High protein compositions comprising yeast oleosomes

Abstract

The present invention relates to compositions comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the total solids content of the composition, to food products or beverages comprising such compositions, to the use of yeast oleosomes to improve the sensory properties of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition and to a process for the preparation of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition.

Description

[0001] PAT8513PC00

[0002] High protein compositions comprising yeast oleosomes

[0003] Technical field

[0004] The present invention relates to compositions comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the total solids content of the composition, to food products or beverages comprising such compositions, to the use of yeast oleosomes to improve the sensory properties of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition and to a process for the preparation of a composition comprising at least 30 wt% exogenous protein based on the total solids content of the composition.

[0005] Background of the invention

[0006] Vegetarian and vegan diets are getting more and more frequent, for sustainability, health and ethical reasons. In this framework, all products traditionally produced using ingredients of animal origin have to be reformulated using only ingredients that do not originate from animals. High protein products, such as those used in performance nutrition products, for example protein bar and protein shakes, are traditionally based on proteins of animal origin such as whey, which originates from dairy milk. It is sought to replace such proteins by alternative suitable for vegetarian and vegan diets.

[0007] For this purpose, plant proteins have been used in the prior art. Various other nonanimal sources of proteins (i.e. proteins that are not isolated from a living animal) are emerging, like algal proteins, microbial proteins (such as yeast or microbial proteins), fungal proteins and even recombinant animal proteins produced in microbes or proteins obtained from animal cells cultures.

[0008] Furthermore, important proteins, such as functional proteins, are produced by precision fermentation and can be used at high concentrations in compositions.

[0009] Unfortunately, any protein used at high concentrations comes with un-pleasant off- tastes and / or off-flavours, such as astringency and / or bitterness. This is particularly the case of the most widely spread type of non-animal proteins, plant proteins. It is therefore sought to provide means to improve the sensory properties of products with high amounts of exogenous proteins.

[0010] Yeast oleosomes have been described in the prior art. For example, Nikiforidis et al., RSC Adv., 2014, 4, 25067 discloses natural emulsions based on oil bodies from plants, which have properties close to dairy milk or cream. Oil bodies are extracted from plant materials by PAT8513PC00 using aqueous media. An oil-in-water emulsion, based on intact or partially disrupted oil bodies is obtained and a protein co-extraction takes place. The oil bodies size can be nanometric, up to a few microns. The oil in such emulsions is however originating from the yeast and is present in the structure of oleosome. This document is however silent with respect to the behaviour of oleosomes in the presence of exogenous proteins, such as plant proteins.

[0011] WO98 / 53698A1 discloses emulsion formulations prepared from oil bodies originating from living cells. The content of the document is focused on plant oil bodies, but other sources of oil bodies are mentioned in a very general way, including fungal sources, such as yeast cells. The oil bodies are obtained from a cell, washed and formulated. All disclosed process aspects are focusing on the preparation of formulations from plant seeds. The size of the oil bodies varies between 0.4 and 1.5 pm. The disclosed emulsions can be used as substitutes for dairy products. Again, this document is silent with respect to the behaviour of oleosomes in the presence of exogenous proteins, such as plant proteins.

[0012] WO2017 / 066569A1 discloses oleosome compositions comprising two different oleosome compositions, preferably originating from two different sources, and having different particles sizes: a first oleosome composition is characterized by a D50 of at least 120 nm and the second oleosome composition is characterized by a D50 of at least 600 nm. Yeast cells are mentioned in very general terms. However, all aspects described in detail are related to plant oleosomes. The use of oleosomes in combination with exogenous proteins, such as plant proteins, is not described in this document.

[0013] WO2021 / 126409A1 discloses roasted oleosome compositions, which may originate from a variety of sources, including yeast, but preferably originating from plant sources. The isolated oleosome composition has a dry-matter content of 30 to 80 wt%, and, based on dry matter, a protein content of 1 to 6 % and an oil content of 94 to 99 %. This document relates to the amelioration of the taste of the oleosomes themselves but is silent with respect to any potential effect of oleosomes on the sensory properties of other ingredients combined therewith.

[0014] It is further desirable to optimize the stability of high protein products, in particular of high protein products in liquid form, such as protein shakes. Particle size and emulsification capacity are features impacting the stability of such product. Small particle size and good emulsification properties lead to products having abetter stability, in turn translating in a longer shelf life, more appealing appearance and texture and possibility to incorporate higher amounts of proteins. It is thus desirable to provide high protein products having a small particle size distribution and a being well emulsified. PAT8513PC00

[0015] The present invention aims at solving the above-mentioned sensory problems associated with the use of high amounts of proteins, such as performance nutrition products or high-protein medical nutrition. It would be further desirable to provide means that improve the sensory properties of compositions comprising high amounts of proteins, while having a beneficial effect on the nutritional properties of such compositions.

[0016] Summary of the invention

[0017] In a first aspect, the invention provides a composition comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition.

[0018] In a second aspect, the invention provides for the use of yeast oleosomes for improving at least one sensory property of a composition comprising at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition.

[0019] In a third aspect, the invention provides a food product or a beverage comprising a composition according to the present invention.

[0020] In a fourth aspect, the invention provides a process for the preparation of a composition comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition comprising the step of: a) providing yeast oleosomes; b) admixing the yeast oleosomes provided in step a) with exogenous proteins and optionally other ingredients, such that the exogenous proteins are present in a final concentration of at least 30 wt%, based on the weight of the total solids content in the composition; and c) homogenising the mixture obtained in step a).

[0021] Brief description of the drawings

[0022] [Fig. 1] is a spider graph representing the respective creaminess, bitterness, sweetness, astringency and off aroma of Samples 1 and 2 (according to the invention, respectively referred to as CB 8 % and CB 15 % in the graph) and Controls 1 and 2 (comparative, respectively referred to as Control 8 % and Control 15 % in the graph), as determined in Example 1. PAT8513PC00

[0023] Detailed description of the invention

[0024] General definitions

[0025] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The publications and applications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0026] In the case of conflict, the present specification, including definitions, will control. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in art to which the subject matter herein belongs. As used herein, the following definitions are supplied in order to facilitate the understanding of the present invention.

[0027] Reference throughout this specification to "one aspect", "an aspect", "another aspect", "a particular aspect", "combinations thereof' means that a particular feature, structure or characteristic described in connection with the invention aspect is included in at least one aspect of the present invention. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same aspect. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more aspects.

[0028] The term "comprise(s)", or "comprising" is generally used in the sense of include(s) / including, that is to say, permitting the presence of one or more features or components. The terms "comprise(s)" and "comprising" also encompass the more restricted ones "consist(s)" and "consisting", respectively.

[0029] As used in the specification and claims, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise.

[0030] The present inventors have used yeast oleosomes to prepare compositions with at least 30 wt% exogenous protein contents.

[0031] "Yeast oleosomes" are defined as oil bodies isolated from yeast, which are formed of oil droplets surrounded by a layer (membrane) of proteins, phospholipids and carbohydrates, (mostly in the form of polysaccharides). Obtaining oleosomes "isolated from yeast" requires the breakage of the yeast cell wall and thus whole yeast cells are excluded from the definition PAT8513PC00 of yeast oleosomes, even though such whole yeast cells contain oleosomes. Also, when yeast oleosomes are provided in the form of a yeast oleosome composition, such as described herein, such composition comprises only a negligible number of remining whole (intact) yeast cells, such as less than 10% by weight of whole yeast cells, based on the total weight of the oleosomes composition.

[0032] Oleosomes also exist in plants. However, plant and yeast oleosomes vary in their composition and properties. Namely, the materials present in the layer separating the oil droplet from the surrounding medium are different between plant oleosomes and yeast oleosomes, which provides yeast oleosomes with a different behaviour compared to plant oleosomes, in diverse applications. As defined herein, yeast oleosomes refer to oleosomes extracted from the yeast cell. Even though the oleosomes can be in the form of a mixture with cell components, such as the yeast cell wall components, such yeast cell wall shall not be intact, so that the oleosomes can freely pass through the yeast cell wall fragments. This excludes oleosomes present inside live yeast cells.

[0033] Yeast oleosomes are preferably present in the form of a mixture (i.e. in the form of a yeast oleosomes composition) of yeast oleosomes with at least one cell wall component, preferably an isolated yeast cell wall component (i.e. one or more compound(s) released from the yeast cell wall). The yeast cell wall component can for example be lipids (such as phospholipids), carbohydrates or proteins. Preferably the yeast cell wall component is selected from the group consisting of chitin, P-D-glucan, mannoprotein and combinations thereof, preferably P-D-glucan. The terms "released" or "isolated" from the yeast cell wall structure herein means that the yeast cell wall component is present in the yeast oleosome composition in free form, i.e. separated from the yeast cell wall structure, and is not part of the structure of a yeast cell wall debris. Thus, in other words, the yeast cell wall component is not fully embedded in the yeast cell wall structure so that it is available for interaction with the surrounding environment. The yeast cell wall component may nevertheless interact or be bound with other components of the yeast oleosome composition, provided it is not embedded in the yeast cell wall structure in a way that it is inaccessible to the yeast oleosomes medium. Indeed, when a yeast cell wall component is present, it is essential that such yeast cell wall component be "released" or "isolated" from the yeast cell wall structure to make the yeast cell wall component available for interaction with other components of the yeast oleosomes composition, with the oral cavity of an individual consuming the yeast oleosomes composition or a high protein composition comprising it or with other components of the high protein composition. PAT8513PC00

[0034] The at least one yeast cell wall component can be present in the yeast oleosomes composition either in solubilized or suspended form, or it can be associated with the yeast oleosome membrane or it can be partly solubilized or suspended and partly associated with the yeast oleosome membrane. In a preferred aspect, the at least one yeast cell wall component is associated with the yeast oleosome membrane, preferably through non-covalent bonds, such as electrostatic interactions, hydrogen bonds or hydrophobic interactions. In an embodiment the yeast oleosomes composition comprises at least two yeast cell wall components selected from chitin, P-D-glucan and mannoprotein, one or two of these yeast cell walls components is (are) in solubilized or suspended form, and the other one or two component(s) is (are) associated with the yeast oleosome membrane. Preferably chitin, P-D-glucan and mannoprotein are all associated with the yeast oleosome membrane.

[0035] In a preferred aspect, the at least one isolated yeast cell wall component is present in an amount of at least 1.5 wt%, preferably at least 2 wt%, more preferably at least 2.5 wt%, more preferably at least 3 wt%, more preferably at least 3.5 wt%, more preferably at least 4 wt%, more preferably at least 4.5 wt%, most preferably at least 5 wt%, preferably at least 7.5 wt%, preferably at least 10 wt%, preferably at least 15 wt%, preferably at least 20 wt% by weight based on the total weight of the solids in of the yeast oleosomes composition. In a particular aspect, the at least one isolated yeast cell wall component is present in the yeast oleosomes composition of the present invention in an amount of up to 25 wt%, or up to 30 wt%, or up to 35 wt% or up to 35 wt%, or up to 40 wt% or up to 45 wt% or up to 50 wt%, based on the total weight of the solids in the yeast oleosomes composition.

[0036] In a further preferred aspect, P-D-glucan is present in an amount of at least 1.5 wt%, preferably at least 2 wt%, more preferably at least 2.5 wt%, more preferably at least 3 wt%, more preferably at least 3.5 wt%, preferably at least 4 wt%, preferably at least 4.5 wt%, preferably at least 5 wt%, preferably at least 7.5 wt%, preferably at least 10 wt%, preferably at least 15 wt%, preferably at least 20 wt% based on the total weight of the solids in the yeast oleosomes composition. In a particular aspect, the at least one isolated yeast cell wall component is present in the yeast oleosomes composition in an amount of up to 25 wt%, or up to 30 wt%, or up to 35 wt% or up to 35 wt%, or up to 40 wt% or up to 45 wt%, based on the total weight of the solids in the yeast oleosomes composition.

[0037] In a particular aspect, the P-D-glucan is in its native form, as obtained after extraction of the oleosomes composition from the yeast cell. In another particular aspect, the P-D-glucan has not been enzymatically degraded in the yeast oleosomes composition. PAT8513PC00

[0038] In another preferred aspect, the yeast oleosomes composition comprises intact oleosomes in an amount of at least 25wt%, such as 25 to 90wt%, or 25 to 80wt% or 25 to 79wt% or 25 to 60wt% based on the total solids in the yeast oleosomes composition.

[0039] In a preferred aspect, the particle size of the yeast oleosomes composition is characterized by a mean diameter by volume of 0.2 to 7 pm as measured by laser diffraction, preferably using a Anton Paar PSA particle size analyser, preferably with Anton Paar PSA 1190, with a measuring time set to 1 min with 6 repetitions per sample, the reconstruction algorithm being MIE, the refractive index for the material being 1.47 and the absorption index being 0.01, wherein the samples are dissolved in distilled water and the refractive index is set to 1.33. For example, the mean diameter by volume can be of 0.2 to 2 pm, or 0.2 to 1 pm, or 0.2 to 0.8 pm, or 0.4 to 0.8 pm or 0.6. to 0.8 pm, or 0.2 to 0.6 pm, such as 0.2 to 0.5 pm.

[0040] In a preferred aspect, the density of the yeast oleosome composition is of 0.8 to 1.1 g / ml.

[0041] In another preferred aspect, the zeta potential at the surface of the oleosomes is in the range of -45 to -5 mV, -45 to -8 mV, -45 to -10 mV, -45 to -20 mV, -40 to -5 mV, -40 to -8 mV, -40 to -10 mV, -40 to -20 mV, or -30 to -20 mV at physiological pH. Such zeta potential is advantageous because within such range, the electrostatic interactions between the oleosomes are sufficiently low to avoid aggregation of the oleosomes and ensure proper physical stability of the composition of the invention. A zeta potential within the present range also contributes to the interaction of the yeast oleosome membrane with the polysaccharides. The zeta potential varies within this range, depending on the proteins present in the oleosome membrane. In a preferred aspect, the zeta potential is measured with a Malvern Zetasizer Nano ZS90 at a sample dilution of 1 : 100 sample:water, with a refractive index of solvent of 1.33, a refractive index of sample of 1.47, an absorption of 0,010, and with a DTS1060 cell

[0042] In a preferred aspect the pH of the yeast oleosomes composition is of 3 to 8, preferably 6 to 7. Such pH range is preferred because it impacts the net charge of the oleosomes and positively impacts the physical stability of the yeast oleosomes composition over time.

[0043] In a particular aspect, the yeast oleosomes composition is in liquid form and has a total solids content of 2 to 45 wt%. Within this range, the total solids content of a yeast oleosomes composition in liquid form can be adjusted, depending on the intended application. The total solids content can be, for example of 2 to 12 wt%, or 2 to 10 wt%, or 2 to 8 wt%, or 2 to 6 wt%, or 2 to 5 wt%, or 15 to 45 wt%, or 16 to 45 wt%, or 20 to 45 wt% or 30 to 45 wt%, or 35 to 45 wt%, or 40 to 45 wt%, or 16.6 wt% or 42 wt%, based on the total weight of the composition of the invention. PAT8513PC00

[0044] In a particular aspect the composition is in solid form, such as in the form of a lyophilized or spray-dried composition. Such solid composition can be obtained by drying any liquid composition of the invention, using any drying technique known to the person skilled in the art. A composition of the invention in solid form preferably has a total solids content of up to 97wt%.

[0045] In a particular aspect, the yeast oleosomes composition has a fat content of 25 to 60wt%, based on the total solids content in the oleosomes composition.

[0046] Compositions of the invention

[0047] The invention provides a composition comprising yeast oleosomes, such as described herein, and at least 30 wt%, preferably more than 30 wt%, preferably at least 31 wt%, more preferably at least 32 wt%, more preferably at least 33 wt%, preferably at least 35 wt%, preferably at least 40 wt% exogenous protein or even at least 50 wt% or 55 wt% or 60 wt%, or 65wt% or 66 wt% or 70 wt%, based on the weight of the total solids content in the composition of the invention. In a preferred aspect, the composition comprises up to 93 wt%, such as 90 wt%, such as up to 85 wt%, or up to 80 wt%, or up to 75 wt%, or up to 70 wt%, or up to 65 wt%, or up to 60 wt% or up to 55 wt% or up to 52.5 wt% of exogenous proteins, based on the weight of the total solids content in the composition of the invention. The amount of protein can be varied depending on the desired protein dose and the intended use of the composition. In a particular aspect, the exogenous protein is present in an amount of 50 to 93, or 50 to 90 wt% or 50 to 85 wt% or 50 to 80 wt% or 50 to 75wt% or 55 to 70 wt%.

[0048] By "exogenous protein" it is referred to herein to any protein that does not form part of the yeast oleosomes or of the yeast oleosomes composition, such as defined above. Any kind of exogenous protein can be used, such as plant proteins, fungal proteins (including yeast proteins), bacterial proteins, algal proteins, animal proteins, such as recombinant animal proteins, or animal proteins produced by animal cell culture, functional proteins, such as those produced by precision fermentation and mixtures thereof. In a particular aspect, the exogenous protein is not yeast protein. In another particular aspect, the exogenous protein is selected from plant proteins, non-yeast fungal proteins, bacterial proteins, algal proteins, functional proteins produced by precision fermentation and mixtures thereof. In another particular aspect, the exogenous protein is selected from plant proteins, non-yeast fungal proteins, bacterial proteins, algal proteins and mixtures thereof. In a further particular aspect, the exogenous protein is selected from plant proteins, fungal proteins, bacterial proteins, algal proteins, functional proteins produced by precision fermentation and mixtures thereof. In still a further particular PAT8513PC00 aspect, the exogenous protein is selected from plant proteins, fungal proteins, bacterial proteins, algal proteins and mixtures thereof.

[0049] Plant proteins include for example cereal proteins, legume proteins, nut proteins, fruit proteins and root proteins and mixtures thereof. In a preferred aspect the plant proteins are selected from oat proteins, almond proteins, pea proteins, rice proteins, soy proteins, potato proteins and mixtures thereof.

[0050] As yeast oleosomes themselves comprise proteins, proteins originating from the oleosomes add up to the exogenous proteins to boost the total protein content in the composition of the invention. Therefore, less exogenous proteins are required to obtain the same overall protein content in the composition. This is advantageous, because the ratio of the digestible amino acid content in yeast protein is higher in yeast proteins compared to, for example, plant proteins, as evidenced by a higher DIASS ratio.

[0051] The yeast oleosomes are included in the composition of the invention in the form of a liquid or solid yeast oleosomes composition, depending on the type of the intended end composition. For example, the oleosomes are used in solid form (such as in lyophilized or spray-dried form) in solid compositions of the invention, such as powder or bars and can be used in solid or liquid form in drinks (such as protein shakes) or other liquid compositions of the invention.

[0052] In a preferred aspect, the yeast oleosomes are provided in an amount such that the solids of the yeast oleosomes composition be present in an amount of at least 1 wt%, preferably at least 3 wt%, or more than 3 wt%, at least 4 wt%, at least 4.5 wt%, at least 5 wt%, at least 5.5 wt%, at least 6 wt%, at least 6.5wt%, at least 6.6 wt%, based on the weight of the total solids content in the composition of the invention. In another preferred aspect, the yeast oleosomes are provided in an amount such that the solids of the yeast oleosomes composition be present in an amount of up to 70 wt%, or up to 65 wt%, or up to 60 wt%, or up to 55 wt%, or up to 50 wt%, or up to 45 wt%, or up to 40 wt%, or up to 35 wt%, or up to 30 wt% or up to 25 wt% or up to 20 wt% or up to 15 wt% or up to 10 wt% of up to 5 wt% or up to 4 wt% or up to 3 wt% or up to 2 wt%, based on the weight of the total solids content in the composition of the invention. The amount of yeast oleosomes can be varied depending on the desired composition properties and the intended use of the composition. In a particular aspect, the yeast oleosomes are provided in an amount such that the solids of the yeast oleosomes composition be present in an amount of 0.5 to 30 wt, preferablyl to 30 wt%, preferably 1 to 25 wt%, preferably 1 to 20 wt%, preferably 1 to 15wt%, preferably 1 to 10 wt%, preferably 1 to 5 wt%, preferably 1 to 4 wt%, preferably 1 to 3 wt%, preferably 1 to 2 wt%. PAT8513PC00

[0053] In still another preferred aspect, the ratio of the total solids in the yeast oleosomes composition to the exogenous protein in the composition of the invention is of 1 :15 to 5: 1, 1: 12 to 5: 1, 1 :10 to 5: 1, 1 :5 to 5: 1, 1 :4.5 to 4.5: 1, 1 :4 to 4: 1, 1 :3 to 3: 1, 1 :2 to 2: 1, 1 :2 to 1.5: 1, or 1 :2 to 1.2: 1. The ratio can be adjusted by the person skilled in the art depending on the desired application and protein content.

[0054] In another aspect, the compositions of the invention have a total protein content of at least 31 wt%, preferably at least 35 wt%, preferably at least 40 wt%, preferably at least 42 wt%, preferably at least 45 wt%, preferably at least 47 wt%, preferably at least 50 wt%, preferably at least 55 wt%, preferably at least 60 wt%, based on the weight of the total solids content in the composition of the invention. In another aspect, the compositions of the invention have a total protein content of up to 99 wt%, or up to 95 wt%, or up to 90 wt%, or up to 85 wt%, or up to 80 wt%, or up to 75 wt%, or up to 70 wt%, or up to 65 wt% or up to 63 wt% or up to 60 wt% or up to 58 wt%, based on the weight of the total solids content in the composition of the invention.

[0055] The compositions of the invention can further comprise other optional ingredients, such as carbohydrates, lipids, micronutrients such as vitamins, minerals or nucleotides, flavours, colorants, preservatives, low calorie sweeteners, spices, taste enhancers, emulsifiers, thickeners, and / or cereals, nuts, legumes, fruits vegetable flour, juice, pieces and the like.

[0056] Examples of carbohydrates include sugars and fibres (including oligo- and polysaccharides). Sugars can be used in any amount, based on the desired sweet taste to be imparted to the composition of the invention. For example, sugar can be present in an amount of 1 to 20 wt%, or 3 to 15 wt%, or 5 to 10 wt% based on the weight of the total solids content in the composition of the invention. Fibres can be present in any desired amount, depending on the desired nutritional value of the composition. Yeast oleosomes are however particularly advantageous in that they contain fibres, in addition to lipids and proteins, thus reducing the need to add exogenous fibres to the composition. For example, fibres can be present in an amount of 5 to 40 wt%, or 20 to 30 wt% based on the weight of the total solids content in the composition of the invention.

[0057] Lipids can be in the form of an oil (i.e. a triglyceride composition that is liquid at room temperature) or a fat (i.e. a triglyceride composition that is in solid form at room temperature). Yeast oleosomes are however particularly advantageous in that they are a source of lipids, thus reducing the need to add exogenous lipids to the composition of the invention. The amount of lipids can be adjusted by the person skilled in the art based on the desired nutritional value of the composition of the invention. For example, lipids can be present in an amount of 5 to 30 PAT8513PC00 wt%, or 10 to 25 wt% based on the weight of the total solids content in the composition of the invention.

[0058] Micronutrients are also added in the amounts necessary to meet the nutritional requirements of the composition, depending on its intended use. Functional or medical nutritional compositions for example require fortification with specific amounts of specific micronutrients. Any desired micronutrient can be used in the composition of the invention, in any desired amount.

[0059] Colorants, preservatives and the like are used as appropriate, depending on the desired taste, visual appearance and shelf life of the composition of the invention.

[0060] Any thickener can be used, such as gellan gum, guar gum, P-carrageenan, xanthan, locust bean gum, agar-agar, starch (such as cornstarch, tapioca starch), methylcellulose, P- glucan, gum Arabic, pectin and combinations thereof. The amount of thickener depends on the intended texture of the composition of the invention. For certain purpose a relatively fluid composition is desired, so that no or little thickener is used, while other applications, such as medical nutrition for individuals having swallowing problems, require thicker compositions, in which higher amounts of thickeners are used. Any desired amount of thickener can be used in the composition of the invention.

[0061] The composition of the invention can further comprise any kind of edible material that is desired, depending on the intended applications. In a particular aspect, the composition of the invention comprises solid particles, preferable vegetal solid particles, such as vegetal flours, in particular cereal or nut flour, for example oat flour, almond flour, rice flour, pea flour, soy flour, coconut flour and mixtures thereof. The solid particle is preferably present in an amount of 0.1 to 20 wt%, preferably 1 to 20 wt%, more preferably 5 to 15 wt%, based on the weight of the total solids content in the composition of the invention.

[0062] In a particular aspect, the composition of the invention comprises an emulsifier, preferably an emulsifier comprising polar lipids, including phospholipids, such as lecithin, mono- and di-glycerides and mixtures thereof. The polar lipids are preferably present in an amount of 0.5 to 10 wt%, preferably 1 to 8 wt%, preferably 3 to 6 wt%, based on the weight of the total solids content in the composition of the invention. As yeast oleosomes, such as defined herein, include polar lipids as part of the oleosome membrane, they already act as an emulsifier. Therefore, addition of an exogenous emulsifier is not essential, but may be useful for applications requiring high emulsification.

[0063] In a particular aspect the composition of the invention does not comprise dairy milk. In another particular aspect the composition of the invention does not comprise dairy PAT8513PC00 ingredients (i.e. components of dairy milk), such as milk fat, milk proteins and / or milk carbohydrates.

[0064] Uses

[0065] The present invention provides for the use of yeast oleosomes for improving at least one sensory property of a composition comprising at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition.

[0066] In a preferred aspect, the yeast oleosomes are present in the form of a yeast oleosome composition as described above and the exogenous protein is as described above.

[0067] In another preferred aspect, the yeast oleosomes are used in combination with any of the optional ingredients recited above in the section dedicated to the composition.

[0068] The use of yeast oleosomes in combination with high amounts of exogenous proteins such as described herein advantageously improves the sensory properties of the composition, compared to a composition comprising the same amount of proteins, but wherein the total amount of the proteins originating from the oleosomes and from the exogenous proteins are provided solely in the form of exogenous proteins.

[0069] Yeast oleosomes are effective in reducing or masking the off-flavours, off-taste and off-aromas of exogenous proteins, such as plant proteins. In particular, the astringency of exogenous proteins, such as plant proteins is effectively masked by the yeast oleosomes. The mouthfeel, in particular the creaminess, of liquid products comprising exogenous proteins, such as plant proteins is also improved. The color imparted by plant proteins is also attenuated when the plant proteins are admixed with yeast oleosomes, providing high-protein products with a blander color. The flowability (viscosity) is also improved when the plant proteins are admixed with yeast oleosomes, in particular in products exposed to heat during their productions, such as for example UHT treated products.

[0070] Thus, the present use is preferably for improving creaminess, reducing bitterness, reducing astringency, reducing off aroma, reducing the intensity of the color (whitening) and / or improving the flowability of a composition comprising at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition, such as herein described.

[0071] In an aspect, the present invention provides for the use of yeast oleosomes for improving the flowability (decreasing the viscosity) of a composition comprising at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition.

[0072] Yeast oleosomes also have the property of reducing the gelling of proteins during thermal treatment, such as UHT treatment and / or during storage. The addition of yeast PAT8513PC00 oleosomes makes it possible to include more proteins in an aqueous composition than in the same aqueous composition without the oleosomes. Also with an identical protein content, a composition with yeast oleosomes has a longer physical shelf life than a composition without oleosomes, because the gelling is delayed / reduced. Thus, in a further aspect, the present invention provides for the use of yeast oleosomes for preventing or reducing gelling of a composition comprising at least 30 wt% protein.

[0073] The sedimentation of high-protein compositions is also reduced when yeast oleosomes are present, compared to the same composition without yeast oleosomes. Thus, in a further aspect, the present invention provides for the use of yeast oleosomes for increasing the physical shelf life of a composition comprising at least 30 wt% protein and for the use of yeast oleosomes for reducing sedimentation in a composition comprising at least 30 wt% protein.

[0074] In all uses described herein, the composition comprising at least 30% protein and the yeast oleosomes are as defined herein above.

[0075] Products

[0076] The invention relates to a food product or beverage comprising a composition of the invention. The composition of the invention can constitute by itself a final product. Alternatively, the composition of the invention can be used in combination with other composition(s) and / or ingredient(s) to form a more complex product, wherein the composition of the invention can either be admixed with the other composition(s) and / or ingredient(s) or used in combination therewith without mixing.

[0077] Examples of food products and beverages of the invention include any product comprising the specified amount of protein. Examples of such products include protein shakes (in solid form for reconstitution or in liquid form), protein powders, protein bars, nutritional supplements, dairy products and dairy alternative products such as cream, milk, cheese (soft or hard) or yogurt (such as protein-enriched dairy products and dairy alternative products), egg products and egg alternative products such as powdered eggs, egg preparations, egg-containing sauces, puddings and egg-based desserts. Such products are of particular interest for performance nutrition especially for athletes and individuals having extensive physical activity or seeking to promote muscle growth, for medical nutrition (for enteral and parenteral consumption) and specialized nutrition, for example for the fortification of individuals in need of concentrated nutrition in a small volume, such as elderly patients that lose appetite and for individuals following special diets (such as ketogenic diet) to lose / maintain body weight. PAT8513PC00

[0078] In a particular aspect, the invention relates to a protein shake comprising yeast oleosomes and at least 50wt%, preferably at least 55wt%, more preferably at least 60wt%, more preferably at least 65wt%, more preferably at least 66wt% of exogenous proteins, based on the total solids content of the protein shake. In an aspect, the protein shake has up to 93 wt%, or up to 90 wt% or up to 85 wt% or up to 80 wt% or up to 75 wt% or up to 70 wt% of exogenous proteins, based on the total solids content of the protein shake. Preferably, the yeast oleosomes are present in the form of composition, such as described herein. Preferably the protein shake comprises 0.5 to 30 wt%, preferably 0.5 to 25 wt%, preferably 0.5 to 20 wt%, preferably 0.5 to 15 wt%, preferably 0.5 to 10 wt%, preferably 0.5 to 7 wt%, preferably 1 to 7 wt%, preferably 1 to 5 wt%, preferably 1 to 4 wt%, preferably 1 to 3 wt%, more preferably 1 to 2 wt% of yeast oleosomes solids, based on the total solids content of the protein shake. In a preferred aspect, the protein shake does not contain vegetable oil. In another preferred aspect, the protein shake does not contain vegetable carbohydrates (such as vegetable flour). In a preferred aspect, the protein shake comprises yeast oleosomes solids and exogenous proteins in a weight ratio of 1 : 1 to 1 : 25, preferably of 1 : 1 to 1 :20, preferably of 1 : 1 to 1 : 17 or 1 :2 to 1 :20 preferably 1 :5 to 1 :20 preferably 1 :8 to 1 :20, preferably 1 :8 to 1 : 17.

[0079] Process

[0080] The invention also relates to a process for the preparation of a composition comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition comprising the step of: a) providing yeast oleosomes; b) admixing the yeast oleosomes provided in step a) with exogenous proteins and optionally other ingredients, such that the exogenous proteins are present in a final concentration of at least 30 wt%, based on the weight of the total solids content in the composition; and c) homogenising the mixture obtained in step a).

[0081] In a preferred aspect, step a) is performed by isolating oleosomes from yeast in any suitable manner. Preferably, the composition is obtained by a process comprising the steps of:

[0082] I. providing oleaginous yeast cells;

[0083] II. lysing the cell wall; and

[0084] III. separating the oleosomes from the other yeast cell components.

[0085] The yeast cells can optionally be subjected to pre-treatment, such as: i. washing with water, preferably warm water; ii. washing with an alkaline solution; PAT8513PC00 iii. washing with an acidic solution; and / or iv. enzymatic digestion of the cell wall.

[0086] Step II. can be performed by chemical or mechanical means. Suitable chemical means include: i. alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours; ii. contacting the yeast cells with an organic solvent such as ethanol, methanol or heptane; iii. autolysis; iv. enzymatic treatment, preferably using one or more hydrolase(s), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers); and combinations thereof.

[0087] The one or more hydrolase(s) is preferably selected from one or more protease(s), glucanase(s), mannanase(s), chitinase(s), nuclease(s), beta-glucosidase(s), cellulase(s), xylanase(s), pectinase(s) and combinations thereof. Preferably, it is one or more protease(s) and / or glucanase(s).

[0088] Preferred chemical means for the cell lysis include: i. alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours; ii. autolysis, by heating the cells in the end of the fermentation to a temperature of 50°C or more, reducing the stirring and stopping the aeration until spontaneous lysis of the yeast cell occur (under the action of the endogenous enzymes); iii. enzymatic treatment, preferably using one or more hydrolases (as defined above), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers); and combinations thereof.

[0089] Such preferred chemical means are particularly advantageous because they are successful in breaking the cell wall, thus releasing the yeast cell wall components chitin, P-D- glucan and mannoproteins, while preserving the structure of the oleosomes. PAT8513PC00

[0090] Suitable mechanical means of the cell wall lysis of step II. include high pressure homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar, more preferably with 2 to 10 passes at a pressure of 700 to 3000 bar

[0091] When high pressure homogenization is used, the number of passes and the homogenization pressure can advantageously be adapted, when high pressure homogenization is combined with chemical means for lysing the membrane. For example, 1 to 10 passes and a pressure of 500 to 3000 bar is sufficient when the high-pressure homogenization is combined with prior enzymatic treatment, whereas it is preferred to perform the high-pressure homogenization with 2 to 10, preferably 2 to 5 passes at a pressure of 700 to 3000 bar, preferably 700 to 2000 bar, in particular when the high pressure homogenization is performed after alkaline treatment. Preferably high-pressure homogenization is performed in high- pressure homogenizer comprising a nozzle, as it contributes to breaking down the cells.

[0092] High-pressure homogenization as disclosed herein is particularly advantageous over other mechanical means of lysing the yeast cell wall, such as bead milling, ultrasound treatment and microwave treatment, in that it is successful in breaking the cell wall, thus releasing the yeast cell wall components chitin, P-D-glucan and mannoproteins, while preserving the structure of the oleosomes. High-pressure homogenization is also advantageous in terms of yield and allows to control the particle size.

[0093] Chemical and mechanical means for lysing the cell wall can advantageously be combined with each other. Particularly advantageous combinations are: i. alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours, followed by high pressure homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar, more preferably with 2 to 10 passes at a pressure of 700 to 3000 bar; ii. enzymatic treatment, preferably using one or more hydrolases (as defined above), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers), followed by high pressure homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar, preferably with prior pre-treatment by washing with water or an alkaline solution; iii. autolysis, by heating the cells in the end of the fermentation to a temperature of 50°C or more, reducing the stirring and stopping the aeration until spontaneous lysis of the yeast cell occur (under the action of the endogenous enzymes), followed by high pressure PAT8513PC00 homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar, preferably with 2 to 10 passes at a pressure of 700 to 3000 bar;

[0094] In a preferred aspect, cell lysis is performed by one of the following means: i. alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours, followed by high pressure homogenization, preferably with 2 to 10 passes at a pressure of 700 to 3000 bar; ii. autolysis, by heating the cells in the end of the fermentation to a temperature of 50°C or more, reducing the stirring and stopping the aeration until spontaneous lysis of the yeast cell occur (under the action of the endogenous enzymes); and iii. enzymatic treatment, preferably using one or more hydrolases (as defined above), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers), followed by high pressure homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar,

[0095] The separation step is preferably performed by i. decantation, preferably using a disk stack at 5000 to 20000 g for 10 seconds to 5 minutes, ii. centrifugation, under any suitable condition, preferably at 3000 to 30000 g, preferably 3000 to 20000 g, preferably 3000 to 10000 g, such as 7000 g in disc centrifuge, such as a continuous disk stack centrifuge, preferably with a residence time of 10 seconds to 10 minutes, preferably 10 seconds to 5 minutes, preferably 30 seconds to 5 minutes for example 1 minute. Preferably, the temperature is of 5 to 85°C, preferably 40 to 65°C. iii. filtration, preferably in cross counter flow, such as by filtration using polymeric or ceramic membranes with pore sizes from 10 kDa to 300 kDa, and from 0.2 pm to 20 pm or iv. Flocculation, preferably by contacting the composition with a flocculant, preferably for 1 to 24 hours at a temperature of 2 to 60°C. Any flocculant known in the art can be used. Preferably the flocculant is selected from inorganic salts such as CaCh, or a natural flocculant such as chitosan, cellulose and the like. The flocculant is preferably used in an amount of 0.1 to 50 mg / L.

[0096] Preferably, the separation step is performed by decantation, centrifugation or filtration, as described above. More preferably, it is performed by decantation, as described above.

[0097] In a particularly preferred aspect, the compositions of the present invention are obtained by a process comprising the steps of: PAT8513PC00

[0098] 1) providing oleaginous yeast cells, wherein the yeast cells are preferably washed and optionally weakening the cell walls by one or more of the following pre-treatments i. alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours; ii. enzymatic treatment, preferably using one or more hydrolases (as defined above), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers); and / or iii. autolysis, by heating the cells at the end of the fermentation to a temperature of 50°C or more, reducing the stirring and stopping the aeration until spontaneous lysis of the yeast cell occurs (under the action of the endogenous enzymes);

[0099] 2) lysing the cell wall by i. Alkaline treatment, preferably by soaking the yeast cells in an aqueous medium at pH 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours; ii. contacting the yeast cells with an organic solvent such as ethanol, methanol or heptane; iii. autolysis, by heating the cells in the end of the fermentation to a temperature of 50°C or more, reducing the stirring and stopping the aeration until spontaneous lysis of the yeast cell occurs (under the action of the endogenous enzymes); iv. enzymatic treatment, preferably using one or more hydrolases (as defined above), applied to the washed biomass and suspended at a concentration of 50 to 200 g / L in water or a suitable pH buffer with an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours with low stirring or shaking, to allow the degradation of cell wall biopolymers); v. high pressure homogenization, preferably with 1 to 10 passes at a pressure of 500 to 3000 bar, more preferably with 2 to 10 passes at a pressure of 700 to 3000 bar; or vi. any combinations of combinations of any two or more of the means provided in items i. to v.; and

[0100] 3) separating the oleosomes from the other yeast cell components by i. decantation, preferably using a disk stack at 5000 to 20000 g for 10 seconds to 5 minutes ii. centrifugation iii. filtration.

[0101] In an optional further step 4), the composition obtained in the end of step 3) is dried by PAT8513PC00 i. spray drying or ii. lyophilization.

[0102] In such cases, the dry oleosome composition shall be reconstituted with water in step a) of the process of the present invention

[0103] In a preferred aspect of the invention, no washing step is performed after the separation step. This aspect is advantageous in that it avoids removing of the "isolated" yeast cell wall component such as described above that are recovered by the separation step.

[0104] In step b) of the process of the present invention, the yeast oleosomes are admixed with the exogenous proteins and any desired optional ingredient. The yeast oleosomes, exogenous protein and optional ingredients are as described above in the section dedicated to the composition.

[0105] In step c), the mixture of step b) is homogenized to provide a homogenous composition. Preferably the mixture is subjected to high-pressure homogenization. More preferably at a pressure of at least 50 bar, preferably at least 100 bar, more preferably at least 200 bar, more preferably at least 300 bar. In an aspect, the high-pressure homogenization is performed at a pressure of up to 1000 bar, or up to 900 bar, or up to 800 bar or up to 700 bar or up to 600 bar or up to 500 bar. The homogenization can be performed in two steps at different pressures.

[0106] Examples

[0107] Example 1: Comparison of the sensory properties of compositions according to the invention and comparative compositions

[0108] The present examples aim at assessing the sensory characteristics of protein shakes with addition of yeast oleosomes compared to plant-based controls. Formulations of protein shakes with different total protein concentrations (Sample 1 and Sample 2, with 31.5 wt% and 44.5 wt% total protein, respectively, based on the weight of the total solids content in the composition) were submitted to sensory analysis. Such amounts of proteins are common in commercial products of the protein shake type. Samples 1 and 2 were compared to corresponding controls (Control 1 and Control 2) having the same total protein amount, but wherein the whole protein content originates from plant proteins. Samples 1 and 2 had respectively 31.5 and 44.5 wt% exogenous protein (pea protein).

[0109] Formulations were prepared to ensure the matching of fat, carbohydrate and protein content between the Samples and the Controls. The inclusion rate of yeast oleosomes PAT8513PC00 composition in Sample 1 and Sample 2 was calculated so that the fat present in Control 1 and Control 2, respectively, would be completely substituted by the yeast oleosomes composition. Protein and carbohydrates were then adapted based on the protein and carbohydrate content of the yeast oleosomes composition. The yeast oleosomes composition is extracted from Yarrowia lipolytica, has a total solids content of 20 wt% and a water content of 80 wt%, based on the total weight of the oleosomes composition (origin: Cultivated Biosciences, Switzerland). The amount of water in the yeast oleosomes composition was used to adjust the amount of water added to match the Controls. All Samples and Controls have the same water content. Gellan gum was not added at higher protein content because of the already high viscosity given by high amounts of proteins.

[0110] Table 1: Nutritional values of the yeast oleosomes composition used in Samples 1 and 2:

[0111] Table 2: Composition Samples 1 and 2 (according to the invention) and of Controls 1 and 2 (comparative) PAT8513PC00

[0112] 1) aqueous yeast oleosomes composition extracted from Yarrowia lipolytica with a total solids content of 20 wt% and 5wt% P-D-glucan (origin: Cultivated Biosciences, Switzerland)

[0113] Table 3: Nutritional values of Samples and Controls:

[0114] The Samples and the Controls were prepared as follows. The water, gellan gum, lecithin, oat flour, pea protein (when present the yeast oleosomes composition) and the sugar were mixed. The mixture was heated to 60°C for 20 minutes under stirring and stirring was then continued until the mixture cooled down to room temperature. The sunflower oil was then added and the composition was pre-homogenized with an overhead stirrer at 20000 rpm for 7 min. The mixture was then subjected to two steps of high-pressure homogenization (device: SPXFLOW, type APV 1000) with high pressure at 500 bar and small pressure at 50 bar.

[0115] The homogenized composition was then pasteurized at 75°C for 2 minutes in a Thermomix. The composition was let to rest overnight in the fridge before analysis.

[0116] The samples were subjected to sensory analysis based on the protocol described by Agorastos et al. (2023) (Agorastos, G., Klosse, B., Hoekstra, A., Meuffels, M., Welzen, J. J. M. J., van E, H., ... & Klosse, P. (2023). Instrumental classification of beer based on mouthfeel. International Journal of Gastronomy and Food Science, 32, 100697). A trained panel assessed the samples’ sensory properties. Experienced tasters (n=20) were selected for the sensory evaluation. The selected panellists were screened by a questionnaire based on age, gender, nonsmoking, and absence of oral disorders. The panellists were trained on relevant sensory attributes based on previous studies. The sensory attributes that were selected are creaminess, bitterness, sweetness, astringency and off aroma. A sample of 50 ml was given to each panellist. Samples were presented in amber vials labelled with random three-digit codes throughout the study to minimize expectation error and to reduce bias from appearance. Panellists were given a 3-minute break among each sample to avoid palate fatigue. During the break, panellists used water to cleanse the palate and minimize sample carry-over. The panellists rated the samples based on a continuous scale with the value 1 representing the lowest and 7 representing the PAT8513PC00 highest sensation perceived by the panellists. The results were analysed via statistical analysis. Analysis of variance (ANOVA) was performed. The data were first checked for normality and homogeneity of variance.

[0117] Sample 1 was significantly less astringent and bitter than Control 1 and Sample 1 was also lower than Control 1 in off aromas. Sample 1 was creamier than Control 1. Sample 2 was the creamiest among the samples. In a similar pattern Sample 2 was less astringent and bitter, and had less off aroma and higher creaminess than Control 2. All identified differences between Sample and corresponding Control were statistically significant (p-value lower than 0.05). The results are provided in the graph of [Fig. 1],

[0118] Example 2: Comparison of protein shakes with variable amounts of yeast oleosomes and fixed amount of pea proteins

[0119] Protein shakes were prepared with the ingredients of Table 4. Formulation of the Samples and of the Control were prepared to ensure the matching of total fat and protein contents in the Samples and Control. The amount of Pea Protein isolate was adapted in each Sample to the yeast oleosome inclusion rate, such as to obtain the same total protein content in each Sample.

[0120] Table 4: Composition Samples 3 and 4 (invention) and of Control 3 (comparative) PAT8513PC00

[0121] 1) aqueous yeast oleosomes composition extracted from Yarrowia lipolytica with a total solids content of 20 wt% and 5wt% P-D-glucan (origin: Cultivated Biosciences, Switzerland)

[0122] Table 5: Nutritional values of Samples and Controls:

[0123] The Samples and the Controls were prepared as follows. The water, pea protein (when present the yeast oleosomes composition or lecithin), the sugar and the aroma were mixed. The mixture was heated to 60°C for 20 minutes under stirring and stirring was then continued until the mixture cooled down to room temperature. The sunflower oil (when present) was then added and the composition was pre-homogenized with an overhead stirrer at 20000 rpm for 7 minutes. The mixture was then subjected to two steps of high-pressure homogenization (device: SPXFLOW, type APV 1000) with high pressure at 500 bar and small pressure at 50 bar. The homogenized composition was then UHT treated at 136°C for 3 seconds with an OMVE HT220 HTST / UHT system. The composition was let to rest overnight in the fridge before analysis.

[0124] In terms of texture, Samples 4 and 5 and Control 3 were flowable . Sample 4 exhibited the best viscosity for a drinkable protein shake and exhibited a good flowability. The proteins in Samples 3 and 4 gelled much less than in Control 3, and this advantage in terms of texture was obtained despite the UHT treatment to which Samples 3 and 4 were subjected. This is surprising as UHT treatment is known to promote gelling of protein, especially at high concentrations. The prevention of protein gelling is due to the combination of the proteins with yeast oleosomes.

[0125] Samples 3 and 4 were tasted. The samples were subjected to sensory analysis based on the protocol described by Agorastos et al. (2023) (Agorastos, G., Klosse, B., Hoekstra, A., Meuffels, M., Welzen, J. J. M. J., van E, H., ... & Klosse, P. (2023). Instrumental classification of beer based on mouthfeel. International Journal of Gastronomy and Food Science, 32, 100697). A trained panel assessed the samples’ sensory properties. Experienced tasters (n=8) PAT8513PC00 were selected for the sensory evaluation. The panellists rated the samples based on a continuous scale with the value 1 representing the lowest and 7 representing the highest sensation perceived by the panellists. The results were analysed via statistical analysis. Analysis of variance (ANOVA) was performed. The data were first checked for normality and homogeneity of variance.

[0126] Both samples presented very smooth and creamy mouthfeel. Plant off aromas and stringency were only slightly perceptible in both Samples, and advantageously significantly less present in Sample 3 than in Sample 4.

[0127] The viscosity of the Samples was measured in an Anton Paar rheometer MCR 301, with cylinder geometry (serial number CC27 9342). The viscosity values were obtained at a shear rate of 50s'1and a temperature of 10°C. Viscosity of sample 4 was lower than viscosity of Sample 3. An as low viscosity as possible is desired in order to facilitate processing and to being able to increase the protein content even more. The results are provided in Table 6 below.

[0128] Table 6: Viscosity measurements for Samples 3 and 4 and Control 3

[0129] The stability of the protein shakes was assessed by measuring the particle size distribution of the samples after UHT and over shelf life with a Anton Paar PSA 1190. Measuring time was set to 1 min with 6 repetitions per sample. The reconstruction algorithm was MIE, the refractive index for the material was 1.47 and the absorption index was 0.01. Samples were dissolved in distilled water, the refractive index was set to 1.33. The average of the D(4,3) of the six measurements was used for the evaluation of the samples. The particle size of samples 4 and 5 was below D4,3 of 15 pm after UHT and stayed below D4,3 of 15 pm after 1 and after 2 months of storage, indicating a good stability over this period. The results after 2 months storage are shown in Table 7. PAT8513PC00

[0130] Table 7: D4,3 of Samples 3 and 4 after two months storage

[0131] Example 3: Effect of p-D-glucan in yeast oleosomes stability

[0132] The objective of this experiment was to compare the stability of yeast oleosomes having a total solids (TS) content of 20 wt% and comprising about 5 wt% P-D-glucan based on TS, to yeast oleosomes comprising degraded P-D-glucan. To achieve this, samples were prepared by degrading the P-D-glucan contained in a yeast oleosomes composition extracted from Yarrowia lipolytica with a total solids content of 20 wt% and 5wt% P-D-glucan (origin: Cultivated Biosciences, Switzerland) (Sample 5). One composition was produced by treating the yeast oleosomes composition with endo-glucanase (Sample 6) and another one by treating the yeast oleosomes composition with exo-glucanase (Sample 7). Endo-glucanase degrades P- D-glucan by cleaving the beta-1,3 bonds, thus cutting the native P-D-glucan into smaller fractions. Exo-glucanase cleaves the non-reducing ends of P-D-glucan, thus releasing alphaglucose.

[0133] Samples 5, 6 and 7 were kept at the same temperature for the same time as the enzymatic treatment. All samples have also been heated to 90°C (temperature used to inactivate the enzymes).

[0134] The size of the particles in Samples 5, 6 and 7 was then assessed. For all samples, particle size distribution (PSD) was measured using a particle size analyzer (PSA 1190, Anton Paar). The measurement parameters were set as follows: equilibration time of 1.5 minutes, measurement time of 30 seconds, stirrer speed of 150 rpm, pump speed of 120 rpm, and obscuration range of 3 - 8%. Each sample was measured six times in sequence to ensure reproducibility. The model used was Mie, with weighting by volume. The D4,3 of all samples is provided in Table 8 below. PAT8513PC00

[0135] Table 8: particle size of Sample 5, 6 and 7

[0136] These results show that the particle size of the yeast oleosomes emulsion is very much increased when P-D-glucan is degraded, thus showing a very advantageous role of P-D-glucan on the structure of the emulsion. It also advantageously improves the stability of the emulsion over time, because the smaller the particles, the more stable the emulsion is over time. It is therefore particularly advantageous to use yeast oleosomes comprising P-D-glucan in products such as high protein liquid products, such as protein shakes, in which stability over time is a very important aspect for product quality and acceptance by the consumers.

[0137] Example 4: effect of P-D-glucan on emulsification capacity

[0138] The emulsification capacity of Samples 5, 6 and 7 used in Example 3 was assessed. An amount of 5 g of Samples 5, 6 or 7, respectively, was added to a 40 mL beaker. Another beaker was filled with oil and weighed. Homogenization of the 5 g of sample using a mediumsized disperser (Ika S25N-10G) at medium speed (13500 rpm) was started. Then 1 mL of oil was added with a pipette to the emulsion, until it was emulsified. Additions of 1 mL of oil were repeated until the emulsion broke (i.e. until the viscosity of the emulsion dropped and the emulsion became visibly heterogeneous). The oil beaker was weighed again and the amount of oil added to the Sample until emulsion broke was calculated as being the difference between the weight of the oil beaker in the beginning of the experiment and the weight of the beaker at the time the emulsion broke.

[0139] The emulsification capacity was calculated by dividing the amount of oil added (in grams) by the amount of yeast oleosomes composition total solids (TS) in the beaker (in grams, here 1 g). Measurements were performed in duplicates. The emulsion viscosity before the breaking point of the emulsion was also assessed on a scale from 1 (very liquid) to 4 (solidlike). The results are provided in Table 9 below. 1

[0140] PAT8513PC00

[0141] Table 9: Emulsification capacity (EC) of Samples 5, 6 and 7

[0142] These results demonstrate that degradation of P-D-glucan significantly decreases the emulsification capacity of the yeast oleosome composition. This shows that P-D-glucan in the form of long enough chains, provide improved emulsification capacity to the yeast oleosome composition making it particularly advantageous in high protein product, in particular those in liquid form such as protein shakes, wherein emulsification capacity is a key parameter impacting product stability over time, product appearance, product texture and the possibility to use high amounts of proteins.

[0143] Example 5: comparison of emulsification capacity of yeast oleosomes over plant-based oleosomes

[0144] The emulsification capacity of the yeast oleosome composition of Sample 5 was compared to that of a 20% TS sunflower oleosomes composition (Sample 8).

[0145] The sunflower oleosomes were extracted using the method disclosed in Karfyllakis et al., Sof Matter, 2019, 15, 4639. First, seeds were soaked overnight in an aqueous solution of pH 8, adjusted with 4 M NaOH, and stored at 4 °C. After overnight soaking (16 hours), the excess aqueous solution was drained off using a strainer. The seeds were then ground with demi water in the ratio 1 : 7 (w / v) for 90 seconds in a laboratory blender at maximum speed. The slurry was then passed through a commercial juicer in order to remove the solid leftovers. The crude extract obtained as filtrate was centrifuged at 10000g at 4 °C for 30 min. The cream layer was then carefully removed and subsequently re-suspended in aqueous solution pH 8 at 1 : 4 ratio (w / v) and centrifuged under the same conditions. This washing cycle was repeated once more. The total solids of the obtained composition was of 20%, thus obtaining the sunflower oleosomes composition of Sample 8.

[0146] The emulsification capacity of Samples 5 and 8 was assessed. An amount of 5 g of Samples 5 and 8, respectively, was added to a 40 mL beaker. Another beaker was filled with oil and weighed. Homogenization of the 5 g of sample using a medium-sized disperser (Ika PAT8513PC00

[0147] S25N-10G) at medium speed (13500 rpm) was started. Then 1 mL of oil was added with a pipette to the emulsion, until it was emulsified. Additions of 1 mL of oil were repeated until the emulsion broke (i.e. until the viscosity of the emulsion dropped and the emulsion became visibly heterogeneous). The oil beaker was weighed again and the amount of oil added to the Sample until emulsion broke was calculated as being the difference between the weight of the oil beaker in the beginning of the experiment and the weight of the beaker at the time the emulsion broke.

[0148] The emulsification capacity was calculated by dividing the amount of oil added (in grams) by the amount of TS in the sample place in the beaker (in grams, here 1 g). Measurements were performed in duplicates. The emulsion viscosity before the breaking point of the emulsion was also assessed on a scale from 1 (very liquid) to 4 (solid-like). The results are provided in Table 10 below.

[0149] Table 10: Emulsification capacity (EC) of Samples 5 and 8

[0150] These results demonstrate that yeast oleosomes have a significantly better emulsification capacity compared to plant-based oleosomes such as sunflower oleosomes. This shows that yeast oleosome are particularly advantageous in high protein products, in particular those in liquid form such as protein shakes, wherein emulsification capacity is a key parameter impacting product stability over time, product appearance, product texture and the possibility to use high amounts of proteins.

[0151] Example 6: Comparison of the stability of yeast oleosomes and sunflower oleosomes

[0152] The objective of this experiment was to compare the stability of yeast oleosomes (Sample 5) with sunflower oleosomes (Sample 8), such as used in Example 5. The size of the particles in Samples 5 and 8 was then assessed as described in Example 4. The D4,3 of all samples is provided in Table 11 below. PAT8513PC00

[0153] Table 11: particle size of Sample 5 and 8

[0154] These results show that the particle size of the yeast oleosomes is smaller than that of plant-based oleosomes, such as sunflower oleosomes, thus showing a very advantageous emulsion structure of yeast oleosomes emulsion. It also advantageously translates in improved stability of yeast oleosomes over plant-based oleosomes, such as sunflower oleosomes, because the smaller the particles, the more stable the emulsion is over time. It is therefore particularly advantageous to use yeast oleosomes in high protein products, particularly in liquid products, such as protein shakes, in which stability over time is a very important aspect for product quality and acceptance by the consumers.

Claims

PAT8513PC00Claims1. A composition comprising yeast oleosomes and at least 30 wt% exogenous protein, preferably at least 31 wt%, more preferably at least 32 wt%, more preferably at least 33 wt% exogenous protein, based on the weight of the total solids content in the composition.

2. The composition according to claim 1, wherein the yeast oleosomes are provided in the form of a yeast oleosomes composition comprising at least one isolated yeast cell wall component, preferably selected from lipids, carbohydrates and proteins, more preferably selected from chitin, P-D-glucan, mannoprotein and combinations thereof, preferably P- D-glucan.

3. The composition according to claim 2, wherein the at least one isolated yeast cell wall component is present in an amount of at least 1 wt%, preferably 1.5 wt%, preferably at least 2 wt%, more preferably at least 2.5 wt%, more preferably at least 3 wt%, more preferably at least 3.5 wt%, preferably at least 4 wt%, preferably at least 4.5 wt%, preferably at least 5 wt%, preferably at least 7.5 wt%, preferably at least 10 wt%, preferably at least 15 wt%, preferably at least 20 wt% based on the total weight of the solids in the yeast oleosomes composition.

4. The composition according to any one of claims 1 to 3, wherein the exogenous protein is selected from plant proteins, non-yeast fungal proteins, bacterial proteins, algal proteins, animal proteins selected from recombinant animal proteins and animal proteins produced by animal cell cultures, functional proteins produced by precision fermentation and mixtures thereof.

5. The composition according to claim 4, wherein the exogenous protein is plant protein.

6. The composition according to any one of claims 1 to 5, wherein the yeast oleosomes are provided in an amount such that the solids of the yeast oleosomes be present in an amount of at least 1 wt%, preferably at least 3 wt%, preferably at least 4 wt%, preferably at least 4.5 wt%, preferably at least 5 wt%, preferably at least 5.5 wt%, preferably at least 6 wt%, preferably at least 6.5 wt%, more preferably at least 6.6 wt%, based on the weight of the total solids content in the composition of the invention.

7. The composition according to any one of claims 1 to 6, wherein the ratio of the total solids in the oleosome composition to the exogenous protein in the composition is of 1 :5 to 5: 1, preferably 1 :4.5 to 4.5: 1, preferably 1 :4 to 4: 1, preferably 1 :3 to 3: 1, preferably 1 :2 to 2: 1, more preferably 1 :2 to 1.5: 1, preferably 1 :2 to 1.2: 1.PAT8513PC008. The composition according to any one of claims 1 to 7, wherein the composition has a total protein content of more than 30 wt%, preferably at least 31 wt%, preferably at least 35 wt%, preferably at least 40 wt%, preferably at least 45 wt%, more preferably at least 47 wt%, based on the weight of the total solids content in the composition of the invention.

9. The composition according to any one of claims 1 to 8, wherein the composition further comprises an additional ingredient selected from carbohydrates, lipids, micronutrients, flavours, colorants, preservatives, low calorie sweeteners, spices, taste enhancers, emulsifiers, thickeners, and / or cereals, nuts, legumes, fruits or vegetables flour, juice or pieces and combinations thereof.

10. Use of yeast oleosomes for improving at least one sensory property of a composition comprising at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition.

11. The use according to claim 10, wherein the yeast oleosomes are used for improving creaminess, reducing bitterness, reducing astringency and / or reducing off-aroma of the composition.

12. A food product or a beverage comprising a composition according to any one of claims 1 to 9.

13. The food product or beverage according to claim 10, in the form of a protein shake, a protein powder, a protein bar, a dairy product or a non-dairy alternative product.

14. A process for the preparation of a composition comprising yeast oleosomes and at least 30 wt% exogenous protein, based on the weight of the total solids content in the composition comprising the step of: d) providing yeast oleosomes; e) admixing the yeast oleosomes provided in step a) with exogenous proteins and optionally other ingredients, such that the exogenous proteins are present in a final concentration of at least 30 wt%, based on the weight of the total solids content in the composition; and f) homogenising the mixture obtained in step a).

15. The process according to claim 13, wherein the homogenisation performed in step c) is high-pressure homogenization.

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