High protein yeast product

Abstract

The present disclosure relates to a method of obtaining a composition containing a yeast protein derived from yeast protoplast. The disclosure also includes compositions comprising yeast proteins derived from yeast protoplasts, edible products derived from the compositions, and uses thereof.

Description

[0001] Cross - reference to related applications and literature

[0002] This patent application claims the priority of U.S. Provisional Patent Application 63 / 381,353, filed on October 28, 2023, and incorporates its entire content herein. Technical Field

[0003] The present disclosure relates to yeast proteins having a neutral taste, compositions thereof, and methods for their production. Background Art

[0004] Proteins are biological macromolecules that are essential components of animal nutrition. Once ingested, proteins are degraded by proteases into polypeptides and then into amino acids. The latter will then be able to reach the bloodstream by crossing the intestinal barrier and then diffuse throughout the body to perform their important functions.

[0005] In human and / or animal nutrition, the main protein sources are animal proteins, plant proteins (including legume proteins), and fungal proteins. Over the past 50 years, the consumption of animal proteins has been steadily increasing, thus causing environmental problems as well as health problems for consumers. Therefore, the current nutritional challenge is to reverse the trend in protein consumption worldwide, namely: reducing the share of animal proteins and increasing the share of plant / legume proteins (including, for example, pea proteins) and fungal proteins (such as those found in yeast extracts).

[0006] However, proteins from yeast extracts as well as plant / legume proteins have a unique taste, which limits their concentration and thus their applications. The present disclosure attempts to provide protein sources having a neutral taste to address these problems. Summary of the Invention

[0007] According to a first aspect, the present disclosure provides a method for obtaining a composition containing yeast protein. Generally speaking, the method includes: a) providing a yeast extract containing yeast; b) inactivating the endogenous enzymes of the yeast to provide an inactivated yeast extract; c) subjecting the inactivated yeast extract to an enzymatic treatment to obtain an insoluble fraction and a soluble fraction containing yeast protoplasts; d) separating the insoluble fraction from the soluble fraction; and e) collecting the insoluble fraction, wherein the collected insoluble fraction is a composition containing yeast protein when dried and has a protein content equal to or higher than 60% based on the total mass of the dried collected insoluble fraction. At step c), the enzymatic treatment includes at least one polypeptide having glucanase activity; and the enzymatic treatment lacks ribonuclease activity. In one embodiment, the inactivated yeast extract is obtained by exposing the yeast extract to a temperature of 65°C to 100°C for a time of 30 seconds to 5 hours. In one embodiment, the enzymatic treatment is carried out at a temperature of 20°C to 80°C for a time of 1 hour to 24 hours. In another embodiment, the composition has a neutral taste. In some embodiments, the method further includes drying the collected insoluble fraction of step e) to provide the composition. In some additional embodiments, the dried insoluble fraction has a lipid content of less than 20% based on the total mass of the dried collected insoluble fraction; a nucleic acid content of higher than 6% based on the total mass of the dried collected insoluble fraction; a carbohydrate content of less than 25% based on the total mass of the dried collected insoluble fraction; a mannan content of less than 6% based on the total mass of the dried collected insoluble fraction; a glucan content of less than 10% based on the total mass of the dried collected insoluble fraction; and / or a glucose content of less than 25% based on the total mass of the dried collected insoluble fraction. In some embodiments, the method may further include, after b) and before c), performing an alkali extraction step on the inactivated yeast extract. In some additional embodiments, the method may further include drying the collected insoluble fraction of step e) (which has undergone the alkali extraction step) to provide the composition. In such embodiments, the dried insoluble fraction may have a lipid content of less than 20% based on the total mass of the dried collected insoluble fraction; a nucleic acid content of less than 3% based on the total mass of the dried collected insoluble fraction; a carbohydrate content of less than 25% based on the total mass of the dried collected insoluble fraction; a mannan content of less than 6% based on the total mass of the dried collected insoluble fraction; a glucan content of less than 10% based on the total mass of the dried collected insoluble fraction; and / or a glucose content of less than 25% based on the total mass of the dried collected insoluble fraction.In another embodiment, the yeast is from the genus Saccharomyces, Komagataella, Pichia, Candida, Kluyveromyces, Yarrowia, Cyberlindera (Torula), Wickerhamomyces, or a combination thereof.

[0008] According to a second aspect, the present disclosure provides a composition that comprises a yeast protein derived from yeast protoplasts and has a protein content equal to or higher than 60% based on the total mass of the composition; a lipid content lower than 20% based on the total mass of the composition; a carbohydrate content lower than 25% based on the total mass of the composition; a mannan content lower than 6% based on the total mass of the composition; a glucan content lower than 10% based on the total mass of the composition; and / or a glucose content lower than 25% based on the total mass of the composition. In some embodiments, the composition may have a nucleic acid content higher than 6% based on the total mass of the composition. In other embodiments, the composition may have a nucleic acid content lower than 3% based on the total mass of the composition. In one embodiment, the composition has a neutral taste. In one embodiment, the composition is obtainable or obtained by a method for obtaining a composition containing a yeast protein.

[0009] According to a third aspect, the present disclosure provides an edible product that comprises the composition and at least one additional ingredient, wherein the composition accounts for at least 1% w / w based on the total mass of the edible product. In one embodiment, the edible product is a beverage, a milkshake, a bar, a meat product, or a baked product.

[0010] According to a fourth aspect, the present invention provides the use of the edible product in human and / or animal nutrition. In one embodiment, the edible product is provided in the form of a substitute for an edible product based on animal or plant / legume protein, or in combination with an edible product based on animal or plant / legume protein. In one embodiment, the edible product is a food / feed supplement or a food / feed additive. In another embodiment, the food / feed supplement or the food / feed additive is for weight control, the elderly, oral / enteral clinical nutrition, sports applications, and / or animal nutrition. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Thus, the nature of the present invention has been generally described, and now, with reference to the drawings, preferred embodiments of the present invention are illustrated by way of example, and in which:

[0012] Figure 1 Shows one embodiment of a method for preparing a composition (Composition C) containing yeast protein and comparative products (Compositions A and B).

[0013] Figure 2A1 、 Figure 2A2 、 Figure 2B1 、 Figure 2B2 、 Figure 2C1 and Figure 2C2 Shows the sensory characteristics (i.e., odor and taste) of the all-applicable items scoring (RATA) method based on Compositions A ( Figure 2A1 and Figure 2A2 ), Composition B ( Figure 2B1 and Figure 2B2 ) and for F85M ( Figure 2C1 and Figure 2C2 ). * Refers to the mandatory fields required for the sensory evaluation of the composition.

[0014] Figure 3 A to Figure 3 E show the texture profile analysis (TPA) of the extrudates (i.e., H0, H5, H10, H15, H25, and H35). The results of (A) chewiness, (B) cohesiveness, (C) hardness, (D) resilience, and (E) springiness are shown. Vertical error bars show the 95% confidence interval, and samples without shared letters are statistically significantly different (Kruskal-Wallis test, p < 0.05). H100 was too strong and not measured.

[0015] Figure 4 A to Figure 4 I show the descriptive sensory analysis (DSA) of the extrudates (i.e., H0, H5, H10, H15, H25, H35, and H100) evaluating the odor (O) and taste (T) attributes. The results of (A) bean odor, (B) meaty and yeasty flavor, (C) off-flavor, and (D) overall intensity, as well as (E) aftertaste intensity, (F) bean taste, (G) off-flavor intensity, (H) overall intensity, and (I) umami are shown. Vertical error bars show the 95% confidence interval, and samples without shared letters are statistically significantly different (Kruskal-Wallis test, p < 0.05).

[0016] Figure 5 A to Figure 5H shows the descriptive sensory analysis (DSA) of extrudates (i.e., H0, H5, H10, H15, H25, H35, and H100) evaluating the texture attribute (X). The results are shown as the attributes of (A) adhesiveness, (B) chewiness, (C) cohesiveness, (D) fibrousness, (E) particle size, (F) hardness, (G) moisture, and (H) springiness. Vertical error bars show the 95% confidence intervals, and samples without shared letters are statistically significantly different (Kruskal-Wallis test, p < 0.05). Detailed Description

[0017] Method for obtaining a composition containing yeast proteins

[0018] In one embodiment, the present disclosure aims to provide a method for obtaining a composition containing yeast protein. The method of the present disclosure includes: a) providing a yeast paste containing yeast; b) inactivating the endogenous enzymes of the yeast to provide an inactivated yeast paste; c) subjecting the inactivated yeast paste to enzymatic treatment to obtain an insoluble fraction and a soluble fraction containing yeast protoplasts; d) separating the insoluble fraction from the soluble fraction; and e) collecting the insoluble fraction.

[0019] In the context of the present disclosure, the term "yeast" refers to eukaryotic single-celled microorganisms belonging to the kingdom Fungi. Suitable yeasts for use in the methods for obtaining the compositions can be, for example, from the genus Saccharomyces, Komagataella, Pichia, Candida, Kluyveromyces, Yarrowia, Cyberlindnera (Torulaspora), Wickerhamomyces or combinations thereof. Suitable yeast species for use in the methods for obtaining the compositions can include, but are not limited to, for example, Saccharomyces cerevisiae, Cyberlindnera jadinii, Komagataella phaffii (Pichia pastoris), Yarrowia lipolytica, Candida glabrata, Kluyveromyces lactis, Kluyveromyces marxianus, Wickerhamomyces anomalus, Debaryomyces hansenii or combinations thereof. In some embodiments, the yeast species is selected from the group consisting of Saccharomyces cerevisiae, Cyberlindnera jadinii or combinations thereof. In one embodiment, the yeast is from the genus Saccharomyces, and in some embodiments, from the species Saccharomyces cerevisiae. In one embodiment, the yeast is from the genus Cyberlindnera, and in some embodiments, from the species Cyberlindnera jadinii. Figure 1 One embodiment of a method for obtaining a composition comprising yeast protein is provided. In Figure 1 Method 100, a yeast paste is provided. In one embodiment, the method may optionally include proliferating the yeast ( Figure 1 , not shown in the figure). Yeast proliferation is carried out by a person of ordinary skill in the art according to suitable methods known in the art. As used in the context of the present disclosure, the expression "yeast proliferation" refers to the amplification stage of a commercial process, in which the yeast proliferates under aerobic conditions to maximize the conversion of the substrate into biomass. The proliferation step can be a continuous process, a batch process or a fed-batch process. The proliferation medium can contain a carbon source (such as molasses, sucrose, glucose, dextran syrup, ethanol, corn, glycerol, corn steep liquor and / or lignocellulosic biomass), a nitrogen source (such as ammonia or another inorganic nitrogen source) and a phosphorus source (such as phosphoric acid or another inorganic phosphorus source). The proliferation medium can also contain additional micronutrients, such as vitamins and / or minerals, to support the proliferation of yeast cells.

[0020] The proliferation process can be carried out under hyperventilation conditions. For example, in some embodiments, the proliferation step can include controlling the ventilation of the container to achieve a specific ventilation rate of, for example, at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 air volumes / container volume / minute.

[0021] The proliferation step can be carried out at a specific pH and / or a specific temperature that is optimal for yeast biomass production. Thus, in embodiments where the yeast is from the genus Saccharomyces or the genus Cyberlindnera, the method can include controlling the pH of the culture medium to be between about 3.0 and about 8.0, about 3.5 and about 7.0, or about 4.0 and about 6.5. In a specific embodiment, the pH is controlled to about 4.5. In another example, in embodiments where the yeast is from the genus Saccharomyces or the genus Cyberlindnera, the method can include controlling the temperature of the culture medium to be between about 20°C and about 40°C, about 25°C and about 30°C, or about 30°C and about 35°C. In a specific embodiment, the temperature is controlled to be between about 30°C and about 35°C (e.g., 32°C).

[0022] At the end of the proliferation step, a specific concentration can be sought or achieved. In some embodiments, the concentration of the proliferated yeast cells in the culture medium is at least about 0.1 wt%, 0.15 wt%, 0.2 wt%, 0.25 wt%, 0.3 wt%, 0.35 wt%, 0.4 wt%, 0.45 wt%, 0.5 wt%, 1.0 wt%, 1.5 wt%, 2.0 wt%, 2.5 wt%, 3.0 wt%, 3.5 wt%, 4.0 wt%, 4.5 wt%, 5.0 wt% or more wt% relative to the volume of the culture medium. In a specific embodiment of proliferating yeast cells using a fed-batch method, the concentration of the proliferated yeast cells in the culture medium is at least about 0.25 wt% relative to the volume of the culture medium.

[0023] In some embodiments, a formulation step is carried out directly on the proliferated yeast to provide yeast extract. For example, prior to the formulation step, no subsequent fermentation step (which can be carried out under anaerobic conditions) is performed on the proliferated yeast.

[0024] In the formulation step 010, the mixture obtained after proliferation (containing proliferated yeast cells) is modified to provide yeast extract. For example, at least one component of the mixture obtained after proliferation is removed from the culture medium to provide a yeast composition (a yeast extract, an embodiment of the yeast composition, as Figure 1Embodiments thereof are provided). The at least one component can be, but is not limited to, water, amino acids, peptides and proteins, nucleic acid residues and nucleic acid molecules, cell debris, fermentation products, etc. In one embodiment, formulation step 010 includes substantially separating the proliferated yeast cells (e.g., biomass) from the components of the culture medium. As used in the context of the present disclosure, the expression "substantially separated" means removing most of the components of the culture medium from the proliferated yeast cells. In some embodiments, "substantially separated" means concentrating the proliferated yeast cells to at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300% or higher compared to the concentration of yeast cells in the culture medium before separation. To provide a yeast composition, the proliferated yeast cells can be centrifuged (and optionally the resulting cell pellet containing the proliferated yeast cells can be washed) and / or filtered. The separated yeast cells can then be formulated into a yeast composition (which can be a yeast paste as provided in Figure 1 the yeast paste provided). In some embodiments, formulation step 010 can maintain the viability (at least in part) of the yeast cells. Thus, the proliferated yeast can be provided in an active or semi-active form. The proliferated yeast can be provided in a liquid or semi-solid form. In one embodiment, the proliferated yeast can be provided in the form of a yeast paste as shown in Figure 1 .

[0025] In the method of the present disclosure, the endogenous enzymes of the yeast contained in the yeast paste are inactivated to provide an inactivated yeast paste. This is shown as Figure 1Step 020 in. Such an endogenous yeast enzyme inactivation step is carried out to limit / avoid yeast autolysis. In the context of the present disclosure, the term "autolysis" means the self-degradation of the cellular components of yeast cells by their own enzymes. Thus, the endogenous yeast enzyme inactivation step 020 is carried out to limit / avoid the degradation of cellular components (especially yeast proteins), which mainly corresponds to the decomposition of proteinaceous substances (i.e., proteolysis). In one embodiment, the endogenous enzyme inactivation does not cause thermo-plasmolysis. In the context of the present disclosure, the expression "thermo-plasmolysis" refers to the denaturation of yeast and the permeabilization of the yeast membrane. In another embodiment, the endogenous enzyme inactivation of the present disclosure does not permeabilize the yeast membrane. In another embodiment, the endogenous enzyme inactivation maintains the integrity of the yeast membrane. In one embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature and / or pH change for a period of time. In one embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature above 4 °C and below 120 °C. In one embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature of at least 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C, 105 °C, 110 °C, 115 °C or 120 °C. In another embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature not exceeding 120 °C, 115 °C, 110 °C, 105 °C, 100 °C, 95 °C, 90 °C, 85 °C, 80 °C, 75 °C, 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C or 5 °C. In yet another embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature of 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C, 105 °C, 110 °C, 115 °C or 120 °C to 120 °C, 115 °C, 110 °C, 105 °C, 100 °C, 95 °C, 90 °C, 85 °C, 80 °C, 75 °C, 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C or 5 °C. In another embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature of 50 °C to 110 °C. In yet another embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a temperature of 60 °C to 100 °C, for example at a temperature of 80 °C. In one embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a specific pH or pH range. In one embodiment, the endogenous enzyme inactivation is obtained by exposing the yeast paste to a pH of 1 to 13.In one embodiment, endogenous enzyme inactivation is obtained by exposing yeast extract to a pH of at least 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1,

[0026] 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0 or higher. In another embodiment, endogenous enzyme inactivation is obtained by exposing yeast extract to a pH not exceeding 13.0, 12.9, 12.8, 12.7, 12.6,

[0027] At a pH of 12.5, 12.4, 12.3, 12.2, 12.1, 12.0, 11.9, 11.8, 11.7, 11.6, 11.5, 11.4, 11.3, 11.2, 11.1, 11.0, 10.9, 10.8, 10.7, 10.6, 10.5, 10.4, 10.3, 10.2, 10.1, 10.0, 9.9, 9.8, 9.7, 9.6, 9.5, 9.4, 9.3, 9.2, 9.1, 9.0, 8.9, 8.8, 8.7, 8.6, 8.5, 8.4, 8.3, 8.2, 8.1, 8.0, 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, 7.1, 7.0, 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, 6.1, 6.0, 5.9, 5.8, 5.7, 5.6, 5.5, 5.4, 5.3, 5.2, 5.1, 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2,4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 or 1.0 to obtain endogenous enzyme inactivation. In yet another embodiment, by exposing the yeast cream to 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1,6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8,

[0028] Endogenous enzyme inactivation is achieved at a pH of 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9 or 13.0 down to 13.0, 12.9, 12.8, 12.7, 12.6, 12.5, 12.4, 12.3, 12.2, 12.1, 12.0, 11.9, 11.8, 11.7, 11.6, 11.5, 11.4, 11.3, 11.2, 11.1, 11.0, 10.9, 10.8, 10.7, 10.6, 10.5, 10.4, 10.3, 10.2, 10.1, 10.0, 9.9, 9.8, 9.7, 9.6, 9.5, 9.4, 9.3, 9.2, 9.1, 9.0, 8.9, 8.8, 8.7, 8.6, 8.5, 8.4, 8.3, 8.2, 8.1, 8.0, 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, 7.1, 7.0, 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, 6.1, 6.0, 5.9, 5.8, 5.7, 5.6, 5.5, 5.4, 5.3, 5.2, 5.1, 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 or 1.0. In a specific example, endogenous enzyme inactivation is achieved by exposing yeast extract to a pH below 5 or above 7, such as at a pH of 3 or 9.In a specific example, endogenous enzyme inactivation is obtained by exposing the yeast extract to a pH below 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 or 1.0 or to a pH above 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9 or 13.0. In one embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to a temperature and / or pH change for a time exceeding 1 second and less than 10 hours. In one embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to a temperature and / or pH change of at least 15 s, at least 30 s, at least 45 s, at least 1 min, at least 2 min, at least 5 min, at least 10 min, at least 15 min, at least 20 min, at least 25 min, at least 30 min, at least 35 min, at least 40 min, at least 45 min, at least 50 min, at least 55 min, at least 1 h, at least 1.5 h, at least 2 h, at least 2.5 h, at least 3 h, at least 3.5 h, at least 4 h, at least 4.5 h, at least 5 h, at least 5.5 h, at least 6 h, at least 6.5 h, at least 7 h, at least 7.5 h, at least 8 h, at least 8.5 h, at least 9 h, at least 9.5 h or at least 10 h. In another embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to a temperature and / or pH change not exceeding 10 h, 9.5 h, 9 h, 8.5 h, 8 h, 7.5 h, 7 h, 6.5 h, 6 h, 5.5 h, 5 h, 4.5 h, 4 h, 3.5 h, 3 h, 2.5 h, 2 h, 1.5 h, 1 h, 55 min, 50 min, 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s, 30 s or 15 s.In yet another embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to temperature and / or pH changes for 15 s, 30 s, 45 s, 1 min, 2 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 5.5 h, 6 h, 6.5 h, 7 h, 7.5 h, 8 h, 8.5 h, 9 h, 9.5 h or 10 h up to 10 h, 9.5 h, 9 h, 8.5 h, 8 h, 7.5 h, 7 h, 6.5 h, 6 h, 5.5 h, 5 h, 4.5 h, 4 h, 3.5 h, 3 h, 2.5 h, 2 h, 1.5 h, 1 h, 55 min, 50 min, 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s, 30 s or 15 s. In another embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to temperature and / or pH changes for 15 s to 7 h. In yet another embodiment, endogenous enzyme inactivation is obtained by exposing the yeast extract to temperature and / or pH changes for 30 s to 2 h, such as 5 minutes. One of ordinary skill in the art will know how to adjust the temperature and / or pH and the exposure time according to the yeast source and the endogenous enzyme to be inactivated. In another embodiment, endogenous enzyme inactivation can be confirmed by using standard techniques in the art such as, but not limited to, protease activity assays (azocasein, fluorescein, thiocarbamyl-κ-casein or curd clot assays) and / or incubation of purified β-glucan with the inactivated yeast extract, and by measuring glucose formation (HPLC).

[0029] The inactivated yeast extract is then subjected to exogenous enzyme treatment. This is shown as Figure 1Step 030 therein. In the context of the present disclosure, the exogenous enzyme treatment lacks ribonuclease activity (i.e., endonucleases and exonucleases; (EC 3.1.4.1)). Still in the context of the present disclosure, the exogenous enzyme treatment comprises at least one polypeptide capable of hydrolyzing oligosaccharides. In one embodiment, the exogenous enzyme treatment comprises at least one polypeptide having glucanase, mannanase, and / or chitinase activity. In one embodiment, the exogenous enzyme treatment comprises at least one polypeptide having endo-β-1,3-glucanase, exo-β-1,3-glucanase, endo-β-1,6-glucanase, and / or exo-β-1,6-glucanase activity (EC 3.2.1). In another embodiment, the exogenous enzyme treatment consists essentially of using a polypeptide having β-1,3-glucanase activity and / or a polypeptide having β-1,6-glucanase activity. In the context of the present disclosure, the expression "consisting essentially of" excludes other enzyme treatments that would substantially affect the properties of protoplast formation and / or the yeast proteins obtainable. In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature above 4 °C and below 120 °C. In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature of at least 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C, 105 °C, 110 °C, 115 °C, or 120 °C. In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature not exceeding 120 °C, 115 °C, 110 °C, 105 °C, 100 °C, 95 °C, 90 °C, 85 °C, 80 °C, 75 °C, 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C, or 5 °C. In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature from 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C, 105 °C, 110 °C, 115 °C, or 120 °C to 120 °C, 115 °C, 110 °C, 105 °C, 100 °C, 95 °C, 90 °C, 85 °C, 80 °C, 75 °C, 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C, or 5 °C.In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature of 20 °C to 110 °C. In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature of 50 °C to 80 °C, for example at a temperature of 60 °C. In one embodiment, the amount of the polypeptide having glucanase activity (EC 3.2.1) is higher than 1 UI and lower than 5000 UI per kg of inactivated yeast paste. In the context of the present disclosure, one unit of enzyme activity (UI) catalyzes the formation of 1 μmol of glucose per minute at 37 °C. In one embodiment, the amount of the polypeptide having glucanase activity (EC 3.2.1) is at least 1 UI, 5 UI, 10 UI, 20 UI, 30 UI, 40 UI, 50 UI, 60 UI, 70 UI, 80 UI, 90 UI, 100 UI, 200 UI, 300 UI, 400 UI, 500 UI, 600 UI, 700 UI, 800 UI, 900 UI, 1000 UI, 1100 UI, 1200 UI, 1300 UI, 1400 UI, 1500 UI, 1600 UI, 1700 UI, 1800 UI, 1900 UI, 2000 UI, 2100 UI, 2200 UI, 2300 UI, 2400 UI, 2500 UI, 2600 UI, 2700 UI, 2800 UI, 2900 UI, 3000 UI, 3100 UI, 3200 UI, 3300 UI, 3400 UI, 3500 UI, 3600 UI, 3700 UI, 3800 UI, 3900 UI, 4000 UI, 4100 UI, 4200 UI, 4300 UI, 4400 UI, 4500 UI, 4600 UI, 4700 UI, 4800 UI, 4900 UI or 5000 UI per kg of inactivated yeast paste.In another embodiment, the amount of the polypeptide having dextranase activity (EC 3.2.1) is not more than 5000 UI, 4900 UI, 4800 UI, 4700 UI, 4600 UI, 4500 UI, 4400 UI, 4300 UI, 4200 UI, 4100 UI, 4000 UI, 3900 UI, 3800 UI, 3700 UI, 3600 UI, 3500 UI, 3400 UI, 3300 UI, 3200 UI, 3100 UI, 3000 UI, 2900 UI, 2800 UI, 2700 UI, 2600 UI, 2500 UI, 2400 UI, 2300 UI, 2200 UI, 2100 UI, 2000 UI, 1900 UI, 1800 UI, 1700 UI, 1600 UI, 1500 UI, 1400 UI, 1300 UI, 1200 UI, 1100 UI, 1000 UI, 900 UI, 800 UI, 700 UI, 600 UI, 500 UI, 400 UI, 300 UI, 200 UI, 100 UI, 90 UI, 80 UI, 70 UI, 60 UI, 50 UI, 40 UI, 30 UI, 20 UI, 10 UI, 5 UI or 1 UI per kg of inactivated yeast extract.In another embodiment, the amount of the polypeptide having glucanase activity (EC 3.2.1) is 1 UI, 5 UI, 10 UI, 20 UI, 30 UI, 40 UI, 50 UI, 60 UI, 70 UI, 80 UI, 90 UI, 100 UI, 200 UI, 300 UI, 400 UI, 500 UI, 600 UI, 700 UI, 800 UI, 900 UI, 1000 UI, 1100 UI, 1200 UI, 1300 UI, 1400 UI, 1500 UI, 1600 UI, 1700 UI, 1800 UI, 1900 UI, 2000 UI, 2100 UI, 2200 UI, 2300 UI, 2400 UI, 2500 UI, 2600 UI, 2700 UI, 2800 UI, 2900 UI, 3000 UI, 3100 UI, 3200 UI, 3300 UI, 3400 UI, 3500 UI, 3600 UI, 3700 UI, 3800 UI, 3900 UI, 4000 UI, 4100 UI, 4200 UI, 4300 UI, 4400 UI, 4500 UI, 4600 UI, 4700 UI, 4800 UI, 4900 UI or 5000 UI per kg of inactivated yeast extract to 5000 UI, 4900 UI, 4800 UI, 4700 UI, 4600 UI, 4500 UI, 4400 UI, 4300 UI, 4200 UI, 4100 UI, 4000 UI, 3900 UI, 3800 UI, 3700 UI, 3600 UI, 3500 UI, 3400 UI, 3300 UI, 3200 UI, 3100 UI, 3000 UI, 2900 UI, 2800 UI, 2700 UI, 2600 UI, 2500 UI, 2400 UI, 2300 UI, 2200 UI, 2100 UI, 2000 UI, 1900 UI, 1800 UI, 1700 UI, 1600 UI, 1500 UI, 1400 UI, 1300 UI, 1200 UI, 1100 UI, 1000 UI, 900 UI, 800 UI, 700 UI, 600 UI, 500 UI, 400 UI, 300 UI, 200 UI, 100 UI, 90 UI, 80 UI, 70 UI, 60 UI, 50 UI, 40 UI, 30 UI, 20 UI, 10 UI, 5 UI or 1 UI. In another embodiment, the amount of the polypeptide having glucanase activity (EC 3.2.1) is 300 UI to 3000 UI per kg of inactivated yeast extract. In yet another embodiment, the amount of the polypeptide having glucanase activity (EC 3.2.1) is 1500 UI to 2500 UI per kg of inactivated yeast extract, for example, the amount of the polypeptide having glucanase activity (EC 3.2.1) is 2000 UI per kg of inactivated yeast extract.In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of from 3 to 9. In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of at least 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0 or higher. In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH not exceeding 9.0, 8.9, 8.8, 8.7, 8.6, 8.5, 8.4, 8.3, 8.2, 8.1, 8.0, 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, 7.1, 7.0, 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, 6.1, 6.0, 5.9, 5.8, 5.7, 5.6, 5.5, 5.4, 5.3, 5.2, 5.1, 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1 or 3.0.In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9 or 9.0 to 9.0, 8.9, 8.8, 8.7, 8.6, 8.5, 8.4, 8.3, 8.2, 8.1, 8.0, 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, 7.1, 7.0, 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, 6.1, 6.0, 5.9, 5.8, 5.7, 5.6, 5.5, 5.4, 5.3, 5.2, 5.1, 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1 or 3.0. In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of 4 to 8. In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of 5 to 7, for example at a pH of 5.5. In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for a time of more than 1 second and less than 10 hours.In one embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for at least 15 s, at least 30 s, at least 45 s, at least 1 min, at least 2 min, at least 5 min, at least 10 min, at least 15 min, at least 20 min, at least 25 min, at least 30 min, at least 35 min, at least 40 min, at least 45 min, at least 50 min, at least 55 min, at least 1 h, at least 1.5 h, at least 2 h, at least 2.5 h, at least 3 h, at least 3.5 h, at least 4 h, at least 4.5 h, at least 5 h, at least 5.5 h, at least 6 h, at least 6.5 h, at least 7 h, at least 7.5 h, at least 8 h, at least 8.5 h, at least 9 h, at least 9.5 h, at least 10 h, at least 11 h, at least 12 h, at least 13 h, at least 14 h, at least 15 h, at least 16 h, at least 17 h, at least 18 h, at least 19 h, at least 20 h, at least 21 h, at least 22 h, at least 23 h or at least 24 h. In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for not more than 24 h, 23 h, 22 h, 21 h, 20 h, 19 h, 18 h, 17 h, 16 h, 15 h, 14 h, 13 h, 12 h, 11 h, 10 h, 9.5 h, 9 h, 8.5 h, 8 h, 7.5 h, 7 h, 6.5 h, 6 h, 5.5 h, 5 h, 4.5 h, 4 h, 3.5 h, 3 h, 2.5 h, 2 h, 1.5 h, 1 h, 55 min, 50 min, 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s, 30 s or 15 s.In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for 15 s, 30 s, 45 s, 1 min, 2 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 5.5 h, 6 h, 6.5 h, 7 h, 7.5 h, 8 h, 8.5 h, 9 h, 9.5 h, 10 h, 11 h, 12 h, 13 h, 14 h, 15 h, 16 h, 17 h, 18 h, 19 h, 20 h, 21 h, 22 h, 23 h or 24 h up to 24 h, 23 h, 22 h, 21 h, 20 h, 19 h, 18 h, 17 h, 16 h, 15 h, 14 h, 13 h, 12 h, 11 h, 10 h, 9.5 h, 9 h, 8.5 h, 8 h, 7.5 h, 7 h, 6.5 h, 6 h, 5.5 h, 5 h, 4.5 h, 4 h, 3.5 h, 3 h, 2.5 h, 2 h, 1.5 h, 1 h, 55 min, 50 min, 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s, 30 s or 15 s. In another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for 1 h to 24 h. In yet another embodiment, the exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for 2 h to 6 h, such as 4 h. Those of ordinary skill in the art will know how to adjust the concentration, temperature, pH and exposure time of the polypeptide having glucanase activity according to the yeast source and the polypeptide having glucanase activity under consideration.

[0030] The inactivated yeast paste is subjected to an enzyme treatment to obtain yeast protoplasts. In the context of the present disclosure, the expression "yeast protoplasts" refers to yeast that has had most of its cell wall components removed. In another embodiment, the enzyme treatment of the method of the present disclosure avoids the use of enzymes having proteolytic activity and thus aims to maintain the integrity of yeast proteins (e.g., secondary, tertiary or quaternary structure). In some additional embodiments, the enzyme treatment of the present disclosure aims to maintain the integrity of intracellular proteins as well as yeast cell membrane proteins. In another embodiment, the enzyme treatment of the present disclosure aims to dissolve the yeast cell wall, leaving a soluble fraction containing mannoproteins and / or β-glucans and an insoluble fraction containing yeast protoplasts. In yet another embodiment, the enzyme treatment of the present disclosure allows some yeast membranes to be dissolved but does not allow yeast proteins to diffuse outside the yeast protoplasts.

[0031] In some embodiments, before the enzyme treatment step 030, an alkali extraction step 025 can be performed on the inactivated yeast paste. The alkali extraction step can be carried out to reduce the amount of nucleotides (including polynucleotides such as RNA) in the final composition. In some embodiments, the alkali extraction step can be performed under conditions that allow extraction of nucleotides from the inactivated yeast paste and prevent or limit degradation of nucleotides from the inactivated yeast paste. The expression "prevent or limit degradation of nucleotides from the inactivated yeast paste" means the fact that the alkali extraction step prevents or limits nucleotide degradation by at least 50%. Without wishing to be bound by theory, it should be understood that alkali extraction of the inactivated yeast paste at least partially dissolves the nucleotides present in the inactivated yeast paste while allowing the formation of yeast protoplasts (after treatment with exogenous enzymes). The alkali extraction step 025 includes placing the inactivated yeast paste under alkaline conditions within a certain temperature range for a certain time range to allow extracellular dissolution of nucleotides (including polynucleotides such as RNA) in the inactivated yeast paste. In one embodiment, the alkali extraction step 025 is performed at a pH of 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9 up to 12.0, 11.9, 11.8., 11.7, 11.6, 11.5, 11.4, 11.3, 11.2, 11.1, 11.0, 10.9, 10.8., 10.7, 10.6, 10.5, 10.4, 10.3, 10.2, 10.1, 10.0, 9.9, 9.8., 9.7, 9.6, 9.5, 9.4, 9.3, 9.2, 9.1, 9.0, 8.9, 8.8., 8.7, 8.6, 8.5, 8.4, 8.3, 8.2 or 8.1. In another embodiment, the alkali extraction step 025 is performed at a pH of 8 to 11. In yet another embodiment, the alkali extraction step 025 is performed at a pH of 8.5 to 9.5, for example at a pH of 9.0. In additional embodiments, the alkali extraction step 025 is performed at a temperature above 4 °C and below 70 °C. In one embodiment, the alkali extraction step 025 is performed at a temperature of at least 4 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C or 70 °C. In another embodiment, the alkali extraction step is performed at a temperature not exceeding 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C, 5 °C or 4 °C.In yet another embodiment, the alkali extraction step 025 is carried out at a temperature from 4 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C or 70 °C to 70 °C, 65 °C, 60 °C, 55 °C, 50 °C, 45 °C, 40 °C, 35 °C, 30 °C, 25 °C, 20 °C, 15 °C, 10 °C, 5 °C or 4 °C. In another embodiment, the alkali extraction step is carried out at a temperature from 50 °C to 70 °C, for example at a temperature of 65 °C. In yet another embodiment, the alkali extraction step 025 is carried out for 15 s, 30 s, 45 s, 1 min, 2 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 11 h, 12 h, 13 h, 14 h, 15 h, 16 h, 17 h, 18 h, 19 h, 20 h, 21 h, 22 h, 23 h or 24 h to 24 h, 23 h, 22 h, 21 h, 20 h, 19 h, 18 h, 17 h, 16 h, 15 h, 14 h, 13 h, 12 h, 11 h, 10 h, 9 h, 8 h, 7 h, 6 h, 5 h, 4 h, 3.5 h, 3 h, 2.5 h, 2 h, 1.5 h, 1 h, 55 min, 50 min, 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s or 30 s. In another embodiment, the alkali extraction step 025 is carried out for 1 h to 4 h. In yet another embodiment, the alkali extraction step 025 is carried out for 1.5 h to 2.5 h, for example 2 h. In yet another embodiment, the alkali extraction step 025 is carried out for 15 s, 30 s, 45 s, 1 min, 2 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min and 45 min to 45 min, 40 min, 35 min, 30 min, 25 min, 20 min, 15 min, 10 min, 5 min, 2 min, 1 min, 45 s or 30 s. In another embodiment, the alkali extraction step 025 is carried out for 10 min to 45 min. In yet another embodiment, the alkali extraction step 025 is carried out for 20 min to 40 min, for example 30 min. One of ordinary skill in the art will know how to optimize the temperature, pH and exposure time of the alkali extraction step to dissolve nucleotides from the inactivated yeast paste.

[0032] In some embodiments, the method may include a step for inactivating the exogenous enzyme used during the enzyme treatment step 030 ( Figure 1(not shown in the figure).

[0033] Then, the insoluble fraction and the soluble fraction obtained during the enzymatic treatment carried out in the methods herein are separated by conventional techniques known to those of ordinary skill in the art. This is shown as Figure 1 step 050. Techniques that can be used include, but are not limited to, sedimentation, centrifugation, and / or filtration of the insoluble fraction. In one embodiment, the obtained insoluble fraction is directly separated. In one embodiment, once separated from the soluble fraction, the insoluble fraction is optionally rinsed and then collected. In another embodiment, once separated from the soluble fraction, the insoluble fraction is directly collected. In one embodiment, the collected insoluble fraction has a neutral taste and can be in a non-dried or dried state. In the context of the present disclosure, the non-dried insoluble fraction can be in a semi-liquid state.

[0034] In one embodiment, the insoluble fraction can be provided in a non-dried state. In one embodiment, the non-dried insoluble fraction can be used immediately after collection to formulate an edible composition. In one embodiment, the non-dried insoluble fraction can be stored. In another embodiment, the non-dried insoluble fraction can be frozen according to conventional methods in the art for later formulation into an edible composition. In another embodiment, non-dried insoluble fractions obtained from different yeast sources can be combined to formulate an edible composition. In one embodiment, the insoluble fraction can be provided in a dried state. In another embodiment, the dried insoluble fraction is obtained by, but not limited to, spray drying, fluidized bed drying, pan drying, drum / drum drying, infrared drying, or freeze drying / lyophilizing the non-dried insoluble fraction. In another embodiment, the dried insoluble fraction can be stored. In yet another embodiment, the dried insoluble fraction can be stored for later formulation into an edible composition. In another embodiment, dried insoluble fractions obtained from different yeast sources can be combined to formulate an edible composition.

[0035] In one embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of greater than 50% and less than 100% based on the total mass of the dried collected insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. In one embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of at least 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% based on the total mass of the dried collected insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. In another embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of no more than 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 76%, 74%, 72%, 70%, 68%, 66%, 64%, 62%, 60%, 58%, 56%, 54%, 52% or 50% based on the total mass of the dried collected insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. In yet another embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 76%, 74%, 72%, 70%, 68%, 66%, 64%, 62%, 60%, 58%, 56%, 54%, 52% or 50% based on the total mass of the dried collected insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25.In another embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of 70% to 100% based on the total mass of the dried collected insoluble fraction as determined by the Kjeldahl method for nitrogen analysis using 6.25 as a conversion factor. In yet another embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of 75% to 85%, such as 80% protein content, based on the total mass of the dried collected insoluble fraction as determined by the Kjeldahl method for nitrogen analysis using 6.25 as a conversion factor. In another embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of at least 80% based on the total mass of the dried collected insoluble fraction as determined by the Kjeldahl method for nitrogen analysis using 6.25 as a conversion factor.

[0036] The collected insoluble portion containing yeast protoplasts may also have at least one of the following characteristics when dried:

[0037] - A lipid content of greater than 0% and less than 20% based on the total mass of the dried collected insoluble fraction, determined by the modified Mojonnier method (AOAC 989.05); alternatively, a lipid content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0% or 20.0% based on the total mass of the dried collected insoluble fraction, determined by the modified Mojonnier method (AOAC 989.05); alternatively, a lipid content of not more than 20.0%, 19.0%, 18.0%, 17.0%, 16.0%, 15.0%, 14.0%, 13.0%, 12.0%, 11.0%, 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.a lipid content of 2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0% or 20.0% to 20.0%, 19.0%, 18.0%, 17.0%, 16.0%, 15.0%, 14.0%, 13.0%, 12.0%, 11.0%, 10.0%, 9.9%, 9.8%, 9.a lipid content of 7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, a lipid content of 6% to 10% based on the total mass of the dried collected insoluble fraction determined by the modified Mojonnier method (AOAC 989.05); alternatively, a lipid content of 7% to 9% based on the total mass of the dried collected insoluble fraction determined by the modified Mojonnier method (AOAC 989.05), such as a lipid content of 8%.

[0038] - When no alkali extraction step is performed, a nucleic acid content of higher than 2% and lower than 30% based on the total mass of the dried collected insoluble fraction; alternatively, a nucleic acid content of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% based on the total mass of the dried collected insoluble fraction; alternatively, a nucleic acid content of not more than 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% based on the total mass of the dried collected insoluble fraction; alternatively, a nucleic acid content of 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% to 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% based on the total mass of the dried collected insoluble fraction; alternatively, a nucleic acid content of 5% to 20% based on the total mass of the dried collected insoluble fraction; alternatively, a nucleic acid content of 7% to 15% based on the total mass of the dried collected insoluble fraction, such as 10% nucleic acid content. In the context of the present disclosure, the expression "nucleic acid" refers to a biopolymer (i.e., polynucleotide) composed of nucleotides, which are composed of a 5-carbon sugar, a phosphate group, and a nitrogenous base. Still in the context of the present disclosure, the expression "nucleic acid" refers to a polynucleotide, such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Still in the context of the present disclosure, the expression "nucleic acid" refers to ribonucleic acid (RNA). In the context of the present disclosure, the term "nucleic acid" does not refer to nucleotide residues that may have been generated during the process. In the present disclosure, the nucleic acid content is measured according to the method of Fish et al. (1991), and the values with and without enzymatic digestion are compared;

[0039] - When an alkali extraction step has been carried out, a nucleic acid content equal to or lower than 3% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content equal to or lower than 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.2% or 0.1% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content of 0.1% to 3% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content of 0.5% to 2% based on the total mass of the dried collected insoluble fraction. In the context of the present disclosure, the expression "nucleic acid" refers to a biopolymer (i.e., polynucleotide) composed of nucleotides, which are composed of a 5-carbon sugar, a phosphate group, and a nitrogenous base. Still in the context of the present disclosure, the expression "nucleic acid" refers to a polynucleotide, such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Still in the context of the present disclosure, the expression "nucleic acid" refers to ribonucleic acid (RNA). In the context of the present disclosure, the term "nucleic acid" does not refer to nucleotide residues that may have been generated during the process. In the present disclosure, the nucleic acid content is measured according to the method of Fish et al. (1991), and the values with and without enzymatic digestion are compared;

[0040] - A carbohydrate content higher than 0% and lower than 25%; alternatively, a carbohydrate content of at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% based on the total mass of the dried collected insoluble fraction; alternatively, a carbohydrate content not exceeding 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the dried collected insoluble fraction; alternatively, a carbohydrate content of 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% to 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the dried collected insoluble fraction; alternatively, a carbohydrate content of 5% to 20% based on the total mass of the dried collected insoluble fraction; alternatively, a carbohydrate content of 7% to 15% based on the total mass of the dried insoluble fraction, such as a carbohydrate content of 10%. In the context of the present disclosure, the term "carbohydrate" refers to the total sugars measured by HPLC after chemical or enzymatic digestion (i.e., AOAC 980.13 method);

[0041] - A mannan content of higher than 0% and lower than 6% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9% or 6.0% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of not more than 6.0%, 5.9%, 5.8, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% based on the total mass of the dried collected insoluble fraction; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.A mannan content of 6%, 5.7%, 5.8%, or 5.9% to 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%; alternatively, a mannan content of 0% to 5.5% or 0% to 4% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of 0.5% to 5.5% or 0.5% to 4.0% based on the total mass of the dried collected insoluble fraction, such as a mannan content of about 2% or 5%. In the context of the present disclosure, the mannan content is measured by HPLC after chemical or enzymatic digestion;.

[0042] - a dextran content that is higher than 0% and lower than 10% based on the total mass of the dried collected insoluble fraction; alternatively, a dextran content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9% or 10.0% based on the total mass of the dried collected insoluble fraction; alternatively, not exceeding 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.a dextran content of 9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9% or 10.0% to 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.a dextran content of 5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, a dextran content of 0% to 8% based on the total mass of the dried collected insoluble fraction; alternatively, a dextran content of 1% to 4% based on the total mass of the dried collected insoluble fraction, such as a dextran content of 2%. In the context of the present disclosure, the dextran content is measured by HPLC after chemical or enzymatic digestion; and / or.

[0043] - a glucose content higher than 0% and lower than 25%; alternatively, a glucose content of at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% based on the total mass of the dried collected insoluble fraction;

[0044] alternatively, a glucose content of not more than 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the dried collected insoluble fraction; alternatively, a glucose content of 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% to 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the dried collected insoluble fraction; alternatively, a glucose content of 5% to 20% based on the total mass of the dried collected insoluble fraction; alternatively, a glucose content of 7% to 15% based on the total mass of the dried insoluble fraction, such as a glucose content of 10%. In the context of the present disclosure, the term "glucose" refers to the monosaccharide having the molecular formula C6H 12 O6. Still in the context of the present disclosure, the glucose content is measured directly by HPLC.

[0045] In some embodiments, the method may include obtaining a soluble fraction of an enzymatically treated yeast extract. This corresponds to Figure 1 step 040 of

[0046] Composition derived from yeast protoplasts

[0047] In one embodiment, the present disclosure aims to provide a composition comprising a yeast protein derived from yeast protoplasts. In one embodiment, the composition comprising a yeast protein derived from yeast protoplasts has at least one of the following characteristics:

[0048] - A protein content that is higher than 50% and lower than 100% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. Alternatively, a protein content of at least 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. Alternatively, a protein content that does not exceed 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 76%, 74%, 72%, 70%, 68%, 66%, 64%, 62%, 60%, 58%, 56%, 54%, 52% or 50% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. Alternatively, a protein content of 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 76%, 74%, 72%, 70%, 68%, 66%, 64%, 62%, 60%, 58%, 56%, 54%, 52% or 50% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. Alternatively, a protein content of 70% to 100% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25. Alternatively, a protein content of 75% to 85% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis with a conversion factor of 6.25, such as 80% protein content;

[0049] - A lipid content of greater than 0% and less than 20% based on the total mass of the dried collected insoluble fraction, determined by the modified Mojonnier method (AOAC 989.05); alternatively, a lipid content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0% or 20.0% based on the total mass of the dried collected insoluble fraction, determined by the modified Mojonnier method (AOAC 989.05); alternatively, a lipid content of not more than 20.0%, 19.0%, 18.0%, 17.0%, 16.0%, 15.0%, 14.0%, 13.0%, 12.0%, 11.0%, 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.a lipid content of 8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0% or 20.0% to 20.0%, 19.0%, 18.0%, 17.0%, 16.0%, 15.0%, 14.0%, 13.0%, 12.0%, 11.0%, 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.A lipid content of 3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, a lipid content of 6% to 10% based on the total mass of the collected insoluble fraction dried as determined by the modified Mojonnier method (AOAC 989.05); alternatively, by the modified Mojonnier method (AOAC 989.05) A lipid content of 7% to 9% by total mass of the collected insoluble fraction based on dry weight, such as 8% lipid content; - when no alkali extraction step is performed, a nucleic acid content of higher than 2% and lower than 30% by total mass of the collected insoluble fraction based on dry weight; alternatively, a nucleic acid content of at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% by total mass of the collected insoluble fraction based on dry weight; alternatively, a nucleic acid content of not exceeding 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% by total mass of the collected insoluble fraction based on dry weight; alternatively, a nucleic acid content of 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% to 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% by total mass of the collected insoluble fraction based on dry weight; alternatively, a nucleic acid content of 5% to 20% by total mass of the collected insoluble fraction based on dry weight; alternatively, a nucleic acid content of 7% to 15% by total mass of the collected insoluble fraction based on dry weight, such as 10% nucleic acid content. In the context of the present disclosure, the expression "nucleic acid" refers to a biopolymer (i.e., polynucleotide) composed of nucleotides, which are composed of a 5-carbon sugar, a phosphate group, and a nitrogenous base. Still in the context of the present disclosure, the expression "nucleic acid" refers to a polynucleotide, such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Still in the context of the present disclosure, the expression "nucleic acid" refers to ribonucleic acid (RNA). In the context of the present disclosure, the term "nucleic acid" does not refer to nucleotide residues that may have been generated during the process. In the present disclosure, the nucleic acid content is measured according to the method of Fish et al. (1991), and the values with and without enzymatic digestion are compared;.

[0050] - When an alkali extraction step has been carried out, a nucleic acid content equal to or lower than 3% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content equal to or lower than 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.2% or 0.1% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content of 0.1% to 3% based on the total mass of the dried collected insoluble fraction. Alternatively, a nucleic acid content of 0.5% to 2.0% based on the total mass of the dried collected insoluble fraction. In the context of the present disclosure, the expression "nucleic acid" refers to a biopolymer (i.e., polynucleotide) composed of nucleotides, which are composed of a 5-carbon sugar, a phosphate group and a nitrogenous base. Still in the context of the present disclosure, the expression "nucleic acid" refers to a polynucleotide, such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Still in the context of the present disclosure, the expression "nucleic acid" refers to ribonucleic acid (RNA). In the context of the present disclosure, the term "nucleic acid" does not refer to nucleotide residues that may have been generated during the process. In the present disclosure, the nucleic acid content is measured according to the method of Fish et al. (1991), and the values with and without enzymatic digestion are compared;

[0051] - a carbohydrate content greater than 0% and less than 25%; alternatively, a carbohydrate content of at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% based on the total mass of the composition; alternatively, a carbohydrate content not exceeding 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the composition; alternatively, a carbohydrate content of 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% to 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% based on the total mass of the composition; alternatively, a carbohydrate content of 5% to 20% based on the total mass of the composition; alternatively, a carbohydrate content of 7% to 15% based on the total mass of the composition, such as a carbohydrate content of 10%. In the context of the present disclosure, the term "carbohydrate" refers to the total sugars measured by HPLC after chemical or enzymatic digestion (i.e., AOAC 980.13 method);

[0052] - A mannan content of more than 0% and less than 6% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9% or 6.0% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of not more than 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% based on the total mass of the dried collected insoluble fraction; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.A mannan content of 7%, 5.8%, 5.9% or 6.0% to 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, a mannan content of 0% to 5.5% or 0% to 4.0% based on the total mass of the dried collected insoluble fraction; alternatively, a mannan content of 0.5% to 5.5% or 0.5% to 3.0% based on the total mass of the dried collected insoluble fraction, such as a mannan content of about 2% or about 5.5%. In the context of the present disclosure, the mannan content is measured by HPLC after chemical or enzymatic digestion;.

[0053] - a dextran content that is higher than 0% and lower than 10% based on the total mass of the dried collected insoluble fraction; alternatively, a dextran content of at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9% or 10.0% based on the total mass of the dried collected insoluble fraction; alternatively, not exceeding 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.a dextran content of 9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9% or 10.0% to 10.0%, 9.9%, 9.8%, 9.7%, 9.6%, 9.5%, 9.4%, 9.3%, 9.2%, 9.1%, 9.0%, 8.9%, 8.8%, 8.7%, 8.6%, 8.5%, 8.4%, 8.3%, 8.2%, 8.1%, 8.0%, 7.9%, 7.8%, 7.7%, 7.6%, 7.5%, 7.4%, 7.3%, 7.2%, 7.1%, 7.0%, 6.9%, 6.8%, 6.7%, 6.6%, 6.5%, 6.4%, 6.3%, 6.2%, 6.1%, 6.0%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, 3.0%, 2.9%, 2.8%, 2.7%, 2.6%, 2.A dextran content of 5%, 2.4%, 2.3%, 2.2%, 2.1%, 2.0%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1%; alternatively, a dextran content of 0% to 8% based on the total mass of the dried collected insoluble fraction; alternatively, a dextran content of 1% to 4% based on the total mass of the dried collected insoluble fraction, such as a dextran content of 2%. In the context of the present disclosure, the dextran content is measured by HPLC after chemical or enzymatic digestion; and / or.

[0054] - A glucose content higher than 0% and lower than 25%; alternatively, at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% glucose content based on the total mass of the composition; alternatively, not more than 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% glucose content based on the total mass of the composition; alternatively, a glucose content of 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24% or 25% to 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% glucose content based on the total mass of the composition; alternatively, a glucose content of 5% to 20% based on the total mass of the composition; alternatively, a glucose content of 7% to 15% based on the total mass of the composition, such as a glucose content of 10%. In the context of the present disclosure, the term "glucose" refers to a monosaccharide having the molecular formula C6H 12 O6. Still in the context of the present disclosure, the glucose content is measured directly by HPLC.

[0055] In one embodiment, the yeast protein derived from yeast protoplasts is intracellular yeast protein, yeast cell membrane protein or a combination thereof.

[0056] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a neutral taste. This is contrary to yeast proteins obtained from yeast extracts (especially those obtained using protease treatment), which typically exhibit a medium taste. In the context of the present disclosure, yeast proteins derived from yeast protoplasts of an edible product have a neutral taste and can thus be used as an ingredient whose taste does not affect / alter the taste of other components of the composition. Thus, the compositions of the present disclosure do not affect / alter the taste of the formulated edible product. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a neutral taste when compared to a plant / legume protein composition. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a neutral taste when compared to a yeast protein composition derived from a method lacking a glucanase treatment step. In yet another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a neutral taste when compared to a composition lacking yeast protoplasts. In the context of the present disclosure, a method for preparing a yeast protein composition (the method lacking a glucanase treatment step) is a method comprising the steps of: i) providing a yeast paste comprising yeast; ii) inactivating the endogenous enzymes of the yeast to provide an inactivated yeast paste; iii) optionally separating an insoluble fraction from a soluble fraction and iv) collecting the inactivated yeast paste or optionally collecting the insoluble fraction of the inactivated yeast paste, excluding a glucanase treatment step. Still in the context of the present disclosure, a method for preparing a yeast protein composition (the method lacking a glucanase treatment step) is a method consisting essentially of the steps of: i) providing a yeast paste comprising yeast; ii) inactivating the endogenous enzymes of the yeast to provide an inactivated yeast paste; iii) optionally separating an insoluble fraction from a soluble fraction and iv) collecting the inactivated yeast paste or optionally collecting the insoluble fraction of the inactivated yeast paste. The transitional phrase "consisting essentially of" limits the method steps to the specified steps "(i.e., without a glucanase treatment step and without an insoluble / soluble fraction separation step) and those steps that do not substantially affect the basic and novel characteristics of the final product".

[0057] In one embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index greater than 1% and less than 15%. In one embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index greater than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15%. In another embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index not exceeding 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1%. In yet another embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15% to 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1%. In another embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index of 1% to 10%. In yet another embodiment, based on the total mass of the dried composition, the composition comprising yeast protein derived from yeast protoplasts has a water solubility index of 3% to 7%, such as a water solubility index of 5. In the context of the present disclosure, the water solubility index represents the weight of the dried solids in the supernatant, expressed as a percentage of the original weight of the sample. In another embodiment, the composition comprising yeast protein derived from yeast protoplasts has a lower water solubility index compared to a yeast protein composition prepared by a similar method lacking a glucanase treatment step.

[0058] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has weak emulsifying properties. In the context of the present disclosure, the expression "emulsifying properties" refers to the emulsifying activity and / or emulsion stability of the composition. Still in the context of the present disclosure, the expression "emulsifying activity" refers to the ability of the composition to form an emulsion, while the expression "emulsion stability" refers to the emulsion remaining after heat treatment. In one embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is 1 to 10 times lower than that of lecithin. In another embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times lower than that of lecithin. In another embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times lower than that of lecithin. In yet another embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times to 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times lower than that of lecithin. In another embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is 1 to 10 times lower than that of lecithin. In yet another embodiment, when measured by the method of Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has an emulsifying activity that is 3 to 7 times lower than that of lecithin, for example, has an emulsifying activity that is 5 times lower than that of lecithin. In one embodiment, when measured by the method of Yasumatsu et al. (1972), the composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is 1 to 15 times lower than that of lecithin. In another embodiment, when measured by the method of Yasumatsu et al. (1972), the composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times lower than that of lecithin. In another embodiment, when measured by the method of Yasumatsu et al. (1972), the composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is not more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times lower than that of lecithin.In yet another embodiment, when measured by the method of Yasumatsu et al. (1972), a composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times lower than that of lecithin, up to 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times lower. In another embodiment, when measured by the method of Yasumatsu et al. (1972), a composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is 5 to 15 times lower than that of lecithin. In yet another embodiment, when measured by the method of Yasumatsu et al. (1972), a composition comprising yeast proteins derived from yeast protoplasts has an emulsion stability that is 8 to 12 times lower than that of lecithin, for example, the composition has an emulsion stability that is 10 times lower than that of lecithin.

[0059] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has weak foaming properties. In the context of the present disclosure, the expression "foaming properties" refers to foam capacity and / or foam stability over time. Still in the context of the present disclosure, the "foam capacity" of a composition refers to the amount of interfacial area that can be generated by the composition, while the expression "foam stability" refers to the ability of the composition to resist gravity and mechanical stress over time. In one embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is 1 to 10 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In another embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In another embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In yet another embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times to 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In another embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is 1 to 6 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In yet another embodiment, when measured by the method described in Chandra et al. (2015), the composition comprising yeast proteins derived from yeast protoplasts has a foam capacity that is 1 to 4 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step, for example, the composition has a foam capacity that is 3 times smaller than that of a yeast protein composition prepared by a similar method lacking the glucanase treatment step. In one embodiment, when measured 30 minutes later by the method described in Brishti et al. (2017), the composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is 1 to 10 times lower than that of a yeast protein composition prepared by a method lacking the glucanase treatment step.In another embodiment, when measured after 30 minutes using the method described in Brishti et al. (2017), a composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold lower than that of a yeast protein composition prepared by a method lacking a glucanase treatment step. In another embodiment, when measured after 30 minutes using the method described in Brishti et al. (2017), a composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is not more than 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold lower than that of a yeast protein composition prepared by a method lacking a glucanase treatment step. In yet another embodiment, when measured after 30 minutes using the method described in Brishti et al. (2017), a composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold to 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold lower than that of a yeast protein composition prepared by a method lacking a glucanase treatment step. In another embodiment, when measured after 30 minutes using the method described in Brishti et al. (2017), a composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is 3-fold to 9-fold lower than that of a yeast protein composition prepared by a method lacking a glucanase treatment step. In yet another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a foam stability that is 4-fold to 8-fold lower than that of a yeast protein composition prepared by a method lacking a glucanase treatment step.

[0060] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of less than 12.2 mPa·s as measured by the method of Onwulata et al. (2014) at room temperature. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of about 10 mPa·s as measured by the method of Onwulata et al. (2014) at room temperature. In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of less than 8.5 mPa·s as measured by the method of Onwulata et al. (2014) when heated at 85 °C for 5 minutes. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of about 7.3 mPa·s as measured by the method of Onwulata et al. (2014) when heated at 85 °C for 5 minutes. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of less than 14.1 mPa·s as measured by the method of Onwulata et al. (2014) when cooled to room temperature after heating at 85 °C for 5 minutes. In yet another embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a viscosity of about 11.2 mPa·s as measured by the method of Onwulata et al. (2014) when cooled to room temperature after heating at 85 °C for 5 minutes.

[0061] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts has a higher Protein Digestibility Corrected Amino Acid Score (PDCAAS) than a yeast protein composition prepared by a method lacking a glucanase treatment step. For example, in some embodiments, the PDCAAS of the compositions of the present disclosure can be equal to or higher than 1.0. In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts is more digestible than a yeast protein composition prepared by a method lacking a glucanase treatment step.

[0062] In one embodiment, a composition comprising yeast proteins derived from yeast protoplasts can be obtained by or through the method for obtaining a composition containing yeast proteins disclosed herein. In another embodiment, a composition comprising yeast proteins derived from yeast protoplasts comprises the collected insoluble fraction containing yeast protoplasts of the method for obtaining a composition containing yeast proteins disclosed herein. In yet another embodiment, a composition comprising yeast proteins derived from yeast protoplasts consists essentially of the collected insoluble fraction containing yeast protoplasts of the method for obtaining a composition containing yeast proteins disclosed herein. In another embodiment, compositions comprising yeast proteins derived from yeast protoplasts obtained from different yeast sources can be combined in order to formulate an edible composition.

[0063] Edible product

[0064] In one embodiment, the present disclosure aims to provide an edible product. In one embodiment, the edible product comprises yeast protein and at least one additional ingredient. In another embodiment, the edible product comprises a composition containing yeast protein derived from yeast protoplasts and at least one additional ingredient. In yet another embodiment, the composition included in the edible product is a composition comprising yeast protein derived from the yeast protoplasts of the present disclosure and at least one additional ingredient.

[0065] In one embodiment, the composition contained in the edible product provides from 1% to 100% of the protein of the edible product based on the total mass of the edible product. In another embodiment, the composition contained in the edible product provides at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% of the protein of the edible product. In yet another embodiment, the composition contained in the edible product provides not more than 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, 70%, 69%, 68%, 67%, 66%, 65%, 64%, 63%, 62%, 61%, 60%, 59%, 58%, 57%, 56%, 55%, 54%, 53%, 52%, 51%, 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the protein of the edible product.In some embodiments, the composition contained in the edible product provides from 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, 70%, 69%, 68%, 67%, 66%, 65%, 64%, 63%, 62%, 61%, 60%, 59%, 58%, 57%, 56%, 55%, 54%, 53%, 52%, 51%, 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the protein of the edible product. In the context of the present disclosure, the proportion of the composition within the edible product will depend on the nature of the edible product. In some specific embodiments, based on the total mass of the cheese analogue product, the proportion of the composition in the cheese analogue can be 2% - 4%. In some additional specific embodiments, based on the total mass of the protein bar, the proportion of the composition in the protein bar can be 30% - 40%. In another embodiment, if the composition of the present disclosure can be eaten as is (i.e., in flakes), the proportion of the composition will be close to 100%.One of ordinary skill in the art will know how to adjust the composition ratios according to the edible product under consideration. In one embodiment, the edible product can be a liquid or a solid. In another embodiment, the edible product can be a beverage ingredient, a beverage product, a food ingredient, a food product, a feed ingredient, and / or a feed product. In some embodiments, the edible product can be a beverage ingredient and / or a beverage product. In some additional embodiments, the edible product can be, but is not limited to, a milkshake, a dairy alternative such as a milk substitute, a liquid food, a soup, a broth, a smoothie, a cream, a gravy, etc. In some embodiments, the edible product can be a food ingredient and / or a food product. In some additional embodiments, the edible product can be a baked product, a steamed / boiled product, and / or a dairy-like product. In yet another embodiment, the baked product can be, but is not limited to, bread, cake, muffin, cookie, pita, tortilla, roll, cracker, brownie, crisp, pastry, pie, tart, cupcake, pizza, etc. In yet another embodiment, the steamed / boiled product can be, but is not limited to, a stick food, a flake food, a breakfast cereal, a meat product (such as bacon, sausage, hamburger / ground steak / ground meat), a meat alternative product, a plant-based meat, pasta, noodles, a condiment, a snack, a confectionery, a fudge, a chocolate product, a ready-to-eat food, etc. In yet another embodiment, the dairy-like product can be, but is not limited to, yogurt, cheese, a cheese alternative product, a cream, a butter, a custard, an ice cream, etc. In some embodiments, the edible product can be a feed ingredient and / or a feed product. In some additional embodiments, the feed ingredient / product can be a concentrate, a roughage, or a mixed feed. In yet another embodiment, the feed ingredient / product can be, but is not limited to, a baked pellet, a feed cake, a powdered feed, an extruded product, a loaf, a chunk, a loaf-with-filling product, a pellet feed, a crumbled feed, etc.

[0066] The present disclosure provides a method for preparing an edible product. The method includes combining the composition of the present disclosure with at least one ingredient to obtain an edible product. The method can include heating the composition and at least one ingredient to obtain an edible product (e.g., a heated edible product). The method can include baking the composition and at least one ingredient to obtain an edible product (e.g., a baked edible product). The method can include blending the composition and at least one ingredient to obtain an edible product (e.g., a blended edible product). The method can include freezing the composition and at least one ingredient to obtain an edible product (e.g., a frozen edible product). The method can include extruding the composition and at least one ingredient to obtain an edible product (e.g., an extruded edible product). The method can include fermenting the composition and at least one ingredient to obtain an edible product (e.g., a fermented edible product).

[0067] Use of an edible product

[0068] In one embodiment, the present disclosure is directed to using the edible products of the present disclosure for human and / or animal nutrition. In one embodiment, the edible product is provided in the form of a substitute for an edible product based on animal or plant / legume protein. In another embodiment, the edible product is combined with an edible product based on animal or plant / legume protein. In yet another embodiment, the edible product is consumed / orally administered.

[0069] In one embodiment, the edible products of the present disclosure are intended to be combined with at least one food ingredient, at least one feed ingredient, and / or at least one beverage ingredient. In some embodiments, the yeast composition can be directly included in a food product, a feed product, and / or a beverage product. In such embodiments, the yeast composition can be mixed with feed additives, food additives, additional beverage additives, and / or binders. When the edible products of the present disclosure are intended to be used as additives, they can be provided in liquid form, and in some additional embodiments, they can be in a spray-dryable liquid form. Alternatively or in combination, the edible product can be provided in powder form, and in some additional embodiments, it can be a free-flowing powder.

[0070] In one embodiment, the edible products of the present disclosure are used as food / feed supplements. In another embodiment, the food / feed supplement is administered enterally and is intended to supplement the diet by increasing the total dietary intake of a subject, or a concentrate, metabolite, ingredient, and / or extract, compared to an unsupplemented subject receiving the same diet. Thus, the food / feed supplement can be co-administered orally with vitamins, minerals, essential fatty acids, natural products, and / or probiotics, but is not limited thereto. When the edible products of the present disclosure are intended to be used as supplements, they can be provided in liquid form, and in some additional embodiments, they can be in the form of an oil, a solution, or a spray-dryable liquid. Alternatively or in combination, the supplement can be provided in powder form, and in some additional embodiments, it can be in the form of a capsule, a pill, a tablet, a confectionery, a gummy, etc.

[0071] In one embodiment, the edible products disclosed herein are for the health, weight control, elderly and / or sports applications of humans and / or animals. In another embodiment, the edible products herein can be incorporated into a diet or a restricted diet. In the context of the present disclosure, the term "diet" should be understood to mean any kind and amount of food and drink ingested by a subject (i.e., a human and / or an animal). Still in the context of the present disclosure, the expression "restricted diet" refers to a restriction imposed on the diet of a subject (i.e., the diet of a human and / or an animal) to limit the kind and / or amount of food prescribed for ingestion. In another embodiment, when administered for a restricted diet, the edible product can be used to reduce the overall weight and / or fat mass of a subject, increase the lean body mass, muscle mass, average power, endurance and / or regulate the hormonal balance of a subject.

[0072] In one embodiment, the edible products disclosed herein can be used as meal replacements and / or medical food products. In another embodiment, the edible products of the present disclosure can be used to limit, avoid or replace the use of animal protein in the formulation of the edible product. In another embodiment, the edible product is intended for use in disease, injury and / or surgical recovery. In another embodiment, the edible products disclosed herein are intended for oral / enteral clinical nutrition.

[0073] Examples

[0074] Example 1: Method for preparing various compositions comprising yeast proteins from the genus Saccharomyces

[0075] Figure 1 A method for preparing various compositions is schematically provided. Composition A (obtained after step 020 in Figure 1 corresponds to a composition containing yeast protein prepared according to the following method. Briefly: Saccharomyces cerevisiae is cultured according to a standard fed-batch fermentation protocol, washed and separated by centrifugation to obtain yeast paste. The endogenous enzymes of the yeast are inactivated in a holding tube with a heat exchanger by heating the yeast paste at 95 °C for 4 minutes. Then the resulting slurry is spray-dried to obtain Composition A. Composition B (obtained after step 050 in Figure 1 corresponds to a composition containing yeast protein. Briefly: Saccharomyces cerevisiae is cultured to obtain the yeast paste as described above. The endogenous enzymes of the yeast are inactivated by heating the yeast paste at 95 °C for 4 minutes. Then the resulting slurry is cooled to 60 °C, and the pH is adjusted to pH 5.5, and then β-1,3-glucanase treatment (i.e., Denazyme GEL TM, Nagase; based on the dry matter of yeast, at 0.5%) for at least 4 hours. The enzymatic reaction was stopped by heating the hydrolysis product to 95 °C for at least 2 minutes (this step also serves as a pasteurization step). The resulting insoluble fraction and soluble fraction were separated by centrifugation, and the insoluble fraction was collected and then spray-dried at 85 °C to obtain Composition B. The soluble fraction thus obtained (obtained after step 040 in Figure 1 was also spray-dried to obtain Composition C.

[0076] Example 2: Comparison of Compositions A, B and C

[0077] The protein content (via Kjeldahl nitrogen analysis, with 6.25 as the coefficient), lipid content (via the modified Mojonnier method; AOAC 989.05), nucleic acid content (via the method of Fish et al. (1991); comparison of values with and without enzymatic digestion of nucleic acids); total carbohydrate content (monosaccharide titration by HPLC after enzymatic / chemical digestion; AOAC 980.13), mannan content (mannose titration by HPLC after enzymatic / chemical digestion), glucan content (glucose titration by HPLC after enzymatic / chemical digestion), and glucose content (direct glucose titration by HPLC) of Compositions A, B, and C were determined. These compositions (i.e., A, B, and C) were obtained from Example 1 and were characterized according to standard methods known to those of ordinary skill in the art in the examples herein. The characterization results are provided in Table 1. Briefly, Composition B contains more protein but less total carbohydrate than Compositions A and C. This observation is consistent with the microscopic analysis of Composition B, revealing protoplast formation. (Data not shown). Neither the lipid content nor the nucleic acid content of Composition C was reported.

[0078] Table 1. Characterization results of Compositions A, B, and C. For each column, the component percentages are based on the total mass of the dried composition.

[0079]

[0080] Example 3: Comparison of Compositions A and B

[0081] The Kjeldahl nitrogen analysis method (i.e., the standard method for the quantitative determination of the protein content in organic substances) was used to determine and compare the protein contents of Compositions A and B obtained from the method described in Example 1. Briefly, in the presence of copper sulfate as a catalyst, Compositions A and B (0.25 g) were heated with concentrated sulfuric acid to 373 °C. The purpose of this oxidation reaction was to decompose the sample while releasing the reduced nitrogen in the form of ammonium sulfate. The resulting solution was then distilled with sodium hydroxide to release ammonia, which was further reacted with boric acid before being dissolved in distilled water. The basic product was titrated with hydrochloric acid. Both the Tashiro indicator and pH were used to determine the equivalence point (i.e., pH 4.9). Based on the titration results, a factor of 6.25 was used to determine the amount of nitrogen in the sample, and thus the protein content was calculated.

[0082] The results of the Kjeldahl nitrogen analysis of Samples A and B are presented in Table 2 below. Composition A had a protein content of approximately 60%, while Composition B had a protein content of approximately 80%, both based on the total mass of the analyzed composition (dry or non-dry). The sample (Composition B) subjected to dextranase treatment had approximately 20% more protein content than its non-enzyme-treated homolog (Composition A).

[0083] Table 2. Results of the Kjeldahl nitrogen analysis of Compositions A and B. Dw b% represents the percentage of protein content based on the total dry composition mass.

[0084]

[0085] The water solubility index (WSI) of two compositions (i.e., A and B) containing yeast protein was determined and compared herein. For each sample, 1 g of the dry powder was suspended in 10 mL of distilled water. The resulting suspension was gently mixed at room temperature for 30 minutes and then centrifuged at 3000 × g for 15 minutes at 25 °C. The remaining supernatant was collected and decanted to obtain the soluble fraction and some insoluble components. The soluble fraction was discarded, and the insoluble components were dried before weighing. WSI represents the mass of the dried insoluble components expressed as a percentage of the original sample mass.

[0086] The results of the WSI determination for both Samples A and B are presented in Table 3 below. Composition A had a WSI of approximately 23%, while Composition B had a WSI of approximately 5%, both based on the dry weight of the test composition. The solubility of the sample (Composition B) subjected to dextranase treatment was approximately 5 times lower than that of its non-enzyme-treated homolog (Composition A).

[0087] Table 3. Results of the water solubility index (WSI) of Samples A and B. Dw b% represents the percentage of protein content based on the total dry composition mass.

[0088] Sample name Dry weight % WSIDwb % A 98.4 23.2 B 96.5 5.3

[0089] The emulsifying activity and emulsion stability of two compositions containing yeast proteins (i.e., Composition A and B) were determined and compared with lecithin (i.e., the standard emulsifier in the art). Emulsion activity (EA) was defined as the maximum amount of oil that could be emulsified per unit amount of the dried composition. Emulsion stability (ES) was defined as the rate of phase separation in water and oil during emulsion storage. The Brishti et al. (2017) and Yasumatsu et al. (1972) methods were used to determine EA and ES, respectively. Briefly: Each sample (i.e., Composition A, Composition B, and lecithin; 0.24 g) was resuspended in 12 mL of distilled water and 12 mL of sunflower oil in a 50 mL centrifuge tube. The resulting mixture was homogenized for 1 minute with a homogenizer and then centrifuged at 1100×g for 5 minutes at 20 °C.

[0090] The emulsifying activity (EA) of two samples was calculated according to the following formula: EA = (H1 / H0) × 100; where H1 refers to the measured emulsion volume and H0 refers to the total volume of the solution in the tube measured.

[0091] The emulsion stability (ES) of two samples was determined after i) heating the above emulsion at 80 °C for 30 minutes, ii) cooling the emulsion with tap water, and iii) centrifuging the emulsion at 1100×g for 5 minutes at 20 °C. ES was calculated using the following formula: ES = (H1 / H0) × 100; where H1 refers to the measured emulsion volume and H0 refers to the total volume of the solution in the tube measured.

[0092] The results of the emulsifying properties (i.e., EA and ES) are presented in Table 4 below. The EA of Composition A was comparable to that of lecithin, while the sample treated with dextranase (Composition B) had an activity 4 to 5 times lower than that of its non - enzyme - treated homolog (Composition A) and lecithin. The ES of Composition A was 3 to 4 times lower than that of lecithin but 3 to 4 times higher than that of its enzyme - treated counterpart (Sample B). Thus, Composition B had an emulsion stability 10 to 11 times lower than that of lecithin.

[0093] Table 4. Results of the emulsifying activity (EA) and emulsion stability (ES) of Compositions A and B compared with lecithin (n = 2).

[0094] Sample name EA % EA standard deviation % ES % ES standard deviation % Sample A 45.7 2.7 16.1 0.0 Sample B 11.5 0.0 4.5 1.9 Lecithin 50.4 1.2 51.4 0.8

[0095] The foaming capacity (FC) and foam stability (FS) of two compositions containing yeast proteins (i.e., Composition A and Composition B) were determined according to the methods provided by Chandra et al. (2015) and Brishti et al. (2017), respectively. Briefly: In a 50 mL centrifuge tube, each sample (Composition A and Composition B; 0.2 g) was resuspended in 20 mL of distilled water. The suspension thus obtained was homogenized using a homogenizer and then whipped for 1 minute before measurement. The foaming capacity (FC) of each sample was calculated according to the following formula: FC = [(V2 - V1) / V1] × 100; where V1 refers to the measured volume of the suspension before the whipping step and V2 refers to the measured volume of the whipped suspension.

[0096] The foam stability (FS) of each sample was measured over time (0 minutes, 15 minutes, 30 minutes, 45 minutes, and 60 minutes after the suspension was whipped) and calculated according to the following formula: FS = (VF t / VF t0 ) × 100; where VF t refers to the foam volume measured at a specific time after whipping, and VF t0 refers to the foam volume measured immediately after whipping (i.e., 0 minutes).

[0097] The results of the foaming properties (i.e., FC and FS) are presented in Table 5 below. The sample treated with dextranase (Composition B) had a foaming capacity (FC) that was approximately 3 to 4 times lower than that of its non-enzyme-treated homolog (Composition A). In other words, Composition A was 3 to 4 times more efficient in forming foam than its dextranase-treated counterpart. The foam stability study showed that the foam based on Composition A retained approximately half of its stability after 30 minutes, while the foam formed by the dextranase-treated composition (Composition B) retained only 10% after the same time. Therefore, the stability of the foam derived from Composition B was approximately 4 to 5 times lower than that of the foam prepared when the composition was not treated with dextranase (Composition A).

[0098] Table 5. Results of the foaming capacity (FC) and foam stability (FS) of Samples A and B (N = 2).

[0099]

[0100] The odor and taste of two compositions containing yeast proteins (i.e., Composition A and Composition B) were evaluated by trained assessors and compared with F85M (pea protein isolate; Roquette). The samples (Composition A, Composition B, and F85M was suspended in hot water to achieve a 2% w / w suspension. The 2% w / w suspension (40 mL) was thermally supplied to each assessor in a glass container for sensory analysis (N = 2, n = 2).

[0101] The all applicable terms scoring (RATA) method was used to determine the sensory characteristics of each sample, allowing participants to select relevant terms from a given list and rate their intensity. During the evaluation of the samples, two main sensory modalities were investigated, namely odor and taste. For this purpose, the following parameters were evaluated: overall intensity (mandatory), sourness, cheesy flavor, creamy flavor, vegetable flavor, chicken / poultry flavor, beef flavor, baked flavor, bread flavor, fermented / alcoholic flavor, and off / altered flavor. In addition, the taste modality also included umami, saltiness, sweetness, bitterness, and astringency attributes. To measure the intensity, a numerical scale of 1 to 9 was applied, where 1 refers to "very low"; 5 refers to "medium" and 9 refers to "very strong". During the evaluation, comments could also be added to all modalities if needed.

[0102] The results of these sensory determinations are depicted in Figure 2. F85M (Figure 2C) can be distinguished by strong vegetable notes (pea odor and taste), bitterness, and astringency. Composition A (Figure 2A) was evaluated as having strong odor and taste characteristics, with strong baked and beef flavors, accompanied by chicken / poultry flavor. Compared to the other samples, Composition B (Figure 2B) was much lower in overall intensity and more neutral (i.e., presented the lowest average score). None of the samples had cheesy flavor (odor), fermented flavor (taste), off-odor, or altered flavor, and therefore, these attributes were excluded from Figure 2A, Figure 2B, and Figure 2C.

[0103] Viscosity measurements of two samples (Composition A and B) were also carried out according to a slightly modified method of Onwulata et al. (2014). Briefly: Each sample (2.7 g) was suspended in 27.3 mL of ddH2O, homogenized, and viscosity measurements were performed using a rheometer Anton Paar Physica MCR301, program RheoCompass. RVA was performed using a temperature ramp as the test type. The temperature profile implemented was as follows: 2 minutes at 25 °C → linear heating from 25 °C to 85 °C for 5 minutes → 5 minutes at 85 °C → linear cooling from 85 °C to 25 °C for 5 minutes → 2 minutes at 25 °C. The results of the viscosity measurements of the two samples are presented in Table 6. Regardless of the test conditions, the viscosity of Composition B was slightly lower than that of its non-enzyme-treated counterpart.

[0104] Table 6.Viscosity measurement results of both Composition A and Composition B using a temperature profile of 2 minutes at 25°C → 5 minutes of linear temperature increase from 25°C to 85°C → 5 minutes at 85°C → 5 minutes of linear temperature decrease from 85°C to 25°C → 2 minutes at 25°C.

[0105]

[0106] Protein Digestibility Corrected Amino Acid Score (PDCAAS) is a method for evaluating the quality of proteins based on human amino acid requirements and their digestibility. Therefore, the PDCAAS method was performed to evaluate the digestibility of Compositions “A” and “B” (data not shown). The PDCAAS of Composition B (which is higher than 1.0) is higher than the PDCAAS of one of its non - enzyme - treated counterparts (i.e., Composition A).

[0107] Example 4: Alkaline extraction optimization

[0108] It was determined whether the alkali extraction step could be used to extract nucleic acids and their derivatives from yeast extract. Various pH values and temperatures were used to determine their effect on dissolving nucleic acids outside yeast cells.

[0109] Yeast extract of Saccharomyces cerevisiae with an initial dry matter of 17.72% and an initial protein content of 60.95% was obtained. The pH of yeast extract samples (40 g) was adjusted to 8.0, 9.0, 10.0, or 11.0. Each sample was incubated at different temperatures (4°C, room temperature, 55°C, or 65°C) for 2 hours to generate different samples.

[0110] Each sample was centrifuged at 4500 rpm for 15 minutes, and the supernatant was further characterized. No washing step was applied. The supernatant was spray - dried. The DW of the supernatant was determined using a halogen dryer (moisture analyzer, MA 37 - 1US, Sartorius). α - Amino nitrogen (AAN) and ribonucleic acid (RNA) were determined using high - performance liquid chromatography.

[0111] Table 7 shows the AAN and RNA yields found in the supernatant after various alkali extractions. As shown in Table 7, one of the compounds that was better extracted under alkaline conditions is RNA, as indicated by the strong correlation between extraction pH and RNA recovery. When compared to extractions at lower temperatures, the RNA content was significantly higher at 55°C and 65°C, reaching 23.5% in the extractions at 55°C and pH 10 and at 65°C and pH 9 (Table 7). In the supernatants produced at 55°C / pH 11, 65°C / pH 10, and 65°C / pH 11, the RNA content was lower than that in the extractions at lower pH at the same temperature (Table 7).

[0112] Table 7.AAN and RNA yields in the supernatant of samples subjected to alkali extraction. Super. = supernatant,

[0113]

[0114] Example 5 Combination of alkaline extraction and dextranase treatment

[0115] Then it was determined whether the alkali extraction step could be used to reduce the nucleic acid content of the protein composition before dextranase treatment.

[0116] Yeast extract of Saccharomyces cerevisiae containing 16.0% solids, 63.47% protein, 4.06% phosphate and 7.90% RNA was collected. The yeast extract was heat-inactivated at 95 °C for 5 minutes in an autolyser (Bailun, 20 L, 200 rpm), diluted to obtain 15% dry matter (DM) and subjected to an alkali extraction process (65 °C, pH 9.0, 2 h). The presence of protoplasts (microscopic method) and dry matter (DM) in the samples obtained after alkali treatment was analyzed using a halogen dryer (moisture analyzer, MA 37-1US, Sartorius). Then the samples obtained after alkali extraction were centrifuged at 4500 rpm for 10 minutes using a Sigma centrifuge (model 4-5L, rotor: 11650) to produce a soluble fraction and an insoluble fraction. The insoluble fraction was washed once to a volume ratio of 1:1 and centrifuged again. The washed insoluble fraction was subjected to dextranase treatment (Denazyme GEL, 60 °C, pH 5.6, 5 h). The soluble fraction and the enzyme-treated insoluble fraction were spray-dried and further analyzed.

[0117] α-Amino nitrogen (AAN) was determined by spectrophotometry (adapted from the EBC-ninhydrin method for the determination of free α-amino nitrogen). High performance liquid chromatography was used to determine ribonucleic acid (RNA), dextran and mannan. The protein yield was determined using the Kjeldahl method. The AN / TN ratio was calculated by dividing the AAN content by the total nitrogen (e.g., protein content divided by 6.25).

[0118] After alkali extraction and dextranase treatment, the presence of protoplasts was confirmed (data not shown). Under the experimental conditions tested, the alkali extraction step was used to reduce the nucleic acid content in the yeast extract to below 2% (see Table 8), while maintaining the protein content above 80% (Table 9).

[0119] Table 8. RNA mass balance (% w / w) in various materials.

[0120]

[0121] Table 9. Characteristics of the insoluble and soluble fractions.

[0122]

[0123] Example 6: Composition B in a vegan cheese American mozzarella style product

[0124] It was decided to characterize the sensory and functional properties of a composition containing yeast protein prepared according to the method claimed in the present disclosure (i.e., Composition B, as in the above examples) as a supplement to pea protein in a vegan cheese type product. Two trials were conducted: i) vegan cheese based on pea protein (3.59%) and ii) partial replacement of pea protein with a vegan cheese based on "2.74% Composition B + 0.85% Pea Protein", while all other components remained unchanged. The complete vegan cheese recipe is provided in Table 10 below. No colorants or opacifiers were included in the recipe.

[0125] Table 10. Firm, shreddable mozzarella style chunks recipe.

[0126]

[0127] Color evaluation was performed by visually analyzing chunks and pieces. The pea protein vegan cheese exhibited a creamy yellow hue, while its partially substituted counterpart (mixture of Composition B and pea protein) was a light earthy beige.

[0128] The sensory and textural properties were evaluated by cutting samples of both cheese analogs into 20 mm cubes after removal from the refrigerator (4°C). The flavor and texture characteristics were determined by trained assessors. The results are provided in Table 11 below.

[0129] Table 11. Flavor and texture characterization of vegan mozzarella cheese analogs, namely pea protein vegan cheese and “pea protein + composition B” vegan cheese.

[0130]

[0131] Partial replacement of pea protein with a mixture of pea protein + composition B did not affect the flavor profile (ie, neutral impact). However, chunks prepared with the claimed composition were slightly softer than their pea protein analogs, even though the chunks were similarly chopped.

[0132] Example 6: High moisture extrusion cooking of meat analog containing Composition B

[0133] It was decided to evaluate the effect of adding Composition B powder on plant-based artificial meat prepared by high-moisture extrusion cooking. During high-moisture extrusion, a powder blend and water (about 50%-70% of the total mass) were continuously metered into an extruder where the material was rapidly heated to 150 °C and the co-rotating screws mechanically mixed and sheared the mass. As a result, the dough was transformed into a flowing melt. At the end of the barrel, the melt was pushed by the screw into a long cooling tunnel that compacted the material and prevented water boiling by cooling the mass to below 100 °C and providing backpressure. Friction between the solidified layers of the flowing material within the cooling tunnel generated long protein fibers similar to the structure of meat.

[0134] As shown in Table 12, different proportions of Composition B were added to the dry blend, thus partially replacing the legume protein (i.e., H5, H10, H15, H25, and H35). H0 did not contain any Composition B, while H100 contained only Composition B.

[0135] Table 12. The amounts of different proteins contained in the samples of various artificial meats.

[0136]

[0137] As described below, artificial meat extrudates were prepared from all the tested blends. Samples were processed in a co-rotating intermeshing twin-screw extruder KETSE 20 / 40 (Brabender GmbH, Duisburg, Germany) with a long cooling die (24×7×700 mm, W×H×L). The screw with a length-to-diameter ratio of 40 was configured to apply medium shear force. The mass flow rate of the material was set to a constant 4 kg h by calibrating a volumetric feeder with the powder. -1 Water was added through a calibrated peristaltic pump through a separate port. The temperature profile was set to 45 °C, 83 °C - 84 °C, 135 °C - 137 °C, and 151 °C - 154 °C. The cooling die temperature was maintained at 65 °C with a tempering unit, but only for the last 300 mm, thus creating a temperature gradient along the die. The moisture content of each blend was adjusted, and when the process reached stability, a sample was collected from each blend, as evidenced by the measured pressure and temperature. The screw speed and temperature were slightly adjusted between blends to maintain process stability. The samples were packed into zipper bags and frozen at -20 °C until analysis.

[0138] Instrument texture profile analysis: The frozen samples were thawed and rehydrated in water at 60 °C for 2 h (without temperature maintenance). Before measurement, these slices were blotted dry with paper towels. Texture profile analysis (TPA) was performed using a TA.XTplusC texture analyzer (Stable Micro-Systems, Godalming, UK) equipped with a 75 mm flat probe and a 5 kg load cell. The extruded slices were cut into a shape of 15×15×7 mm and compressed twice at a probe speed of 3 mm s -1 and a pre-test speed of 1.5 mm s -1 with a hold time between compressions of 1 s, 70% each time. The load cell was 50 kg. Hardness, chewiness, cohesiveness, springiness, and resilience were calculated by the texture analyzer software.

[0139] TPA of the extrudates revealed differences in the mechanical properties of the extrudates, which are summarized in Figure 3 . Sample H100 was not measured because its hardness was too high for the testing equipment. All tested extrudate parameters (i.e., hardness, chewiness, cohesiveness, springiness, and resilience) of the extrudates based on composition B (H5, H10, H15, H25, and H35) were higher than those of the control (H0).

[0140] Descriptive sensory analysis (DSA): The frozen samples were thawed and rehydrated in water at 60 °C for 2 h (without temperature maintenance). Before sensory evaluation, these slices were blotted dry with paper towels and cut into slices 4 cm long. Sensory analysis was performed by eight expert assessors who had been previously trained and had experience with such extruded plant-based samples. The analysis was carried out in a standard (ISO 6668:2008) sensory room. The samples were coded with three-digit codes and placed in cups. The order of the samples was randomized according to the Williams Latin square experimental design. The samples were presented at room temperature. Water and crackers were provided between samples for palate cleansing. The samples were evaluated on a scale of 0 - 9 ("0" - none; "1" - very weak; "5" - moderate; "9" - very strong). A total of three modalities were evaluated: odor, taste, and texture. For odor and taste, the sensory analysis included attributes such as overall intensity, bean odor, yeast flavor (odor) / umami (taste), and off-flavors. The taste modality additionally included aftertaste. Texture characteristics included evaluations of fibrousness, elasticity, hardness, chewiness, adhesiveness, particle size, cohesiveness, and moisture. Fibrousness was evaluated only by hand, and the others were evaluated by mouthfeel. Assessors could also add comments to the voluntary text boxes for each modality. The DSA results of different extrudates are depicted in Figure 4 and Figure 5 . When the yeast addition was up to 35%, the observed differences were very small. However, according to all attributes, the 100% composition B extrudate (H100) was significantly different from the other extrudates. As Figure 4As shown, Composition B has a mild flavor, and when added to samples H5 - H35, it only slightly increases the meaty / yeasty and umami flavors compared to the control (H0), but this increase is much greater in H100. On the other hand, it has been commented that H35 has a subtle roasted odor, which may be related to a higher yeast content compared to H5 - H25, although the meaty intensity is similar to that of H25. The bean odor and flavor, as well as the overall odor and flavor intensity, are not different among H0 - H35. However, H100 has a stronger overall odor and flavor. Off - flavors, taints, and aftertastes are detected only in H100. The off - flavors described for H100 include "earthy" and "paper - like" flavors. As Figure 5 shown, there are no differences in the texture properties (such as adhesiveness, cohesiveness, fibrousness, and elasticity) among samples H0 - H35, except for H100, which is more cohesive and elastic but less fibrous. Other properties have more distinct differences: compared to the control (H0), the chewiness, hardness, and granularity are increased in samples H5 - H35.

[0141] Example 7: Method for preparing a composition comprising yeast proteins from the genus Cyberlindnera

[0142] Saccharomyces cerevisiae var. boulardii was cultured to obtain yeast extract paste. The endogenous enzymes of the yeast were inactivated by heating the yeast extract paste at 95 °C for 5 minutes. Then the resulting slurry was cooled to 65 °C, and the pH was adjusted to pH 9.0. These conditions were maintained for 30 minutes before reducing the pH to 8.0 for alkaline extraction. The resulting insoluble fraction and soluble fraction were separated by centrifugation, and the insoluble fraction was collected and then treated with β - 1,3 - glucanase (i.e., Denazyme GELTM, Nagase; 0.5% based on dry yeast matter) at 55 °C and pH 5.5 for 5 hours. The enzyme reaction was stopped by heating the hydrolysate to 95 °C for at least 2 minutes (this step also serves as a pasteurization step). The resulting insoluble fraction and soluble fraction were separated by centrifugation, and the insoluble fraction was collected and then spray - dried at 85 °C. The protein content was determined by Kjeldahl nitrogen analysis with 6.25 as the coefficient. The obtained yeast extract contains 83.97% protein.

[0143] Although the present invention has been described in connection with specific embodiments of the invention, it should be understood that the scope of the claims should not be limited by the preferred embodiments described in the examples, but rather should be given the broadest interpretation consistent with the overall description.

[0144] References

[0145] [1]AOAC 980.13 - 1980; Name: Fructose, Glucose, Lactose, Maltose, and Sucrose in Milk Chocolate - Liquid Chromatography Method

[0146] [2]AOAC 989.05 - 1992; Name: Fat in Milk Modified Mojonnier Ether Extraction Method

[0147] [3]Brishti F.H. et al., (2017) Evaluation of the functional properties of mung bean protein isolate for development of textured vegetable protein. International Food Research Journal, 24(4). pp. 1595 - 1605. ISSN 1985 - 4668; ESSN: 2231 - 7546

[0148] [4]Chandra S. et al., (2015) Evaluation of functional properties of composite flours and sensorial attributes of composite flour biscuits. J Food Sci Technol. 52(6). pp. 3681 - 3688. doi:10.1007 / s13197 - 014 - 1427 - 2. Electronic publication June 10, 2014. PMID: 26028751; PMCID: PMC4444897

[0149] [5]Fish W.W. et al., (1991) A method for the quantitation of 5'-mononucleotides in foods and food ingredients. Journal of Agricultural and Food Chemistry. 39(6), pp. 1098 - 1101. DOI: 10.1021 / jf00006a019

[0150] [6]Onwulata C.I. et al., (2014) Rapid visco analysis of food protein pastes. Journal of Food Processing and Preservation. 38, pp. 2083 - 2089

[0151] [7]Yasumatsu K. et al., (1972) Whipping and Emulsifying Properties of Soybean Products, Agricultural and Biological Chemistry, 36(5), pp. 719 - 727, DOI: 10.1080 / 00021369.1972.10860321。

Claims

1. A method for obtaining a composition containing yeast protein, wherein the method comprises the following steps: a) Provide a yeast extract containing yeast; b) Inactivate the endogenous enzymes of the yeast to provide an inactivated yeast extract; c) Subject the inactivated yeast extract to an enzymatic treatment to obtain an insoluble fraction and a soluble fraction containing yeast protoplasts; wherein the enzymatic treatment comprises at least one polypeptide having glucanase activity; and wherein the enzymatic treatment lacks ribonuclease activity; d) Separate the insoluble fraction from the soluble fraction; and e) Collect the insoluble fraction, wherein the collected insoluble fraction is a composition containing the yeast protein when dried and has a protein content equal to or higher than 60% based on the total mass of the dried collected insoluble fraction.

2. The method according to claim 1, wherein the inactivated yeast paste is obtained by exposing the yeast paste to a temperature of 65°C to 100°C for a time of 30 seconds to 5 hours.

3. The method according to claim 1 or 2, wherein the enzymatic treatment is carried out at a temperature of 20°C to 80°C for a time of 1 hour to 24 hours.

4. The method according to any one of claims 1 to 3, wherein the composition has a neutral taste.

5. The method according to any one of claims 1 to 4, the method further comprising drying the collected insoluble fraction in step e) to provide the composition.

6. The method according to claim 5, wherein the dried insoluble matter has: · a lipid content of less than 20% based on the total mass of the dried collected insoluble fraction; · a nucleic acid content of more than 6% based on the total mass of the dried collected insoluble fraction; · a carbohydrate content of less than 25% based on the total mass of the dried collected insoluble fraction; · a mannan content of less than 6% based on the total mass of the dried collected insoluble fraction; · a glucan content of less than 10% based on the total mass of the dried collected insoluble fraction; and / or · a glucose content of less than 25% based on the total mass of the dried collected insoluble fraction.

7. The method according to any one of claims 1 to 4, the method further comprising, after b) and before c), an alkali extraction step on the inactivated yeast paste.

8. The method according to claim 7, the method further drying the collected insoluble fraction in step e) to provide the composition.

9. The method according to claim 8, wherein the dried insoluble fraction has: · a lipid content of less than 20% based on the total mass of the dried collected insoluble fraction; · A nucleic acid content of less than 3% based on the total mass of the dried collected insoluble fraction; · A carbohydrate content of less than 25% based on the total mass of the dried collected insoluble fraction; · A mannan content of less than 6% based on the total mass of the dried collected insoluble fraction; · A glucan content of less than 10% based on the total mass of the dried collected insoluble fraction; and / or · A glucose content of less than 25% based on the total mass of the dried collected insoluble fraction.

10. The method according to any one of claims 1 to 9, wherein the yeast is from the genus Saccharomyces, Komagataella, Pichia, Candida, Kluyveromyces, Yarrowia, Cyberlindnera (Torulaspora), Wickerhamomyces or a combination thereof.

11. A composition comprising yeast protein derived from yeast protoplasts and having: · A protein content of equal to or higher than 60% based on the total mass of the composition; · A lipid content of less than 20% based on the total mass of the composition; · A carbohydrate content of less than 25% based on the total mass of the composition; · A mannan content of less than 6% based on the total mass of the composition; · A glucan content of less than 10% based on the total mass of the composition; and / or · A glucose content of less than 25% based on the total mass of the composition.

12. The composition according to claim 11, wherein the composition has a nucleic acid content of higher than 6% based on the total mass of the composition.

13. The composition according to claim 11, wherein the composition has a nucleic acid content of less than 3% based on the total mass of the composition.

14. The composition according to any one of claims 11 to 13, wherein the composition has a neutral taste.

15. The composition according to any one of claims 11 to 14, wherein the composition can be obtained by or is obtained by the method according to any one of claims 1 to 10.

16. An edible product comprising the composition according to any one of claims 11 to 13 and at least one additional ingredient, wherein the composition accounts for at least 1% w / w based on the total mass of the edible product.

17. The edible product according to claim 16, wherein the edible product is a beverage, a milk shake, a bar food, a meat product or a baked product.

18. Use of the edible product according to claim 16 or 17 in human and / or animal nutrition.

19. Use according to claim 18, wherein the edible product is provided in the form of a substitute for an edible product based on animal or plant / legume protein, or in combination with an edible product based on animal or plant / legume protein.

20. Use according to claim 16 or 17, wherein the edible product is a food / feed supplement or a food / feed additive.

21. Use according to claim 20, wherein the food / feed supplement or the food / feed additive is for weight control, the elderly, oral / enteral clinical nutrition, sports applications and / or animal nutrition.

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