Yeast oleosome composition

Abstract

The present invention relates to a yeast oleosome composition comprising yeast oleosomes and at least one isolated yeast cell wall component, preferably having a specific particle size, density, and zeta potential. This composition is characterized by good physical stability and functional properties. The present invention also relates to a process for producing the yeast oleosome composition of the present invention, a product containing the yeast oleosome composition of the present invention, and the use of the composition to improve the functional properties of food compositions or beverages or edible components. Furthermore, the composition of the present invention has been proven useful in reducing the intensity of astringency in foods and beverages.

Description

Technical Field

[0001] The present invention relates to a yeast oleosome composition comprising a yeast oleosome and at least one isolated yeast cell wall component, preferably having a specific particle size, density and zeta potential. This composition is characterized by good physical stability and functional properties. The present invention also relates to a process for producing the yeast oleosome composition of the present invention, a product comprising the yeast oleosome composition of the present invention, and the use of said composition for improving the functional properties of a food composition or beverage or edible ingredient. Furthermore, the composition of the present invention has been proven to be useful for reducing the intensity of astringency (astringency) in foods and beverages.

Background Art

[0002] As the diets of vegetarians and vegans become increasingly common, various types of dairy alternatives have been developed. Most dairy alternatives are plant-based, for example based on oil bodies or oleosomes. Most of the prior art relates to compositions based on plant oleosomes.

[0003] Oleosomes, also commonly referred to as fat droplets or oil bodies, are stable oil bodies having a core of triglycerides surrounded by a membrane composed of phospholipids and proteins. Oleosomes are present in large amounts in plants, particularly seeds, and are also known to be found in other organisms such as yeast and other microorganisms. The composition and properties of oleosomes vary depending on the source.

[0004] For example, Nikiforidis et al., RSC Adv., 2014, 4, 25067 disclose natural emulsions based on plant-derived oil bodies having properties similar to milk or cream. The oil bodies are extracted from plant material by using an aqueous medium. An oil-in-water emulsion based on intact or partially disrupted oil bodies is obtained, and protein co-extraction occurs. The size of the oil bodies can be in nanometers and can be several microns.

[0005] International Publication No. 98 / 53698A1 discloses emulsion formulations prepared from oils derived from living cells. While the document focuses on plant oils, other oil sources, including fungal sources such as yeast cells, are mentioned very generally. The oils are obtained from cells, washed, and formulated. All disclosed process aspects focus on the preparation of formulations from plant seeds. The size of the oils varies between 0.4 and 1.5 μm. The disclosed emulsions can be used as dairy substitutes.

[0006] International Publication No. 2017 / 066569A1 discloses an oleosome composition comprising two different oleosome compositions having different particle sizes, preferably derived from two different sources, wherein the 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 very broadly. However, all embodiments described in detail relate to plant oleosomes.

[0007] International Publication No. 2021126409A1 discloses roasted oleosome compositions which may be derived from various sources, including yeast, but preferably from plant sources. The isolated oleosome compositions have a dry matter content of 30-80% by weight, a protein content of 1-6% and an oil content of 94-99% based on the dry matter.

[0008] Despite extensive research into developing dairy alternatives, there is a need to further improve the mouthfeel of non-dairy products. Indeed, consumers still perceive differences in mouthfeel and mouth coating between dairy products and their non-dairy substitutes for milk and cream. To further stimulate consumption of these more environmentally friendly alternatives, it is desirable to improve consumer acceptance of non-dairy products. Therefore, further fine-tuning of oleosome compositions is necessary to improve physical and sensory properties, taking into account the production of dairy substitutes that more closely meet consumer expectations in terms of creaminess and mouthfeel.

[0009] Furthermore, plant production requires a large land surface area and consumes a considerable amount of water, which is becoming an increasingly scarce resource. Therefore, it is desirable to provide reliable, finely tuned alternatives to dairy products that are not derived from plants. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] The present invention aims to solve these problems. [Means for solving the problem]

[0011] In a first aspect, the present invention provides a yeast oleosome composition comprising yeast oleosomes and at least one yeast cell wall component selected from chitin, β-D-glucan, and mannoprotein, wherein such yeast cell wall component is released from the cell wall structure.

[0012] In certain embodiments, the yeast oleosome composition is in the form of a yeast oleosome-based dairy substitute.

[0013] In a second aspect, the present invention relates to a process (method) for producing a yeast oleosome composition according to the present invention, a) A step of providing oilseed yeast cells, b) A step of dissolving the cell wall, c) A step to separate oleosomes from other yeast cell components. This relates to processes (methods), including steps.

[0014] In a third aspect, the present invention relates to a product obtained or obtainable by the process of the present invention.

[0015] In a fourth aspect, the present invention relates to a food, beverage, feed product, or cosmetic composition comprising the yeast oleosome composition according to the present invention.

[0016] In a fifth aspect, the present invention relates to the use of the yeast oleosome composition of the present invention for improving the sensory properties and / or physical stability of a food, beverage or edible ingredient.

[0017] In a sixth aspect, the present invention relates to a lyophilized product of the yeast oleosome composition according to the present invention (i.e., a yeast oleosome composition in solid form).

[0018] In a seventh aspect, the present invention has a volume average diameter of 0.2 to 7 μm, a density of 0.8 to 1.1 g / ml, a pH of 5 to 8, and a zeta potential of -40 to -5 mV at physiological pH measured by laser diffraction, preferably the particle size (particle diameter), density, pH and zeta potential, total solids content, fat content, saturated fat content, carbohydrate content, protein content, yeast definition and coefficient of friction are as disclosed in any of the preferred embodiments of the following detailed description. It relates to a yeast oleosome composition characterized by being.

[0019] In an eighth aspect, the present invention relates to the use of the yeast oleosome composition of the present invention for reducing the astringency intensity of a food, beverage or edible ingredient.

[0020] [[ID=…]]

[0021] <0000…]] In a tenth aspect, the present invention relates to a food or beverage comprising the yeast oleosome composition of the present invention and at least one astringent (astringent) component.

Brief Description of Drawings

[0022] [Figure 1] Results of comparative stability tests of sample A1 and A2 according to the present invention and comparative coconut, oatmeal, almond, cashew and soybean oleosome milks evaluated in Example 6.

[0023] ​ [Figure 2] Friction measurements as a function of the slip rate of sample A1 and A2 according to the present invention and comparative coconut, oat, almond, cashew and soybean oleosome milks evaluated in Example 6.

[0024] [Figure 3] Viscosity measurements as a function of the shear rate of an oleosome composition suitable as an alternative to dairy cream according to the present invention and a comparative plant-based cream (cultured sample) evaluated in Example 7.

[0025] [Figure 4] Viscosity measurements as a function of the shear rate for commercially available products (dairy and plant-based) and an oleosome composition (cultured cream sample) suitable as an alternative to dairy cream according to the present invention evaluated in Example 8.

[0026] [Figure 5] Friction measurements as a function of the slip rate of a composition according to the present invention (cultured cream 20%) suitable as an alternative to dairy cream and commercially available plant-based creams (Vlove cream, Soja cuisine and Oat cuisine) evaluated in Example 9.

[0027] [Figure 6] Results of soft tribology measurements performed in Example 10, representing the delta friction coefficient (Δμ) of a sample containing a pea protein solution in saliva, a sample in which the pea protein solution is combined with the yeast oleosome composition and saliva of the present invention, a sample containing only the yeast oleosome of the present invention in saliva, and a control consisting of water in saliva.

[0028] [Figure 7]Graph showing the definition of the delta friction coefficient (Δμ): Schematic diagram of the friction coefficient μ as a function of time obtained by the dynamic protocol used in Example 10. Saliva is added at time 0 of the measurement. After obtaining a constant baseline value (AvCoF1) as a result of saliva addition, the model solution (MS) is added. Av.CoF2 is calculated using only the data points after the stabilization of the interaction between saliva and MS. Δμ is the difference between Av.CoF2 and Av.CoF1.

[0029] [Figure 8] Results of sensory evaluation of Example 11. Average astringency scores by panelists on a scale of 1-5, where 1 is the lowest score and 5 is the highest.

[0030] [Figure 9] Particle size distribution by volume of both samples analyzed in Example 13.

[0031] [Figure 10] Particle size distribution by number of both samples analyzed in Example 13. [Modes for carrying out the invention]

[0032] Yeast oleosome composition The inventors have developed a yeast oleosome composition characterized by particularly favorable physical stability and functional properties, especially from the viewpoint of use as an emulsion in either food applications (e.g., substitutes for dairy products, sauces, soups, and meat or egg products) or cosmetic applications (e.g., creams, lotions, etc.). A yeast oleosome is defined as a stable oil body (also called a lipid droplet) derived from yeast, having a triglyceride core surrounded by a membrane composed of phospholipids and proteins.

[0033] The inventors have optimized the composition and properties of the yeast oleosome composition to obtain a pleasant mouthfeel, particularly preferably one that matches the mouthfeel and other sensory characteristics of milk or cream. Such advantageous properties are imparted to the composition by a combination of oleosomes and at least one isolated yeast cell component selected from chitin, β-D-glucan, and mannoprotein, wherein this at least one yeast cell wall component is released (or, in other words, isolated) from the cell wall structure. In a preferred embodiment, the yeast oleosome composition of the present invention comprises chitin, β-D-glucan, and mannoprotein. The terms “released” or “isolated” from the yeast cell wall structure as used herein mean that the yeast cell wall component is present in the composition in a free form, i.e., separated from the yeast cell wall structure and not part of the structure of the yeast cell wall debris. The characteristic that yeast cell wall components are "released" or "isolated" from the yeast cell wall structure is important for making the yeast cell wall components available for interaction with other components of the composition, such as oleosomes, and / or with the environment to which the composition is exposed, such as the oral cavity and / or other molecules to which the composition of the present invention is mixed (such as other components of a food containing the composition of the present invention).

[0034] At least one yeast cell wall component may be present in the yeast oleosome composition in either a solubilized or suspended form, or may be associated with the yeast oleosome membrane, or may be partially solubilized or suspended and partially associated with the yeast oleosome membrane. In a preferred embodiment, at least one yeast cell wall component is preferably associated with the yeast oleosome membrane via non-covalent bonds such as electrostatic interactions, hydrogen bonds, or hydrophobic interactions. In one embodiment, the composition comprises at least two yeast cell wall components selected from chitin, β-D-glucan, and mannoprotein, one or two of these yeast cell wall components being in a solubilized or suspended form, and the other one or two components being associated with the yeast oleosome membrane. Preferably, chitin, β-D-glucan, and mannoprotein are all associated with the yeast oleosome membrane.

[0035] In a preferred embodiment, at least one isolated yeast cell wall component is present in the composition in an amount of at least 0.3% by weight, preferably at least 0.4% by weight, more preferably at least 0.5% by weight, more preferably at least 0.6% by weight, more preferably at least 0.7% by weight, more preferably at least 0.8% by weight, more preferably at least 0.9% by weight, and most preferably at least 1% by weight, based on the total weight of the composition of the present invention.

[0036] In a more preferred embodiment, the composition of the present invention contains, based on the total weight of the composition, at least 0.3% by weight, preferably at least 0.4% by weight, more preferably at least 0.5% by weight, more preferably at least 0.6% by weight, more preferably at least 0.7% by weight, more preferably at least 0.8% by weight, more preferably at least 0.9% by weight, and most preferably at least 1% by weight of β-glucan.

[0037] The compositions of the present invention are the result of a process that ensures the efficient lysis of yeast cell walls, enables the release of chitin, β-D-glucan, and mannoprotein from the yeast cell walls, and preserves the structure of oleosomes. An overly gentle process either fails to destroy the cell wall or fails to release chitin, β-D-glucan, and mannoprotein. In contrast, an overly harsh process destroys the cell wall but also destroys the structure of oleosomes, thus adversely affecting the emulsifying properties and mouthfeel of the composition.

[0038] The presence of at least one isolated yeast cell wall component selected from chitin, β-D-glucan, and mannoprotein favorably affects the viscosity of the composition. In fact, the viscosity of a composition containing oleosomes in combination with such yeast cell wall components is characterized by a higher viscosity than the same composition without such yeast cell wall components.

[0039] In certain embodiments, the compositions of the present invention are in the form of dairy substitute compositions. These compositions may be provided in liquid or solid form. If the composition is in liquid form, it is ready for use as a substitute for milk or cream. If the composition is in solid form, it will be reconstituted with water before consumption as a substitute for milk or cream, similar to the case of dehydrated dairy products.

[0040] In a preferred embodiment, the particle size of the oleosome composition of the present invention is preferably characterized by a volume-average diameter of 0.2 to 7 μm, measured by dynamic light scattering using Zetasize Ultra (Malvern Instruments). The particle size may vary within the above range depending on the type of composition. Preferably, for milk alternative applications, the volume-average diameter is 0.2 to 2 μm, preferably 0.2 to 1 μm, more preferably 0.2 to 0.8 μm, more preferably 0.4 to 0.8 μm, even more preferably 0.6 to 0.8 μm, or 0.2 to 0.6 μm, for example, 0.2 to 0.5 μm, while for cream alternative applications, the volume-average diameter is 0.4 to 7 μm. Larger particles are more suitable for cream as they give the composition a creamier mouthfeel.

[0041] Particle size is a determining parameter that imparts advantageous physical and functional properties to the composition, such as advantageous physical stability, wettability, and improved mouth coating properties, as shown in the following examples, resulting in optimal lubrication in the mouth when the product is ingested.

[0042] In a preferred embodiment, the density of the yeast oleosome composition is 0.8 to 1.1 g / ml. The density of the composition also plays an important role in fine-tuning the functional properties of the yeast oleosome composition of the present invention. Within the above range, it is preferable that the density of the composition differs slightly between the composition for use as a milk substitute and the composition for use as a cream substitute. For milk substitutes, the density is preferably 0.8 to 1.1 g / ml, more preferably 0.85 to 1.0 g / ml. For cream substitutes, the density is preferably 0.85 to 1.2 g / ml, more preferably 0.9 to 1.1 g / ml.

[0043] In a preferred embodiment, the zeta potential at the surface of oleosomes in the composition is in the range of -40 to -5 mV, preferably -40 to -8 mV, more preferably -40 to -10 mV, even more preferably -40 to -20 mV, and most preferably -30 to -20 mV at physiological pH. Such a zeta potential is advantageous because, within such a range, the electrostatic interactions between oleosomes are low enough to avoid oleosome aggregation and ensure the proper physical stability of the composition. A zeta potential within this range also contributes to the interaction between the yeast oleosome membrane and polysaccharides. The zeta potential varies within this range depending on the proteins present in the oleosome membrane.

[0044] In a preferred embodiment, the pH of the oleosome composition is 5 to 8, preferably 6 to 7. This pH range is preferred because it affects the net charge of the oleosomes and has a positive (favorable) effect on the physical stability of the composition over time.

[0045] In certain embodiments, the yeast oleosome composition of the present invention is in liquid form and has a total solids content of 2 to 45% by weight. Within this range, the total solids content of the yeast oleosome composition of the present invention in liquid form can be adjusted depending on the intended use. The total solids content is higher, for example, in the case of a cream substitute than in the case of a milk substitute. Preferably, the total solids content of the yeast oleosome composition of the present invention, which is particularly suitable as a milk substitute, is 2 to 12% by weight, preferably 2 to 10% by weight, more preferably 2 to 8% by weight, more preferably 2 to 6% by weight, and even more preferably 2 to 5% by weight, based on the total weight of the composition. On the other hand, the total solids content of the yeast oleosome composition of the present invention, which is suitable as a cream substitute, is 15 to 45% by weight, more preferably 16 to 45% by weight, more preferably 20 to 45% by weight, preferably 30 to 45% by weight, more preferably 35 to 45% by weight, and more preferably 40 to 45% by weight, for example, 16.6% by weight or 42% by weight, based on the total weight of the composition. The wide range of total solids content applicable to cream substitutes makes it possible to manufacture a variety of cream substitutes for various types of dairy products, such as half cream, whole cream (unprocessed cream), double cream, and coffee cream, by varying the total solids content.

[0046] The yeast oleosome composition of the present invention in solid form (i.e., a composition that is a dried version of the yeast oleosome composition of the present invention in liquid form, which can be obtained by spray drying, freeze-drying, freeze-drying, or any other suitable drying method, preferably freeze-drying, to form a lyophilized product) preferably has a residual moisture content of 3-4% by weight, i.e., a total solids content of 96-97% by weight. Typically, such a composition in solid form or a lyophilized product is reconstituted with water before ingestion. The amount of water used for reconstitution may vary depending on the dairy substitute to be produced. The composition is preferably reconstituted with an amount of water suitable for obtaining the above-mentioned final total solids content for each dairy substitute and cream substitute, respectively.

[0047] In certain embodiments, the yeast oleosome composition of the present invention in liquid form has a fat content of 1.25 to 60% by weight, preferably 1.25 to 50% by weight, for example 1.25 to 38% by weight, based on the total weight of the composition. Within this range, the fat content of the yeast oleosome composition of the present invention can be adjusted depending on the intended use. The fat content is higher, for example, in the case of a cream substitute than in the case of a milk substitute. Preferably, the yeast oleosome composition of the present invention, which is particularly suitable as a milk substitute, has a total fat content of 1.25 to 4% by weight, more preferably 1.25 to 3% by weight, even more preferably 1.25 to 2% by weight, even more preferably 1.25 to 1.6% by weight, and most preferably 1.3 to 1.5% by weight, for example 1.4% by weight, based on the total weight of the composition. Preferably, the fat content of the yeast oleosome composition according to the present invention, which is suitable as a cream substitute, is preferably 17-60% by weight, preferably 17-50% by weight, preferably 17-38% by weight, preferably 20-35% by weight, more preferably 25-35% by weight, even more preferably 28-32% by weight, and most preferably 30% by weight, based on the total weight of the composition. The wide range of fat content applicable to cream substitute applications makes it possible to produce a variety of cream substitutes that are conventionally available in dairy product applications, such as half cream, whole cream, double cream, and coffee cream, by varying the fat content.

[0048] In a preferred embodiment, the yeast oleosome composition of the present invention has a saturated fat content of 0.35 to 40% by weight, preferably 0.35 to 30% by weight, and preferably 0.35 to 19% by weight, based on the total weight of the composition. Within this range, the saturated fat content of the yeast oleosome composition varies depending on the total amount of fat. The total amount of fat, and therefore the saturated fat content, is higher, for example, in the case of a cream substitute than in the case of a milk substitute. Preferably, the yeast oleosome composition of the present invention, which is particularly suitable as a milk substitute, has a total saturated fat content of 0.35 to 2% by weight, more preferably 0.35 to 1.5% by weight, even more preferably 0.35 to 1% by weight, and even more preferably 0.35 to 0.5% by weight, for example 0.4% by weight, based on the total weight of the composition. Preferably, the fat content in the yeast oleosome composition according to the present invention, which is suitable as a cream substitute, is preferably 4.5 to 40% by weight, preferably 4.5 to 30% by weight, preferably 4.5 to 19% by weight, preferably 4.5 to 15% by weight, more preferably 4.5 to 10% by weight, and even more preferably 4.5 to 9% by weight, for example, 8.5% by weight, based on the total weight of the composition.

[0049] In a preferred embodiment, the yeast oleosome composition of the present invention has a carbohydrate content of up to 15% by weight based on the total weight of the composition. Within this range, the carbohydrate content of the yeast oleosome composition varies depending on the intended use. The total amount of carbohydrates is higher, for example, in the case of a cream substitute than in the case of a milk substitute. In a preferred embodiment, the yeast oleosome composition of the present invention, which is particularly suitable as a milk substitute, has a carbohydrate content of up to 3% by weight, preferably up to 2% by weight, more preferably up to 1.5% by weight, more preferably up to 1% by weight, even more preferably up to 0.5% by weight, and most preferably 0.2% by weight, based on the total weight of the composition. The carbohydrate content in the yeast oleosome composition of the present invention, which is suitable as a cream substitute, is preferably 0.5 to 15% by weight, preferably 1 to 10% by weight, more preferably 1 to 8% by weight, even more preferably 1 to 6% by weight, and most preferably 4% by weight, based on the total weight of the composition.

[0050] In a preferred embodiment, the yeast oleosome composition of the present invention has a protein content of 0.1 to 7% by weight based on the total weight of the composition. Within this range, the protein content of the yeast oleosome composition varies depending on the intended use. The total protein content is higher, for example, in the case of a cream substitute than in the case of a milk substitute. Preferably, the yeast oleosome composition of the present invention, particularly suitable as a milk substitute, has a protein content of 0.1 to 1.5% by weight, preferably 0.1 to 1% by weight, more preferably 0.1 to 0.8% by weight, more preferably 0.1 to 0.5% by weight, and most preferably 0.3% by weight, based on the total weight of the composition. The total protein content in the yeast oleosome composition of the present invention, suitable as a cream substitute, is preferably 2.5 to 7% by weight, preferably 3 to 7% by weight, more preferably 4 to 7% by weight, even more preferably 5 to 7% by weight, and most preferably 6.5% by weight, based on the total weight of the composition.

[0051] In a preferred embodiment, the yeast oleosome composition according to the present invention, which is particularly suitable as a milk substitute, has, based on the total weight of the composition, a total solids content of 2 to 10% by weight, a total fat content of 1.25 to 4% by weight, a saturated fat content of 0.35 to 2% by weight, a total carbohydrate content of up to 3% by weight, and a total protein content of 0.1 to 1.5% by weight.

[0052] In another preferred embodiment, a yeast oleosome composition according to the present invention, particularly suitable as a cream substitute, has, based on the total weight of the composition, a total solids content of 30-40% by weight, a total fat content of 17-38% by weight, a saturated fat content of 4.5-19% by weight, a total carbohydrate content of 0.5-15% by weight, and a total protein content of 2.5-7% by weight.

[0053] Yeast oleosomes are preferably from the genera Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, Cutaneotrichosporon, Lipomyces, and Papiliotrema. It can be derived from any oilseed yeast selected from the genera *Apiliotrema*, *Hannaella*, *Sporidiobolus*, *Kodamaea*, *Pichia*, *Saitozyma*, *Cyberlindnera*, *Meyerozyma*, *Piskurozyma*, and mixtures thereof. The oilseed yeasts are preferably Yarrowia lipolytica, Candida 107, Candida tropicalis, Candida utilis, Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodotorula babjevae, Rhodotorula sphaerocarpa, Rhodotorula graminis, Rhodosporidium toruloides, Rhodosporidium fluviale, and Cryptococcus curbatus. Trichosporon curvatus, Trichosporon pullulan, Trichosporon asahii, Cutaneotrichsporon oleaginosus, Cutaneotrichsporon curvatumCurvatum) and Lipomyces lipofer, Lipomyces starkeyi, Lipomyces tretrasporus, Lipomyces mesembrius, Schwanniomyces occidentalis, Papiliotrema terrestris, Papiliotrema flavescens, Papiliotrema laurentii, Sporidiobolus ruineniae, Kodamaea ohmeri, Pichia manshurica manshurica), Saitozyma podzolica, Cyberlindnera saturnus, Meyerozyma guilliermondii, Metschnikowia pulcherrimaThe oilseed yeast is selected from the genera Yarowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, Kutaneotrichosporon, Lipomyces, Papiliotrema, Hannaela, Sporidiovorus, Kodamaea, Pichia, Cytojima, Sivarindonella, Meyerojima, Piscragonia and mixtures thereof. The oilseed yeasts are preferably Yarowia liporitica, Candida 107, Candida tropicalis, Candida uchilis, Rhodotorula glutinis, Rhodotorula musiladinasa, Rhodotorula bavjebae, Rhodotorula sphaerocarpa, Rhodotorula graminis, Rhodosporidium toroides, Rhodosporidium fulviale, Cryptococcus curbatus, Trichosporon pullulan, Trichosporon asahi, Cutaeotrichosporon oleaginosus, Cutaeotrichosporon The species are selected from *Curbatum* and *Lipomyces lipofa*, *Lipomyces starkey*, *Lipomyces tretrasporus*, *Lipomyces mesembryus*, *Swaniomyces occidentalis*, *Papiliotrema terrestris*, *Papiliotrema flavecens*, *Papiliotrema laurentii*, *Sporidioborus ruinenie*, *Kodamaea omerii*, *Pichia manshurica*, *Cytojima podozorica*, *Syvalindonella saturunus*, *Meyerosima gilliermondii*, and mixtures thereof. Most preferably, the oilseed yeast is *Yarrowia liporitica*.

[0054] The yeast oleosome compositions of the present invention are advantageously physically stable. Typically, the yeast oleosomes disclosed above, which are suitable as milk substitutes, are stable at room temperature for at least two weeks, and the yeast oleosome compositions disclosed above, which are suitable as cream substitutes, are stable at room temperature for at least two days.

[0055] The yeast oleosome composition of the present invention is also characterized by advantageous sensory properties, such as a pleasant mouthfeel that successfully mimics dairy products. Mouthfeel is the result of several parameters, including the viscosity (rheology) and surface properties (tribology) of the composition. The inventors have found that the composition of the present invention is characterized by viscosity and surface properties closer to dairy milk / cream than compositions based on plant oleosomes, as shown in the following examples. In particular, the inventors have found that the composition has a higher viscosity and a lower coefficient of friction as a function of sliding speed than commercially available dairy substitutes based on plant oleosomes. Therefore, the composition has been found to provide both rheological and tribological properties that offer improved mouthfeel and mouth coating compared to formulations based on plant oleosomes.

[0056] Viscosity is preferably defined as the viscosity measured by any known method, for example, using an Anton Paar rheometer MCR302 having a plate configuration or any similar apparatus.

[0057] The coefficient of friction is a measure of friction and lubrication between interacting surfaces in relative motion, and is therefore a strong indicator of friction generated between food particles and the oral surface, friction between the tongue and the palate, and adhesion of food to the oral cavity, all of which play an important role in the mouthfeel and mouth coating properties of food products. The coefficient of friction is measured as a function of the velocity of the movement of the surface in question. Slow movements, such as the movement of an object within the oral cavity, are called boundary lubrication regimes. Under such conditions, the wetting and adsorption properties of the product to the oral surface are important parameters. The higher the wetting properties, the lower the coefficient of friction. Conversely, the lower the coefficient of friction, the less friction there is in the oral cavity, and therefore the smoother and more lubricating the product. Therefore, the coefficient of friction is an important parameter that determines the consumer acceptability of dairy products, especially products intended to be perceived as creamy, such as cream.

[0058] In a preferred embodiment, the yeast oleosome composition of the present invention is characterized by a coefficient of friction of 0.01 to 0.5, preferably 0.01 to 0.2, as measured by the method described in the following examples. The friction is preferably 0.02 to 0.5, more preferably 0.02 to 0.2 when the yeast oleosome composition is a milk substitute, and preferably 0.01 to 0.5, more preferably 0.01 to 0.2 when the yeast oleosome composition is a cream substitute.

[0059] process The composition of the present invention, preferably, a) A step of providing oilseed yeast cells, b) A step of dissolving the cell wall, c) A step to separate oleosomes from other yeast cell components. It is obtained by a process that includes this.

[0060] Yeast cells were selected and pretreated, for example, i. Washing with water, preferably warm water. ii. Washing with an alkaline solution, iii. Washing with an acidic solution, and / or iv. Enzymatic digestion of the cell wall It can be used for...

[0061] Step b) can be carried out by chemical or mechanical means. Suitable chemical means include: i. Preferably, alkaline treatment by immersing yeast cells in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours. ii. Contacting yeast cells with an organic solvent, such as ethanol, methanol, or heptane. iii. Self-decomposition; iv. Preferably, the enzyme treatment is applied to the washed biomass and uses one or more hydrolases suspended in water or an appropriate pH buffer at a concentration of 50-200 g / L, with low stirring or low shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, enabling the degradation of cell wall biopolymers. and combinations of these It includes.

[0062] The one or more hydrolases are preferably selected from one or more proteases, glucanases, mannanases, chitinases, nucleases, β-glucosidases, cellulases, xylanases, pectinases, and combinations thereof. Preferably, the one or more hydrolases are one or more proteases and / or glucanases.

[0063] Preferred chemical means for cell lysis include: i. Preferably, alkaline treatment by immersing yeast cells in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours. ii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of yeast cells occurs (under the action of endogenous enzymes). iii. Preferably, an enzymatic treatment is performed on washed biomass using one or more hydrolases (as defined above) suspended in water or a suitable pH buffer at a concentration of 50-200 g / L, with low stirring or shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, which enables the degradation of cell wall biopolymers. and combinations of these It includes.

[0064] Such preferred chemical means are particularly advantageous because they successfully disrupt the cell wall, thereby releasing chitin, β-D-glucan, and mannoprotein, which are components of the yeast cell wall, while simultaneously preserving the structure of oleosomes.

[0065] A suitable mechanical means for cell wall lysis in step b) preferably includes high-pressure homogenization using a pressure of 500 to 3000 bar for 1 to 10 passes, more preferably 700 to 3000 bar for 2 to 10 passes.

[0066] When high-pressure homogenization is used and combined with chemical means for dissolving the membrane, the number of passes and the homogenization pressure can be advantageously adapted. For example, when high-pressure homogenization is combined with prior enzymatic treatment, 1 to 10 passes and a pressure of 500 to 3000 bar are sufficient, but especially when high-pressure homogenization is performed after alkaline treatment, it is preferable to perform high-pressure homogenization for 2 to 10 passes, preferably 2 to 5 passes, at a pressure of 700 to 3000 bar, preferably 700 to 2000 bar. Preferably, high-pressure homogenization is performed in a high-pressure homogenizer that includes a nozzle, as the nozzle contributes to cell destruction.

[0067] The high-pressure homogenization disclosed herein is particularly advantageous over other mechanical means of dissolving yeast cell walls, such as bead milling, sonication, and microwave treatment, in that it successfully disrupts the cell wall, as shown in Example 13 below, thereby releasing yeast cell wall components such as chitin, β-D-glucan, and mannoprotein, while simultaneously preserving the structure of oleosomes. High-pressure homogenization is also advantageous in terms of yield and allows for control of particle size.

[0068] Chemical and mechanical means for dissolving cell walls can be advantageously combined with each other. Particularly advantageous combinations are: i. Preferably, alkaline treatment by immersing yeast cells in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours, followed by high-pressure homogenization using 1 to 10 passes at a pressure of preferably 500 to 3000 bar, more preferably 2 to 10 passes at a pressure of 700 to 3000 bar. ii. Preferably, an enzymatic treatment that enables the degradation of cell wall biopolymers, applied to the washed biomass and using one or more hydrolases (as defined above) suspended in water or a suitable pH buffer at a concentration of 50-200 g / L, with low stirring or low shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, followed by high-pressure homogenization using 1-10 passes at a pressure of preferably 500-3000 bar, preferably with prior pretreatment by washing with water or an alkaline solution. iii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of yeast cells occurs (under the action of endogenous enzymes), followed by high-pressure homogenization using 1 to 10 passes at a pressure of preferably 500 to 3000 bar, and preferably 2 to 10 passes at a pressure of preferably 700 to 3000 bar. That is the case.

[0069] In a preferred embodiment, cell lysis is performed by the following means i. Preferably, the yeast cells are subjected to alkaline treatment by immersing them in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours, followed by high-pressure homogenization using 2 to 10 passes at a pressure of preferably 700 to 3000 bar. ii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of yeast cells occurs (under the action of endogenous enzymes), and iii. Preferably, an enzymatic treatment applied to the washed biomass, using one or more hydrolases (as defined above) suspended in water or a suitable pH buffer at a concentration of 50-200 g / L, with low stirring or shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, which enables the degradation of cell wall biopolymers, followed by high-pressure homogenization using 1-10 passes at a pressure of 500-3000 bar. It is carried out by one of the following.

[0070] The separation process is preferably carried out as follows: i. Preferably, decantation using a disk stack of 5000-20000g for 10 seconds to 5 minutes. ii. Centrifugation in a disk centrifuge such as a continuous disk stack centrifuge, preferably in a disk centrifuge such as a continuous disk stack centrifuge, with a volume of 3000 to 30000 g, preferably 3000 to 20000 g, preferably 3000 to 10000 g, for example 7000 g, preferably for 10 seconds to 10 minutes, preferably 10 seconds to 5 minutes, preferably 30 seconds to 5 minutes, for example 1 minute, under any suitable conditions. Preferably, the temperature is 5 to 85°C, preferably 40 to 65°C. iii. Filtration using a polymer or ceramic membrane having a pore size of 10kDa to 300kDa and a pore size of 0.2μm to 20μm, preferably by cross-counterflow filtration, or iv. Coagulation, preferably by contacting the composition with a coagulant, preferably at a temperature of 2 to 60°C for 1 to 24 hours. Any coagulant known in the art can be used. Preferably, the coagulant is selected from inorganic salts such as CaCl2, or natural coagulants such as chitosan or cellulose. The coagulant is preferably used in an amount of 0.1 to 50 mg / L.

[0071] Preferably, the separation step is carried out by decantation, centrifugation, or filtration as described above. More preferably, the separation step is carried out by decantation as described above.

[0072] In a particularly preferred embodiment, the composition of the present invention, a) A step of providing oilseed yeast cells, wherein the yeast cells are preferably washed and optionally subjected to the following pretreatments. i. Preferably, alkaline treatment by immersing yeast cells in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours. ii. Preferably, an enzymatic treatment that enables the degradation of cell wall biopolymers, applied to washed biomass and using one or more hydrolases (as defined above) suspended in water or a suitable pH buffer at a concentration of 50-200 g / L, with low stirring or low shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, and / or iii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of yeast cells occurs (under the action of endogenous enzymes). A process in which the cell wall is weakened by one or more of the following, b) The cell wall, i. Preferably, alkaline treatment by immersing yeast cells in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours. ii. Contacting yeast cells with an organic solvent, such as ethanol, methanol, or heptane. iii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of yeast cells occurs (under the action of endogenous enzymes). iv. Preferably, the enzymatic treatment is applied to the washed biomass and uses one or more hydrolases (as defined above) suspended in water or a suitable pH buffer at a concentration of 50-200 g / L, with low stirring or low shaking, at an incubation temperature of 30-50°C and an incubation time of 0.5-8 hours, enabling the degradation of cell wall biopolymers. v. High-pressure homogenization using 1 to 10 passes at a pressure of preferably 500 to 3000 bar, more preferably 2 to 10 passes at a pressure of 700 to 3000 bar, or vi. Any combination of any two or more combinations of the means provided in items i. to v. The process of dissolving by, c) Oleosomes, i. Preferably, decantation using a disk stack of 5000-20000g for 10 seconds to 5 minutes. ii. Centrifugal separation iii. Filtration The process of separating it from other yeast cell components and It is obtained by a process that includes this.

[0073] In an optional further step d), the composition obtained at the end of step c) is i. Spray drying or ii. Freeze-drying It is dried by [method].

[0074] In a preferred embodiment of the present invention, the washing step is not performed after the separation step. This embodiment is advantageous in that it avoids the removal of "isolated" yeast cell wall components, such as those described above, that are recovered by the separation step.

[0075] Process-derived products In another aspect, the present invention relates to a yeast oleosome composition obtained or obtainable by the process of the present invention.

[0076] Freeze-dried product In one embodiment, the present invention provides a freeze-dried product of any of the compositions of the present invention. In such a case, the compositions of the present invention described above are dried by any known method such as spray drying, freeze-drying, or lyophilization.

[0077] product In another aspect, the present invention relates to a product comprising the yeast oleosome composition of the present invention or a lyophilized product of the present invention. Such a product is preferably a food, beverage, or cosmetic comprising the yeast oleosome composition of the present invention. Preferably, the product is a food or beverage.

[0078] Such foods or beverages may be of any type, preferably foods containing emulsions, and more preferably foods that can be produced from or contain milk and serve as substitutes for foods and beverages. For example, such foods or beverages include yogurt, cheese, curd, cream, mousse, sauce, soup, mayonnaise, smoothies, juices, desserts, bread, pastries, cakes, infant formula, growing-up milk, cream coffee or milk coffee, tea or chocolate, ice cream, or confectionery products (such as chocolate or candy).

[0079] Ice cream products prepared using the oleosome composition of the present invention are advantageously very stable and exhibit properties that closely mimic the characteristics of dairy-based ice cream, particularly with respect to melting behavior, tribology (i.e., mouthfeel), and rheology. Such advantageous properties can be obtained advantageously using a very simple ice cream formulation (Ice Cream C) containing only the oleosome composition and sugars, or a more diluted formulation containing water and optionally flavorings, additional fats, and / or stabilizers such as gum. The oleosome composition of the present invention provides a highly advantageous vegan alternative to real (with clean label) dairy ice cream.

[0080] In a preferred embodiment, the food or beverage contains at least one astringent component, preferably a polyphenol.

[0081] Suitable cosmetics include creams and lotions.

[0082] use The present invention also relates to the use of the yeast oleosome composition of the present invention for improving the sensory properties of food or beverage products or edible components, for example, improving mouthfeel, improving flavor, improving mouth coating, reducing off-flavors, and / or reducing off-taste. In a preferred embodiment, the sensory properties of the food or beverage product or edible component are improved compared to the same food or beverage product or edible component that does not contain the oleosome composition. In a preferred embodiment, the off-taste to be reduced is selected from astringency and / or bitterness.

[0083] In particular, the compositions of the present invention have been proven useful in reducing the intensity of astringency in foods or beverages, especially foods or beverages containing at least one astringent component. Therefore, the present invention relates to the use of the above-described yeast oleosome composition for reducing the astringency of foods or beverages, preferably foods or beverages containing at least one astringent component. The yeast oleosome composition of the present invention can also be used to reduce the perception of astringency by subjects when consuming foods containing at least one astringent component.

[0084] In a preferred embodiment, such use includes adding the above-mentioned yeast oleosome composition to a food or beverage containing at least one astringent component, or ingesting the yeast oleosome composition before, after, or simultaneously with the consumption of the food or beverage. When the yeast oleosome composition of the present invention is ingested before or after the consumption of a food or beverage containing at least one astringent component, the yeast oleosome composition of the present invention is preferably ingested within 15 minutes (or less), preferably 10 minutes, more preferably 8 minutes, more preferably 6 minutes, more preferably 5 minutes, more preferably 4 minutes, more preferably 3 minutes, even more preferably 2 minutes, and most preferably 1 minute before or after the consumption of the food or beverage containing at least one astringent component. In the most preferred embodiment, the yeast oleosome composition is added to a food or beverage containing at least one astringent component, or the yeast oleosome composition is ingested simultaneously with a food or beverage containing at least one astringent component.

[0085] Astringency is often defined as a sensory attribute described in terms of a dry mouthfeel, a mouthfeel that makes one pucker their lips, or a rough mouthfeel. At least one astringent component is preferably selected from the group consisting of salts of polyvalent metal cations, ethanol, organic acids and / or polyphenols, and mixtures thereof. In a preferred embodiment, a food or beverage containing at least one astringent component includes edible materials containing polyphenols such as tea (especially green tea and black tea), wine (especially red wine), cocoa, coffee, fruits (e.g., elderberries, cherries, blueberries, strawberries, blackberries, blackcurrants, raspberries, olives, plums, and apples), vegetables (e.g., spinach, shallots, artichokes, red chicory, red onions, and green chicory), legumes (e.g., peas, black beans, and white beans), herbs and spices (e.g., cloves, star anise, turmeric, peppermint, oregano, sage, rosemary, thyme, and curry powder), seeds and nuts (e.g., flaxseed, chestnuts, hazelnuts, pecans, almonds, walnuts, and celery seeds), and / or grains (e.g., soybeans or tempeh).

[0086] Astringency is thought to arise from increased friction in the mouth when consuming astringent foods or beverages. Astringent compounds such as polyphenols have been found to bind to proteins in saliva, causing protein precipitation or changes in their three-dimensional structure. This reduces the lubrication provided by saliva, leading to increased friction in the mouth and the characteristic sensation of astringency (Sarkar, A., Andablo-Reyes, E., Bryant, M., Dowson, D., and Neville, A. (2019), Lubrication of soft oral surfaces, Current Opinion in Colloid & Interface Science, 39, 61-75). The yeast oleosome composition of the present invention can surprisingly reduce friction in the mouth when combined with astringent components, as shown in the following examples. [Examples]

[0087] Example 1: Process for preparing a yeast oleosome composition according to the present invention First, Yarowia liporitica cells were separated from the broth by centrifugation. The resulting biomass was washed with water in a 1:2 biomass-to-water ratio, and the cells were recovered by centrifugation.

[0088] Next, to dissolve the cell walls, the cells were subjected to alkaline treatment by immersing them twice in an aqueous solution of biomass with a pH in the range of 8 to 12 for 2 to 36 hours. Then, the biomass was subjected to homogenization, passing through a homogenizer 2 to 10 times at a pressure of 700 to 3000 bar.

[0089] Next, the oleosome composition was separated from the lysed cell composition by decantation using a 7000g disk stack for 1 minute, and the oleosomes and the oleosome composition containing chitin, β-D-glucan, and mannoprotein were recovered.

[0090] Example 2: Process for preparing a yeast oleosome composition according to the present invention Yarowia liporitica cells were cultured in a suitable broth. At the end of fermentation, the bioreactor was heated to 50°C, stirring was reduced, and aeration was stopped. These conditions were maintained until the yeast cells spontaneously lysed under the action of endogenous enzymes. The resulting medium containing the lysed cells was then introduced into a disc stack decanter and decanted at 7000 g for 1 minute to recover oleosomes and the oleosome composition containing chitin, β-D-glucan, and mannoprotein.

[0091] Example 3: Process for preparing the yeast oleosome composition according to the present invention Yarowia liporitica cells were cultured in a suitable broth. At the end of fermentation, the bioreactor was heated to 50°C for 12 hours, stirring was reduced, and aeration was stopped. These conditions were maintained until the yeast cells spontaneously weakened under the action of endogenous enzymes.

[0092] Next, the yeast cell walls were dissolved and extracted by first performing high-pressure homogenization. The cells were then passed through a homogenizer 2 to 5 times at a pressure of 700 to 3000 bar. Subsequently, the cells were subjected to enzymatic treatment with a neutral protease (2% dw) for 2 hours.

[0093] After lysing the cell walls, the oleosome composition was separated from the fragmented walls by centrifugation at 50°C with 5000g for 1 hour.

[0094] Example 4: Process for preparing a yeast oleosome composition according to the present invention Yarowia liporitica cells were cultured in a suitable broth. At the end of fermentation, the cell walls were weakened by enzymatic treatment of the cells with pectinase (4% dw) for 2 hours.

[0095] Next, the yeast cell walls were lysed, and the weakened cells were extracted by heating them to 50°C until the yeast cells spontaneously lysed under the action of endogenous enzymes. The cells were then subjected to further high-pressure homogenization (passed twice at 1500 bar using a high-pressure homogenizer equipped with a nozzle).

[0096] After lysing the cell walls, the oleosome composition was separated from the fragmented walls by centrifugation at 5000g for 1 hour at 60°C.

[0097] Example 5: Yeast oleosome composition of the present invention [Table 1(1)] 1 Average diameter measured by dynamic light scattering using Zetasizer Ultra (Malvern Instruments)

[0098] [Table 1(2)] 2 Based on the total weight of the composition

[0099] The characteristics of the sample were measured as follows.

[0100] fat determination The fat content in the sample was determined by GC after derivatization of the fat into fatty acid methyl esters, and then expressed as methyl oleate equivalents. The lyophilized sample was suspended in methanolic boron trifluoride at 20 g / L and incubated at 85°C for 10 minutes. The mixture was then allowed to return to room temperature, and four times the volume of hexane was added. Next, 1 microliter of the organic phase was injected into a 6860N gas chromatograph (Agilent Technologies AG) equipped with an HP-5MS column (5% phenyl, polymethylsiloxane, 30 m × 250 mm i.d. × 0.25 μm). Fat content was determined based on chromatographic peak area using an external standard solution of methyl oleate at a known concentration.

[0101] Protein quantification The protein content in the sample was quantified using the nitrogen content of the freeze-dried sample. Nitrogen content was determined using a Vario Micro Cube organic elemental analyzer (Elementar Analysensysteme GmbH). A 3 mg sample was placed in a tin dish, then transferred to the analyzer and burned at X°C, converting it to nitrogen, water vapor, and carbon dioxide through a reaction with copper. The resulting gases were separated and quantified using an integrated thermal conductivity detector. The protein content was obtained by multiplying the detected N percentage by 6.25.

[0102] pH measurement A Metrohm 780 pH meter was used for each pH measurement. Measurements were performed after calibration.

[0103] Example 6: Comparison of properties between the composition of the present invention and a plant oleosome composition Plant oleosome compositions were prepared from coconut, oat, almond, cashew, and soybean. Each plant-based oleosome (oil droplet or oil body) was prepared by grinding the seeds after soaking them overnight at room temperature. The milky phase was separated via a cheesecloth filter. This method of extracting oleosomes is a practice in scientific research and industry. The total solids content was measured for all samples and adjusted by dilution so that different milky phases could be compared. All oleosome milk phases had a total solids content of 2%, and as a result, other properties of these oleosome compositions can be compared with samples A1 and A2 of Example 5, which have the same total solids content.

[0104] Table 2 provides the properties of the comparative plant oleosome compositions prepared. Particle size and pH were measured as described in Example 5.

[0105] The coefficient of friction was measured as follows. The lubrication behavior of the samples was evaluated according to the soft tribology protocol described in Rudge, RED, Fuhrmann, PL, Scheermeijer, R., van der Zanden, EM, Dijksman, JA, and Scholten, E. (2021). A tribological approach to astringency perception and astringency prevention. Food Hydrocolloids, 121, 106951. A commercially available glass ball was used with a rheometer (Anton Paar rheometer, MCR302) with a 3-pin PDMS configuration. The load was 1N, and friction was measured at sliding speeds of 0.001 to 1 m / s. All measurements were performed using 1 ml of sample.

[0106] The results are shown in [Figure 2]. Samples A1 and A2 of the present invention tend to have low coefficients of friction across the entire measurement range, i.e., at low and high sliding speeds. In contrast, cashews, which have a coefficient of friction close to that of the present invention samples at low speeds, have a high coefficient of friction at high speeds. Coconuts, which have a coefficient of friction between A1 and A2 at a speed of 0.01 m / s, have a much higher coefficient of friction at lower and higher speeds. Finally, oats, which have a coefficient of friction similar to A2 at 0.1 m / s, have a much higher coefficient of friction at speeds of 0.0001 and 0.001 m / s.

[0107] The coefficient of friction at a sliding speed of 0.1 m / s is particularly relevant to the evaluation of mouthfeel, so to clarify such values, they are shown in [Table 2] below.

[0108] These results demonstrate that the compositions according to the present invention have an advantageously low coefficient of friction, and a low coefficient of friction implies a better mouthfeel than compositions with a higher coefficient of friction. In particular, sample A1 has a lower coefficient of friction than all other samples across the entire measurement range and therefore has the best mouthfeel among all the samples tested.

[0109] [Table 2]

[0110] The stability of all samples was also tested. Samples were placed in 15 mL containers. To track the physical stability of the emulsions as a function of time, the samples were stored at room temperature for 4 days. Photographs of the containers were taken to characterize the physical stability of oleosomes based on time [Figure 1]. All plant-based samples separated, but samples A1 and A2 of the present invention remained stable.

[0111] Example 7: Comparison of properties of the present invention composition suitable as a cream substitute with a comparative plant oleosome composition. Plant-based oleosome creams were prepared from cashews, almonds, hemp, and soybeans by grinding the seeds after soaking them overnight at room temperature. The emulsion phase was separated via a cheesecloth filter. This method of extracting oleosomes is a practice in scientific research and industry. The total solids content was measured for all samples and adjusted by dilution so that different emulsion phases could be compared. All oleosome cream phases had a total solids content of 20% by weight and were compared to the oleosome composition according to the present invention (designated as "Culture" in [Figure 3]) having a total solids content of 20%.

[0112] The viscosity of the "cultured" sample and the comparative plant-based cream was measured as follows. The sample was measured using an Anton Paar rheometer MCR302 with a plate configuration. Viscosity values ​​were obtained under shear rates ranging from 0.1 to 100 1 / s.

[0113] The results are shown in [Figure 3]. Advantageously, the composition of the present invention is characterized by the highest viscosity, which implies a better mouthfeel than the comparative composition.

[0114] Example 8: Comparison of viscosity between the present invention's composition, suitable as a cream substitute, and a commercially available plant-based cream. The plant-based creams were obtained from the Swiss supermarket Migros: V-Love® Plant-based cuisine (designated as "V love Migros" in Figure 4), Alnatura® Soja Cuisine (designated as "Alnatura Soy" in Figure 4), and Alnatura® Hafer Cuisine (designated as "Alnatura oat" in Figure 4). The composition of Alnatura Soja Cuisine was water, 15% sunflower oil, 8% soybean glucose syrup, xanthan gum, salt, and vanilla extract. The composition of Alnatura Hafer Cuisine was water, 9% whole grain oats, 7% sunflower oil, carob gum, guar gum, and salt. The composition of V-Love® Plant-based cuisine was 68.5% soy composition (consisting of 95.6% water and 4.4% soy flour), 26.5% hydrogenated rapeseed oil, glucose, maltodextrin, lactic acid esters of fatty acid monoglycerides and diglycerides, methylcellulose, guar gum, carrageenan, sodium phosphate, and salts.

[0115] The viscosity of the above-mentioned commercially available sample, the oleosome composition according to the present invention having a total solids content of 20% (designated as "cultured cream" in [Figure 4]), and the milk cream having a total solids content of 20% was measured as described in Example 7. The results are shown in [Figure 4]. The sample of the present invention (cultured cream) is advantageous in that it has a viscosity of 1 s -1The composition exhibits a higher viscosity than both Alnatura samples, a higher viscosity than dairy cream, and a viscosity similar to that of the V-love sample at shear rates up to a certain point. This implies a better mouthfeel of the composition of the present invention compared to the Alnatura samples, and a mouthfeel similar to that of the V-love sample compared to the comparative compositions. This is particularly surprising, as all the commercially available compositions tested contained texture-improving ingredients such as thickeners and / or emulsifiers intended to improve viscosity and mouthfeel. The composition of the present invention contains no texture-improving ingredients such as thickeners and emulsifiers, yet the cream substitute composition of the present invention still exhibits a higher viscosity than most of the comparative samples and a viscosity similar to that of the most viscous comparative sample. This demonstrates that the composition of the present invention does not require texture-improving agents and, therefore advantageously, is a more authentic non-dairy cream characterized by a good mouthfeel.

[0116] Example 9: Comparison of the coefficient of friction between the composition of the present invention, suitable as a cream substitute, and a commercially available plant-based cream. The plant-based creams were obtained from the Swiss supermarket Migros: V-Love® Plant-based cuisine (designated as "Vlove cream" in Figure 5), Alnatura® Soja Cuisine (designated as "Soja cuisine" in Figure 5), and Alnatura® Hafer Cuisine (designated as "Oat cuisine" in Figure 5). The composition of Alnatura Soja Cuisine was water, 15% sunflower oil, 8% soybean glucose syrup, xanthan gum, salt, and vanilla extract. The composition of Alnatura Hafer Cuisine was water, 9% whole grain oats, 7% sunflower oil, carob gum, guar gum, and salt. The composition of V-Love® Plant-based cuisine was 68.5% soy composition (consisting of 95.6% water and 4.4% soy flour), 26.5% hydrogenated rapeseed oil, glucose, maltodextrin, lactic acid esters of fatty acid monoglycerides and diglycerides, methylcellulose, guar gum, carrageenan, sodium phosphate, and salts.

[0117] The coefficient of friction as a function of sliding velocity of the above-mentioned commercially available composition and the oleosome composition according to the present invention having a total solids content of 20% (designated as "Culture Cream 20%") was measured as described in Example 6. The results are shown in Figure 5.

[0118] In the region corresponding to the boundary regime, the compositions of the present invention exhibit the lowest coefficient of friction as a function of sliding speed at a sliding speed of 0.1 m / s, which best mimics the conditions that occur in the oral cavity, particularly when ingesting foods such as cream. Only at higher sliding speeds does the coefficient of friction of the compositions of the present invention increase significantly. However, such high speeds are largely irrelevant to the behavior of the compositions in the oral cavity. This demonstrates that the cream compositions of the present invention feature a better mouthfeel than commercially available creams, despite the presence of texture enhancers in commercially available products. This is even more true because the cream compositions of the present invention also exhibit high viscosity, as demonstrated in Examples 7 and 8.

[0119] Example 10: Reduction of bitterness in pea protein A dynamic tribological approach was used to measure the change in the coefficient of friction of saliva upon addition of the yeast oleosome composition according to the present invention. All tribological measurements were performed using an Anton Paar rheometer MCR302 (Republic of Austria). The lubrication properties of the sample combined with saliva were measured using a tribology cell (BC12.7 / SS52837). Polydimethylsiloxane (PDMS) was used as the dominant material currently used in soft tribology, and therefore PDMS pins were used. Friction was measured using a commercially available (glass) ball with a three-PDMS pin setup. The glass ball had a diameter of 12.7 mm, the PDMS pins had a diameter of 6 mm and a height of 6 mm, and their modulus of elasticity was approximately 2 MPa. The glass ball and PDMS pins were obtained from the rheometer manufacturer.

[0120] Measurements were performed three times per condition. A normal force Fn of 1 N was applied. The experiment was conducted at a constant rotation speed of 1 mm / s to obtain the friction profile of the boundary regime. This is because this regime is thought to be closely related to the perception of astringency in humans. Measurements were performed within 10 minutes, with the first 5 minutes used to allow salivary proteins to cover the PDMS pin (extravitreal salivary pellicle). The salivary layer allowed the glass probe to slide against the PDMS pin while being lubricated by 0.5 mL of saliva. After 5 minutes, when a constant coefficient of friction was obtained, model solutions (MS) were added in a 1:1 ratio (saliva:MS).

[0121] An example of measurement (unrelated to any of the MSs described here) is shown in [Figure 7]. Using such a graph (data not shown), the difference in friction coefficients (Δμ) is calculated as Av.CoF1-Av.CoF2, where Av.CoF1 is the friction coefficient obtained when salivary protein completely covers the PDMS surface, and Av.CoF2 is the friction coefficient after MS addition. All friction coefficients represent the mean value obtained by taking the average value based on five points.

[0122] Three different aqueous model solutions were prepared for tribological measurements, as summarized in [Table 3] below, where "parts" are defined by weight based on the total weight of the model solution. Water was used as a control.

[0123] [Table 3]

[0124] The selected astringent was pea protein isolate (PPI; KoRo Handels GmbH, Koppenplatz 9, 10115 Berlin), which typically provides a dry oral sensation. A 5% pea protein solution, consisting of 5% by weight of PPI and 95% by weight of water, was prepared as follows: To completely solubilize the PPI in water, the solution was stirred at a temperature of 43°C for 30 minutes. After this, the solution was cooled to 4°C.

[0125] The yeast oleosome composition (referred to as the "culture extract") was as disclosed in [Table 4].

[0126] [Table 4(1)] 1 Average diameter measured by dynamic light scattering using Zetasizer Ultra (Malvern Instruments)

[0127] [Table 4(2)] 2 Based on the total weight of the composition

[0128] For the preparation of MS2, the above 5% pea protein solution was combined with the culture extract in a 1:1 ratio, and a new solution was effectively generated using equal parts of the yeast oleosome composition and the pea protein solution.

[0129] The results are shown in [Figure 6]. A decrease in salivary lubrication was observed in MS2, which contained only pea protein. In contrast, in MS3, the yeast oleosome composition was able to completely mask the increased friction induced by pea protein, effectively creating a smoother texture. The friction observed in MS3 was even lower than that of the control without pea protein.

[0130] Example 11: Sensory evaluation of astringency reduction The purpose of this evaluation was to assess the sensory characteristics of different dairy analog formulations and to determine their astringency-masking properties. The samples tested are listed in [Table 5].

[0131] [Table 5] 1 KoRo Handels GmbH, Koppenplatz 9, 10115 Berlin.

[0132] The samples were prepared by dissolving pea protein isolate in either water or a culture extract. To completely solubilize the PPI in water, the solution was stirred at 43°C for 30 minutes. After that, the solution was cooled to 4°C.

[0133] A panel of 10-15 panelists was assembled. The panelists had no known allergies or intolerances to the dairy analog formulations being tested. The panelists did not suffer from a cold or other sensory impairment during the evaluation. The panelists did not eat or smoke within one hour of tasting.

[0134] All samples were maintained at approximately 10°C and placed in identical containers (20g) labeled with randomized 3-digit codes to ensure anonymity. Samples were pre-weighed (approximately 10ml). Panelists were given a standardized evaluation form to score the samples based on specific sensory attributes. The sensory attributes to be evaluated included astringency. Each attribute was scored on a scale of 1 to 5, with 1 being the lowest score and 5 being the highest score. The results are shown in [Figure 8]. The sample containing the yeast oleosome composition of the present invention (CB A14) was found to have significantly lower astringency than the sample without the yeast oleosome composition of the present invention (PPI solution).

[0135] Example 12: Ice cream composition An ice cream composition containing the ingredients shown in [Table 6] below was prepared.

[0136] The ice cream was prepared by mixing all the ingredients in a Termomix, maintaining a temperature of 80°C for 10 minutes, and then homogenizing the resulting solution in a Termomix at maximum speed for 2 minutes. The solution was stored in a refrigerator overnight, and the ice cream was prepared the following morning.

[0137] [Table 6] 1) The fat content is 13% by weight, the fiber content is 3.5% by weight, and the protein content is 4.5% by weight.

[0138] Example 13: Comparison of product properties related to extraction method A comparative study was conducted to investigate the effect of specific steps in the extraction method on product characteristics. Two samples of oleosome composition were prepared from the same biomass. One sample was prepared using the process of Example 4 (the present invention), and the second sample was prepared using a process similar to that used in Han et al., Tunable nano-oleosomes derived from engineered Yarrowia lipolytica, Biotechnology and Bioengineering, 110(3), 702-710, i.e. a) Wash the biomass with PBS buffer. b) Cell lysis using glass beads and a vortex mixer (bead milling), c) and centrifugation It was manufactured using a process that includes [a specific component / method].

[0139] The characteristics of both samples were analyzed as follows.

[0140] The total solid content of the sample was determined by gravimetric analysis, specifically the difference between the fresh weight and the weight after complete freeze-drying.

[0141] Fat quantification of the samples was performed by a modified Folch gravimetric assay. Briefly, hydrophobic compounds were extracted from 300 mg aliquots of lyophilized material with a mixture of methanol and tert-butyl ether, followed by the addition of water. Fat remained in the organic fraction, which was collected, dried, and weighed. The analysis was performed twice for each condition.

[0142] Protein content was measured by total combustion using a CHNS elemental analyzer (Vario EL Cube, Elementar Analysensysteme GmbH). Briefly, a 3 mg aliquot of the freeze-dried material was loaded into a tin tray and incinerated in the analyzer's furnace. The combustion gas was eluted on its chromatographic column, and nitrogen gas was quantified using an accompanying thermal conductivity detector. The protein content was then obtained by multiplying the nitrogen content by a conventional conversion factor of 6.25. The analysis was performed twice for each condition.

[0143] Protein content was measured by total combustion using a CHNS elemental analyzer (Vario EL Cube, Elementar Analysensysteme GmbH). Briefly, a 3 mg aliquot of the freeze-dried material was loaded into a tin tray and incinerated in the analyzer's furnace. The combustion gas was eluted on its chromatographic column, and nitrogen gas was quantified using an accompanying thermal conductivity detector. The protein content was then obtained by multiplying the nitrogen content by a conventional conversion factor of 6.25. The analysis was performed twice for each condition.

[0144] The individual droplet sizes of yeast-extracted oleosomes were measured by dynamic light scattering (Malvern Instruments Zetasizer, UK). Droplet size was expressed as volume-average diameter and number-average diameter. Samples were measured using a refractive index of 1.47. After determining the particle size, the zeta potential was measured in a foldable capillary zeta cell (DTS1070, Marvern, UK) by applying a maximum of 40 mV.

[0145] The results are shown in [Table 7] below.

[0146] [Table 7(1)]

[0147] [Table 7(2)]

[0148] The method described by Han et al. yielded a very liquid-like composition with a very low solids content, essentially fat-free and essentially β-glucan-free, and exhibited a very small particle size, suggesting that the oleosome structure was not preserved. In contrast, the composition obtained according to the present invention has a much higher fat and solids content (resulting in a favorable, thicker, and more viscous texture), a larger particle size, and a non-negligible amount of free β-glucan released from the cell wall structure. These differences are due to the fact that bead milling, in contrast to the high-pressure homogenization used in the present invention, cannot break down the oleosome structure, cannot dissolve the cell wall, and cannot extract the yeast cell wall component β-glucan from the yeast cell wall structure.

[0149] The process characteristics were as shown in [Table 8] below.

[0150] [Table 8]

[0151] These results demonstrate that the process of the present invention (including high-pressure homogenization instead of bead milling) achieves a much better yield in terms of total solids and fat content, more efficiently breaks down the cell wall, and thus releases β-glucan from the yeast cell wall structure.

[0152] The particle sizes of both samples are further shown in [Figure 9] and [Figure 10]. The average diameter and zeta potential of the droplet size based on the number distribution are clearly different between the product obtained by Han et al.'s process and the product obtained by the process of the present invention.

[0153] Microscopic images of both samples are shown in [Figure 11]. The image on the right, corresponding to the sample of the present invention, shows more intact oleosome structure compared to the image on the left, corresponding to Han et al. This confirms the conclusion drawn above from the analytical results.

[0154] [Figure 12] shows both samples after centrifugation. For each sample, the solid phase is identified by a dashed black triangle, and the light phase (cream phase) containing oleosomes is identified by a solid black triangle. The composition obtained by Han et al. has a much larger solid phase and a smaller cream phase, but the process of the present invention provides a larger cream phase and a smaller solid phase. This photograph demonstrates the better extraction of oleosomes and lipids achieved by the process of the present invention.

[0155] In conclusion, the products obtained by Han et al.'s process and the products of the present invention have significantly different chemical compositions. Samples extracted by the process of the present invention have different amounts of solids, fats, proteins, and glucans. Han et al.'s method results in a small amount of oleosomes that cannot be recovered by the separation step, and polysaccharides from the cell wall are shown to be unbranched, resulting in a very small amount of β-glucan (isolated β-glucan) extracted from the yeast cell wall. In contrast, the composition obtained by the process of the present invention results in a large amount of intact oleosomes along with isolated soluble fibers such as β-glucan, and a high fat content of 60-80% by weight based on the total dry weight of the composition.

Claims

1. A yeast oleosome composition comprising yeast oleosomes and at least one isolated yeast cell wall component selected from chitin, β-D-glucan, and mannoprotein.

2. The yeast oleosome composition according to claim 1, having a particle size characterized by a volume-average diameter of 0.2 to 7 μm as measured by dynamic light scattering.

3. A yeast oleosome composition according to claim 1 or claim 2, having a density of 0.8 to 1.1 g / ml.

4. The yeast oleosome composition according to any one of claims 1 to 3, wherein the zeta potential at the surface of the oleosomes in the composition is -40 to -20 mV, and / or the pH of the yeast oleosome composition is 5 to 8.

5. The yeast is preferably an oilseed yeast selected from the genera Yarowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, Cthaneotrichosporon, Lipomyces, Papiliotrema, Hannaela, Sporidioborus, Kodamaea, Pichia, Cytojima, Sivarindonella, Meyerojima, Piscurojima and mixtures thereof, more preferably Yarowia liporitica, Candida 107, Candida tropicalis, Candida uchiris, Rhodotorula glutinis, Rhodotorula musiladinasa, Rhodotorula bavjebae, Rhodotorula sphaerocarpa, Rhodotorula graminis, Rhodosporidium toroides, Rhodosporidium fulviale, Cryptococcus curbatus, A yeast oleosome composition according to any one of claims 1 to 4, wherein the selected species is an oilseed yeast, most preferably Jarrowia lipotica, selected from the species of Lycosporon pullulan, Trichosporon asahi, Cthaneotrichosporon oleaginosus, Cthaneotrichosporon curvatum, and Lipomyces lipofa, Lipomyces starkey, Lipomyces tretrasporus, Lipomyces mesembryus, Swaniomyces occidentalis, Papiliotrema terrestris, Papiliotrema flavecens, Papiliotrema laurentii, Sporidioborus ruinenie, Kodamaea omerii, Pichia manshurica, Cytojima podozorica, Sivarindonella saturunus, Meyerosima gilliermondii, and mixtures thereof.

6. The yeast oleosome composition according to any one of claims 1 to 5, wherein the composition is in the form of a milk substitute composition having a particle size characterized by a volume average diameter of 0.2 to 2 μm, a density of 0.8 to 1.1 g / mL, a total solids content of 2 to 10% by weight based on the total weight of the composition, a fat content of 1.25 to 4% by weight, a carbohydrate content of up to 3%, and a protein content of 0.1 to 1.5% by weight.

7. The yeast oleosome composition according to any one of claims 1 to 5, wherein the composition is in the form of a cream substitute composition having a particle size characterized by a volume average diameter of 0.4 to 7 μm, a density of 0.85 to 1.2 g / mL, a total solids content of 30 to 45% by weight, a fat content of 17 to 38% by weight, a carbohydrate content of 0.5 to 15% by weight, and a protein content of 2.5 to 7% by weight.

8. A yeast oleosome composition according to any one of claims 1 to 7, characterized by a coefficient of friction in the boundary regime of 0.01 to 0.5, preferably a coefficient of friction at a sliding speed of 0.1 m / s in the range of 0.01 to 0.

5.

9. A method for preparing a yeast oleosome composition, a) A step of providing washed yeast cells, b) A step of dissolving the cell wall, c) A step to separate oleosomes from other yeast cell components. Including step b), i. Preferably, the yeast cells are subjected to alkaline treatment by immersing them in an aqueous medium with a pH of 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. self-decomposition, iv. Preferably, the enzyme treatment is applied to the washed biomass and uses one or more hydrolases (e.g., protease, glucanase, mannanase, chitinase, nuclease, β-glucosidase, cellulase, xylanase, pectinase, and combinations thereof) suspended in water or a suitable pH buffer at a concentration of 50 to 200 g / L, with low stirring or low shaking, at an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours, enabling the degradation of cell wall biopolymers. v. Preferably, high-pressure homogenization using 1 to 10 passes at a pressure of 500 to 3000 bar, more preferably 2 to 10 passes at a pressure of 700 to 3000 bar. and these mixed forms A method carried out by means selected from among.

10. Step b) is, i. Preferably, the yeast cells are subjected to alkaline treatment by immersing them in an aqueous medium with a pH of 8 to 12 for 1 to 36 hours, preferably 2 to 36 hours, followed by high-pressure homogenization using 2 to 10 passes at a pressure of preferably 700 to 3000 bar. ii. Autolysis by heating the cells at the end of fermentation to a temperature of 50°C or higher, reducing stirring, and stopping aeration until spontaneous lysis of the yeast cells occurs (under the action of endogenous enzymes), and iii. Preferably, the enzyme treatment is applied to the washed biomass and uses one or more hydrolases (e.g., protease, glucanase, mannanase, chitinase, nuclease, β-glucosidase, cellulase, xylanase, pectinase, and combinations thereof) suspended in water or a suitable pH buffer at a concentration of 50 to 200 g / L, with low stirring or low shaking, at an incubation temperature of 30 to 50°C and an incubation time of 0.5 to 8 hours, enabling the degradation of cell wall biopolymers, followed by high-pressure homogenization using 1 to 10 passes at a pressure of preferably 500 to 3000 bar. The method according to claim 9, carried out by means selected from.

11. The method according to claim 9 or 10, wherein step c) is carried out by decantation, centrifugation, filtration, or flocculation.

12. A yeast oleosome composition that can be obtained or obtained by the method described in any one of claims 9 to 11.

13. A yeast oleosome composition in solid form obtained or obtainable by drying the yeast oleosome composition according to any one of claims 1 to 8 and claim 12.

14. A product comprising the yeast oleosome composition according to any one of claims 1 to 8 and claim 12, or the solid-state yeast oleosome composition according to claim 13.

15. The product according to claim 14, which is a food, beverage, or cosmetic.

16. Use of a yeast oleosome composition according to any one of claims 1 to 8 and 12 or a yeast oleosome composition in solid form according to claim 13 for improving the sensory properties of a food or beverage or food component, preferably improving mouthfeel, improving flavor, improving mouth coating, reducing unpleasant flavors, and / or reducing off-flavors.

17. The use according to claim 16 for reducing the intensity of astringency in a food or beverage containing at least one astringency component.

18. The use according to claim 17 for reducing the perception of astringency by an object when consuming a food or beverage containing at least one astringent component.

19. The food or beverage according to claim 16, wherein the food or beverage comprises at least one astringent component.

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