the liquid pre-emulsion comprises at most 30wt.%, preferably at most 18wt.% of soluble yeast proteins.
[0342] In some embodiment, the liquid pre-emulsion comprises less comprises less than 5wt.%, preferably less than 4wt.%, more preferably less than 3wt.%, even more preferably less than 2wt.% mannoproteins.
[0343] In some embodiment, the liquid pre-emulsion may comprise 0.4wt.% to 5wt.%, preferably 0.4wt.% to 4wt.%, more preferably 0.4wt.% to 3wt.%, even more 0.4wt.% to 2wt.% mannoproteins. In some other embodiment, the liquid pre-emulsion may comprise lwt.% to 5wt.%, preferably lwt.% to 4wt.%, more preferably lwt.% to 3wt.%, even more lwt.% to 2wt.% mannoproteins. The advantage of low mannoprotein content is provided in the first aspect of the invention.
[0344] Less than 20%, less than 10%, preferably less than 5%, more preferably less than 4%of the proteins of the liquid pre-emulsion are mannoproteins.
[0345] In some embodiment, before step (c) or (d), the liquid pre-emulsion obtained in step (a) may be mixed with at least one ingredient selected from the list consisting of vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, herbs and combination thereof. The vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, symbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices and herbs may be as provided in the first aspect of the invention.
[0346] The process comprises a step (c) of adding at least one non-proteic thickening agent to the liquid pre-emulsion, in particular the liquid pre-emulsion obtained in step (b). The non- proteic thickening agent may be selected from the list consisting of alginate, xanthan gum, pectin, locust bean gum, gellan gum, carrageenan, guar gum, cellulose, carboxymethylcellulose, agar, gum arabic, starch, konjac gum, tara gum, chitosan, tragacanth gum, psyllium husk and mixture thereof. In a preferred embodiment, the non-proteic thickening agent is gellan gum or guar gum or a combination of gellan gum and guar gum.
[0347] In some embodiment, the non-proteic thickening agent may be added to the yeast protein dispersion in an amount of at least 0.01wt.%, preferably from 0.01wt.% to 2wt.%, more preferably 0.05wt.% to 1.5wt.%, even more preferably 0.1 to 1.5wt.%.
[0348] The process comprises a step (d) of homogenizing and heat-treating the liquid preemulsion to form a liquid composition, preferably a liquid emulsion. The heat treatment is before and / or after the homogenization.
[0349] The homogenization step of step (d) may be performed at a pressure above 50 bar. Preferably, the homogenizing step may be performed at a pressure of 50 bar to 700 bar. Further preferably, the homogenizing step may be performed at a pressure of 50 bar to 500 bar. More preferably, the homogenizing step may be performed at a pressure of 50 to 300 bar, from 100 to 300 bar or from 150 to 300 bar. In a preferred embodiment, the homogenization step may be performed at a temperature from 50°C to 70°C. More preferably, the step may be performed at a temperature from 55°C to 65°C.
[0350] The heat treatment of step (d) may be performed a temperature of at least 75°C, preferably of at least 80°C, more preferably at least 90°C. In addition, the heat treatment of step e) may be performed at a temperature of at most 140°C, preferably at most 135°C, more preferably at most 125°C. In addition, the heat treatment of step e) may be performed for a time of at least 3 seconds, preferably 3 seconds to 20 minutes, more preferably 3 seconds to 90 seconds.
[0351] The process may further comprise a step (e) of filling the liquid composition of step (d) in a container.
[0352] In some embodiment, in step (e), the filling may be performed by aseptic filling and the container may be an aseptic container.
[0353] When the filling is performed by aseptic filling, the aseptic filling process involves the use of thereof and the sterilized filling equipment and container to ensure that the liquid composition is not contaminated by any unwanted microorganisms during the filling process. The filling equipment and the packaging may be typically sterilized through the use of heat, chemical, sterilant, and / or radiation. The term "aseptic container" refers to a container which has been sterilized. In other words, "aseptic container" refers to a packaging which has been treated such that it is not contaminated by any unwanted microorganisms.
[0354] Once the container and filling equipment have been sterilized, the aseptic filling process can begin. This is typically done in a controlled environment to further minimize the risk of contamination. The liquid composition is transferred into the aseptic container using the sterilized filling equipment, ensuring that at no point the liquid composition come into contact with any non-sterilized surfaces.
[0355] Aseptic filling is generally performed at low temperature. In particular, the aseptic filling may be performed at a temperature of 25°C or less, preferably at a temperature of 3°C to 25°C.
[0356] In some embodiment, in step (e), the filling may be performed by hot filling. In this embodiment, the container may be aseptic container as disclosed above.
[0357] When the filling is performed by hot filling, the hot filling process involves the filling of the liquid composition at a high temperature in a container to ensure microbial safety and extend shelf life. The hot filling is generally performed at elevated temperature. In particular, the hot filling may be performed at a temperature above 75°C, preferably above 85°C, more preferably of 85°C to 140°C, even more preferably 85°C to 100°C. The high temperature of the liquid composition and the container helps to kill or inhibit the growth of unwanted microorganisms, ensuring the safety and preservation of the packaged liquid composition.
[0358] After step (e), the container may be sealed. It may be sealed using sterilized equipment to ensure that the liquid composition in the container remains free from contamination during storage and distribution. In some embodiment, the container may be sealed with a lid and / or a cap.
[0359] In a sixth aspect, the invention relates to a process for preparing a powder composition which comprises the step of drying the liquid composition obtained in step (d) of the fifth aspect of the invention or the liquid composition of the first aspect of the invention.
[0360] In a preferred embodiment, the powder composition is a powdered emulsion, preferably powdered oil-in-water emulsion.
[0361] The powder composition may be stored in a sachet and then suspended in an aqueous liquid such as water for use.
[0362] The drying of the liquid composition may be performed by any drying method suitable for food, nutritional and / or pharmaceutical product. For example, the drying of the liquid composition may be performed by freeze drying, roller drying, spray drying, vacuum drying or drum drying.
[0363] The powder composition may have the same features as the liquid composition of the first aspect of the invention, except it is not liquid and does not comprise liquid components. Or, if any liquid components are used in the preparation of the powder composition, they are no longer in liquid state in the powder composition due to drying.
[0364] In particular, the powder composition, preferably powdered emulsion of the invention provides a liquid composition, preferably liquid emulsion high in protein, in particular yeast protein, upon reconstitution in an aqueous liquid, preferably water. The obtained liquid composition, preferably liquid emulsion has the advantage of the liquid composition, preferably liquid emulsion of the first aspect. In particular, upon reconstitution, the obtained liquid composition, preferably liquid emulsion is stable: remains liquid, and exhibits limited or no phase separation, such as protein sedimentation and creaming..
[0365] Those skilled in the art will understand that they can freely combine all features of the present invention disclosed herein. In particular, features described for the compositions and products of the present invention may be combined with the processes, methods and uses of the present invention and vice versa. Further, features described for different embodiments of the present invention may be combined.
[0366] Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred in this specification. Further advantages and features of the present invention are apparent from the figures and non-limiting examples.
[0367] EXAMPLES
[0368] Example 1: Nutrient composition and physicochemical properties of the protein ingredients.
[0369] Yeast protein ingredients-The yeast protein ingredients used included two commercial protein concentrates from Saccharomyces cerevisiae, referred to hereinafter as YPC 1 and YPC 2. The two yeast protein concentrates, YPC1 and YPC2, are spray dried concentrates. The protein ingredients used to produce the reference RTD beverage were commercial milk protein concentrate (MPC) and soy protein isolate (SPI). Other ingredients-Other ingredients used included commercial high oleic sunflower oil, white sugar, glucose syrup and gellan gum. pH adjustment of the emulsions was performed using commercial sodium hydroxide.
[0370] The proximate composition of the protein ingredients was determined using the standard methods of the Association of Analytical Chemists (AOAC), 2023 as described below.
[0371] Protein concentration measurement- Total nitrogen was determined by the Kjeldahl method and a nitrogen-protein conversion factor of 6.25 was used to calculate the protein concentration of the protein ingredients.
[0372] Moisture concentration measurement-Moisture concentration was determined by oven drying at 103°C for 5 hours.
[0373] Ash concentration measurement-Ash concentration was determined by dry ashing in a muffle furnace at 500°C for 5 hours.
[0374] Fat concentration measurement-Fat concentration was determined by acid hydrolysis using a Hydrotherm (Gerhardt Analytical Systems, Kbnigswinter, Germany) followed by extraction with petroleum ether using a Soxtherm (Gerhardt Analytical Systems, Kbnigswinter, Germany).
[0375] Total dietary fiber measurement-Total dietary fiber was determined using the enzymatic kit K-TDFR (Megazyme, Bray, Co. Wicklow, Ireland).
[0376] Total carbohydrate measurement-Total carbohydrate (excluding fibre) was calculated by difference (100 - sum of protein, moisture, ash, fat and fibre). T
[0377] PDCAAS measurement-the Protein Digestibility-Corrected Amino Acid Score (PDCAAS) was determined using the Megazyme K-PDCAAS assay kit, an in vitro digestion method, according to the protocol suggested by the supplier (Megazyme, Megazyme, Bray, Co. Wicklow, Ireland). The proximate composition and PDCAAS of the protein ingredients are reported in Table 1. Table 1: Proximate composition (g / 100 g) and PDCAAS of yeast protein concentrate 1 (YPC 1), yeast protein concentrate 2 (YPC 2) and soy protein isolate (SPI). Both protein concentrates had a relatively high protein concentration due to the removal of most of the cell wall material during the extraction process, as can be inferred from the considerably lower fiber concentration when compared to their unrefined counterparts (i.e., biomass, ~40 g / lOOg). Interestingly, both ingredients were characterized by a high biological value and good protein quality, as they displayed a PDCAAS of 1.0, that is the same as that of animal proteins such as those derived from milk and eggs, as well as soy proteins.
[0378] Amino acid profil determination-The amino acid profile of the protein ingredients was determined by acid hydrolysis followed by ion-exchange chromatography.
[0379] Tryptophan concentration measurement- Tryptophan concentration was determined by alkaline hydrolysis followed by ion-exchange chromatography (IEC). The non-essential and essential amino acid profiles of the protein ingredients are reported in Table 2 and 3, respectively.
[0380] Table 2: Non-essential amino acid profile (g / 100 g protein) of yeast protein concentrate 1 (YPC 1), yeast protein concentrate 2 (YPC 2) and soy protein isolate (SPI). Table 3: Essential amino acid profile (g / 100 g protein) of yeast protein concentrate 1 (YPC 1), yeast protein concentrate 2 (YPC 2) and soy protein isolate (SPI).
[0381] The two YPCs displayed a similar amino acid profile, even though they were produced by different suppliers, and thus the processing conditions may have differed. Interestingly, both ingredients displayed a significantly higher concentration of the essential amino acid lysine compared to SPI.
[0382] Mineral profile assessment-The mineral profile of the protein ingredients was determined using inductively coupled plasma-emission spectroscopy (ICP-ES). The mineral profile of the protein ingredients is reported in Table 4. Table 4: Mineral profile (mg / 100 g) of yeast protein concentrate 1 (YPC 1), yeast protein concentrate 2 (YPC 2) and soy protein isolate (SPI).
[0383] The main difference in terms of nutrient composition between the YPCs was related to their mineral profile, with the concentrations of calcium and magnesium being significantly higher in YPC 1 and YPC 2, respectively. Both ingredients displayed a relatively high concentration of phosphorous, the main component of the yeast biomass ash, and which exerts several important physiological functions, being a constituent of phospholipids, coenzymes and nucleic acids. However, its concentration was lower compared to that of SPI. Interestingly, the latter also had a considerably higher concentration of sodium, which is typically due to the sodium hydroxide used for the alkaline treatment during the extraction process to obtain plant protein isolates.
[0384] Protein profile analysis-The protein profile of the yeast protein ingredients was determined by ultra performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS / MS). The analysis was performed using a Vanquish UPLC system coupled with an Orbitrap Elite mass spectrometer (Thermo Fisher Scientific, Waltham, MA, US). The yeast protein ingredients were reconstituted in ultrapure water (1 wt% protein) under low-speed magnetic stirring for 1 hour at room temperature followed by high pressure homogenization using an Emulsiflex C5 (Avestin, Mannheim, Germany) operating at 1000 bars (two passes), as this treatment was shown to facilitate complete protein solubilization upon sample preparation before UPLC-MS / MS. The protein dispersions were diluted five times with a solution of urea (final concentration 6 M), mixed with ammonium bicarbonate buffer (pH 8.5, final concentration 100 mM), reduced with dithiothreitol (final concentration 4 mM) for 30 min at 60°C in a ThermoMixer C (Eppendorf, Hamburg, Germany), and alkylated with iodoacetamide (final concentration 10 mM) for 30 min in the dark at room temperature. A further dilution of the dispersions with 100 mM ammonium bicarbonate buffer (pH 8.5) was performed to obtain a urea concentration of 2 M. Yeast proteins were then digested using sequencing grade trypsin (Promega Corporation, Madison, Wl, US) (enzyme-to-substrate ratio 1:50, w / w) for 4 h at 37°C. An aliquot of each digesta (6 pg protein) was then loaded onto an Acquity BEH C18 column (130 A, 1.7 pm, 3 mm X 100 mm) (Waters Corporation, Milford, MA, US) for peptide separation. Elution was performed at 0.75 mL min1. A mobile phase of two solvents was used, i.e., solvent A, consisting of 0.1% v / v formic acid and 2% v / v acetonitrile in ultrapure LC-MS grade water, and solvent B, consisting of 0.1% v / v formic acid and 80% v / v acetonitrile in ultrapure LC-MS grade water. The samples were eluted with a gradient from 2 to 50% B over 45 min. The mass spectrometer operated in a data-dependent ToplO setup to acquire full scan (MSI) and peptide fragment (MS2) spectra over the entire chromatographic run. The raw data were processed with the PEAKS X+ software (Bioinformatics Solutions Inc., Waterloo, Canada) using the Swiss-Prot database for protein identification.
[0385] The most abundant proteins in both yeast protein concentrates were enzymes involved in functions such as metabolism of carbohydrates (i.e., G3P2, G3P3, PGK, ENO1, ENO2, KPYK1, PDC1), amino acids (i.e., METE), proteins (i.e., EF1A, EF2) and fatty acids (i.e., FAS1, FAS2) (Figure 1). The mannoproteins represent a minor fraction in YPC1 and YPC2 and are present at a content much lower than 20wt.%.
[0386] Microstructure analysis-Microstructure of the yeast protein dispersions (3 wt% protein) was analyzed with a LSM 710 confocal laser scanning microscope (CLSM) upgraded with an Airyscan detector (Carl Zeiss, Oberkochen, Germany) and using Plan-APOCHROMAT objectives (10x / 0.45, 20x / 0.8, 63x / 1.4). Proteins were fluorescently labelled by adding 10 pL of 1% (w / v) Fast Green FCF (Sigma-Aldrich, Saint Louis, MO, USA) in deionized water into 1 mL of heated dispersion. The fluorescently labelled samples (100 pL) were placed inside a 1 mm deep plastic chamber closed by a glass slide coverslip to prevent compression and drying artefacts. Imaging of the proteins was performed at an excitation wavelength of 633 nm and an emission wavelength of 645 nm. Acquisition and treatment of the images were performed using the Zen 2.1 software (Carl Zeiss, Oberkochen, Germany). The protein dispersions were analyzed before and after homogenization using a PandaPLUS 2000 (GEA, Parma, Italy) with first and second stage pressures of 250 and 50 bars, respectively. The CLSM images are shown in Figure 2. The two YPCs displayed a similar microstructure, both consisting of roughly spherical, densely packed aggregates, which might have been induced by the spray drying process, with diameters ranging from ~10 to >20 pm. These were broken down by high pressure homogenization into their individual subunits, also roughly spherical and with diameters of ~3 pm.
[0387] In order to further understand the composition and microstructure of the yeast particles, CLSM analysis of a non-homogenized YPC 2 dispersion (3 wt% protein) was performed upon fluorescent labelling of proteins, lipids and fibers (i.e., chitin and cellulose) by adding 10 pL of 1% Fast Green FCF (Sigma-Aldrich, Saint Louis, MO, USA) in Milli-Q water, 10 pL of 2.5% Nile Red (Sigma-Aldrich, Saint Louis, MO, USA) in ethanol and 10 pL of Calcofluor White (Sigma-Aldrich, Saint Louis, MO, USA), respectively, into 1 mL of heated dispersion. The fluorescently labelled samples (100 pL) were placed inside a 1 mm deep plastic chamber closed by a glass slide coverslip to prevent compression and drying artefacts. Imaging of the proteins was performed at an excitation wavelength of 633 nm and an emission wavelength of 645 nm, imaging of the lipids was performed at an excitation wavelength of 488 nm and an emission wavelength of 570-620 nm, while imaging of the fibers was performed at an excitation wavelength of 405 nm and an emission wavelength of 475 nm. The images show that while proteins were dominant (green), lipid inclusions (red) and fibers (blue) were also present in the outer shell of the particles (Figure 3a). The presence of fibers, potentially both chitin and cellulose, was even more evident when only the filter for Calcofluor White was used (Figure 3b). Therefore, it is safe to assume that the particles observed were yeast cells comprising only the inner cell wall portion, where chitin is found, which could explain their resistance to high pressure homogenization.
[0388] Protein sol ubi lity-For the determination of protein solubility, the protein ingredients were reconstituted in ultrapure water (1 wt% protein) under low-speed magnetic stirring for 1 hour at room temperature and their pH was adjusted to values in the range 2-9 (at 1 pH unit intervals) using 0.1-1 M HCI and / or NaOH, as required. Samples were centrifuged at 1000 rpm for 15 min using a Sorvall evolution RC centrifuge (Thermo Fischer, Waltham, MA) equipped with a fixed angle rotor SS-34. The protein concentration of each supernatant was determined by the Kjeldahl method according to the AOAC Official Method 930.29 (AOAC, 2005) using the nitrogen-protein conversion of 6.25. Solubility was calculated as the protein concentration of each supernatant expressed as a percentage of the protein concentration of the initial dispersion. The protein solubility curves are shown in Figure 4. Both YPCs were characterized by low protein solubility across the whole pH range, with values of 12-16% and 8-12% for YPC
[0389] 1 and YPC 2, respectively.
[0390] The YPC 1 has a soluble yeast protein to insoluble yeast protein concentration ratio of 12:88. The YPC 2 has a soluble yeast protein to insoluble yeast protein concentration ratio of 10:90.
[0391] Example 2: Production of the yeast protein-based RTD beverage prototypes at pilot scale.
[0392] The protein ingredients, white sugar and glucose syrup were hydrated in demineralized water at 65°C for 30 min under continuous shear mixing using a Stephan Mixer UMSK 24 E (Stephan Food Service Equipment GmbH, Hameln, Germany) equipped with a blade whose rotational speed was set to 750 rpm. High oleic sunflower oil was added to the protein dispersion and a pre-emulsification step was performed by increasing the rotational speed to 1,500 rpm for 5 minutes. The pH was then adjusted to a value corresponding to 6.8 at 20°C using 1 M NaOH. Subsequently, gellan gum was added and the dispersion was kept under continuous shear mixing at a rotational speed of 750 rpm for 10 min. The product was sterilized via direct steam injection at 145°C for 5 s using a HT320 HTST / UHT System (OMVE, Utrecht, Netherlands) equipped with a tubular heat exchanger operating at a flow rate of 20 L h-1and with a flash cooling temperature of 80°C. Different homogenization configurations were tested, including upstream (i.e., before UHT treatment), downstream (i.e., after UHT treatment) and combination of upstream and downstream. Upstream and downstream homogenization were performed using a PandaPLUS 2000 (GEA, Parma, Italy) and an in-line TriplexPanda Lab Homogenizer 600 (GEA, Parma, Italy), respectively, with first and second stage pressures of 250 and 50 bars, respectively. The recipes of the prototypes produced are shown in Table 5. It should be highlighted that inclusion of gellan gum does not stabilize fat droplets but was required in order to increase viscosity of the continuous phase and provide colloidal stability to the emulsions, as prototypes produced without the stabilizer displayed sedimentation upon storage, although no creaming was observed (results not shown). Interestingly, despite the relatively high protein concentration, the yeast protein-based prototypes withstood UHT treatment without any processing issues (e.g., fouling, gelation and / or line blockages). Table 5: Recipes of the ready-to-drink beverage prototypes produced at pilot scale.
[0393] Example 3: Characterization of the RTD beverage prototypes.
[0394] Particle size distribution (PSD) of the prototypes was analyzed by static light scattering using a Mastersizer 3000 (Malvern Instruments Ltd, Malvern, Worcestershire, UK) equipped with a Reverse Fourier lens with an effective confocal length of 300 mm, a He-Ne red light source (633 nm) and a LED blue light source (470 nm). Particle and dispersant refractive indices of 1.47 and 1.33, respectively, were selected. The emulsions were added dropwise to the Hydro SM sample dispersion unit containing demineralized water until a laser obscuration of 10% (± 0.5%) was reached. For the analysis of the oil droplet size distribution, the emulsions were diluted 1:10 in a 1% solution of the denaturing agent sodium dodecyl sulfate (SDS) before particle size analysis so as to break non-covalent interactions between the oil droplets. Results were calculated using the Mie theory and presented as PSD and volume-weighted mean particle diameter (D [4,3] ). As shown in Figure 5, the homogenization configuration had little influence on the PSD of the yeast protein-based prototypes, which were characterized by a narrow monomodal distribution, in the range 1-10 pm, with a slightly broader distribution being observed only for the YPC 1-based prototype obtained by upstream homogenization. Conversely, the reference was characterized by a bimodal distribution, with peaks in the range 0.1-1 and 1-10 pm, respectively. Interestingly, SDS had no influence on the particle size of the yeast protein-based prototypes, while it promoted an increase in D[4,3] from ~4 to 7 pm for the reference (Figure 5c). This could be ascribed to oil droplet coalescence resulting from SDS- induced displacement of milk and / or soy proteins from the oil-water interface, thus indicating a relatively weak interfacial film.
[0395] The microstructure of the prototypes was analyzed using a LSM 710 confocal laser scanning microscope (CLSM) upgraded with an Airyscan detector (Carl Zeiss, Oberkochen, Germany). Proteins and lipids were fluorescently labelled by adding 10 pL of 1% Fast Green FCF (Sigma-Aldrich, Saint Louis, MO, USA) in Milli-Q water and 10 pL of 2.5% Nile Red (Sigma- Aldrich, Saint Louis, MO, USA) in ethanol, respectively, into 1 mL of heated sample. The fluorescently labelled samples (100 pL) were placed inside a 1 mm deep plastic chamber closed by a glass slide coverslip to prevent compression and drying artefacts. Imaging of the proteins was performed at an excitation wavelength of 633 nm and an emission wavelength of 645 nm, while imaging of the lipids was performed at an excitation wavelength of 488 nm and an emission wavelength of 570-620 nm. Acquisition and treatment of the images were performed using the software Zen 2.1 (Carl Zeiss, Oberkochen, Germany). Confocal laser scanning microscopy images of the prototypes are shown in Figure 6. The yeast protein-based prototypes were characterized by the presence of the individual spherical particles described in Example 1 and aggregates thereof, which explains the PSD observed in Figure 5. The oil droplets were found either on the surface of such particles or separated from them, possibly stabilized by the soluble yeast protein fractions. Conversely, the reference displayed oil droplets with diameters <1 pm coexisting with protein aggregates of various sizes, which is in line with its bimodal PSD.
[0396] Viscosity of the prototypes was analyzed using a Physica MCR 501 controlled shear stress rheometer (Anton Paar GmbH, Graz, Austria) equipped with a concentric cylinder geometry (CC27 / S, Anton Paar GmbH, Graz, Austria) characterized by a rough (sandblasted) surface to prevent wall slip and a gap with the outer cup (C-CC27 / T200 / SS / S, Anton Paar GmbH, Graz, Austria) of 1.13 mm. An aliquot of the samples (25 mL) was poured into the outer cup. The outer cup and the concentric cylinder were mounted above a Peltier plate (C-PTD200, Anton Paar GmbH, Graz, Austria) whose temperature was set to 20°C. Samples were sheared from 0.1 to 100 s“1in 4.8 min, their viscosity being recorded using the Rheoplus software (Anton Paar GmbH, Graz, Austria). The viscosity recorded at 10 s-1was used to compare the samples (i.e., close to the oral shear rate during oral processing and swallowing). This physicochemical property was the most affected by the homogenization configuration used, the yeast protein-based prototypes produced using upstream homogenization displaying higher values compared to those obtained by downstream or a combination of upstream and downstream homogenization (Figure 7). This was particularly evident for the YPC 1-based prototypes (i.e., ~38 vs 11-13 mPa.s), which could be explained by the broader PSD, as observed in Figure 5a. The reference displayed a viscosity of ~23 mPa.s, that is intermediate between the values observed for the various yeast protein-based prototypes. The colloidal stability of the prototypes was monitored using a Turbiscan Lab Expert (Formulaction, Toulouse, France), comprising a pulsed near infrared light source (A. = 880 nm) and two synchronous optical sensors, which receive the light transmitted through and backscattered by the emulsions at an angle of 180° and 45° with respect to the incident beam, respectively. The prototypes were loaded into cylinder glass tubes and scanned throughout their height (~50 mm) upon production and after 30 days of refrigerated storage. The stability of the prototypes is presented as the variation of their backscattering profiles in Figure 8. Neither the yeast protein-based prototypes nor the soy / milk reference displayed any signs of destabilization, being that creaming or sedimentation.
[0397] The stability of the prototypes was also evaluated visually by taking pictures using the DigiEye (Carl von Gehlen Spezialmaschinen und Zubehbr GmbH & Co. KG, Mbnchengladbach, Germany) equipped with an integrated digital camera (D-7500, Nikon, Tokyo, Japan). The pictures confirmed that, regardless of the homogenization configuration employed, the prototypes were colloidally stable after 30 days of refrigerated storage at 4°C (Figure 9).
[0398] Example 4: Sensory evaluation of the RTD beverage prototypes
[0399] Sensory evaluation of the prototypes produced at pilot scale was performed with a panel (n = 8-10) in sensory booths and under red light. The panelists attended two training sessions on commercial products to train them on the glossary, including "yeast" (i.e., fresh and dry baker's yeast extract dispersed in water), "fermented" (i.e., miso paste dispersed in water), "umami" (i.e., glutamic acid solution), "earthy" (i.e., l-octen-3-one solution) and "cereal" (i.e., oat bran and texturized soy protein) odour and flavour. "Yeast", "fermented", "earthy" and "umami" references were provided to the panelists before every sensory evaluation.
[0400] The yeast protein-based prototypes were assessed with the RATA (Rate All That Apply) methodology, either with or without reference:
[0401] • Without reference: o Overall intensity: the panelists were asked to score the intensity of the overall odour or flavour (Figure 10a); not applicable to texture. o RATA: the panelists were asked to taste the samples and check / tick off all attributes that they would use to describe the sensory profile, and to rate each selected attribute on the corresponding scale (Figure 10b).
[0402] With reference: o Overall intensity difference: the panelists were asked to score the intensity of the difference in "overall odour" or "overall flavour" intensity between the tested sample and the reference (Figure 10c); not applicable to texture. o RATA: the panelists were asked to taste the samples and check / tick off all attributes that they would use to describe the differences between prototypes, and to rate each selected attribute on the corresponding scale (Figure lOd).
[0403] Comments were also recorded. The panelists were allowed a two-minute break between the evaluation of different prototypes, during which they were requested to drink freshly opened Acqua Panna water as palate cleaner.
[0404] The prototypes were taken out of the fridge at least 30 minutes before the sensory evaluation, presented in black ceramic glasses in random order across panellists and identified using a 3-digit random code. A total of twenty-six attributes was evaluated, including seven for the odour, twelve for the flavour and taste, and seven for the texture (Table 6 and 7).
[0405] Computerized data acquisition was performed using the EyeQuestion® software (Logic 8, Elst, Netherlands), while sensory data processing was performed using the EyeOpenR® software (Logic 8, Elst, Netherlands). An analysis of variance (ANOVA) was performed for each sensory attribute to determine significant differences among products (P<0.05). Post-hoc comparisons between individual factor levels were done with Fisher's Least Significant Difference (LSD).
[0406] Table 6: Odour, flavour and taste attributes used for the sensory evaluation of the ready-to-drink beverage prototypes. Table 7: Texture attributes used for the sensory evaluation of the ready-to-drink beverage prototypes.
[0407] With few exceptions, the homogenization configuration had no significant influence on the sensory properties of both YPC 1- and YPC 2-based prototypes (Figure 11a and lib). Furthermore, no major differences were observed between the sensory properties of YPC 1- and YPC 2-based prototypes, the latter being perceived as being characterized by a slightly, but not significantly, lower overall odour intensity as well as a significantly lower thickness (Figure 11c). Interestingly, the yeast protein-based prototypes displayed relatively low yeast notes, as the average perceived intensity of this attribute in terms of odour and flavour was in most cases <2, the intensity scale ranging from 0 to 5. Furthermore, the intensity of perceived fermented and umami notes was <1 for all the prototypes, potentially indicating that (i) most of the compounds responsible for the typical notes associated with yeast biomass and derived products, which would not be desirable in RTD beverages, are removed during the extraction processes employed to obtain yeast protein concentrates; (iii) these compounds are removed during the process employed to obtain the yeast protein-based prototypes (e.g., direct UHT treatment in combination with flash cooling); (iii) the yeast, fermented and umami notes are partially covered by the other ingredients present in the recipe (i.e., high oleic sunflower oil, white sugar and glucose syrup); or a combination of these hypotheses. The YPC-2 based prototype was selected for the comparison with the milk and soy protein-based reference (Figure lid). The reference was perceived as being characterized by a significantly lower yeast odour and sweet flavour intensity, as well as a higher astringency and thickness, although absolute values were quite low, that is ~1, thus it is safe to assert that no major differences in sensory properties were perceived between these prototypes. These outcomes are particularly important from a product development perspective as they highlight that yeast proteins could potentially replace an hybrid protein blend, comprising milk and soy proteins, in high protein RTD beverages without any negative impact on the sensory properties of these products.
[0408] Example 5: Influence of mineral fortification on the physicochemical properties of yeast protein dispersions.
[0409] YPC 1 and YPC 2 stock dispersions were prepared by dispersing the protein ingredients in ultrapure water (10 wt% protein) at 20°C for 1 h under low-speed magnetic stirring to promote protein hydration. The pH of the YPC 1 and YPC 2 dispersions after hydration was 5.7 and 6.3, respectively, and it was adjusted to 6.8 using 1 M NaOH. This was followed by their homogenization using a PandaPLUS 2000 (GEA, Parma, Italy) with first and second stage pressures of 250 and 50 bars, respectively, to break large protein aggregates into their individual subunits. The stock dispersions were split into sub-dispersions to which different aliquots of calcium and / or potassium salts were added, including calcium chloride (CaCI), tricalcium phosphate (CaP) and tripotassium citrate (KCit). Ultrapure water was added to reach the desired concentrations of protein (8.5 wt%, that is within the range of the typical protein concentrations of commercial high protein RTD beverages), calcium (0-40 mM) and potassium (0-70 mM). When blends of CaCI and KCit or CaP and KCit were added, target concentrations of calcium and potassium were 40 and 70 mM, respectively. The dispersions (100 mL) were transferred to 250 mL sealed glass bottles which were placed in a water bath where they were heated at 95°C for 5 min (from the moment that the dispersions reached the desired temperature) under low-speed magnetic stirring to mimic a pasteurization treatment, and then cooled down in ice to 20°C. The protein dispersions to which CaCI alone and a blend of CaCI and KCit were added were homogenized after the heat treatment using a PandaPLUS 2000 (GEA, Parma, Italy) with first and second stage pressures of 250 and 50 bars, respectively, to investigate whether the aggregates formed could be broken down through mechanical processing.
[0410] The solubility and particle size of the yeast protein dispersions with and without added salts was determined using the methods described in Example 1 and 3, respectively.
[0411] Addition of the insoluble calcium salt CaP and / or KCit had no significant influence on the physicochemical properties of the YPC 1 and YPC 2 dispersions, neither before nor after heat treatment (Figure 12 and 13). Conversely, the soluble calcium salt CaCI promoted a significant increase in particle size upon heat treatment, that is from ~3 pm to 7-9 pm and 11- 13 pm for YPC 1 and YPC 2 dispersions, respectively, thus indicating the occurrence of protein aggregation (Figure 12). The blend of CaCI and KCit promoted a similar particle size increase as CaCI alone upon heat treatment. However, downstream homogenization was sufficient to break the aggregates formed into their individual subunits. Mineral fortification had no major impact on the solubility of the yeast protein dispersions, regardless of the salts and salt blends tested, with values ranging from 2.9 to 5.8 and from 2.5 to 4.2 for YPC 1 and YPC 2, respectively (Figure 13). Example 6: Influence of mineral fortification on the physicochemical and sensory properties of yeast protein-based RTD beverage prototypes produced at pilot scale.
[0412] The yeast protein concentrate YPC 2, white sugar, glucose syrup and mineral salts, including blends of CaCI and KCit or CaP and KCit, were hydrated in demineralized water at 65°C for 30 min under continuous shear mixing using a Stephan Mixer UMSK 24 E (Stephan Food Service Equipment GmbH, Hameln, Germany) equipped with a blade whose rotational speed was set to 750 rpm. High oleic sunflower oil was added to the protein dispersion and a pre-emulsification step was performed by increasing the rotational speed to 1,500 rpm for 5 minutes. The pH was then adjusted to a value corresponding to 6.8 at 20°C using 1 M NaOH. Subsequently, gellan gum was added and the dispersion was kept under continuous shear mixing at a rotational speed of 750 rpm for 10 min. All the ingredients were added at the same concentrations as those listed in Table 5 for Prototype 2, while the mineral salts were added at concentrations required to reach the target calcium and potassium concentrations, that is 40 and 70 mM, respectively. The product was sterilized via direct steam injection at 145°C for 5 s using a HT320 HTST / UHT System (OMVE, Utrecht, Netherlands) equipped with a tubular heat exchanger operating at a flow rate of 20 L h-1and with a flash cooling temperature of 80°C. The prototypes underwent a combination of upstream and downstream homogenization, which were performed using a PandaPLUS 2000 (GEA, Parma, Italy) and an in-line TriplexPanda Lab Homogenizer 600 (GEA, Parma, Italy), respectively, with first and second stage pressures of 250 and 50 bars, respectively.
[0413] The particle size, viscosity and colloidal stability of the YPC 2-based prototypes with and without added salts were determined using the methods described in Example 3. Mineral fortification had no significant impact on the PSD of the prototypes, the only difference observed being a slightly broader distribution for the one with added CaCI and KCit (Figure 14a). Interestingly, addition of calcium and potassium salts promoted a decrease in viscosity, that is from 26 to 17-18 mPa.s, as determined at 10 s’1, although these values are all within a relatively narrow range (Figure 14b). Furthermore, the mineral-fortified prototypes displayed no signs of destabilization after 30 days of refrigerated storage (Figure 14c).
[0414] The sensory properties of the mineral-fortified yeast protein-based prototypes were benchmarked against those of the prototype without added salts using the comparative RATA methodology described in Example 4 (Figure 15). No significant differences were observed, the prototype with added CaCI and KCit displaying only a slightly higher salty flavour and thickness. Therefore, it is safe to assume that mineral fortification had no negative impact on the functional and sensory properties of yeast protein-based high protein RTD beverages.
[0415] Although the invention has been described by way of example, it should be appreciated that variations and modifications may be made without departing from the scope of the invention as defined in the claims.
[0416] BIBLIOGRAPHY
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Claims
1. 74CLAIMS1. A liquid composition which comprises a fat component, at least one non-proteic thickening agent and a yeast protein source, wherein the yeast protein source comprises at least 40wt.% yeast proteins, and wherein the total protein content of the liquid composition is at least 5.01wt.%.
2. A liquid composition according to claim 1, wherein the total protein content of the liquid composition is from 5.01wt.% to 20wt.%.
3. A liquid composition according to claim 1 or 2, wherein the yeast protein source is selected from the list consisting of: yeast biomass, yeast protein concentrate, yeast protein isolate, and mixture thereof.
4. A liquid composition according to any one of the preceding claims, wherein the yeast protein source comprises an insoluble yeast protein to soluble yeast proteins ratio of 95:5 to 70:30, preferably 95:5 to 80:20.
5. A liquid composition according to any one of the preceding claims, which is a liquid emulsion, in particular the fat component is in the form of fat droplets in the liquid emulsion and the fat droplets formed by the fat component are surrounded at their surface by yeast proteins.
6. A liquid composition according to any one of the preceding claims, wherein the fat component comprises at least 80wt.% fat and / or the fat of the fat component is not absorbed to any solids and / or the fat component is in liquid form.
7. A liquid composition according any one of the preceding claims, wherein the liquid composition is beverage, preferably ready-to-drink beverage, more preferably ambient storage ready-to-drink beverage.
8. A liquid composition according to any one of the preceding claims, wherein the liquid composition comprises at least 0.5wt.%, preferably 0.5 to 20wt% of said fat component.
759. A liquid composition according to any one of the preceding claims, wherein the yeast protein source comprises less than 20wt.%, preferably less than 15wt.%, more preferably less than 10wt.%, even more preferably less than 5wt.% mannoproteins.
10. A liquid composition according to any one of the preceding claims, which further comprises at least one milk protein source and / or at least one plant protein source and / or at least one collagen source and / or at least one collagen peptide source and / or at least one gelatin source and / or at least one fungal protein source and / or at least one bacterial protein source.
11. A liquid composition according to any one of the preceding claims, wherein at least 10%; at least 20%; at least 30 %; at least 40%; at least 50%; at least 60%; at least 70%; at least 80%; at least 90%; at least 95%; or at least 98% of the proteins of the liquid composition are yeast proteins.
12. A liquid composition according to any one of the claims 1 to 9, wherein the proteins of the liquid composition consist only of yeast proteins.
13. A liquid composition according to any one the preceding claims, which further comprises at least one ingredient selected from the list consisting of vitamin, mineral, free amino acid, carbohydrate, prebiotic, probiotic, postbiotic, synbiotic, low molecular weight surfactant, pharmaceutically acceptable carrier, bioactive agent, flavour agent, colorant, cocoa, coffee, malt extract, spices, herbs and combination thereof.
14. A powder composition which is obtained by drying the liquid composition of any one of claims 1 to 13.
15. Use of the liquid composition of any one of claims 1 to 13 or the powder composition of claim 14 in the preparation and / or formulation of a product selected from the list consisting of: beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake,76 powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement (ONS), tube feeding and combinations thereof.
16. A product comprisingthe liquid composition of any one of claims Ito 13 orthe powder composition of claim 14, wherein the product is selected from the list consisting of beverage, sauce, dressing, bouillon, coffee and / or tea-containing beverage, coffee, cocoa and / or malt containing-beverage, confectionery, ice cream, spread, creamer, baked food, dairy product, plant-based dairy product analogue, snack bar, pudding, dessert, protein shake, powder suitable for sport nutrition, infant formula, dietary supplement, complete nutritional composition, incomplete nutritional composition, growing-up milk, baby food, infant cereal composition, fortifier, supplement or nutritional composition for pregnant or lactating women, paediatric supplement, pharmaceuticals, medical food, nutraceuticals, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, food for special medical purpose (FSMP), medicaments, tablet, oral nutritional supplement (ONS), tube feeding and combinations thereof.
17. Process for preparing a liquid composition comprising the steps of:(a) Dispersing a yeast protein source comprising at least 40wt.% yeast proteins in an aqueous liquid to form a yeast protein dispersion, wherein the yeast protein dispersion comprises at least 5.01wt.% protein,(b) Adding a fat component to the yeast protein dispersion to form a liquid pre-emulsion,(c) Adding at least one non-proteic thickening agent to the liquid pre-emulsion,(d) Homogenizing and heat-treating the liquid pre-emulsion of step (c) to form a liquid composition, wherein the heat treatment is before and / or after the homogenization.
18. A process according to claim 17, wherein the yeast protein source is selected from the list consisting of yeast biomass, yeast protein concentrate, yeast protein isolate, and mixture thereof.7719. A process for preparing a powder composition which comprises the step of drying the liquid composition obtained in step (d) of any one of claim 17 or 18 or the liquid composition of any one of claims 1 to 13.
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