High-protein yeast products

Abstract

This disclosure relates to a method for obtaining a composition having yeast protein derived from yeast protoplasts. This disclosure also includes compositions containing yeast protein derived from yeast protoplasts, edible products derived from such compositions, and their uses.

Description

[Technical Field]

[0001] Cross-referencing of related applications (multiple) and related documents (multiple) This patent application claims priority to U.S. Provisional Patent Application No. 63 / 381,353, filed on 28 October 2023, which is incorporated herein by reference in its entirety.

[0002] This disclosure relates to a yeast protein having a neutral taste, a composition thereof, and a method for producing the same. [Background technology]

[0003] Proteins are biomacromolecules that are essential components of animal nutrition. When protein is ingested, it is broken down by proteases into polypeptides, and then into amino acids. These amino acids then pass through the intestinal barrier to reach the bloodstream, and are then distributed throughout the body to perform their vital functions.

[0004] In human and / or animal nutrition, the main sources of protein are animal protein, plant protein (including legume protein), and fungal protein. Consumption of animal protein has steadily increased over the past 50 years, leading to environmental and consumer health problems. Therefore, the current nutritional challenge is to reverse the global trend in protein consumption, namely, to reduce the proportion of animal protein and increase the proportion of plant / legume protein (e.g., pea protein) and fungal protein, such as that found in yeast extracts.

[0005] However, proteins derived from yeast extracts, as well as plant / legume proteins, have a distinctive taste, which limits their concentration and, consequently, their uses. This disclosure aims to address these issues by providing a source of protein with a neutral taste. [Overview of the project]

[0006] According to a first aspect, the present disclosure provides a method for obtaining a composition having yeast protein. Broadly speaking, the method comprises: a) providing a yeast cream containing yeast; b) inactivating the endogenous enzymes of yeast to provide an inactivated yeast cream; c) subjecting the inactivated yeast cream to enzymatic treatment to obtain an insoluble fraction containing yeast protoplasts and a soluble fraction; d) separating the insoluble fraction from the soluble fraction; and e) collecting the insoluble fraction, wherein the collected insoluble fraction, when dried, is a composition containing yeast protein and has a protein content of 60% or more based on the total mass of the collected and dried insoluble fraction. In step c), the enzymatic treatment comprises at least one polypeptide having glucanase activity, and the enzymatic treatment lacks ribonuclease activity. In one embodiment, an inactivated yeast cream is obtained by exposing the yeast cream to a temperature of 65 to 100°C for a period of time from 30 seconds to 5 hours. In one embodiment, the enzymatic treatment is carried out at a temperature of 20-80°C for a period of 1-24 hours. In another embodiment, the composition has a neutral taste. In some embodiments, the method further comprises drying the insoluble fraction collected in step e) to provide the composition. In some other embodiments, the dried insoluble fraction has a lipid content of less than 20% based on the total mass of the collected and dried insoluble fraction, a nucleic acid content of more than 6% based on the total mass of the collected and dried insoluble fraction, a carbohydrate content of less than 25% based on the total mass of the collected and dried insoluble fraction, a mannan content of less than 6% based on the total mass of the collected and dried insoluble fraction, a glucan content of less than 10% based on the total mass of the collected and dried insoluble fraction, and / or a glucose content of less than 25% based on the total mass of the collected and dried insoluble fraction. In some embodiments, the method may further comprise subjecting the inactivated yeast cream to an alkaline extraction step after b) and before c). In some further embodiments, the method may further include drying the insoluble fraction collected in step e) (subjected to the alkaline extraction step) to provide a composition.In such embodiments, the dried insoluble fraction may have a lipid content of less than 20% based on the total mass of the collected and dried insoluble fraction, a nucleic acid content of less than 3% based on the total mass of the collected and dried insoluble fraction, a carbohydrate content of less than 25% based on the total mass of the collected and dried insoluble fraction, a mannan content of less than 6% based on the total mass of the collected and dried insoluble fraction, a glucan content of less than 10% based on the total mass of the collected and dried insoluble fraction, and / or a glucose content of less than 25% based on the total mass of the collected and dried insoluble fraction. In further embodiments, the yeast is derived from the genera Saccharomyces, Komagataella, Pichia, Candida, Kluyveromyces, Yarrowia, Cyberlindera (Torula), Wickerhamomyces, or a combination thereof.

[0007] According to a second aspect, the disclosure provides a composition comprising yeast protein derived from yeast protoplasts, and having a protein content of 60% or more 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. In some embodiments, the composition may have a nucleic acid content of more than 6% based on the total mass of the composition. In other embodiments, the composition may have a nucleic acid content of less than 3% based on the total mass of the composition. In one embodiment, the composition has a neutral taste. In one embodiment, the composition may be obtained by or has been obtained by a method for obtaining a composition having yeast protein.

[0008] According to a third aspect, the disclosure provides an edible product comprising a composition and at least one further ingredient, wherein the composition provides at least 1% w / w based on the total mass of the edible product. In one embodiment, the edible product is a beverage, a shake, a bar, a meat product, or a baked product.

[0009] According to a fourth aspect, the disclosure provides the use of edible products in human and / or animal nutritional supplementation. In one embodiment, the edible product is provided as a substitute for or in combination with animal protein or plant / legume protein-based edible products. In one embodiment, the edible product is a food / feed supplement or food / feed additive. In another embodiment, the food / feed supplement or food / feed additive is for weight management, elderly, oral / enteral clinical nutritional supplementation, sports use and / or animal nutritional supplementation. [Brief explanation of the drawing]

[0010] Having described the properties of the present invention in general terms, we now refer to the attached drawings which illustrate preferred embodiments thereof. [Figure 1] Embodiments of a method for producing a composition containing yeast protein (Composition C) and comparative products (Composition A and Composition B) are shown. [Figure 2A1] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 2A2] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 2B1] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 2B2] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 2C1] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 2C2] The sensory characteristics (i.e., smell and taste) based on the Rate-All-That-Apply (RATA) method applied to Composition A (Figures 2A1 and 2A2), Composition B (Figures 2B1 and 2B2), and Nutralys® F85M (Figures 2C1 and 2C2) are shown. * indicates essential items obtained in the sensory evaluation of the composition. [Figure 3] Figures 3A–3E show the texture profile analysis (TPA) of extruded materials (i.e., H0, H5, H10, H15, H25, and H35). Results for (A) chewiness, (B) cohesiveness, (C) hardness, (D) resilience, and (E) elasticity are shown. Vertical error bars indicate 95% confidence intervals, and statistically significant differences are observed between samples that do not share the same letter (Kruskal-Wallis test, p<0.05). H100 was too strong to be measured. [Figure 4]Figures 4A to 4I show the descriptive sensory analysis (DSA) of extruders (i.e., H0, H5, H10, H15, H25, H35, and H100) evaluated for odor (O) and taste (T). Results are shown for (A) bean-like odor, (B) meat-like and yeast-like odor, (C) off-odor and (D) overall intensity, as well as (E) aftertaste intensity, (F) bean-like taste, (G) off-odor intensity, (H) overall intensity, and (I) umami. Vertical error bars indicate 95% confidence intervals, and statistically significant differences are observed between samples that do not share the same letter (Kruskal-Wallis test, p<0.05). [Figure 5] Figures 5A to 5H show the descriptive sensory analysis (DSA) of extruded materials (i.e., H0, H5, H10, H15, H25, H35, and H100) whose texture attribute (×) was evaluated. The results are shown as attributes: (A) adhesion, (B) chewiness, (C) cohesiveness, (D) fibrous texture, (E) granular texture, (F) hardness, (G) wettability, and (H) elasticity. Vertical error bars indicate 95% confidence intervals, and statistically significant differences are observed between samples that do not share the same letter (Kruskal-Wallis test, p<0.05). [Modes for carrying out the invention]

[0011] Method for obtaining a composition containing yeast protein In one embodiment, the present disclosure aims to provide a method for obtaining a composition having yeast protein. The method of the present disclosure includes a) providing a yeast cream containing yeast; b) inactivating the endogenous enzymes of yeast to provide an inactivated yeast cream; c) subjecting the inactivated yeast cream to enzymatic treatment to obtain an insoluble fraction containing yeast protoplasts and a soluble fraction; d) separating the insoluble fraction from the soluble fraction; and e) collecting the insoluble fraction.

[0012] In the context of this disclosure, the term “yeast” refers to a eukaryotic single-celled microorganism belonging to the Kingdom of Fungi. Suitable yeasts that can be used in the method for obtaining the composition may be derived, for example, from the genera Saccharomyces, Komagataella, Pichia, Candida, Kluyveromyces, Yarrowia, Cyberlindnera (Torula), Wickerhamomyces, or combinations thereof. Suitable yeast species that can be used in the method for obtaining the composition include, but are not limited to, budding yeast (Saccharomyces cerevisiae), Torula yeast (Cyberlindnera jadinii), Komagataella yeast (Pichia pastoris) (methanol-assimilating yeast), 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 budding yeast (Saccharomyces cerevisiae), Torula yeast (Cyberlindnera jadinii), or a combination thereof. In one embodiment, the yeast is derived from the genus Saccharomyces, and in some embodiments, from the budding yeast species Saccharomyces cerevisiae. In one embodiment, the yeast is derived from the genus Cyberlindnera, and in some embodiments, from the Torula yeast species Cyberlindnera jadinii. An embodiment of a method for obtaining a composition containing yeast protein is provided in Figure 1.Method 100 in Figure 1 provides a yeast cream. In one embodiment, the method may optionally include growing yeast (not shown in Figure 1). Yeast growth is carried out by those skilled in the art according to suitable methods known in the art. Where used in the context of this disclosure, the expression “yeast growth” refers to the growth phase of a commercial method of growing yeast under aerobic conditions to maximize the conversion of substrate to biomass. The growth step may be continuous, batch, or fed-batch. The growth medium may include a carbon source (e.g., molasses, sucrose, glucose, dextrose syrup, ethanol, maize, glycerol, maize maceration, and / or lignocellulosic biomass), a nitrogen source (e.g., ammonia or another inorganic nitrogen source), and a phosphorus source (e.g., phosphoric acid or another inorganic phosphorus source). The growth medium may further include additional micronutrients such as vitamins and / or minerals to support the growth of yeast cells.

[0013] The propagation process can be carried out under highly aerated conditions. For example, in some embodiments, the growth step may include controlling the ventilation of the container to achieve a specific ventilation rate such as, for example, at least 0.2 air volume / container volume / min, 0.3 air volume / container volume / min, 0.4 air volume / container volume / min, 0.5 air volume / container volume / min, 0.6 air volume / container volume / min, 0.7 air volume / container volume / min, 0.8 air volume / container volume / min, 0.9 air volume / container volume / min, 1.0 air volume / container volume / min, 1.1 air volume / container volume / min, 1.2 air volume / container volume / min, 1.3 air volume / container volume / min, 1.4 air volume / container volume / min, 1.5 air volume / container volume / min, 1.6 air volume / container volume / min, 1.7 air volume / container volume / min, 1.8 air volume / container volume / min, 1.9 air volume / container volume / min, or 2.0 air volume / container volume / min.

[0014] The growth process 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 process can include controlling the pH of the culture medium to about 3.0 to about 8.0, about 3.5 to about 7.0 or about 4.0 to about 6.5. In certain embodiments, 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 process can include controlling the temperature of the culture medium to about 20°C to about 40°C, about 25°C to about 30°C or about 30°C to about 35°C. In certain embodiments, the temperature is controlled to about 30°C to about 35°C (e.g., 32°C).

[0015] At the end of the growth process, a specific concentration can be determined or achieved. In some embodiments, the concentration of the grown 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, based on the volume of the culture medium. In certain embodiments where a fed-batch process is used to grow the yeast cells, the concentration of the grown yeast cells in the culture medium is at least about 0.25 wt% based on the volume of the culture medium.

[0016] In some embodiments, the grown yeast is directly subjected to a formulation step to provide yeast cream. For example, the grown yeast is not subjected to a subsequent fermentation step (which can be carried out under anaerobic conditions) before the formulation step.

[0017] In formulation step 010, the mixture obtained after growth (including the grown yeast cells) is modified to provide a yeast cream. For example, at least one component of the mixture obtained after growth is removed from the culture medium to provide a yeast composition (a yeast cream, which is one embodiment of the yeast composition, is described as an embodiment in Figure 1). This at least one component may 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 isolating the grown yeast cells (e.g., biomass) from the components of the culture medium. As used in the context of this disclosure, the expression “substantially isolate” means removing most of the components of the culture medium from the grown yeast cells. In some embodiments, “substantially isolated” means concentrating the grown 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 more, compared to the concentration of yeast cells in the culture medium before isolation. To provide a yeast composition, the grown yeast cells can be centrifuged (and the resulting cell pellet containing the grown yeast cells can optionally be washed) and / or filtered. The isolated yeast cells can then be formulated into a yeast composition (which may be a yeast cream as shown in Figure 1). The compounding step 010 can preserve (at least partially) the viability of the yeast cells in some embodiments. Therefore, the grown yeast can be provided in an active or semi-active form. The grown yeast can be provided in a liquid or semi-solid form. In one embodiment, the grown yeast can be provided in the form of a yeast cream shown in Figure 1.

[0018] The method of this disclosure provides an inactivated yeast cream by inactivating the endogenous enzymes of yeast contained in the yeast cream. This is shown as step 020 in Figure 1. Such yeast endogenous enzyme inactivation steps are performed to limit / avoid yeast autolysis. In the context of this disclosure, the term “autolysis” refers to the autolysis of its own cellular components by the enzymes of the yeast cell. Therefore, yeast endogenous enzyme inactivation step 020 is performed to limit / avoid the degradation of cellular components (particularly yeast proteins), which corresponds primarily to the degradation of proteinaceous substances (i.e., proteolysis). In one embodiment, inactivation of endogenous enzymes does not cause thermal plasmolysis. In the context of this disclosure, the expression “thermal plasmolysis” refers to the denaturation of yeast and permeabilization of the yeast membrane. In another embodiment, the inactivation of endogenous enzymes of this disclosure does not permeabilize the yeast membrane. In yet another embodiment, the inactivation of endogenous enzymes maintains the integrity of the yeast membrane. In one embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to changes in temperature and / or pH over a period of time. In another embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to a temperature greater than 4°C and less than 120°C. In yet another embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to temperatures 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, inactivation of endogenous enzymes is achieved by exposing the yeast cream to temperatures of 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 or lower. In yet another embodiment, the yeast cream is heated 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,Inactivation of endogenous enzymes is achieved by exposure to temperatures between 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 a further embodiment, inactivation of endogenous enzymes is achieved by exposure of the yeast cream to temperatures between 50 and 110°C. In yet another embodiment, inactivation of endogenous enzymes is achieved by exposure of the yeast cream to temperatures between 60 and 100°C, for example, 80°C. In one embodiment, inactivation of endogenous enzymes is achieved by exposure of the yeast cream to a specific pH or pH range. In one embodiment, inactivation of endogenous enzymes is achieved by exposure of the yeast cream to a pH of 1 to 13. In one embodiment, the yeast cream is made up 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, and 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 0.6, 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, 10. Inactivation of endogenous enzymes is achieved by exposing the yeast cream to pH 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, yeast cream is exposed to pH 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. Inactivation of endogenous enzymes is achieved by exposure to pH levels 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, or 1.0 or lower. In yet another embodiment, the yeast cream is expressed in amounts of 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, 1 0.6, 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 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. Inactivation of endogenous enzymes is achieved by exposure to pH levels between 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 specific cases, inactivation of endogenous enzymes is achieved by exposing the yeast cream to pH levels below 5 or above 7, for example, pH 3 or 9. In certain cases, yeast cream is prepared with pH values ​​of 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 less than 1.0. Or 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,Inactivation of endogenous enzymes is achieved by exposure to a pH greater than 12.9 or 13.0. In one embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to changes in temperature and / or pH for a period of time greater than 1 second but less than 10 hours. In one embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to changes in temperature and / or pH for at least 15 seconds, at least 30 seconds, at least 45 seconds, at least 1 minute, at least 2 minutes, at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 1 hour, at least 1.5 hours, at least 2 hours, at least 2.5 hours, at least 3 hours, at least 3.5 hours, at least 4 hours, at least 4.5 hours, at least 5 hours, at least 5.5 hours, at least 6 hours, at least 6.5 hours, at least 7 hours, at least 7.5 hours, at least 8 hours, at least 8.5 hours, at least 9 hours, at least 9.5 hours, or at least 10 hours. In another embodiment, inactivation of endogenous enzymes is achieved by exposing the yeast cream to a certain temperature and / or pH change for 10 hours, 9.5 hours, 9 hours, 8.5 hours, 8 hours, 7.5 hours, 7 hours, 6.5 hours, 6 hours, 5.5 hours, 5 hours, 4.5 hours, 4 hours, 3.5 hours, 3 hours, 2.5 hours, 2 hours, 1.5 hours, 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 45 seconds, 30 seconds, or 15 seconds or less. In yet another embodiment, the yeast cream is steeped for 15 seconds, 30 seconds, 45 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, 8.5 hours, 9 hours, 9.5 hours or 10 hours, 10 hours, 9.5 hours, 9 hours, 8.5 hours, 8 hours, 7.5 hours, 7 hours, 6.5 hours, 6 hours, 5.5 hours, 5 hours, 4.5 hours, 4 hours, 3.5 hours, 3 hours, 2.5 hours, 2 hours, 1.5 hours,Inactivation of endogenous enzymes is achieved by exposing them to a temperature and / or pH change for a period of 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 45 seconds, 30 seconds, or 15 seconds. In a further embodiment, the yeast cream is exposed to a temperature and / or pH change for a period of 15 seconds to 7 hours to achieve inactivation of endogenous enzymes. In yet a further embodiment, the yeast cream is exposed to a temperature and / or pH change for a period of 30 seconds to 2 hours, for example 5 minutes, to achieve inactivation of endogenous enzymes. One skilled in the art will know how to adapt the temperature and / or pH, and the exposure time, depending on the yeast source and the endogenous enzymes to be inactivated. In a further embodiment, without limitation, standard techniques in the art such as protease activity assays (azoalbumin assay, fluorescein assay, thiocarbamoyl-kappa-casein assay or curdle assay) and / or incubation of purified beta-glucan with inactivated yeast cream, and by measuring glucose formation (HPLC), can be used to confirm inactivation of endogenous enzymes.

[0019] ​Next, the inactivated yeast cream is subjected to exogenous enzymatic treatment, which is shown as step 030 in Figure 1. In the context of this disclosure, the exogenous enzymatic treatment lacks ribonuclease activity (i.e., endonuclease and exonuclease; or nuclease activity; (EC 3.1.4.1)). Further in the context of this disclosure, the exogenous enzymatic treatment comprises at least one polypeptide capable of hydrolyzing oligosaccharides. In one embodiment, the exogenous enzymatic treatment comprises at least one polypeptide having glucanase activity, mannanase activity and / or chitinase activity. In one embodiment, the exogenous enzymatic treatment comprises at least one polypeptide having endo-β-1,3-glucanase activity, exo-β-1,3-glucanase activity, endo-β-1,6-glucanase activity and / or exo-β-1,6-glucanase activity (EC 3.2.1). In another embodiment, the exogenous enzymatic treatment consists of using polypeptides having β-1,3-glucanase activity and / or polypeptides having β-1,6-glucanase activity. In the context of this disclosure, the expression "consists of" excludes other enzymatic treatments that substantially affect protoplast formation and / or the characteristics of the yeast protein that can be obtained. In one embodiment, the exogenous enzymatic treatment, comprising at least one polypeptide having glucanase activity (EC 3.2.1), is carried out at a temperature greater than 4°C and less than 120°C. In one embodiment, exogenous enzyme treatment containing 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, exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at temperatures of 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 or lower.In yet another embodiment, exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at temperatures ranging 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 further embodiments, exogenous enzymatic treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature of 20 to 110°C. In yet another embodiment, exogenous enzymatic treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a temperature of 50 to 80°C, for example, 60°C. In one embodiment, the amount of polypeptide having glucanase activity (EC 3.2.1) is greater than 1 UI and less than 5000 UI per kg of inactivated yeast cream. In the context of this disclosure, 1 unit of enzyme activity (UI) catalyzes the formation of 1 μmol of glucose per minute at 37°C.In one embodiment, the amount of 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, 2 The values ​​are 000UI, 2100UI, 2200UI, 2300UI, 2400UI, 2500UI, 2600UI, 2700UI, 2800UI, 2900UI, 3000UI, 3100UI, 3200UI, 3300UI, 3400UI, 3500UI, 3600UI, 3700UI, 3800UI, 3900UI, 4000UI, 4100UI, 4200UI, 4300UI, 4400UI, 4500UI, 4600UI, 4700UI, 4800UI, 4900UI, or 5000UI. In another embodiment, the glucanase activity (EC) is used. 3.2.1) The amount of polypeptide having is 5000UI, 4900UI, 4800UI, 4700UI, 4600UI, 4500UI, 4400UI, 4300UI, 4200UI, 4100UI, 4000UI, 3900UI, 3800UI, 3700UI, 3600UI, 3500UI, 3400UI, 3300UI, 3200UI, 3100UI, 3000UI, 2900UI, 2800UI, 2700UI, 2600UI, 2500UI, The values ​​are 2400UI, 2300UI, 2200UI, 2100UI, 2000UI, 1900UI, 1800UI, 1700UI, 1600UI, 1500UI, 1400UI, 1300UI, 1200UI, 1100UI, 1000UI, 900UI, 800UI, 700UI, 600UI, 500UI, 400UI, 300UI, 200UI, 100UI, 90UI, 80UI, 70UI, 60UI, 50UI, 40UI, 30UI, 20UI, 10UI, 5UI, or 1UI or less.In another embodiment, the amount of polypeptide having glucanase activity (EC 3.2.1) per 1 kg of inactivated yeast cream 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, 19 00UI, 2000UI, 2100UI, 2200UI, 2300UI, 2400UI, 2500UI, 2600UI, 2700UI, 2800UI, 2900UI, 3000UI, 3100UI, 3200UI, 3300UI, 3 400UI, 3500UI, 3600UI, 3700UI, 3800UI, 3900UI, 4000UI, 4100UI, 4200UI, 4300UI, 4400UI, 4500UI, 4600UI, 4700UI, 4800UI, From 4900UI or 5000UI, to 5000UI, 4900UI, 4800UI, 4700UI, 4600UI, 4500UI, 4400UI, 4300UI, 4200UI, 4100UI, 4000UI, 3900UI, 3800UI, 3700UI, 3600UI, 3500UI, 3400UI, 3300UI, 3200UI, 3100UI, 3000UI, 2900UI, 2800UI, 2700UI, 2600UI, 2500UI, 2400UI, 23 The amounts are between 00UI, 2200UI, 2100UI, 2000UI, 1900UI, 1800UI, 1700UI, 1600UI, 1500UI, 1400UI, 1300UI, 1200UI, 1100UI, 1000UI, 900UI, 800UI, 700UI, 600UI, 500UI, 400UI, 300UI, 200UI, 100UI, 90UI, 80UI, 70UI, 60UI, 50UI, 40UI, 30UI, 20UI, 10UI, 5UI, or 1UI. In a further embodiment, the amount of polypeptide having glucanase activity (EC 3.2.1) is 300 to 3000UI per kg of inactivated yeast cream.In further embodiments, the amount of polypeptide having glucanase activity (EC 3.2.1) is 1500 to 2500 UI per kg of inactivated yeast cream, for example, 2000 UI per kg of inactivated yeast cream. In one embodiment, exogenous enzyme treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is carried out at a pH of 3 to 9. In one embodiment, glucanase activity (EC Exogenous enzyme treatment including at least one polypeptide having 3.2.1) is performed 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 The procedure is carried out at pH 0.7, 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 glucanase activity (EC) is measured. Exogenous enzyme treatment containing at least one polypeptide having 3.2.1) was performed on 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. The procedure should be performed at pH levels of 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 or lower.In yet another embodiment, exogenous enzymatic treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is performed as follows: 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 From 0.8, 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, The treatment is carried out at a pH between 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 a further embodiment, the exogenous enzyme treatment containing 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 containing 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, an exogenous enzyme treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for a period of time greater than 1 second and less than 10 hours. In one embodiment, an exogenous enzyme treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for at least 15 seconds, at least 30 seconds, at least 45 seconds, at least 1 minute, at least 2 minutes, at least 5 minutes, at least 10 minutes, at least 15 minutes, and at least. The process shall be carried out for 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 1 hour, at least 1.5 hours, at least 2 hours, at least 2.5 hours, at least 3 hours, at least 3.5 hours, at least 4 hours, at least 4.5 hours, at least 5 hours, at least 5.5 hours, at least 6 hours, at least 6.5 hours, at least 7 hours, at least 7.5 hours, at least 8 hours, at least 8.5 hours, at least 9 hours, at least 9.5 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, or at least 24 hours. In another embodiment, exogenous enzyme treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is performed for 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9.5 hours, 9 hours, 8.5 hours, 8 hours, 7.5 hours, 7 hours, 6.5 hours, 6 hours, 5.5 hours, 5 hours, 4.5 hours, 4 hours, 3.5 hours, 3 hours, 2.5 hours, 2 hours, 1.5 hours, 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 45 seconds, 30 seconds, or 15 seconds or less.In yet another embodiment, exogenous enzyme treatment comprising at least one polypeptide having glucanase activity (EC 3.2.1) is performed for 15 seconds, 30 seconds, 45 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, The process is carried out over a period of 23 or 24 hours, or over a period of 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9.5 hours, 9 hours, 8.5 hours, 8 hours, 7.5 hours, 7 hours, 6.5 hours, 6 hours, 5.5 hours, 5 hours, 4.5 hours, 4 hours, 3.5 hours, 3 hours, 2.5 hours, 2 hours, 1.5 hours, 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 45 seconds, 30 seconds, or 15 seconds. In a further embodiment, exogenous enzyme treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is carried out over a period of 1 to 24 hours. In further embodiments, an exogenous enzymatic treatment containing at least one polypeptide having glucanase activity (EC 3.2.1) is carried out for 2 to 6 hours, for example, 4 hours. Those skilled in the art will know how to adapt the concentration, temperature, pH and exposure time of the glucanase-active polypeptide depending on the yeast source and the glucanase-active polypeptide considered.

[0020] An inactivated yeast cream is subjected to enzymatic treatment to obtain yeast protoplasts. In the context of this disclosure, the term "yeast protoplast" refers to yeast that has lost most of its cell wall components. In another embodiment, the enzymatic treatment of the method of this disclosure aims to avoid the use of enzymes having proteolytic activity and thus to preserve the integrity of yeast proteins (e.g., secondary, tertiary, or quaternary structures). In some other embodiments, the enzymatic treatment of this disclosure aims to preserve the integrity of intracellular proteins and yeast cell membrane proteins. In a further embodiment, the enzymatic treatment of this disclosure aims to solubilize the yeast cell wall, leaving a soluble fraction containing mannoprotein and / or β-glucan and an insoluble fraction containing yeast protoplasts. In yet another embodiment, the enzymatic treatment of this disclosure allows for the solubilization of a portion of the yeast membrane but does not allow for the diffusion of yeast proteins outside the yeast protoplasts.

[0021] In some embodiments, the inactivated yeast cream may be subjected to an alkaline extraction step 025 prior to the enzymatic treatment step 030. The alkaline extraction step can be performed to reduce the amount of nucleotides (including polynucleotides such as RNA) in the final composition. In some embodiments, the alkaline extraction step may be performed under conditions that allow for the extraction of nucleotides from the inactivated yeast cream and prevent or limit the degradation of nucleotides from the inactivated yeast cream. The expression "prevent or limit the degradation of nucleotides from the inactivated yeast cream" refers to the fact that the alkaline extraction step prevents or limits the degradation of nucleotides by at least 50%. While we do not wish to be bound by theory, it is understood that subjecting the inactivated yeast cream to alkaline extraction will at least partially solubilize the nucleotides present in the inactivated yeast cream, while simultaneously enabling the formation of yeast protoplasts (after exogenous enzymatic treatment). The alkaline extraction step 025 includes placing the inactivated yeast cream under alkaline conditions for a certain period of time within a certain temperature range to enable the solubilization of extracellular nucleotides (including polynucleotides such as RNA) in the inactivated yeast cream. In one embodiment, the alkaline extraction step 025 is performed at 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 The process is carried out at pH levels between 0.9, 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 a further embodiment, the alkaline extraction step 025 is carried out at pH levels between 8 and 11.In further embodiments, the alkaline extraction step 025 is carried out at a pH of 8.5 to 9.5, for example, a pH of 9.0. In even further embodiments, the alkaline extraction step 025 is carried out at a temperature greater than 4°C and less than 70°C. In one embodiment, the alkaline extraction step 025 is carried out 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 alkaline extraction step is carried out at a temperature of 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 or lower. In yet another embodiment, the alkaline extraction step 025 is carried out at temperatures ranging 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 yet another embodiment, the alkaline extraction step is carried out at temperatures ranging from 50 to 70°C, for example, at 65°C. In yet another embodiment, the alkaline extraction step 025 is performed for 15 seconds, 30 seconds, 45 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, and 23 hours. Alternatively, the process may be carried out over a period of 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3.5 hours, 3 hours, 2.5 hours, 2 hours, 1.5 hours, 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 45 seconds, or 30 seconds. In a further embodiment, the alkaline extraction process 025 may be carried out over a period of 1 to 4 hours. In yet another embodiment, the alkaline extraction process 025 may be carried out over a period of 1.5 to 2.5 hours, for example, 2 hours.In yet another embodiment, the alkaline extraction step 025 is carried out over a period of 15 seconds, 30 seconds, 45 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, or 45 minutes. In yet another embodiment, the alkaline extraction step 025 is carried out over a period of 10 to 45 minutes. In yet another embodiment, the alkaline extraction step 025 is carried out over a period of 20 to 40 minutes, for example, 30 minutes. Those skilled in the art will know how to optimize the temperature, pH, and exposure time of the alkaline extraction step to solubilize nucleotides from inactivated yeast cream.

[0022] In some embodiments, this method may include a step to inactivate the exogenous enzyme used during the enzyme treatment step 030 (not shown in Figure 1).

[0023] The insoluble fraction obtained by the enzymatic treatment carried out by the method herein is then separated from the soluble fraction by a conventional technique known to those skilled in the art. This is shown as step 050 in Figure 1. Techniques that may 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 subjected directly to separation. In one embodiment, after separating the insoluble fraction from the soluble fraction, it is optionally rinsed and then collected. In another embodiment, after separating the insoluble fraction from the soluble fraction, it is collected directly. In one embodiment, the collected insoluble fraction has a neutral taste and may be in a non-dried or dried state. In the context of this disclosure, a non-dried insoluble fraction may be in a semi-liquid state.

[0024] In one embodiment, the insoluble fraction may be provided in a non-dried state. In one embodiment, the undried insoluble fraction can be collected, formulated into an edible composition, and used immediately. In one embodiment, the undried insoluble fraction can be stored. In another embodiment, the undried insoluble fraction can be frozen according to the usual methods of the art for later formulation into an edible composition. In a further embodiment, undried insoluble fractions obtained from different yeast sources can be combined for formulation into an edible composition. In one embodiment, the insoluble fraction may be provided in a dried state. In another embodiment, a dried insoluble fraction can be obtained by spray drying, fluidized bed drying, tray drying, roller / drum drying, infrared drying, or freeze-drying of the undried 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 a further embodiment, dried insoluble fractions obtained from different yeast sources can be combined for formulation into an edible composition.

[0025] In one embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of more than 50% and less than 100% based on the total mass of the collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using 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 collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25. In another embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, has a protein content of 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% or less, based on the total mass of the collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25.In yet another embodiment, the collected insoluble fraction containing yeast protoplasts, when dried, is determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25, based on the total mass of the collected and dried insoluble fraction, comprising 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%, and 90%. The protein content ranges from %, 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%. In a further embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of 70-100% based on the total mass of the collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25 when dried. In yet another embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of 75-85%, for example, 80%, based on the total mass of the collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25 when dried. In yet another embodiment, the collected insoluble fraction containing yeast protoplasts has a protein content of at least 80% based on the total mass of the collected and dried insoluble fraction, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25 when dried.

[0026] The collected insoluble fraction containing yeast protoplasts may also have at least one of the following characteristics when dried: - Lipid content of more than 0% and less than 20% based on the total mass of the collected and dried insoluble fraction as determined by the modified Majonier method (AOAC 989.05), or 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% based on the total mass of the collected and dried insoluble fraction as determined by the modified Majonier method (AOAC 989.05) , 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 0.5%, 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% Lipid content of %, 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% or modified Majornie method (AOAC) Based on the total mass of the collected and dried insoluble fractions as determined by 989.05), the following percentages were obtained: 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 0.2%, 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% Lipid content of 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% or less, or modified Majornie method (AOAC) Based on the total mass of the collected and dried insoluble fraction as determined by 989.05), the percentages are 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 From 0.1%, 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%, 9.3%, 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. Lipid content between 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%, or a lipid content of 6-10% based on the total mass of the collected and dried insoluble fraction determined by the modified Majornier method (AOAC 989.05), or a lipid content of 7-9% based on the total mass of the collected and dried insoluble fraction determined by the modified Majornier method (AOAC 989.05), for example, an 8% lipid content. - When the alkaline extraction process is not performed, the nucleic acid content is greater than 2% and less than 30% based on the total mass of the collected and dried insoluble fraction, or at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, and 24% based on the total mass of the collected and dried insoluble fraction. nucleic acid content of 25%, 26%, 27%, 28%, 29%, or 30%, or nucleic acid content of 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% or less, based on the total mass of the collected and dried insoluble fraction. The amounts, or the total mass of the collected and dried insoluble fractions, range from 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%. A nucleic acid content between %, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2%, or a nucleic acid content of 5–20% based on the total mass of the collected and dried insoluble fraction, or a nucleic acid content of 7–15% based on the total mass of the collected and dried insoluble fraction, for example, a 10% nucleic acid content. In the context of this disclosure, the term “nucleic acid” refers to a biomacromolecule composed of nucleotides (i.e., polynucleotides), the latter composed of pentoses, phosphate groups, and nitrogen-containing bases. In the context of this disclosure, the term “nucleic acid” further refers to polynucleotides such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). In the context of this disclosure, the term “nucleic acid” further refers to ribonucleic acid (RNA). In the context of this disclosure, the term “nucleic acid” does not refer to nucleotide residues that may be generated during the process. This disclosure measures nucleic acid content according to the method of Fish et al. (1991) and compares the values ​​with and without enzymatic digestion. - When an alkaline extraction process is performed, nucleic acid content of 3% or less based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 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% or less based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 0.1 to 3% based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 0.5 to 2% based on the total mass of the collected and dried insoluble fraction. In the context of this disclosure, the term "nucleic acid" refers to a biomacromolecule composed of nucleotides (i.e., polynucleotides), the latter composed of pentoses, phosphate groups, and nitrogen-containing bases. Further in the context of this disclosure, the term "nucleic acid" refers to polynucleotides such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Further in the context of this disclosure, the term "nucleic acid" refers to ribonucleic acid (RNA). In the context of this disclosure, the term "nucleic acid" does not refer to nucleotide residues that may be generated during the process. This disclosure measures nucleic acid content according to the method of Fish et al. (1991) and compares the values ​​with and without enzymatic digestion. - Carbohydrate content greater than 0% and less than 25%, or 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 collected and dried insoluble fraction. Carbohydrate content of 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% or less based on the total mass of the collected and dried insoluble fraction, or the collected and dried insoluble fraction Based on the total mass, from 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% Carbohydrate content between %, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5%, or a carbohydrate content of 5–20% based on the total mass of the collected and dried insoluble fraction, or a carbohydrate content of 7–15% based on the total mass of the dried insoluble fraction, for example, a 10% carbohydrate content. In the context of this disclosure, the term "carbohydrate" refers to total sugars measured by HPLC after chemical or enzymatic digestion (i.e., AOAC 980.13 method). - Mannan content of more than 0% and less than 6% based on the total mass of the collected and dried insoluble fraction, or 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% based on the total mass of the collected and dried insoluble fraction. Mannan content of 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 collected and dried insoluble fraction, or 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% Mannan content of 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% or less based on the total mass of the collected and dried insoluble fraction. 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.From 2%, 5.3%, 5.4%, 5.5%, 5.6%, 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% Mannan content between %, 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%, or mannan content of 0–5.5% or 0–4% based on the total mass of the collected and dried insoluble fraction, or mannan content of 0.5–5.5% or 0.5–4.0% based on the total mass of the collected and dried insoluble fraction, for example, about 2% or 5% mannan content. In the context of this disclosure, mannan content is measured by HPLC after chemical or enzymatic digestion. - Glucan content of more than 0% and less than 10% based on the total mass of the collected and dried insoluble fraction, or 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% based on the total mass of the collected and dried insoluble fraction. %, 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 10.0% glucan content of 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 collected and dried insoluble fraction, or 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 0.2%, 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 0.7%, 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.Glucan content of 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% or less based on the total mass of the collected and dried insoluble fraction: 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. From 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.Glucan content between 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%, or glucan content of 0-8% based on the total mass of the collected and dried insoluble fraction, or glucan content of 1-4% based on the total mass of the collected and dried insoluble fraction, for example, 2% glucan content. In the context of this disclosure, glucan content is measured by HPLC after chemical or enzymatic digestion, and / or. - Glucose content greater than 0% and less than 25%, or 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 collected and dried insoluble fraction. A glucose content of 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% or less based on the total mass of the collected and dried insoluble fraction, or the collected and dried insoluble fraction. Based on the total mass, from 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%, Glucose content between 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5%, or glucose content of 5–20% based on the total mass of the collected and dried insoluble fraction, or carbohydrate content of 7–15% based on the total mass of the dried insoluble fraction, for example, 10% glucose content. In the context of this disclosure, the term "glucose" refers to the molecular formula C6H 12This refers to monosaccharides containing O6. In the context of this disclosure, the glucose content is further measured directly by HPLC.

[0027] In some embodiments, the method may include obtaining a soluble fraction of an enzyme-treated yeast cream, which corresponds to step 040 in Figure 1.

[0028] Yeast protoplast-derived composition In one embodiment, the present disclosure aims to provide a composition comprising yeast protein derived from yeast protoplasts. In one embodiment, the composition comprising yeast protein derived from yeast protoplasts has at least one of the following characteristics: - A protein content of more than 50% and less than 100% based on the total mass of the composition, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25. Or, 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, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25. Alternatively, a protein content of 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% or less, based on the total mass of the composition, as determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25. Alternatively, the percentages based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25, are 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%, and 96%. Protein content between 100%, 99%, 98%, 99%, or 100%, and between 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%. Alternatively, protein content between 70% and 100% based on the total mass of the composition, determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25.Alternatively, the protein content of 75-85% based on the total mass of the composition, for example, 80% protein content, is determined by the Kjeldahl method for nitrogen analysis using a conversion factor of 6.25. - Lipid content of more than 0% and less than 20% based on the total mass of the collected and dried insoluble fraction as determined by the modified Majonier method (AOAC 989.05), or 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% based on the total mass of the collected and dried insoluble fraction as determined by the modified Majonier method (AOAC 989.05) , 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 0.5%, 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% Lipid content of %, 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% or modified Majornie method (AOAC) Based on the total mass of the collected and dried insoluble fractions as determined by 989.05), the following percentages were obtained: 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 0.2%, 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% Lipid content of 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% or less, or modified Majornie method (AOAC) Based on the total mass of the collected and dried insoluble fraction as determined by 989.05), the percentages are 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 From 0.1%, 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%, 9.3%, 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. Lipid content between 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%, or a lipid content of 6-10% based on the total mass of the collected and dried insoluble fraction determined by the modified Majornier method (AOAC 989.05), or a lipid content of 7-9% based on the total mass of the collected and dried insoluble fraction determined by the modified Majornier method (AOAC 989.05), for example, an 8% lipid content. - When the alkaline extraction process is not performed, the nucleic acid content is greater than 2% and less than 30% based on the total mass of the collected and dried insoluble fraction, or at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, and 24% based on the total mass of the collected and dried insoluble fraction. nucleic acid content of 25%, 26%, 27%, 28%, 29%, or 30%, or nucleic acid content of 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% or less, based on the total mass of the collected and dried insoluble fraction. The amounts, or the total mass of the collected and dried insoluble fractions, range from 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%. A nucleic acid content between %, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2%, or a nucleic acid content of 5–20% based on the total mass of the collected and dried insoluble fraction, or a nucleic acid content of 7–15% based on the total mass of the collected and dried insoluble fraction, for example, a 10% nucleic acid content. In the context of this disclosure, the term “nucleic acid” refers to a biomacromolecule composed of nucleotides (i.e., polynucleotides), the latter composed of pentoses, phosphate groups, and nitrogen-containing bases. In the context of this disclosure, the term “nucleic acid” further refers to polynucleotides such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). In the context of this disclosure, the term “nucleic acid” further refers to ribonucleic acid (RNA). In the context of this disclosure, the term “nucleic acid” does not refer to nucleotide residues that may be generated during the process. This disclosure measures nucleic acid content according to the method of Fish et al. (1991) and compares the values ​​with and without enzymatic digestion. - When an alkaline extraction process is performed, nucleic acid content of 3% or less based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 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% or less based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 0.1 to 3% based on the total mass of the collected and dried insoluble fraction. Or, nucleic acid content of 0.5 to 2.0% based on the total mass of the collected and dried insoluble fraction. In the context of this disclosure, the term "nucleic acid" refers to a biomacromolecule composed of nucleotides (i.e., polynucleotides), the latter composed of pentoses, phosphate groups, and nitrogen-containing bases. Further in the context of this disclosure, the term "nucleic acid" refers to polynucleotides such as deoxyribonucleic acid (DNA) and / or ribonucleic acid (RNA). Further in the context of this disclosure, the term "nucleic acid" refers to ribonucleic acid (RNA). In the context of this disclosure, the term "nucleic acid" does not refer to nucleotide residues that may be generated during the process. This disclosure measures nucleic acid content according to the method of Fish et al. (1991) and compares the values ​​with and without enzymatic digestion. - A carbohydrate content greater than 0% and less than 25%, or 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, or a carbohydrate content of 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% or less based on the total mass of the composition Carbohydrate content ranging from 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%, or a carbohydrate content of 5-20% based on the total mass of the composition, or a carbohydrate content of 7-15% based on the total mass of the composition, for example, a 10% carbohydrate content. In the context of this disclosure, the term "carbohydrate" refers to total sugars measured by HPLC after chemical or enzymatic digestion (i.e., AOAC 980.13 method). - Mannan content of more than 0% and less than 6% based on the total mass of the collected and dried insoluble fraction, or 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% based on the total mass of the collected and dried insoluble fraction. Mannan content of 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 collected and dried insoluble fraction, or 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% Mannan content of 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% or less based on the total mass of the collected and dried insoluble fraction. 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.From 3%, 5.4%, 5.5%, 5.6%, 5.7%, 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 Mannan content between 0.1%, 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%, or mannan content of 0–5.5% or 0–4.0% based on the total mass of the collected and dried insoluble fraction, or mannan content of 0.5–5.5% or 0.5–3.0% based on the total mass of the collected and dried insoluble fraction, for example, about 2% or about 5.5% mannan content. In the context of this disclosure, mannan content is measured by HPLC after chemical or enzymatic digestion. - Glucan content of more than 0% and less than 10% based on the total mass of the collected and dried insoluble fraction, or 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% based on the total mass of the collected and dried insoluble fraction. %, 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 10.0% glucan content of 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 collected and dried insoluble fraction, or 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 0.2%, 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 0.7%, 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.Glucan content of 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% or less based on the total mass of the collected and dried insoluble fraction: 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. From 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.Glucan content between 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%, or glucan content of 0-8% based on the total mass of the collected and dried insoluble fraction, or glucan content of 1-4% based on the total mass of the collected and dried insoluble fraction, for example, 2% glucan content. In the context of this disclosure, glucan content is measured by HPLC after chemical or enzymatic digestion, and / or. - A glucose content greater than 0% and less than 25%, or 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 composition, or a glucose content of 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% or less based on the total mass of the composition A glucose content between 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%, or between 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%, or a glucose content of 5-20% based on the total mass of the composition, or a carbohydrate content of 7-15% based on the total mass of the composition, for example, a 10% glucose content. In the context of this disclosure, the term "glucose" refers to the molecular formula C6H 12 This refers to monosaccharides containing O6. In the context of this disclosure, the glucose content is further measured directly by HPLC.

[0029] In one embodiment, the yeast protein derived from yeast protoplasts is an intracellular yeast protein, a yeast cell membrane protein, or a combination thereof.

[0030] In one embodiment, the composition comprises yeast protein derived from yeast protoplasts, having a neutral taste. This is in contrast to yeast proteins obtained from yeast extracts, which typically exhibit a moderate taste (particularly those obtained using protease treatment). In the context of this disclosure, yeast protein derived from yeast protoplasts in food products has a neutral taste and can therefore be used as an ingredient whose taste does not affect / modify the taste of other components of the composition. Thus, the compositions of this disclosure do not affect / modify the taste of formulated food products. In another embodiment, the composition comprises yeast protein derived from yeast protoplasts, having a neutral taste when compared to plant / legume protein compositions. In yet another embodiment, the composition comprises yeast protein derived from yeast protoplasts, having a neutral taste when compared to yeast protein compositions obtained by methods that do not involve a glucanase treatment step. In yet another embodiment, the composition comprises yeast protein derived from yeast protoplasts, having a neutral taste when compared to compositions that do not contain yeast protoplasts. In the context of this disclosure, a method for preparing a yeast protein composition, which does not involve a glucanase treatment step, is a method comprising: i) providing a yeast cream containing yeast; ii) inactivating the endogenous enzymes of yeast to provide an inactivated yeast cream; iii) optionally separating an insoluble fraction from a soluble fraction; and iv) collecting the inactivated yeast cream, or optionally collecting the insoluble fraction of the inactivated yeast cream, excluding the glucanase treatment step. Further in the context of this disclosure, a method for preparing a yeast protein composition, which does not involve a glucanase treatment step, is a method essentially comprising: i) providing a yeast cream containing yeast; ii) inactivating the endogenous enzymes of yeast to provide an inactivated yeast cream; iii) optionally separating an insoluble fraction from a soluble fraction; and iv) collecting the inactivated yeast cream, or optionally collecting the insoluble fraction of the inactivated yeast cream.The transitional phrase "essentially derived from" limits the steps of this method to specific steps of the final product (i.e., steps that do not substantially affect the glucanase treatment step or the separation of the insoluble / soluble fractions) and its basic and novel characteristics.

[0031] In one embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index of more than 1% and less than 15% based on the total mass of the dry composition. In another embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index of more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% based on the total mass of the dry composition. In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index of 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% or less based on the total mass of the dry composition. In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index between 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% based on the total mass of the dry composition, and between 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%. In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index of 1 to 10% based on the total mass of the dry composition. In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a water solubility index of 3 to 7% based on the total mass of the dry composition, for example, a water solubility index of 5. In the context of this disclosure, the water solubility index represents the weight of the dry solid in the supernatant, expressed as a percentage of the original weight of the sample. In another embodiment, a composition containing yeast protein derived from yeast protoplasts has a lower water solubility index compared to a yeast protein composition prepared by a similar method without a glucanase treatment step.

[0032] In one embodiment, a composition containing yeast protein derived from yeast protoplasts has weak emulsifying properties. In the context of this disclosure, the term "emulsifying properties" refers to the emulsifying activity and / or emulsion stability of the composition. Further in the context of this disclosure, the term "emulsifying activity" refers to the ability of the composition to form an emulsion, while the term "emulsion stability" refers to the emulsion that remains after heat treatment. In one embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity 1 to 10 times lower than that of lecithin, as measured by Brishti et al. (2017). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity at least 1, 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of that of lecithin, as measured by Brishti et al. (2017). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity of 1 / 10, 1 / 9, 1 / 8, 1 / 7, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, or 1 times that of lecithin, as measured by Brishti et al. (2017). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity between 1, 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of lecithin, as measured by Brishti et al. (2017). In further embodiments, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity 1 to 1 / 10 times that of lecithin, as measured by Brishti et al. (2017). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsifying activity 1 / 3 to 1 / 7 times that of lecithin, as measured by Brishti et al. (2017), for example, 1 / 5 times that of lecithin.In one embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsion stability 1 to 1 / 15 times greater than that of lecithin, as measured by the method of Yasumatsu et al. (1972). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsion stability at least 1, 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, 1 / 10, 1 / 11, 1 / 12, 1 / 13, 1 / 14, or 1 / 15 greater than that of lecithin, as measured by the method of Yasumatsu et al. (1972). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsion stability of 1 / 15, 1 / 14, 1 / 13, 1 / 12, 1 / 11, 1 / 10, 1 / 9, 1 / 8, 1 / 7, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, or 1 / 1 compared to lecithin, as measured by the method of Yasumatsu et al. (1972). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsion stability between 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, 1 / 10, 1 / 11, 1 / 12, 1 / 13, 1 / 14, or 1 / 15 of lecithin, as measured by Yasumatsu et al. (1972). In further embodiments, a composition containing yeast protein derived from yeast protoplasts has emulsion stability one-fifth to one-fifteenth that of lecithin, as measured by Yasumatsu et al. (1972). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has emulsion stability one-eighth to one-twelfth that of lecithin, as measured by Yasumatsu et al. (1972). For example, the composition has emulsion stability one-tenth that of lecithin.

[0033] In one embodiment, a composition containing yeast protein derived from yeast protoplasts exhibits weak foaming properties. In the context of this disclosure, the term “foaming properties” refers to foaming ability and / or foam stability over time. Further in the context of this disclosure, the term “foaming ability” of a composition refers to the amount of interfacial area that can be formed by the composition, while the term “foam stability” refers to the ability of the composition to stabilize over time against gravitational and mechanical stress. In one embodiment, a composition containing yeast protein derived from yeast protoplasts has a foaming ability of 1 to 1 / 10 times that of a yeast protein composition prepared by a similar method without a glucanase treatment step, as measured by the method described by Chandra et al. (2015). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has, as measured by the method described by Chandra et al. (2015), at least 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of the foaming ability of a yeast protein composition prepared by a similar method without a glucanase treatment step. In another embodiment, a composition containing yeast protein derived from yeast protoplasts has, as measured by the method described by Chandra et al. (2015), at least 1 / 10, 1 / 9, 1 / 8, 1 / 7, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, or 1 of the foaming ability of a yeast protein composition prepared by a similar method without a glucanase treatment step. In yet another embodiment, a composition comprising yeast protein derived from yeast protoplasts has foaming ability ranging from 1 / 1, 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of a yeast protein composition prepared by a similar method without a glucanase treatment step, as measured by the method described by Chandra et al. (2015), to 1 / 10, 1 / 9, 1 / 8, 1 / 7, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, or 1.In further embodiments, a composition containing yeast protein derived from yeast protoplasts has a foaming ability of 1 to 1 / 6 times that of a yeast protein composition prepared by a similar method without a glucanase treatment step, as measured by the method described by Chandra et al. (2015). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a foaming ability of 1 to 1 / 4 times that of a yeast protein composition prepared by a similar method without a glucanase treatment step, as measured by the method described by Chandra et al. (2015). For example, the composition has a foaming ability of 1 / 3 times that of a yeast protein composition prepared by a similar method without a glucanase treatment step. In one embodiment, a composition containing yeast protein derived from yeast protoplasts has foam stability of 1 to 1 / 10 times that of a yeast protein composition prepared by a similar method without a glucanase treatment step, as measured after 30 minutes by the method described by Brishti et al. (2017). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has foam stability at least 1, 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of that of a yeast protein composition prepared by a similar method without a glucanase treatment step, when measured after 30 minutes using the method described by Brishti et al. (2017). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has foam stability at least 1 / 10, 1 / 9, 1 / 8, 1 / 7, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, or 1 or less of that of a yeast protein composition prepared by a similar method without a glucanase treatment step, when measured after 30 minutes using the method described by Brishti et al. (2017).In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has foam stability between 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 6, 1 / 7, 1 / 8, 1 / 9, or 1 / 10 of a yeast protein composition prepared by a method that does not include a glucanase treatment step, when measured after 30 minutes by the method described by Brishti et al. (2017). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has foam stability between 1 / 3 and 1 / 9 of a yeast protein composition prepared by a similar method that does not include a glucanase treatment step, when measured after 30 minutes by the method described by Brishti et al. (2017). In further embodiments, a composition containing yeast protein derived from yeast protoplasts has a foam stability that is one-quarter to one-eighth that of a yeast protein composition prepared by a similar method without a glucanase treatment step.

[0034] In one embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of less than 12.2 mPa.s at room temperature, as determined by the method of Onwulata et al. (2014). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of about 10 mPa.s at room temperature, as determined by the method of Onwulata et al. (2014). In one embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of less than 8.5 mPa.s when heated at 85°C for 5 minutes, as determined by the method of Onwulata et al. (2014). In another embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of about 7.3 mPa.s when heated at 85°C for 5 minutes, as determined by the method of Onwulata et al. (2014). In a further embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of less than 14.1 mPa.s when heated at 85°C for 5 minutes and then cooled to room temperature, as determined by the method of Onwulata et al. (2014). In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts has a viscosity of approximately 11.2 mPa.s when heated at 85°C for 5 minutes and then cooled to room temperature, as determined by the method of Onwulata et al. (2014).

[0035] In one embodiment, a composition containing yeast protein derived from yeast protoplasts has a higher Protein Digestibility-Corrected Amino Acid Score (PDCAAS) than a yeast protein composition prepared by a similar method without a glucanase treatment step. For example, in some embodiments, the PDCAAS of the compositions of this disclosure may be 1.0 or higher. In one embodiment, a composition containing yeast protein derived from yeast protoplasts is more digestible than a yeast protein composition prepared by a method without a glucanase treatment step.

[0036] In one embodiment, a composition containing yeast protein derived from yeast protoplasts can be obtained by or is obtained by the method for obtaining a composition having yeast protein disclosed herein. In another embodiment, a composition containing yeast protein derived from yeast protoplasts contains a collected insoluble fraction, including yeast protoplasts, from the method for obtaining a composition having yeast protein disclosed herein. In yet another embodiment, a composition containing yeast protein derived from yeast protoplasts essentially consists of a collected insoluble fraction, including yeast protoplasts, from the method for obtaining a composition having yeast protein disclosed herein. In further embodiments, compositions containing yeast protein derived from yeast protoplasts obtained from different yeast sources can be combined for formulation into an edible composition.

[0037] edible products 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 further ingredient. In another embodiment, the edible product comprises a composition comprising yeast protein derived from yeast protoplasts and at least one further ingredient. In yet another embodiment, the composition contained in the edible product is a composition comprising yeast protein derived from yeast protoplasts of the present disclosure and at least one further ingredient.

[0038] In one embodiment, the composition contained in the edible product provides 1 to 100% of the protein in 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%, and 41% of the protein in the edible product, based on the total mass of the edible product. We offer 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%. In yet another embodiment, the composition contained in the edible product is the protein of the edible product, based on the total mass of the edible product, in the following proportions: 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%, 5 We offer 7%, 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% or less.In some embodiments, the composition contained in the edible product is the protein of the edible product, based on the total mass of the edible product, in the following proportions: 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 1 00%, 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%, The percentages provided are between 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%. In the context of this disclosure, the proportion of a composition in an edible product may depend on the properties of the edible product. In some specific embodiments, the proportion of a composition in a cheese analogue may be between 2% and 4% based on the total mass of the cheese analogue product. In some other specific embodiments, the proportion of the composition in the protein bar may be 30-40% based on the total mass of the protein bar.In further embodiments, the compositions of the present disclosure can be consumed as is (i.e., in flake form), and therefore the proportion of the composition is approximately 100%. Those skilled in the art will know how to adapt the proportion of the composition depending on the edible product considered. In one embodiment, the edible product may be a liquid or a solid. In another embodiment, the edible product may be a beverage ingredient, a beverage product, a food ingredient, a food, a feed ingredient, and / or a feed product. In some embodiments, the edible product may be a beverage ingredient and / or a beverage product. In some further embodiments, the edible product may be, but is not limited to, a shake, a dairy substitute such as a milk substitute, a liquid food, a soup, a broth, a smoothie, a cream, a gravie, and the like. In some embodiments, the edible product may be a food ingredient and / or a food. In some further embodiments, the edible product may be a baked product, a cooked / boiled product, and / or a dairy-like product. In yet another embodiment, baked products may include, but are not limited to, bread, cakes, muffins, cookies, pita, tortillas, buns, flatbreads, brownies, crackers, pastries, pies, tarts, torts, pizzas, etc. In yet another embodiment, cooked / boiled products may include, but are not limited to, bars, flakes, breakfast cereals, bacon, sausages, meat products such as hamburgers / sliced / ground steaks, meat substitutes, plant-based meats, pasta, noodles, condiments, snacks, sweets, gummies, chocolate-based products, ready-meal products, etc. In yet another embodiment, dairy-like products may include yogurt, cheese, cheese substitutes, cream, butter, custard, ice cream, etc. In some embodiments, edible products may be feed ingredients and / or feed products. In some other embodiments, feed ingredients / products may be concentrates, roughage, or mixed feeds. In further embodiments, the feed ingredients / products may include, but are not limited to, baked kibble, biscuits, powdered feed, extruded products, loaf, chunks, chunk-in-loaf products, pellets, crumbles, and the like.

[0039] This disclosure provides methods for producing edible products. The methods include obtaining an edible product by combining a composition of the disclosure with at least one raw material. The methods may include obtaining an edible product by heating the composition and at least one raw material (e.g., heated edible product). The methods may include obtaining an edible product by baking the composition and at least one raw material (e.g., baked edible product). The methods may include obtaining an edible product by blending the composition and at least one raw material (e.g., blended edible product). The methods may include obtaining an edible product by freezing the composition and at least one raw material (e.g., frozen edible product). The methods may include obtaining an edible product by extruding the composition and at least one raw material (e.g., extruded edible product). The methods may include obtaining an edible product by fermenting the composition and at least one raw material (e.g., fermented edible product).

[0040] Use of food products In one embodiment, the Disclosure is intended for the use of the edible products of the Disclosure for nutritional supplementation of humans and / or animals. In one embodiment, the edible products are provided as a substitute for edible products based on animal or plant / legume protein. In another embodiment, the edible products are combined with edible products based on animal or plant / legume protein. In yet another embodiment, the edible products are consumed / administered orally.

[0041] In one embodiment, the edible product of the present disclosure is intended to be combined with at least one food ingredient, at least one feed ingredient, and / or at least one beverage ingredient. The yeast composition may, in some embodiments, be directly incorporated into the food, feed product, and / or beverage product. In such embodiments, the yeast composition may be mixed with feed additives, food additives, further beverage additives, and / or binders. When the edible product of the present disclosure is intended to be used as an additive, it may be provided in liquid form, which in some further embodiments may be a spray-dryable liquid form. Alternatively, or in combination therewith, the edible product may be provided in powder form, which in some further embodiments may be a free-flowing powder.

[0042] In one embodiment, the edible product of the Disclosure is used as a food / feed supplement. 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 concentrates, metabolites, components and / or extracts, compared to a subject that does not take the supplement and consumes 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 to these. When the edible product of the Disclosure is intended to be used as a supplement, it may be provided in liquid form, which in some further embodiments may be an oil, solution or a spray-dryable liquid form. Alternatively, or in combination therewith, the supplement may be provided in powder form, which in some further embodiments may be capsules, pills, tablets, confectionery, gummies, etc.

[0043] In one embodiment, the edible products disclosed herein are for human and / or animal health, weight management, elderly and / or sports use. In another embodiment, the edible products herein may be incorporated into a diet or dietary restriction. In the context of this disclosure, the term “diet” is understood to mean any type and amount of food and beverages ingested by the subject (i.e., human and / or animal). Further in the context of this disclosure, the expression “dietary restriction” means restrictions imposed on the subject’s diet (i.e., human and / or animal’s diet) to limit the type and / or amount of food prescribed for ingestion. In another embodiment, when an edible product is administered for dietary restriction, it may be used to reduce the subject’s total weight and / or body fat mass, to increase the subject’s lean body mass, muscle mass, average capacity, endurance, and / or to regulate the subject’s hormone balance.

[0044] In one embodiment, the edible products disclosed herein can be used as meal replacements and / or medical foods. In another embodiment, the edible products disclosed herein can be used to limit, avoid, or replace the use of animal protein in the formulation of the edible products. In another embodiment, the edible products are intended for use in the recovery from illness, injury, and / or surgery. In another embodiment, the edible products disclosed herein are intended for use in oral / enteral clinical nutritional supplementation. [Examples]

[0045] Example 1: Process for producing various compositions containing yeast protein derived from Saccharomyces. Figure 1 schematically shows the processes for producing various compositions. Composition A (obtained after step 020 in Figure 1) corresponds to a composition containing yeast protein prepared according to the following method. Briefly, budding yeast (Saccharomyces cerevisiae) was cultured according to a standard fed-batch fermentation protocol, washed, and separated by centrifugation to obtain yeast cream. The yeast cream was heated at 95°C for 4 minutes using a heat exchanger in a holding tube to inactivate the yeast endogenous enzymes. The resulting slurry was then spray-dried to obtain composition A. Composition B (obtained after step 050 in Figure 1) corresponds to a composition containing yeast protein. Briefly, budding yeast (Saccharomyces cerevisiae) was cultured to obtain yeast cream as described above. The yeast cream was heated at 95°C for 4 minutes to inactivate the yeast endogenous enzymes. Next, the obtained slurry was cooled to 60°C, the pH was adjusted to pH 5.5, and then subjected to β-1,3-glucanase treatment (i.e., Denazyme GEL®, Nagase; 0.5% based on yeast dry material) at 60°C and pH 5.5 for at least 4 hours. The enzymatic reaction was stopped by heating the hydrolysate to 95°C for at least 2 minutes (this step also functioned as a pasteurization step). The obtained insoluble and soluble fractions were separated by centrifugation, the insoluble fraction was collected and then spray-dried at 85°C to obtain composition B. The soluble fraction obtained therefrom (after step 040 in Figure 1) was also spray-dried to obtain composition C.

[0046] Example 2: Comparison of Composition A, Composition B, and Composition C The protein content (by Kjeldahl method of nitrogen analysis using a coefficient of 6.25), lipid content (by the modified Majoria method; according to AOAC 989.05), nucleic acid content (by the method of Fish et al. (1991); by comparing values ​​with and without enzymatic digestion of nucleic acids), total carbohydrate content (monosaccharide quantification by HPLC after enzymatic / chemical digestion; according to AOAC 980.13), mannan content (by mannose quantification by HPLC after enzymatic / chemical digestion), glucan content (by glucose quantification by HPLC after enzymatic / chemical digestion), and glucose content (by direct glucose quantification by HPLC) were determined for compositions A, B, and C. These compositions (i.e., A, B, and C) were obtained from Example 1 and characterized in the examples herein according to standard methods known to those skilled in the art. The results of the characterization are shown 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 that reveals protoplast formation. (Data not included). For composition C, neither the lipid content nor the nucleic acid content has been reported.

[0047] [Table 1]

[0048] Example 3: Comparison of Composition A and Composition B The protein content of compositions A and B, obtained from the method described in Example 1, was determined and compared using the Kjeldahl method for nitrogen analysis (i.e., a standard method for quantifying the protein content of organic substances). Briefly, both compositions A and B (0.25 g) were heated to 373°C with concentrated sulfuric acid in the presence of copper sulfate as a catalyst. This oxidation reaction aims to decompose the sample and simultaneously release the reduced nitrogen as ammonium sulfate. The resulting solution was then distilled with sodium hydroxide to release ammonia, which was further reacted with boric acid and then dissolved in distilled water. The alkaline product was titrated with hydrochloric acid. The equivalence point (i.e., pH 4.9) was determined using both Tashiro indicator and pH. Based on the titration results, the amount of nitrogen in the sample was determined using a coefficient of 6.25, enabling the calculation of the protein content.

[0049] Table 2 below shows the results of the Kjeldahl method for nitrogen analysis of both sample A and composition B. Composition A has a protein content of approximately 60%, while composition B has a protein content of approximately 80%, both based on the total mass of the analyzed (dried or undried) compositions. The sample subjected to glucanase treatment (composition B) has a protein content approximately 20% higher than its non-enzymatically treated homologue (composition A).

[0050] [Table 2]

[0051] In this specification, the water solubility index (WSI) of two compositions (i.e., A and B) containing yeast protein was determined and compared. 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 drained to obtain a soluble fraction and several insoluble components. The soluble fraction was discarded, and the insoluble components were dried and weighed. The WSI represents the mass of the dried insoluble components, expressed as a percentage of the mass of the original sample.

[0052] The results of WSI determination for both Sample A and Sample B are shown in Table 3 below. Composition A has a WSI of approximately 23%, while Composition B has a WSI of approximately 5%, both based on the dry weight of the tested compositions. The sample subjected to glucanase treatment (Composition B) has a solubility of approximately one-fifth that of its non-enzymatically treated homologue (Composition A).

[0053] [Table 3]

[0054] The emulsifying activity and emulsion stability of two compositions containing yeast protein (i.e., composition A and composition B) were determined and compared with lecithin (i.e., a standard emulsifier in the art). Emulsion activity (EA) is defined as the maximum amount of oil that can be emulsified per unit volume of dry composition. Emulsion stability (ES) is defined as the rate of phase separation of water and oil during emulsion storage. Both EA and ES were determined using the methods of Brishti et al. (2017) and Yasumatsu et al. (1972), 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 mixtures were homogenized for 1 minute in a homogenizer and then centrifuged at 1100 × g for 5 minutes at 20°C.

[0055] The emulsification activity (EA) of both samples was calculated according to the following formula, EA = (H1 / H0) x 100, where H1 refers to the volume of the measured emulsion and H0 refers to the total volume of the solution in the measured tube.

[0056] i) The emulsion was heated at 80°C for 30 minutes, ii) the emulsion was cooled with tap water, and iii) the emulsion was centrifuged at 1100 × g for 5 minutes at 20°C. The emulsion stability (ES) of both samples was then determined. ES was calculated using the following formula: ES = (H1 / H0) × 100, where H1 represents the volume of the measured emulsion and H0 represents the total volume of the solution in the measured tube.

[0057] The results for emulsification properties (i.e., EA and ES) are shown in Table 4 below. The EA of composition A is comparable to that of lecithin, while the glucanase-treated sample (composition B) has an activity that is one-quarter to one-fifth that of both its non-enzymatically treated homolog (composition A) and lecithin. The ES of composition A is one-third to one-fourth that of lecithin, but three to four times that of its enzymatically treated counterpart (sample B). Therefore, composition B has an emulsion stability that is one-tenth to one-eleventh that of lecithin.

[0058] [Table 4]

[0059] The foaming capacity (FC) and foam stability (FS) of two compositions containing yeast protein (i.e., Composition A and Composition B) were determined according to the methods described by Chandra et al. (2015) and Brishti et al. (2017), respectively. Briefly, each sample (Composition A and Composition B; 0.2 g) was resuspended in 20 mL of distilled water in a 50 mL centrifuge tube. The resulting suspension was homogenized using a homogenizer and then foamed 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 is the measured volume of the suspension before the foaming step and V2 is the measured volume of the foamed suspension.

[0060] The foaming stability (FS) of each sample was measured over time (at 0, 15, 30, 45, and 60 minutes after suspension foaming), and the following formula was used: FS = (VF t / VF t0 Calculate according to ) × 100, and in the formula, VF t VF refers to the volume of foam measured at a specific time after foaming. t0 This refers to the measured foam volume immediately after whisking (i.e., 0 minutes).

[0061] Table 5 below shows the results of the foaming properties (i.e., FC and FS). The sample subjected to glucanase treatment (composition B) had a foaming ability approximately one-third to one-fourth that of its non-enzyme-treated homologue (composition A). In other words, composition A was 3 to 4 times more efficient at forming foam compared to its glucanase-treated counterpart. From the foam stability test, it was revealed that foam from composition A maintained about half of its stability after 30 minutes, while foam formed from the glucanase-treated composition (composition B) remained at only 10% after the same amount of time. Therefore, the stability of foam derived from composition B is approximately one-fourth to one-fifth that of foam prepared when the composition was not subjected to glucanase treatment (composition A).

[0062] [Table 5]

[0063] The odor and taste of two compositions containing yeast protein (i.e., Composition A and Composition B) were evaluated by trained evaluators and compared to Nutralys® F85M (pea protein isolate; Roquette). The samples (Composition A, Composition B, and Nutralys® F85M) were suspended in hot water to make a 2% w / w suspension. For sensory analysis, the 2% w / w suspension (40 mL) was placed in a glass container and served hot to each evaluator (N=2, n=2).

[0064] The Rate-All-That-Apply (RATA) method was used to determine the sensory characteristics of each sample. Participants were asked to select relevant terms from a given list and rate their intensity. In evaluating the samples, two main sensory modes were considered: smell and taste. To do this, the following parameters were evaluated: overall intensity (mandatory), sourness, cheese-like, creamy, vegetable, chicken / poultry, beef, roast-like, bread-like, fermented / alcoholic, and off-odor / taste. Furthermore, the taste modes included attributes such as umami, saltiness, sweetness, bitterness, and astringency. A numerical scale from 1 to 9 was applied to measure intensity, where 1 was "very low," 5 was "moderate," and 9 was "very strong." During the evaluation, participants were allowed to add comments for all modes as desired.

[0065] The results of these sensory evaluations are shown in Figure 2. Nutralys® F85M (Figure 2C) is identifiable by its strong vegetable flavor (pea smell and taste), bitterness, and astringency. Composition A (Figure 2A) had a strong smell and taste profile and was rated as having a chicken / poultry flavor along with strong roasted and beefy notes. Composition B (Figure 2B) was much less intense overall and more neutral than the other samples (i.e., showed the lowest average score). No samples had a cheese-like (smell), fermented (taste), off-odor, or off-flavor, so these attributes were excluded from Figures 2A, 2B, and 2C.

[0066] Viscosity measurements were also performed on both samples (Composition A and Composition B) 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 subjected to viscosity measurement using an Anton Paar Physica MCR301 rheometer with the RheoCompass program. RVA was performed using a temperature gradient as one test type. The temperature profile performed was as follows: a linear gradient of 25°C for 2 minutes → 25 to 85°C for 5 minutes → 85°C for 5 minutes → a linear gradient of 85 to 25°C for 5 minutes → 25°C for 2 minutes. The viscosity measurement results for both samples are shown in Table 6. Composition B was slightly less viscous than its non-enzymatically treated counterpart, regardless of the test conditions.

[0067] [Table 6]

[0068] The Protein Digestibility-Corrected Amino Acid Score (PDCAAS) is a method for evaluating protein quality based on both human amino acid requirements and human amino acid digestibility. Therefore, the PDCAAS method was used to evaluate the digestibility of both composition "A" and composition "B" (data not shown). The PDCAAS score for composition B (higher than 1.0) was higher than that of its non-enzymatically treated counterpart (i.e., composition A).

[0069] Example 4: Optimization of alkaline extraction We determined whether it is possible to extract nucleic acids and their derivatives from yeast cream using an alkaline extraction process. We also determined the effect of these processes on solubilizing nucleic acids outside of yeast cells using various pH and temperature settings.

[0070] A yeast cream of budding yeast (Saccharomyces cerevisiae) with an initial dry matter content of 17.72% and an initial protein content of 60.95% was obtained. The yeast cream sample (40g) was pH-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 produce different samples.

[0071] 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 dry weight (DW) of the supernatant was determined using a halogen dryer (hygrometer, MA37-1US, Sartorius). Alpha-amino nitrogen (AAN) and ribonucleic acid (RNA) were determined by high-pressure liquid chromatography.

[0072] Table 7 shows the yields of AAN and RNA observed in the supernatants after various alkaline extractions. As shown in Table 7, RNA is one of the compounds that is extracted better under alkaline conditions, as evidenced by the strong correlation between extraction pH and RNA recovery rate. RNA content at 55°C and 65°C was considerably higher compared to extractions at lower temperatures, reaching 23.5% at 55°C with pH 10 and at 65°C with pH 9 (Table 7). The supernatants produced at 55°C / pH 11, 65°C / pH 10, and 65°C / pH 11 had lower RNA content than those extracted at the same temperature but lower pH (Table 7).

[0073] [Table 7]

[0074] Example 5: Combination of alkaline extraction and glucanase treatment Next, we determined whether it was possible to reduce the nucleic acid content of the protein composition by using an alkaline extraction step before glucanase treatment.

[0075] Yeast cream from budding yeast (Saccharomyces cerevisiae) with a solid content of 16.0%, protein of 63.47%, phosphate of 4.06%, and RNA of 7.90% was collected. The yeast cream was heat-inactivated at 95°C for 5 minutes in an autolyser (Bailun, 20L, 200rpm), diluted to 15% dry matter (DM), and subjected to an alkaline extraction process (65°C, pH 9.0, 2 hours). The samples obtained after alkaline treatment were analyzed for the presence or absence of protoplasts (microscope) and for the dry matter (DM) using a halogen dryer (hygrometer, MA37-1US, Sartorius). Subsequently, the samples obtained after alkaline extraction were centrifuged at 4500 rpm for 10 minutes using a Sigma centrifuge (model 4-5L, rotor: 11650) to generate soluble and insoluble fractions. The insoluble fraction was washed once to a volume ratio of 1:1 and centrifuged again. The washed insoluble fraction was subjected to glucanase treatment (Denazyme GEL, 60°C, pH 5.6, 5 hours). The soluble fraction and the enzyme-treated insoluble fraction were spray-dried and further analyzed.

[0076] Alpha amino nitrogen (AAN) was determined by spectrophotometric analysis (adapted from the EBC-ninhydrin method used for determining free alpha amino nitrogen). Ribonucleic acid (RNA), glucans, and mannans were determined using high-pressure liquid chromatography. Protein yields were 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).

[0077] After alkaline extraction and glucanase treatment, the presence of protoplasts was confirmed (data not shown). Under the experimental conditions tested, the use of the alkaline extraction process maintained the protein content in the yeast extract at over 80% (Table 9), while reducing the nucleic acid content to less than 2% (Table 8).

[0078] [Table 8]

[0079] [Table 9]

[0080] Example 6: Composition B in a vegan American mozzarella-style cheese product For compositions having yeast protein prepared according to the method claimed in this disclosure (i.e., Composition B in the above examples), we decided to characterize the sensory and functional properties of a vegan cheese-type product as a complement to pea protein. Two tests were conducted: i) a pea protein-based vegan cheese (3.59%), and ii) a vegan cheese in which a portion of the pea protein was replaced with a "2.74% Composition B + 0.85% pea protein"-based vegan cheese, while keeping all other components unchanged. The formulations of the complete vegan cheese are shown in Table 10 below. The formulations contain no colorants or milking agents.

[0081] [Table 10]

[0082] Color evaluation was performed by visual observation and analysis of blocks and shredded cheese. The vegan cheese made from pea protein exhibited a yellowish hue typical of dairy products, while its partially substituted counterpart (a mixture of composition B and pea protein) was a pale, earthy beige color.

[0083] After being removed from the refrigerator (4°C), both cheese-like samples were cut into 20mm cubes to evaluate their sensory and textural properties. Flavor and texture profiles were determined by trained evaluators. The results are shown in Table 11 below.

[0084] [Table 11]

[0085] Partially replacing pea protein with a mixture of pea protein + composition B did not affect the flavor profile (i.e., neutral effect). However, blocks prepared with the claimed composition, when similarly shredded, were slightly softer than those prepared with its pea protein analogue.

[0086] Example 6: High-moisture extrusion cooking of meat-like material containing composition B We decided to evaluate the effect of adding Composition B powder to a plant-based meat analogue prepared using high-moisture extrusion. In the high-moisture extrusion process, the powder blend and water (approximately 50-70% of the total mass) were continuously fed into the extruder, where the material was rapidly heated to 150°C, and a co-rotating screw mechanically mixed and sheared the mass. As a result, the material was transformed into a fluid molten material. At the end of the barrel, the molten material was pushed by the screw into a long cooling tunnel, which compressed the material and cooled the mass to below 100°C, preventing the water from boiling by applying back pressure. Friction between the solidified layers of the fluid material in the cooling tunnel created long protein fibers similar to the structure of meat.

[0087] As shown in Table 12, different proportions of composition B were added to the dry blend, thereby partially replacing the legume proteins (i.e., H5, H10, H15, H25, and H35). H0 contains no composition B at all, while H100 contains only composition B.

[0088] [Table 12]

[0089] Meat analogue extrudates were produced from all the blends tested as follows. Samples were processed using a co-rotating intermeshing twin-screw extruder KETSE 20 / 40 (Brabender, Duisburg, Germany) equipped with a long cooling die (24×7×700 mm, W×H×L). A screw with a length-to-diameter ratio of 40 was configured to apply moderate shear. By calibrating the volumetric feeder using powder, the mass flow rate of the material was set constant at 4 kg h -1 . Water was added from a separate port using a calibrated peristaltic pump. The temperature profile was set at 45 °C, 83 - 84 °C, 135 - 137 °C and 151 - 154 °C. The temperature of the cooling die was maintained at 65 °C using a temperature control unit, but only for the last 300 mm, thereby creating a temperature gradient along the die. The water content was adjusted for each blend, and when the process reached stability as indicated by the measured pressure and temperature, one sample was collected from each blend. The screw speed and temperature were adjusted slightly between blends to keep the process stable. Samples were packed in zip-lock (registered trademark) bags and stored frozen at -20 °C until analysis.

[0090] Texture profile analysis by instrument: The frozen samples were thawed and rehydrated in water at 60 °C (temperature not maintained) for 2 hours. Before measurement, the moisture of the sample pieces was blotted with tissue paper. 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 sample pieces were cut into a shape of 15××7 mm, and compressed twice at 70% with a probe speed of 3 mm s -1 , a pre-test speed of 1.5 mm s -1 and a holding time of 1 s between compressions. The load cell was 50 kg. Hardness, chewiness, cohesiveness, elasticity and resilience were calculated using the software of the texture measuring instrument.

[0091] The differences in the mechanical properties of the extruded materials were revealed by the TPA (Total Performance Assessment) of the extruded materials, which are summarized in Figure 3. Sample H100 was not measured because its hardness was too high for the test apparatus. All extruded parameters (i.e., hardness, chewiness, cohesiveness, elasticity, and resilience) tested for extruded materials based on composition B (H5, H10, H15, H25, and H35) were higher than those of the control (H0).

[0092] Descriptive Sensory Analysis (DSA): Frozen samples were thawed and rehydrated in 60°C water (temperature not maintained) for 2 hours. Before sensory evaluation, moisture was absorbed from the sample pieces with tissue paper, and they were cut into 4 cm long pieces. Sensory analysis was performed by eight professional evaluators who had previously been trained and experienced with such plant-based extruded samples. The analysis was performed in a standard (ISO 6668:2008) sensory laboratory. Samples were coded with a 3-digit code and placed in cups. The order of the samples was randomized according to the experimental design using the Williams-Latin square method. Samples were served at room temperature. Water and crackers were provided between samples for palate irrigation. Samples were evaluated on a scale from 0 to 9 ("0" - none; "1" - very weak; "5" - moderate; "9" - very strong). A total of three aspects were evaluated: odor, taste, and texture. For aroma and taste, sensory analysis included attributes such as overall intensity, bean-like, yeast-like (smell) / umami (taste), and off-odor. Taste profile further included aftertaste. Texture profile included evaluations of fibrousness, elasticity, hardness, chewiness, adhesion, granularity, cohesiveness, and wetness. Only fibrousness was evaluated manually; all others were evaluated by mouthfeel. Evaluators were also allowed to add comments in the optional fields for each attribute. DSA results for different extruders are shown in Figures 4 and 5. The observed differences were extremely small up to 35% yeast addition. However, the 100% composition B extruder (H100) showed significant differences from the others in all attributes. As shown in Figure 4, composition B had a mild flavor, and when added to samples H5-H35, the meat-like / yeast-like aroma and umami only increased slightly compared to the control (H0), whereas in H100, this increase was much greater. On the other hand, while the intensity of the meaty smell of H35 was similar to that of H25, there was a comment that it had a faint roasted smell, which is thought to be related to its higher yeast content compared to H5-H25. There was no difference in the intensity of the bean-like smell and taste, or the overall smell and taste, among H0-H35, but H100 had a stronger overall smell and taste than the others. Off-odor, off-flavor, and aftertaste were detected only in H100.Off-notes described for H100 included "powdery" and "paper-like." As shown in Figure 5, regarding texture attributes such as adhesion, cohesiveness, fibrousness, and elasticity, no differences were observed among samples H0 to H35, except for H100, which had high cohesiveness and elasticity but low fibrousness. More pronounced differences were observed in other attributes, with samples H5 to H35 showing increased chewiness, hardness, and granularity compared to the control (H0).

[0093] Example 7: Process for producing a composition containing yeast protein derived from Cyberlindera. Yeast of Cyberlindera jardinii was cultured to obtain a yeast cream. The yeast endogenous enzymes were inactivated by heating the yeast cream at 95°C for 5 minutes. The resulting slurry was then cooled to 65°C and the pH was adjusted to pH 9.0. After maintaining these conditions for 30 minutes, the pH was lowered to 8.0 and alkaline extraction was performed. The resulting insoluble and soluble fractions were separated by centrifugation, and the insoluble fraction was collected and subjected to β-1,3-glucanase treatment (i.e., Denazyme GEL®, Nagase; 0.5% based on the yeast dry material) at 55°C and pH 5.5 for 5 hours. The enzymatic reaction was stopped by heating the hydrolysate at 95°C for at least 2 minutes (this step also functioned as a pasteurization step). The resulting insoluble and soluble fractions were separated by centrifugation, and the insoluble fraction was collected and then spray-dried at 85°C. The protein content was determined by the Kjeldahl method of nitrogen analysis using a coefficient of 6.25. The resulting yeast extract contained 83.97% protein.

[0094] While the present invention has been described in relation to its specific embodiments, it will be understood that 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.

[0095] References [1]AOAC 980.13 - 1980; Title: Fructose, Glucose, Lactose, Maltose, and Sucrose in Milk Chocolate - Liquid Chromatography Method [2]AOAC 989.05 - 1992; Title: Fat in Milk Modified Mojonnier Ether Extraction Method [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 [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. Epub 2014 Jun 10. PMID: 26028751; PMCID: PMC4444897 [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 [6]Onwulata C.I.et al.,(2014)Rapid visco analysis of food protein pastes.Journal of Food Processing and Preservation.38,pp.2083-2089 [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 having yeast protein, wherein the method comprises the following steps, namely, a) A process of providing yeast cream containing yeast, b) A step of inactivating the endogenous enzymes of the yeast to provide an inactivated yeast cream, c) A step of subjecting the inactivated yeast cream to enzymatic treatment to obtain an insoluble fraction containing yeast protoplasts and a soluble fraction, wherein the enzymatic treatment comprises at least one polypeptide having glucanase activity and the enzymatic treatment lacks ribonuclease activity. d) A step of separating the insoluble fraction from the soluble fraction, e) A step of collecting the insoluble fraction, wherein the collected insoluble fraction, when dried, is a composition containing the yeast protein and has a protein content of 60% or more based on the total mass of the collected and dried insoluble fraction; Methods that include...

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

3. The method according to claim 1 or 2, wherein the enzyme treatment is carried out at a temperature of 20 to 80°C for a period of 1 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, further comprising drying the insoluble fraction collected in step e) to provide the composition.

6. The method according to claim 5, wherein the dried insoluble material is - Lipid content of less than 20% based on the total mass of the collected and dried insoluble fraction, • A nucleic acid content exceeding 6% based on the total mass of the collected and dried insoluble fraction. - Carbohydrate content of less than 25% based on the total mass of the collected and dried insoluble fraction, - Mannan content of less than 6% based on the total mass of the collected and dried insoluble fraction, - Glucan content of less than 10% based on the total mass of the collected and dried insoluble fraction and / or - Glucose content of less than 25% based on the total mass of the collected and dried insoluble fraction. A method having.

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

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

9. The method according to claim 8, wherein the dried insoluble fraction is - Lipid content of less than 20% based on the total mass of the collected and dried insoluble fraction, - A nucleic acid content of less than 3% based on the total mass of the collected and dried insoluble fraction. - Carbohydrate content of less than 25% based on the total mass of the collected and dried insoluble fraction, - Mannan content of less than 6% based on the total mass of the collected and dried insoluble fraction, - Glucan content of less than 10% based on the total mass of the collected and dried insoluble fraction and / or - Glucose content of less than 25% based on the total mass of the collected and dried insoluble fraction. A method having.

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

11. It contains yeast proteins derived from yeast protoplasts, and - Protein content of 60% or more based on the total mass of the above composition, Lipid content of less than 20% based on the total mass of the above composition, - Carbohydrate content of less than 25% based on the total mass of the above composition, - Mannan content of less than 6% based on the total mass of the above composition, - Glucan content of less than 10% based on the total mass of the composition and / or - Glucose content of less than 25% based on the total mass of the above composition A composition having the following characteristics.

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

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

14. A composition according to any one of claims 11 to 13, having a neutral taste.

15. A composition according to any one of claims 11 to 14, which can be obtained by the method described in any one of claims 1 to 10, or which has been obtained by such method.

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

17. The edible product according to claim 16, which is a beverage, shake, bar, meat product or baked product.

18. Use of the edible product according to claim 16 or 17 in the nutritional supplementation of humans and / or animals.

19. The use according to claim 18, wherein the edible product is provided as a substitute for an edible product based on animal protein or plant / legume protein, or is combined with an edible product based on animal protein or plant / legume protein.

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

21. The use according to claim 20, wherein the food / feed supplement or food / feed additive is for weight management, elderly, oral / enteral clinical nutritional supplementation, sports use and / or animal nutritional supplementation.

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