Composition containing yeast cell wall-derived decomposition product, method for producing the same, and use thereof

Abstract

To provide a production method of a composition containing a yeast cell wall-derived decomposition product capable of improving the solubilization rate and dispersion stability of a yeast cell wall, having high working efficiency and production efficiency without requiring an operation of separating and removing insoluble fractions of the yeast cell wall at low cost; and to provide a composition containing a yeast cell wall-derived decomposition product with excellent solubility and dispersion stability, a beer-like aroma-imparting composition containing the composition containing the yeast cell wall-derived decomposition product, food / beverages containing the composition containing the yeast cell wall-derived decomposition product, and a palatability improver for pet food.SOLUTION: Provided is a beer yeast cell wall-derived decomposition product containing an ethyl caproate and an ethyl caprylate, and further containing at least one compound selected from an ethyl caprate, a phenylethyl alcohol and a capric acid, and having a volume of deposit when measured under specific measurement conditions, of 50 mL or less.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a composition containing a yeast cell wall-derived degradation product and a method for producing the same, as well as a composition for imparting a beer-like aroma, a composition for imparting a roast flavor, a composition for imparting a roast color, a food or beverage, and a palatability improver for pet food, each containing the yeast cell wall-derived degradation product. [Background technology]

[0002] Yeast cells contain various nutritional components such as proteins, vitamins, minerals, nucleic acids, glutathione, and dietary fiber, and are therefore widely used in various fields such as the food, bio, and cosmetics industries.

[0003] Examples of uses of yeast cells include yeast extracts extracted from components contained in yeast cells, yeast cell walls generated as residues after separating yeast extracts during production of yeast extracts, etc. Among these, yeast extracts are often used because they are highly soluble in water and can be highly concentrated to about 65% to 70% by mass, making them easy to process. On the other hand, yeast cell walls are insoluble in water, have very stable physical properties against heat and the like, and further contain a large amount of polysaccharides. Therefore, even in the state of a slurry containing yeast cell walls, when the concentration of the slurry reaches about 15% by mass or more, the viscosity increases significantly, making it impossible to achieve high concentration, and processing is difficult, so that yeast cell walls are not currently being used effectively.

[0004] In recent years, there has been a growing global demand for organic, natural, additive-free products in the food and cosmetics fields, and product development is underway to meet these demands. In particular, in the European Union (EU), the CIAA (Confederation of the European Food and Beverage Industry) has established standards for attaching "clean labels" indicating that products are natural, according to its guidelines on traceability in general food law (see Non-Patent Document 1).

[0005] As a product using yeast extract that can be labeled with a "clean label," for example, a reaction flavor having a beef flavor, roast flavor, or the like has been proposed (see Patent Document 1). This reaction flavor is obtained as a product of the Maillard reaction. The Maillard reaction is a reaction in which reducing monosaccharides (monomers) and amino acids are heated to produce brown substances. In the EU, when reducing monosaccharides such as glucose and amino acids are added externally as secondary ingredients, they must be labeled as additives, and a "clean label" cannot be attached, which does not meet the above demand and is undesirable.

[0006] Therefore, the proposed reaction flavor is produced by stepwise treating yeast extract or yeast autolysate with enzymes such as endoprotease, α-α-trehalase, glucoamylase, endoglucanase, and exoglucanase to obtain a yeast-derived product that produces yeast-derived amino acids and reducing sugars, which is then centrifuged to remove insoluble components (i.e., yeast cell walls), and incubating the yeast extract under specified conditions to cause a Maillard reaction between the yeast-derived amino acids and reducing sugars.

[0007] In the proposed method, a yeast extract or yeast autolysate containing yeast cell walls is treated with an endoprotease and then with glucanase, which is thought to produce a certain amount of amino acids and reducing sugars from the yeast cell walls. However, as mentioned above, the solubilization rate of yeast cell walls is very low, so in the proposed method, only a portion of the components contained in the yeast cell walls is solubilized, resulting in a large amount of precipitate derived from the yeast cell walls, which requires centrifugation to remove the insoluble components.

[0008] As described above, since yeast cell walls can only be partially solubilized, the insoluble components must be removed, resulting in problems of poor work efficiency and production efficiency. Furthermore, yeast cell walls are difficult to process, and are very poor in reducing monosaccharides and amino acids, making it impossible to obtain the reaction flavor proposed above. On the other hand, yeast cell walls are obtained as a residue after separating yeast extract, and therefore have the advantage that they can be used at low cost if their processing properties, such as solubilization, are improved.

[0009] A flavor composition that utilizes yeast cell walls and can enhance or impart a fatty and creamy texture has also been proposed (see Patent Document 2). This flavor composition is produced by contacting a yeast cell wall slurry with endoglucanase (laminaripentaose-producing β-1,3-glucanase) and endoprotease, removing insoluble components by solid-liquid separation, and isolating the liquid fraction. Therefore, the solubilization rate is not fully satisfactory. Furthermore, although this method produces amino acids derived from yeast cell walls, it does not produce reducing monosaccharides as sugars, and instead produces non-reducing polysaccharides (pentasaccharide oligosaccharides consisting of glucose β-1,3 glucoside bonds) due to the action of endoglucanase. Therefore, it is not possible to obtain Maillard reaction products using this flavor composition.

[0010] Maillard reaction products having flavors such as meat flavor and roast flavor are known to contribute not only to improving palatability for humans but also to improving palatability for pets (see Patent Document 3, Non-Patent Document 2, etc.).

[0011] Therefore, there is currently a strong demand for a method for producing a composition containing a yeast cell wall-derived degradation product that can improve the solubilization rate and dispersion stability of yeast cell walls, does not require the separation and removal of the insoluble fraction of yeast cell walls, and has high work efficiency and production efficiency, as well as a method for providing a composition containing a yeast cell wall-derived degradation product that can be produced using only raw materials derived from yeast cell walls and has a good flavor such as meat flavor or roast flavor. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] Patent No. 5982696 [Patent Document 2] Special Publication No. 2018-531586 [Patent Document 3] Japanese Patent Application Laid-Open No. 2017-86079 [Non-patent literature]

[0013] [Non-Patent Document 1] CIAA GUIDELINES, on Regulation (EC) No. 1334 / 2008, on Flavourings and Certain Food Ingredients with Flavouring Properties for Use in and on Foods [Non-patent document 2] The latest trends in pet food and pet medicines, "Chapter 4: Palatability of pet food", December 16, 2013, pp. 29-35, CMC Publishing Summary of the Invention [Problem to be solved by the invention]

[0014] The present invention aims to solve the above-mentioned problems in the prior art and to achieve the following objects. In other words, the present invention aims to provide a method for producing a composition containing a yeast cell wall-derived degradation product, which can improve the solubilization rate and dispersion stability of yeast cell walls, does not require the separation and removal of the insoluble fraction of yeast cell walls, has high work efficiency and production efficiency, and is low cost.

[0015] An object of the present invention is to provide a composition containing a cell wall-derived degradation product that has excellent solubility and dispersion stability.

[0016] An object of the present invention is to provide a composition containing a degradation product derived from yeast cell walls, which has excellent solubility and dispersion stability, a good roast flavor and oily feel, and a good roast color.

[0017] An object of the present invention is to provide a composition for imparting beer-like aroma, which has excellent solubility and dispersion stability, little off-flavor, and a good beer-like aroma.

[0018] An object of the present invention is to provide a composition for imparting roast flavor, which has excellent solubility and dispersion stability and has a good roast flavor and oily feel.

[0019] An object of the present invention is to provide a roasted color-imparting composition that has excellent solubility and dispersion stability and has a good roasted color.

[0020] Another object of the present invention is to provide a food or drink containing the composition containing the yeast cell wall-derived degradation product.

[0021] An object of the present invention is to provide a palatability improver for pet food, which contains the composition containing the yeast cell wall-derived degradation product. [Means for solving the problem]

[0022] The means for solving the above problems are as follows: <1> This is a method for producing a composition containing a degradation product derived from yeast cell walls, which is characterized by including an enzyme treatment step in which yeast cell walls are treated with exoglucanase. <2> The enzyme treatment step further comprises treating the yeast cell wall with a protease. <1> This is a method for producing a composition containing the yeast cell wall-derived degradation product described in the above. <3> The enzyme treatment step further comprises exopeptidase treatment of the yeast cell wall. <2> This is a method for producing a composition containing the yeast cell wall-derived degradation product described in the above. <4> The method further comprises a heat treatment step of heat-treating the enzyme-treated product obtained in the enzyme treatment step. <1> from <3> This is a method for producing a composition containing a yeast cell wall-derived degradation product described in any one of the above. <5> The method further comprises a pH adjustment step of adjusting the enzyme-treated product obtained in the enzyme treatment step to alkaline, The heat treatment step is a step of heat-treating the alkaline enzyme-treated product obtained in the pH adjustment step. <4> This is a method for producing a composition containing the yeast cell wall-derived degradation product described in the above. <6> A yeast cell wall-derived decomposition product-containing composition containing a yeast cell wall-derived decomposition product, The yeast cell wall-derived decomposition product-containing composition is characterized in that when 100 mL of the yeast cell wall-derived decomposition product-containing composition having a solid content concentration of 10% by mass is left standing at 25°C and normal pressure for 48 hours, the volume of the sediment is 50 mL or less. <7> The composition containing a yeast cell wall-derived degradation product has a total free amino acid content of 7% by mass or more and a reducing sugar content of 1% by mass or more relative to the solid content of the composition. <6> The composition contains the yeast cell wall-derived decomposition product described in 1. <8> The composition containing a yeast cell wall-derived degradation product has a total free amino acid content of 12% by mass or more and a reducing sugar content of 10% by mass or more relative to the solid content of the composition. <6> from <7> The present invention relates to a composition containing a yeast cell wall-derived decomposition product according to any one of the above. <9> The composition contains at least one compound selected from ethyl caproate, ethyl caprylate, ethyl caprate, phenylethyl alcohol, and capric acid. <6> from <8> The present invention relates to a composition containing a yeast cell wall-derived decomposition product according to any one of the above. <10> The above-mentioned decomposition product derived from yeast cell wall is a decomposition product derived from brewer's yeast cell wall. <9> The composition contains the yeast cell wall-derived decomposition product described in 1. <11> A yeast cell wall-derived decomposition product-containing composition containing a yeast cell wall-derived decomposition product, The composition contains at least one compound selected from pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol. <12> The aforementioned <9> from <10> 1. A composition for imparting a beer-like aroma, comprising the yeast cell wall-derived decomposition product-containing composition according to any one of the above. <13> The aforementioned <11> The present invention relates to a composition for imparting roast flavor, characterized by containing the yeast cell wall-derived degradation product-containing composition described in the above. <14> The aforementioned <11> The present invention relates to a composition for imparting roasted color, which comprises the yeast cell wall-derived decomposition product-containing composition described in the above. <15> The aforementioned <6> from <11> The food and drink product is characterized by containing the yeast cell wall-derived decomposition product-containing composition described in any one of the above. <16> The aforementioned <6> from <11> The present invention is a pet food palatability improver characterized by containing the yeast cell wall-derived decomposition product-containing composition according to any one of the above. [Effects of the Invention]

[0023] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objectives can be achieved, and a method for producing a composition containing a yeast cell wall-derived degradation product can be provided, which can improve the solubilization rate and dispersion stability of yeast cell walls, does not require the separation and removal operation of the insoluble fraction of yeast cell walls, has high work efficiency and production efficiency, and is low cost.

[0024] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objects can be achieved, and a composition containing a cell wall-derived degradation product having excellent solubility and dispersion stability can be provided.

[0025] According to the present invention, the above-mentioned problems of the prior art can be solved and the above-mentioned object can be achieved, and a composition containing a yeast cell wall-derived decomposition product can be provided which has excellent solubility and dispersion stability, a good roasted flavor and oily feel, and a good roasted color.

[0026] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objects can be achieved, and a composition for imparting a beer-like aroma can be provided which is excellent in solubility and dispersion stability, has little off-flavor, and has a good beer-like aroma.

[0027] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned object can be achieved, and a roast flavor imparting composition having excellent solubility and dispersion stability and having a good roast flavor and oily feel can be provided.

[0028] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objects can be achieved, and a roasted color-imparting composition having excellent solubility and dispersion stability and having a good roasted color can be provided.

[0029] According to the present invention, the above-mentioned conventional problems can be solved, the above-mentioned object can be achieved, and a food or drink containing the above-mentioned composition containing a degradation product derived from yeast cell walls can be provided.

[0030] According to the present invention, the above-mentioned conventional problems can be solved and the above-mentioned object can be achieved, and a palatability improver for pet food containing the above-mentioned yeast cell wall-derived degradation product-containing composition can be provided. [Brief explanation of the drawings]

[0031]

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[0032] (Composition containing yeast cell wall-derived decomposition product and method for producing the same) The method for producing a composition containing a yeast cell wall-derived degradation product of the present invention includes at least an enzyme treatment step, and preferably further includes a pH adjustment step and a heat treatment step, and may further include other steps as necessary. In this specification, the composition containing a yeast cell wall-derived degradation product obtained in the enzyme treatment step may be referred to as an "enzyme-treated product." Also, in this specification, the composition containing a yeast cell wall-derived degradation product obtained in the heat treatment step may be referred to as a "heat-treated product."

[0033] The yeast cell wall-derived decomposition product-containing composition of the present invention contains at least a yeast cell wall-derived decomposition product, and when 100 mL of the slurry yeast cell wall-derived decomposition product-containing composition having a solids concentration of 10% by mass is left standing at 25°C and normal pressure for 48 hours, the volume of the sediment is 50 mL or less. The yeast cell wall-derived decomposition product-containing composition of the present invention can be suitably produced by the method for producing a yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) of the present invention.

[0034] The yeast cell wall-derived decomposition product-containing composition of the present invention contains at least a yeast cell wall-derived decomposition product and at least one compound selected from pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol. The yeast cell wall-derived decomposition product-containing composition of the present invention can be suitably produced by the method for producing a yeast cell wall-derived decomposition product-containing composition (heat-treated product) of the present invention.

[0035] The composition containing a degradation product derived from a yeast cell wall of the present invention will be described below together with a method for producing the composition containing a degradation product derived from a yeast cell wall of the present invention.

[0036] <Enzyme treatment process> The enzyme treatment step is a step of treating yeast cell walls (hereinafter sometimes abbreviated as "yeast cell walls") with exoglucanase. The enzyme treatment step preferably further includes at least one of protease treatment and exopeptidase treatment of the yeast cell walls, and more preferably includes both protease treatment and exopeptidase treatment. In the enzyme treatment step, the yeast cell walls are decomposed, whereby a composition containing a decomposition product derived from the yeast cell walls (enzyme-treated product) having excellent solubility can be obtained.

[0037] Here, the solubilization rate in the present invention is calculated by the following formula (1). Solubilization rate (%)=solid mass A / solid mass B×100...Formula (1) In the formula (1), "solid mass B" represents the mass of the dried product obtained by drying X g of the target sample at 105°C for 5 hours. In addition, in the formula (1), "solid mass A" indicates the mass of the dried product obtained by centrifuging X g of the target sample at 5,000 G for 5 minutes and drying the resulting supernatant at 105°C for 5 hours. Examples of the target sample include the yeast cell wall, the enzyme-treated product, and the heat-treated product.

[0038] <<Yeast cell wall>> In the present invention, the yeast cell wall is an insoluble fraction obtained as a heavy liquid after treatment to extract yeast extract from yeast cells (sometimes referred to as "yeast" or "yeast cells") and separation of the resulting yeast cell extract. The soluble fraction obtained as a supernatant after separation of the extract is the yeast extract. Therefore, the yeast cell wall in the present invention does not include yeast cell wall forms containing the yeast extract.

[0039] The type of yeast that is the raw material for the yeast cell wall is not particularly limited and can be appropriately selected depending on the purpose. Examples include baker's yeast, brewer's yeast, wine yeast, sake yeast, soy sauce yeast, Torula yeast, and yeast for bioethanol.

[0040] The genus of the yeast is not particularly limited and can be appropriately selected depending on the purpose. For example, Saccharomyces Saccharomyces ) genus, Candida ( Candida ) genus, Kluyveromyces ( Kluyveromyces Among these, Saccharomyces ( Saccharomyces ) genus is preferred. The Saccharomyces Saccharomyces ) yeasts include Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) etc. The yeast cell walls used may be prepared from one type of yeast, or may be prepared from two or more types of yeast in combination.

[0041] The yeast cell walls used may be commercially available products, or may be prepared by a known method such as appropriate cultivation for the purpose of using them in the method for producing the composition containing a degradation product derived from yeast cell walls. Furthermore, from the viewpoint of utilizing waste materials and reducing waste disposal costs, yeast cell walls obtained as excess waste from the brewing industry of beer, whiskey, wine, shochu, sake, miso, soy sauce, etc. may be used.

[0042] The treatment method for extracting yeast extract from the yeast cells to prepare the yeast cell walls is not particularly limited and can be appropriately selected from known yeast extract extraction methods, such as autolysis, hot water extraction, enzymatic hydrolysis, alkaline hydrolysis, freeze-thawing, physical disruption, etc. These extraction methods may be carried out alone or in combination of two or more.

[0043] Specifically, the autolysis method is a method in which yeast cells are solubilized by utilizing proteolytic enzymes and the like that are naturally present in the yeast cells. Specifically, the hot water extraction method is a method in which yeast cells are solubilized by immersing them in hot water for a certain period of time. The hot water temperature and immersion time in the hot water extraction method are not particularly limited and can be appropriately selected depending on the purpose. Specifically, the enzymatic degradation method is a method in which yeast cells are solubilized by adding an enzyme preparation derived from a microorganism or a plant. The enzyme preparation used in the enzymatic degradation method is not particularly limited and can be appropriately selected from known enzyme preparations depending on the purpose. Specifically, the acid or alkali decomposition method is a method in which yeast cells are solubilized by adding an acid or alkali. Specifically, the freeze-thaw method is a method in which yeast cells are disrupted by freezing and thawing at least once. Specifically, the physical disruption method disrupts yeast cells by applying a physical stimulus. The physical stimulus used in the physical disruption method is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include ultrasonic waves, homogenization under high pressure, grinding with a microgrinder, and grinding by mixing with a solid material such as glass beads.

[0044] Among these, the yeast cell walls are preferably yeast cell walls obtained by an autolysis method (hereinafter sometimes referred to as "autolysis-type yeast cell walls"), yeast cell walls obtained by a hot water extraction method (hereinafter sometimes referred to as "hot water extraction-type yeast cell walls"), or yeast cell walls obtained by an enzymatic degradation method (hereinafter sometimes referred to as "enzymatic degradation-type yeast cell walls").

[0045] The method for separating the yeast cell walls from the yeast cell extract is not particularly limited and can be appropriately selected depending on the purpose. For example, centrifugation may be used. The conditions for the centrifugation are not particularly limited and can be appropriately selected depending on the purpose, but conditions of 4,700 G to 5,500 G for 5 minutes are preferred, and conditions of 5,000 G for 5 minutes are more preferred.

[0046] The form of the yeast cell walls used in the enzyme treatment step is not particularly limited and can be selected appropriately depending on the purpose. Examples include a slurry (mud-like) obtained by separating the yeast cell walls from an extract, a suspension in which the slurry is suspended in a solvent, a paste-like form obtained by compressing and concentrating the slurry, a dried product obtained by drying the paste-like form and further concentrating it, and a powder-like form obtained by pulverizing the dried product. Among these, the yeast cell walls are preferably in the form of a slurry (mud) or a suspension of the slurry in a solvent, since they do not require processing such as squeezing or drying and have good working efficiency and production efficiency. A suspension of the slurry in a solvent is more preferred, since it has a lower viscosity than the slurry and has good working efficiency and production efficiency.

[0047] The concentration of the slurry in the suspension is not particularly limited and can be appropriately selected depending on the viscosity of the slurry, etc. However, the yeast cell wall slurry usually has a very high viscosity. Therefore, in the case of autolyzed yeast cell walls, the concentration of the slurry in the suspension is often limited to about 15% to 16% by volume, and in the case of hot water-extracted yeast cell walls, the concentration of the slurry in the suspension is often limited to about 15% to 20% by volume.

[0048] The protein content in the yeast cell wall is not particularly limited and can be appropriately selected depending on the extraction method of the yeast extract used to prepare the yeast cell wall. The protein content in the autolytic yeast cell wall is generally about 20% by mass to 30% by mass. The protein content in the hot water-extracted yeast cell walls is generally about 45% to 55% by mass. The protein content in the enzymatically decomposed yeast cell walls varies depending on the type of enzyme used, but is often close to the protein content in the autolyzed yeast cell walls. The protein content in the yeast cell wall is the content relative to the mass of the solid content of the yeast cell wall, and can be measured by a combustion method (modified Dumas method).

[0049] In the present invention, the solid content mass of the yeast cell wall refers to the mass of the dried product obtained by drying the yeast cell wall at 105°C for 5 hours.

[0050] The carbohydrate content in the yeast cell wall is not particularly limited and can be appropriately selected depending on the extraction method of the yeast extract used to prepare the yeast cell wall. The carbohydrate content in the autolytic yeast cell wall is generally 45% by mass or more, with the upper limit being approximately 65% ​​by mass. The carbohydrate content in the hot water-extractable yeast cell walls is generally 50% by mass or less, with the lower limit being approximately 20% by mass. The carbohydrate content in the enzymatically decomposed yeast cell walls varies depending on the type of enzyme used, but is often close to the carbohydrate content in the autolytic yeast cell walls. The carbohydrate content in the yeast cell wall is the content relative to the mass of the solid content of the yeast cell wall, and can be calculated by the following formula (2). Carbohydrate content in yeast cell wall = 100 - (A + B + C + D) Equation (2) In the formula (2), "A" represents the protein content in the yeast cell wall measured by the combustion method (modified Dumas method). "B" represents the moisture content in the yeast cell wall measured by the atmospheric pressure heat drying method. "C" represents the lipid content in the yeast cell wall measured by the acid decomposition method. "D" represents the ash content in the yeast cell wall measured by the direct ashing method.

[0051] <<Exoglucanase>> The exoglucanase is an exoenzyme that cleaves the ends of cellulose chains in the yeast cell wall. The exoglucanase is not particularly limited and can be selected appropriately, but it is preferably one that can be used for food applications or is of a grade that can be used for food applications. The type of exoglucanase is not particularly limited and can be selected appropriately, but glucanases including at least one of exo-1,3-β-glucanase, exo-1,4-β-glucanase, and exo-1,6-β-glucanase are preferred.

[0052] The source of exoglucanase is not particularly limited and can be selected appropriately depending on the purpose. Talaromyces ) genus; Lasamsonia emersonii ( Rasamsonia emersonii ) and other Rasamsonia ( Rasamsonia ) genus; Dysporotrichum zymophosporum ( Disporotrichum dimorphosporum ) and other Dysporotrichum ( Disporotrichum ) origin; Streptomyces violaceolvera ( Streptomyces violaceoruber ) and other species from the genus Streptomyces; Trichoderma longibrachiatum , Trichoderma ressei Trichoderma ( Trichoderma ) genus, and preferably Lasamsonia emersonii ( Rasamsonia emersonii ), Dysporotrichum zymophosporum ( Disporotrichum dimorphosporum ), Talaromyces ( Talaromyces ) genus is more preferred, and Talaromyces ( Talaromyces It is particularly preferable that the genus be derived from these. These may be used alone or in combination of two or more.

[0053] The exoglucanase to be used may be a commercially available product, or may be prepared appropriately by a known method from a bacterium or the like having exoglucanase. Examples of commercially available exoglucanases include FILTRASE (registered trademark) BRX (manufactured by DSM Japan Co., Ltd.), FILTRASE (registered trademark) BR-XL (manufactured by DSM Japan Co., Ltd.), Denazyme GEL-L1 / R (manufactured by Nagase ChemteX Corporation), and Sumiteam TG (manufactured by Shin-Nihon Chemical Industry Co., Ltd.). FILTRASE (registered trademark) BRX (manufactured by DSM Japan Co., Ltd.), FILTRASE (registered trademark) BR-XL (manufactured by DSM Japan Co., Ltd.), and Denazyme GEL-L1 / R (manufactured by Nagase ChemteX Corporation) are examples of commercially available products containing only pure glucanase activity, while Sumiteam TG (manufactured by Shin-Nihon Chemical Industry Co., Ltd.) is an example of a commercially available product containing multiple activities such as protease activity.

[0054] Generally, commercially available exoglucanase products often also contain endoglucanase, but in the enzyme treatment step, a commercially available product containing endoglucanase in addition to exoglucanase may also be used. Endoglucanases are endo-type enzymes that randomly cleave the interior of the cellulose chains in the yeast cell wall. The type of endoglucanase is not particularly limited and can be appropriately selected, and examples thereof include endo-1,3-β-glucanase, endo-1,4-β-glucanase, endo-1,6-β-glucanase, etc. These may be used alone or in combination of two or more types. The origin of the endoglucanase is not particularly limited, and examples thereof include those similar to those of the exoglucanase.

[0055] The lower limit of the amount of exoglucanase to be added to the yeast cell wall is not particularly limited as long as it is not 0% by mass, and can be selected appropriately depending on the purpose. However, the amount is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, and particularly preferably 0.5% by mass or more, relative to the solid mass of the yeast cell wall. There is no particular upper limit to the amount of exoglucanase to be added to the yeast cell wall, and it can be selected appropriately depending on the purpose. However, it is preferably 5% by mass or less, more preferably 2% by mass or less, and particularly preferably 1% by mass or less, relative to the solid mass of the yeast cell wall. The lower and upper limits of the amount of exoglucanase to be added to the yeast cell wall can be combined as appropriate, but the amount of exoglucanase to be added to the yeast cell wall is preferably 0.05% by mass to 5% by mass, more preferably 0.1% by mass to 2% by mass, and particularly preferably 0.5% by mass to 1% by mass, relative to the solid mass of the yeast cell wall.

[0056] Treating the yeast cell walls with exoglucanase is advantageous in that the enzyme-treated product in which the yeast cell walls are decomposed has an improved solubilization rate. Furthermore, treating the yeast cell walls with exoglucanase is advantageous in that it allows the production of a large amount of reducing monosaccharides derived from the yeast cell walls, which can function, for example, in the heat treatment step described below, preferably in the Maillard reaction.

[0057] The reducing monosaccharide is not particularly limited and can be appropriately selected depending on the purpose. Examples include ribose, xylose, glucose, mannose, fructose, galactose, arabinose, and rhamnose. Among these, glucose is a monosaccharide inherent in yeast that can be produced from sugars in the yeast cell wall. Therefore, glucose is preferable in that it does not require the subsequent addition of sugars to the enzyme-treated product. Another advantage is that foods produced using such enzyme-treated products can be labeled as clean label and natural in the EU.

[0058] <<Protease>> The protease is an enzyme that acts on and hydrolyzes peptide bonds in proteins or polypeptide chains in the yeast cell wall. There are no particular limitations on the protease and it can be selected appropriately, but it is preferable that the protease is suitable for food applications or is of a grade suitable for food applications. The type of protease is not particularly limited and can be selected appropriately, but an endo-protease that randomly cleaves the inside of the protein or polypeptide chain in the yeast cell wall is preferred.

[0059] The origin of the protease is not particularly limited and can be selected appropriately depending on the purpose. Bacillus ) genus is preferred.

[0060] The protease to be used may be a commercially available product, or may be appropriately prepared from a bacterium having protease by a known method. Commercially available proteases include, for example, Alcalase (registered trademark) 2.4 L FG (manufactured by Novozymes Japan Co., Ltd.).

[0061] The lower limit of the amount of protease to be added to the yeast cell walls is not particularly limited and can be selected appropriately depending on the purpose, but is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, and particularly preferably 0.2% by mass or more, based on the solid mass of the yeast cell walls. The upper limit of the amount of protease to be added to the yeast cell walls is not particularly limited and can be selected appropriately depending on the purpose, but is preferably 5% by mass or less, more preferably 2% by mass or less, and particularly preferably 1% by mass or less, based on the solid mass of the yeast cell walls. The lower limit and upper limit of the amount of protease to be added to the yeast cell walls can be combined as appropriate, but the amount of protease to be added to the yeast cell walls is preferably 0.05% by mass to 5% by mass, more preferably 0.1% by mass to 2% by mass, and particularly preferably 0.2% by mass to 1% by mass, relative to the solid mass of the yeast cell walls.

[0062] The timing of adding the protease to the yeast cell wall is not particularly limited and can be selected appropriately depending on the purpose. The protease may be added so that it acts at the same time as the exoglucanase, or may be added so that it acts at a different time from the exoglucanase. The method for adding the protease and the exoglucanase so that they act at the same time is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include a method in which they are added simultaneously. There are no particular limitations on the method for adding the protease so that it acts at a different time from the exoglucanase, and it can be selected appropriately depending on the purpose. However, it is preferable to act the protease after the exoglucanase has acted, and examples of such methods include adding the exoglucanase and then the protease. The yeast cell walls are treated simultaneously with exoglucanase and protease, which is advantageous in terms of time efficiency.

[0063] <<Exopeptidase>> Exopeptidase is an enzyme that acts on and hydrolyzes peptide bonds near the termini of proteins or polypeptide chains in the yeast cell wall, and thus treating the yeast cell wall with exopeptidase can produce large amounts of amino acids derived from the yeast cell wall. Therefore, it is preferable to use exopeptidase when the method for producing the composition containing the yeast cell wall-derived degradation product includes a heat treatment step as described below.

[0064] The exopeptidase is not particularly limited and can be selected appropriately, but it is preferably one that can be used for food applications or is of a grade that can be used for food applications.

[0065] The source of the exopeptidase is not particularly limited and can be selected appropriately depending on the purpose. Aspergillus ) genus is preferred.

[0066] The exopeptidase to be used may be a commercially available product, or may be appropriately prepared by a known method from bacteria or the like having exopeptidase. An example of a commercially available exopeptidase is Flavorzyme (registered trademark) 1000L (manufactured by Novozymes Japan Co., Ltd.).

[0067] The lower limit of the amount of exopeptidase to be added to the yeast cell wall is not particularly limited and can be selected appropriately depending on the purpose, but is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, and particularly preferably 0.2% by mass or more, relative to the solid mass of the yeast cell wall. There is no particular upper limit to the amount of exopeptidase to be added to the yeast cell wall, and it can be selected appropriately depending on the purpose. However, it is preferably 5% by mass or less, more preferably 2% by mass or less, and particularly preferably 1% by mass or less, based on the solid mass of the yeast cell wall. The lower limit and upper limit of the amount of exopeptidase to be added to the yeast cell wall can be combined as appropriate, but the amount of exopeptidase to be added to the yeast cell wall is preferably 0.05% by mass to 5% by mass, more preferably 0.1% by mass to 2% by mass, and particularly preferably 0.2% by mass to 1% by mass, relative to the solid mass of the yeast cell wall.

[0068] In the enzyme treatment step, the timing of adding the exopeptidase to the yeast cell walls is not particularly limited, but it is preferable to add the exopeptidase at the same time as the protease or after the protease. It is preferable to carry out exopeptidase treatment after protease treatment, since this improves the efficiency of production of the yeast cell wall-derived amino acids and improves work efficiency.

[0069] The free or low-molecular-weight sugars and amino acids derived from the yeast cell wall produced by treating the yeast cell wall with exopeptidase can be used in culture media as a nutrient source for microorganisms and can also be used as seasonings. Furthermore, they can preferably function during the Maillard reaction in the heat treatment step described below. To efficiently induce the Maillard reaction industrially, it is preferable that both the reducing monosaccharides and the amino acids are monomers. Therefore, when the method for producing a composition containing a yeast cell wall-derived degradation product includes the heat treatment step, the Maillard reaction can be induced using only the yeast cell wall-derived amino acids and the yeast cell wall-derived reducing monosaccharides produced by enzymatic degradation without the need for externally added auxiliary raw materials such as amino acids and glucose. Furthermore, the heat-treated product derived from the yeast cell wall can be obtained with a good roasted flavor and roasted color. This is particularly advantageous in the EU, where clean label and natural product labeling can be used. However, in the method for producing a composition containing a degradation product derived from yeast cell walls, at least one of an amino acid and a reducing sugar may be added externally as appropriate depending on the purpose to cause the Maillard reaction.

[0070] The amino acid is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include free amino acids such as lysine, arginine, histidine, aspartic acid, glutamic acid, serine, threonine, aspartic acid, glutamine, alanine, glycine, valine, isoleucine, leucine, phenylalanine, tyrosine, tryptophan, methionine, cysteine, and proline; and some peptides and tripeptides such as glutathione.

[0071] The pH of the enzyme treatment step is not particularly limited and can be appropriately selected depending on the optimum pH of each enzyme. In the enzyme treatment step, when exoglucanase and protease are added so as to act simultaneously, the pH is preferably 4 to 9, and more preferably 5 to 7. In the enzyme treatment step, when exoglucanase, protease, and exopeptidase are added so as to act simultaneously, the pH is preferably 4 to 9, and more preferably 5 to 7.

[0072] The temperature in the enzyme treatment step is not particularly limited and can be appropriately selected depending on the optimum temperature for each enzyme. In the enzyme treatment step, when exoglucanase and protease are added so as to act simultaneously, the temperature is preferably 30°C to 70°C, more preferably 40°C to 60°C. Furthermore, in the enzyme treatment step, when exoglucanase, protease, and exopeptidase are added so as to act simultaneously, the temperature is preferably 30°C to 70°C, more preferably 40°C to 60°C.

[0073] The time for the enzyme treatment step is not particularly limited and can be appropriately selected depending on the degree of decomposition, the purpose, the amount of each enzyme added, and the like. In the enzyme treatment step, when exoglucanase and protease are added so as to act simultaneously, the time is preferably 1 hour to 40 hours, more preferably 12 hours to 30 hours. Furthermore, in the enzyme treatment step, when exoglucanase, protease, and exopeptidase are added so as to act simultaneously, the time is preferably 1 hour to 40 hours, more preferably 12 hours to 30 hours.

[0074] The enzyme treatment step may be carried out while standing or with stirring, but is preferably carried out with stirring since the efficiency of decomposing the yeast cell walls by each enzyme is high. The stirring speed is not particularly limited as long as it allows the yeast cell walls to react with the enzymes, and can be appropriately selected depending on the purpose.

[0075] -Composition containing yeast cell wall-derived decomposition product (enzyme-treated product)- The yeast cell wall-derived degradation product-containing composition (enzyme-treated product) obtained by the enzyme treatment step has a significantly reduced viscosity compared to the yeast cell wall slurry. The yeast cell wall slurry has a very high viscosity due to its high content of glucan, a thickening polysaccharide in yeast cell walls. However, the yeast cell wall-derived degradation product-containing composition (enzyme-treated product) has a lower viscosity because the enzyme treatment in the enzyme treatment step decomposes and eliminates most of the glucan in the yeast cell walls, resulting in solubilization and monomerization. Therefore, the yeast cell wall-derived degradation product-containing composition (enzyme-treated product) can be solubilized in a solvent or concentrated, which is advantageous in that it significantly improves processability, fluidity, and handleability. Furthermore, the composition containing a degradation product derived from yeast cell walls (enzyme-treated product) has significantly improved dispersion stability compared to a slurry of yeast cell walls.

[0076] The yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) has a sediment volume of 50 mL or less, preferably 40 mL or less, more preferably 30 mL or less, even more preferably 20 mL or less, and particularly preferably 15 mL or less, when 100 mL of the yeast cell wall-derived decomposition product-containing composition having a solids concentration of 10% by mass is allowed to stand at 25°C and normal pressure for 48 hours. When the sediment volume of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is 50 mL or less, most of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is solubilized, but the dispersion stability of the insoluble fraction is good. The volume of the sediment of the yeast cell wall-derived decomposition product-containing composition can be measured, for example, using a measuring cylinder. In the present invention, normal pressure means a pressure when no particular pressure is applied, and usually means a pressure equivalent to atmospheric pressure (101.325 kPa).

[0077] Here, the solid content concentration in the present invention is calculated by the following formula (3). Solid content concentration (%) = solid content mass / enzyme-treated product mass × 100 Equation (3) In the formula (3), the "mass of solid content" indicates the mass of the dried product obtained by drying X g of the enzyme-treated product at 105°C for 5 hours. In addition, in the formula (3), the "mass of the enzyme-treated product" indicates the mass of X g of the enzyme-treated product.

[0078] The particle size distribution (d10) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 4 μm or less, more preferably 3 μm or less, and particularly preferably 2 μm or less.

[0079] The particle size distribution (d50) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 7 μm or less, more preferably 5 μm or less, and particularly preferably 3 μm or less.

[0080] The particle size distribution (d90) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 15 μm or less, more preferably 10 μm or less, even more preferably 5 μm or less, and particularly preferably 4 μm or less.

[0081] The volume average particle size (MV) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 9 μm or less, more preferably 7 μm or less, even more preferably 5 μm or less, and particularly preferably 3 μm or less.

[0082] The number average particle size (MN) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 5 μm or less, more preferably 4 μm or less, even more preferably 3 μm or less, and particularly preferably 2.5 μm or less.

[0083] The area average particle size (MA) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 7 μm or less, more preferably 5 μm or less, even more preferably 3 μm or less, and particularly preferably 2.5 μm or less.

[0084] The standard deviation (SD) of the insoluble fraction of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but from the standpoint of solubility and dispersion stability, it is preferably 4 μm or less, more preferably 3 μm or less, even more preferably 2 μm or less, and particularly preferably 1 μm or less. The standard deviation (SD) indicates a measure of the distribution width of the particle size distribution.

[0085] The particle size distribution (d10, d50, and d90), volume average particle size (MV), number average particle size (MN), area average particle size (MA), and standard deviation (SD) of the insoluble fraction of the yeast cell wall-derived degradation product-containing composition (enzyme-treated product) in the present invention are values ​​measured using a Microtrac particle size distribution meter (MT3300EX, manufactured by Nikkiso Co., Ltd.).

[0086] The composition (enzyme-treated product) containing a degradation product derived from a yeast cell wall preferably contains at least one compound selected from ethyl caproate (hexanoic acid, ethyl ester), ethyl caprylate (octanoic acid, ethyl ester), ethyl caprate (decanoic acid, ethyl ester), phenylethyl alcohol, and capric acid (n-decanoic acid). These compounds are components related to beer-like aroma, and are preferably contained when the composition (enzyme-treated product) containing a degradation product derived from a yeast cell wall of brewer's yeast. The compounds in the composition containing the yeast cell wall-derived decomposition product can be measured by solid phase microextraction (SPME)-gas chromatography mass spectrometry under the measurement conditions described in the Examples (Test Example 7-1 or Test Example 7-2).

[0087] The yeast cell wall-derived degradation product-containing composition (enzyme-treated product) obtained by the enzyme treatment step can be roughly divided into two types depending on whether or not an exopeptidase is used in the enzyme treatment step.

[0088] A first aspect of the enzyme-treated product is an enzyme-treated product (hereinafter sometimes referred to as enzyme-treated product (1)) obtained by treating the yeast cell wall with exoglucanase and, if necessary, a protease in the enzyme treatment step. Therefore, the enzyme-treated product (1) contains at least a yeast cell wall-derived degradation product and exoglucanase, and, if necessary, further contains a protease. The enzyme-treated product (1) has significantly improved solubilization rate and dispersion stability compared to the yeast cell wall, and further contains a large amount of reducing monosaccharide (glucose) derived from the yeast cell wall.

[0089] The yeast cell wall-derived degradation product in the enzyme-treated product (1) is obtained by decomposing the yeast cell wall with exoglucanase and, if necessary, protease. The content of the yeast cell wall-derived degradation product in the enzyme-treated product (1) is not particularly limited and can be selected appropriately depending on the amount of the yeast cell wall used in the enzyme treatment step, etc.

[0090] The content of glucose derived from the yeast cell walls in the enzyme-treated product (1) is not particularly limited and can be selected appropriately depending on the conditions of the enzyme treatment step, etc., but is preferably 1% by mass or more, more preferably 10% by mass or more, and even more preferably 20% by mass or more, based on the solid content mass of the enzyme-treated product (1). When the glucose content is 1% by mass or more, the solubilization rate increases, making it easier for the Maillard reaction to occur in the heat treatment step. The upper limit of the content of glucose derived from the yeast cell walls in the enzyme-treated product (1) is not particularly limited and can be selected appropriately depending on the content of glucan contained in the yeast cell walls used as the raw material, the degree of enzymatic decomposition, etc., and is usually 35% by mass or less.

[0091] The solid content mass of the enzyme-treated product in the present invention refers to the mass of a dried product obtained by drying the enzyme-treated product at 105°C for 5 hours. In the present invention, the glucose content in the enzyme-treated product is a value measured by an enzyme-catalyzed reaction and a hydrogen peroxide electrode detection method using a multifunctional biosensor (BF-7D, manufactured by Oji Scientific Instruments Co., Ltd.) Specifically, the glucose content can be measured using a glucose electrode and a sucrose electrode in the multifunctional biosensor at 30°C using a dedicated buffer solution.

[0092] The content of total free amino acids derived from the yeast cell walls in the enzyme-treated product (1) is not particularly limited, but is often equivalent to the content of amino acids in the yeast cell walls, specifically, about 5% by mass to 8% by mass relative to the solid content mass of the enzyme-treated product (1), and depending on the method of enzyme treatment, for example, 5% by mass to 13% by mass.

[0093] In the present invention, the total free amino acid content in the enzyme-treated product can be measured by derivatizing the amino acids in the enzyme-treated product using an AccQ-Tag Ultra Derivatization kit (manufactured by Waters) according to the product protocol, and then measuring the content by ultra-high performance, high-resolution liquid chromatography under the measurement conditions described in the Examples (Test Example 5).

[0094] The contents of exoglucanase and protease in the enzyme-treated product (1) are not particularly limited and can be selected appropriately depending on the amounts of exoglucanase and protease used in the enzyme treatment step.

[0095] A second embodiment of the enzyme-treated product is an enzyme-treated product (hereinafter sometimes referred to as enzyme-treated product (2)) obtained by treating the yeast cell wall with exoglucanase and exopeptidase, and optionally with a protease in the enzyme treatment step. Therefore, the enzyme-treated product (2) contains at least a yeast cell wall-derived degradation product, exoglucanase, and exopeptidase, and optionally further contains a protease. The enzyme-treated product (2), like the enzyme-treated product (1), has improved solubilization rate and dispersion stability compared to the yeast cell wall. Furthermore, the enzyme-treated product (2) further contains large amounts of total free amino acids derived from the yeast cell wall and reducing monosaccharides derived from the yeast cell wall. Therefore, the enzyme-treated product (2) is suitable for use as a heat-treated target (raw material) in the heat treatment step.

[0096] The yeast cell wall-derived degradation product in the enzyme-treated product (2) is obtained by decomposing the yeast cell wall with exoglucanase and exopeptidase, and further with protease as needed. The content of the yeast cell wall-derived degradation product in the enzyme-treated product (2) is not particularly limited and can be appropriately selected depending on the amount of the yeast cell wall used in the enzyme treatment step.

[0097] The content of glucose derived from the yeast cell walls in the enzyme-treated product (2) is not particularly limited and can be selected appropriately depending on the conditions of the enzyme treatment step, etc., but is preferably 1% by mass or more, more preferably 10% by mass or more, and even more preferably 20% by mass or more, based on the solid content of the enzyme-treated product (2). When the glucose content is 1% by mass or more, the solubilization rate increases, making it easier for the Maillard reaction to occur in the heat treatment step. The upper limit of the content of glucose derived from the yeast cell walls in the enzyme-treated product (2) is not particularly limited and can be selected appropriately depending on the content of glucan contained in the yeast cell walls used as the raw material, the degree of enzymatic decomposition, etc., and is usually 35% by mass or less.

[0098] The content of the total free amino acids derived from the yeast cell walls in the enzyme-treated product (2) is not particularly limited and can be appropriately selected depending on the conditions of the enzyme treatment step, etc. However, it is preferably 7% by mass or more, more preferably 10% by mass or more, and even more preferably 12% by mass or more, based on the solid content of the enzyme-treated product (2). When the content of the total free amino acids is 7% by mass or more, the Maillard reaction can be efficiently induced in the heat treatment step. The upper limit of the content of the total free amino acids derived from the yeast cell walls in the enzyme-treated product (2) is not particularly limited and can be appropriately selected depending on the protein content of the yeast cell walls used as a raw material. Typically, the protein content of the yeast cell walls used as a raw material is 60% by mass or less, and the amino acid decomposition rate is not as high as that of sugars. Therefore, the upper limit of the content of the total free amino acids derived from the yeast cell walls in the enzyme-treated product (2) is approximately 30% by mass. Furthermore, if the total free amino acid content is 12% by mass or more and the reducing sugar content is 10% by mass or more relative to the solid mass of the enzyme-treated product, this is preferable in that the enzyme-treated product can be used as a raw material for a wider range of applications.

[0099] The contents of exoglucanase, protease, and exopeptidase in the enzyme-treated product (2) are not particularly limited and can be selected appropriately depending on the amounts of exoglucanase, protease, and exopeptidase used in the enzyme treatment step.

[0100] The solubilization rates of the enzyme-treated products (1) and (2) are not particularly limited and can be appropriately selected depending on the purpose, but the solubilization rate calculated by the formula (1) is preferably 60% or more, more preferably 80% or more. Since the solubilization rate of the yeast cell wall calculated by the formula (1) is generally about 18% to 27% or less, the enzyme-treated products (1) and (2) are advantageous in that they have significantly improved solubilization rates compared to the yeast cell wall.

[0101] The enzyme-treated product (1) and the enzyme-treated product (2) may further contain other components, if necessary. The other components are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include solvents, pH adjusters, antiseptics such as salts, sugars, reducing monosaccharides, thiamine, ascorbic acid (vitamins), amino acids, peptides, acids, alkalis, emulsifiers, other food additives, sulfur-containing compounds, etc. These may be used alone or in combination of two or more. The content of the other components in the enzyme-treated product is not particularly limited and can be appropriately selected depending on the purpose.

[0102] The form of the enzyme-treated product is not particularly limited and can be selected appropriately depending on the purpose. Examples include a liquid obtained in the enzyme treatment step, a suspension in which the liquid enzyme-treated product is suspended in a solvent, a paste in which the liquid enzyme-treated product is compressed and concentrated, a dried product in which the paste is dried and further concentrated, and a powder in which the dried product is pulverized.

[0103] The use of the enzyme-treated product is not particularly limited and can be appropriately selected according to the purpose. Since the enzyme-treated product has a high solubilization rate and dispersion stability, it can be widely used in various fields such as the food field, the bio field, and the cosmetics field, similar to the uses of general yeast extracts. Specifically, it can be suitably used for food and beverages, alcohols, raw materials for pet food (especially palatability improvers for pet food), culture media, and the like.

[0104] <pH adjustment step> When the method for producing the composition containing the yeast cell wall-derived decomposition product includes a heat treatment step described later, the method for producing the composition containing the yeast cell wall-derived decomposition product preferably includes a pH adjustment step. The pH adjustment step is a step of adjusting the pH of the enzyme-treated product obtained in the enzyme treatment step to a pH suitable for the purpose. In the pH adjustment step, the enzyme-treated product to be adjusted in pH is the enzyme-treated product (2) obtained by treating the yeast cell wall with exoglucanase and exopeptidase, and further, if necessary, protease.

[0105] The timing of performing the pH adjustment step is not particularly limited as long as it is after the enzyme treatment step and before the heat treatment step, and can be appropriately selected according to the purpose. Performing the pH adjustment step before the heat treatment step is advantageous in that the solubilization rate and dispersion stability of the yeast cell wall are further improved. In addition, performing the pH adjustment step before the heat treatment step is also advantageous in that the consumption (Maillard reaction) of total free amino acids in the subsequent heat treatment step is further increased, so that a yeast cell wall-derived decomposition product composition with a better roasted flavor, greasiness, and roasted color can be obtained.

[0106] The method for adjusting the pH of the enzyme-treated product is not particularly limited and can be appropriately selected from known methods according to the purpose. Examples include the method of adding a pH adjuster to the enzyme-treated product.

[0107] The pH adjuster is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include alkalis such as sodium hydroxide, potassium hydroxide, sodium citrate, sodium acetate, and sodium phosphate. These may be used alone or in combination of two or more.

[0108] The pH of the enzyme-treated product in the pH adjustment step is preferably 8 or more and 10 or less. This pH range is preferable in that it improves the efficiency of the Maillard reaction in the subsequent heat treatment step for the purposes of imparting roast flavor or coloring.

[0109] <Heat treatment process> When the objective is to obtain a composition containing a yeast cell wall-derived decomposition product that has a good roast flavor and oily feel, as well as a good roast color, it is preferable that the method for producing the composition containing a yeast cell wall-derived decomposition product includes a heat treatment step. The heat treatment step is a step of heat-treating the enzyme-treated product obtained in the enzyme treatment step. When the method for producing a composition containing a yeast cell wall-derived degradation product includes the pH adjustment step, the heat treatment step is a step of heat-treating the alkaline enzyme-treated product obtained in the pH adjustment step. The heat treatment target in the heat treatment step is the enzyme-treated product (2) obtained by treating the yeast cell wall with exoglucanase and exopeptidase, and further with protease as needed.

[0110] By the heat treatment step, the total free amino acids derived from the yeast cell walls produced by enzymatic degradation in the enzyme-treated product (2) and the reducing monosaccharides derived from the yeast cell walls produced by enzymatic degradation undergo a Maillard reaction, thereby obtaining a composition (heat-treated product) containing a yeast cell wall-derived degradation product with a good roast flavor and roast color. The exoglucanase, protease, and exopeptidase used in the enzyme treatment step are inactivated by the heat treatment step. Therefore, the enzyme-treated product (1) may be used as the heat treatment target in the heat treatment step in order to inactivate these enzymes. Furthermore, the heat treatment target in the heat treatment step may be one that has been subjected to the enzyme inactivation step described below.

[0111] In the heat treatment step, in order to cause the Maillard reaction, both the reducing monosaccharides and the amino acids must be monomers. Generally, in the Maillard reaction, when the amount of reducing monosaccharides is greater than the amount of amino acids, it is advantageous in that it is easier to obtain a stronger roast flavor and roast color. Furthermore, when the amount of amino acids is greater than the amount of reducing monosaccharides, it is advantageous in terms of meat flavor and taste. In the heat treatment step, the preferred contents of the total free amino acids derived from the yeast cell wall and the reducing monosaccharides (glucose) derived from the yeast cell wall in the enzyme-treated product (2) are as described in the section "-Composition containing yeast cell wall-derived decomposition product (enzyme-treated product)-".

[0112] The pH in the heat treatment step is not particularly limited and can be appropriately selected depending on the purpose, but is preferably a pH at which the Maillard reaction can occur. The pH at which the Maillard reaction can occur is preferably 4 to 12, and more preferably 8 to 10. If the pH in the heat treatment step is 4 or higher, the rate of the Maillard reaction increases, and if it is 12 or lower, off-flavors are less likely to occur, resulting in a heat-treated product with a good roast flavor and roast color. On the other hand, if the pH in the heat treatment step is lower than 4, the rate of the Maillard reaction may decrease, and if it exceeds 12, off-flavors may occur, making it difficult to obtain a heat-treated product with a good roast flavor and roast color. In the EU, a pH of 8 or lower is preferred in view of the ability to label products as clean label and natural, as well as other EU regulations.

[0113] The heating temperature in the heat treatment step is not particularly limited and can be appropriately selected depending on the purpose, but is preferably a heating temperature at which the Maillard reaction can occur. The heating temperature at which the Maillard reaction can be industrially efficiently induced in a short time is preferably 60°C to 180°C, and more preferably 90°C to 120°C. When the heating temperature in the heat treatment step is 60°C or higher, the rate of the Maillard reaction increases, while when it is 180°C or lower, side reactions are less likely to occur, resulting in a heat-treated product with a good roast flavor and roast color. On the other hand, when the heating temperature in the heat treatment step is lower than 60°C, the rate of the Maillard reaction may be slowed, and when it exceeds 180°C, other reactions or decomposition may occur, making it difficult to obtain a heat-treated product with a good roast flavor and roast color. Note that when the heating temperature in the heat treatment step is in the more preferred range, it is advantageous in that it can be labeled clean in the EU. Furthermore, in view of other EU regulations, a temperature of 180°C or lower is preferred.

[0114] The heating time in the heat treatment step is not particularly limited and can be appropriately selected depending on the heating temperature in the heat treatment step, etc., but it is preferably a heating time that allows the Maillard reaction to occur efficiently in a short period of time. The time for which the Maillard reaction can occur is preferably 1 minute to 360 minutes, and more preferably 30 minutes to 180 minutes. If the heating time in the heat treatment step is less than 1 minute, the Maillard reaction may not occur sufficiently, and if it exceeds 360 minutes, an off-flavor may occur, and a long time may be disadvantageous in terms of production efficiency.

[0115] The heat treatment step may be carried out by leaving the mixture stationary or by stirring, but stirring is preferred since the components in the yeast cell wall-derived degradation product-containing composition (heat-treated product) obtained by the heat treatment are stable and uniform. The stirring speed is not particularly limited and can be appropriately selected depending on the purpose.

[0116] The heat treatment step may be carried out using an apparatus that is not particularly limited and can be appropriately selected from known apparatuses as long as it can heat-treat the yeast cell walls, and examples of the apparatus include general food manufacturing machines such as kneaders capable of stirring at normal pressure and stirring devices capable of pressurizing and depressurizing. Specific examples of the apparatus that can be used in the heat treatment step include an autoclave, a cooking mixer (manufactured by Kajiwara Co., Ltd.), and a Flex-Mix Processor (manufactured by SPX FLOW Technology). Furthermore, the tank used in the enzyme treatment step may be used as is in the heat treatment step.

[0117] -Composition containing yeast cell wall-derived decomposition product (heat-treated product)- The yeast cell wall-derived degradation product-containing composition (heat-treated product) obtained by the heat treatment step contains at least a yeast cell wall-derived degradation product. The yeast cell wall-derived degradation product in the heat-treated product is obtained by decomposing the yeast cell walls by the heat treatment. The heat-treated product has an improved solubilization rate compared to the yeast cell walls, and has a good roasted flavor, oily texture, and roasted color.

[0118] The composition (heat-treated product) containing a degradation product derived from a yeast cell wall contains at least one compound selected from pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol, and the roasted flavor of the heat-treated product is attributable to these compounds. The compounds in the heat-treated product can be measured by solid phase microextraction (SPME)-gas chromatography mass spectrometry under the measurement conditions described in the Examples (Test Example 7-1 or Test Example 7-2).

[0119] The heat-treated product obtained from yeast cell walls having a high carbohydrate content, such as the autolyzed yeast cell walls, is advantageous in that it has an even higher solubilization rate and can produce a good roasted flavor and a roasted color with a high degree of browning due to a strong Maillard reaction.

[0120] Heat-treated products obtained from protein-rich yeast cell walls, such as the hot water-extractable yeast, are also advantageous in that they have a higher solubilization rate than heat-treated products obtained from carbohydrate-rich yeast cell walls. Furthermore, heat-treated products obtained from protein-rich yeast cell walls are rich in peptides and amino acids derived from the yeast cell walls, and therefore have a lower ratio of reducing sugars than heat-treated products obtained from carbohydrate-rich yeast cell walls, resulting in a weaker roast, which is advantageous in that it can produce a meat-like roast flavor.

[0121] Thus, the flavor and roast color of the heat-treated product can be appropriately adjusted by adjusting the component composition of the yeast cell wall as the raw material, preferably the content ratio (mass ratio) of the total free amino acids derived from the yeast cell wall to the reducing monosaccharides derived from the yeast cell wall in the enzyme-treated product (2), the conditions of the Maillard reaction in the heat-treatment step, etc. Therefore, the method for producing a composition containing a yeast cell wall-derived degradation product is also advantageous in that it allows the preparation of desired heat-treated products according to the purpose.

[0122] The roast flavor of the heat-treated product includes at least one of roast flavor, meat flavor, and oily / fatty feel. In the present invention, the term "roast flavor" refers to a roasted taste (a fragrant taste), a roasted aroma or nutty aroma (mainly an aroma due to pyrazines), a cereal aroma, a coffee aroma, and a bitter taste. In the present invention, the term "oily and fat feeling" refers to a flavor that imparts a so-called fatty feeling, such as fats and oils, phospholipids, etc.

[0123] More specifically, the roast flavor of the heat-treated product has a flavor derived from plants such as cocoa mass, cocoa butter, cocoa powder, chocolate, peanuts, roasted peanuts, and coffee; a flavor derived from birds such as chicken, turkey, pheasant, goose, swan, and duck; a flavor derived from mammals such as beef, pork, lamb, sheep, goat, and horse; and a cooked aroma reminiscent of these.

[0124] The solubilization rate of the heat-treated product is not particularly limited and can be appropriately selected depending on the purpose, but the solubilization rate calculated by the formula (1) is preferably 80% or more, more preferably 85% or more. As described above, the solubilization rate of the yeast cell wall calculated by the formula (1) is generally about 18% to 27% or less. Therefore, the heat-treated product is advantageous in that it has a significantly improved solubilization rate compared to the yeast cell wall. Furthermore, the heat-treated product is even more advantageous in that it has an even improved solubilization rate compared to the enzyme-treated product.

[0125] The content of the yeast cell wall-derived degradation product in the heat-treated product is not particularly limited and can be appropriately selected depending on the amount of the enzyme-treated product (2) used in the heat-treated product, etc.

[0126] The heat treatment step consumes the total free amino acids derived from the yeast cell walls in the enzyme-treated product (2) through the Maillard reaction, and therefore the content of the total free amino acids derived from the yeast cell walls in the heat-treated product is lower than the content of the total free amino acids derived from the yeast cell walls in the enzyme-treated product (2). If the content of total free amino acids derived from the yeast cell walls in the heat-treated product is approximately the same as the content of total free amino acids in the yeast cell walls, this means that most or all of the total free amino acids derived from the yeast cell walls produced in the enzyme treatment step have been consumed in the heat treatment step.

[0127] It is not preferable that the content of the total free amino acids derived from the yeast cell walls in the heat-treated product is entirely consumed by the Maillard reaction, and can be appropriately selected depending on the purpose. For example, when the purpose is to use the product as a seasoning or to impart a flavor, the heat treatment may be performed so as to leave the total free amino acids. The method for performing heat treatment so as to leave the total free amino acids is not particularly limited and can be selected appropriately depending on the purpose. For example, the method can be performed by appropriately adjusting various conditions in the heat treatment step, such as pH, heating temperature, and heating time.

[0128] By the heat treatment step, the yeast cell wall-derived reducing monosaccharides in the enzyme-treated product (2) are consumed by the Maillard reaction together with the total free amino acids, and therefore the content of the yeast cell wall-derived reducing monosaccharides in the heat-treated product is lower than the content of the yeast cell wall-derived reducing monosaccharides in the enzyme-treated product (2). If the content of reducing monosaccharides derived from the yeast cell walls in the heat-treated product is approximately the same as the content of reducing monosaccharides in the yeast cell walls, this means that most or all of the reducing monosaccharides derived from the yeast cell walls produced in the enzyme treatment step have been consumed in the heat treatment step, resulting in the production of various flavors and coloration.

[0129] It is not preferable that the content of the yeast cell wall-derived reducing monosaccharides in the heat-treated product be entirely consumed by the Maillard reaction, and the content can be appropriately selected depending on the purpose. For example, when the purpose is to use the product as a culture medium or a nutrient source for microorganisms, the heat treatment may be performed so as to leave the reducing monosaccharides. The method for carrying out the heat treatment so as to leave the reducing monosaccharides is not particularly limited and can be selected appropriately depending on the purpose. For example, the method can be carried out by appropriately adjusting various conditions in the heat treatment step, such as pH, heating temperature, and heating time.

[0130] The color of the enzyme-treated product (2) browns during the heat treatment step, and can therefore be used as an index for evaluating the degree of roasting of the heat-treated product. The method for evaluating the color of the heat-treated product is not particularly limited, and can be appropriately selected from known methods that can evaluate the degree of browning. For example, * a * b * (L-star, A-star, B-star) color system, or by evaluating absorbance at 470 nm (OD470).

[0131] L * a * b * Specifically, the evaluation method using the color system is based on JIS Z-8722, and involves measuring the L of the heat-treated product using a color difference meter under the measurement conditions described in Example (Test Example 2). * value, a * value, and b * The values ​​can be calculated respectively.

[0132] Said L * a * b * A color system is an index used to represent the color of an object, and was standardized by the International Commission on Illumination (CIE) in 1976. * a * b * In the color system, lightness is L * It is expressed as a value, and the chromaticity (hue and saturation) is a * value and b * It is expressed as a value of L * The larger the value, the brighter the light. * value and b * The values ​​indicate the color direction, and * indicates the red direction, and -a * indicates the green direction, and b * indicates the yellow direction, and -b * indicates the blue direction. In the present invention, L *The L value is a value for evaluating the degree of roasting of the heat-treated product by color (brightness). * A value of 0 is black, L * A value of 100 indicates white.

[0133] The heat-treated product may further contain other components, if necessary. The other components are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include solvents, pH adjusters, antifungals such as salts, glycerin, propylene glycol, dextrin, etc. These may be used alone or in combination of two or more. The content of the other components in the heat-treated product is not particularly limited and can be appropriately selected depending on the purpose.

[0134] The form of the heat-treated product is not particularly limited and can be appropriately selected depending on the purpose. Examples include a liquid obtained in the heat treatment step, a suspension in which the liquid heat-treated product is suspended in a solvent, a paste in which the liquid heat-treated product is compressed and concentrated, a dried product in which the paste is dried and further concentrated, and a powder in which the dried product is pulverized.

[0135] The use of the heat-treated product is not particularly limited and can be appropriately selected depending on the purpose. However, since the heat-treated product has excellent solubility and dispersion stability, a good roasted flavor and oily feel, and a good roasted color, it can be used for at least one of enhancing roasted flavor, imparting roasted flavor, enhancing roasted color, imparting roasted color, enhancing oily feel, and imparting oily feel in foods and beverages, pet foods, palatability improvers for pet foods, and the like. It can also be used as an additive or raw material for flavorings and flavoring ingredients for at least one of enhancing roast flavor, imparting roast flavor, enhancing roast color, imparting roast color, enhancing oily feel, and imparting oily feel.

[0136] <Other processes> The other steps are not particularly limited and can be appropriately selected depending on the purpose. Examples of the other steps include a sterilization step, a neutralization step, an enzyme inactivation step, a cooling step, a centrifugation or filtration step, a concentration or drying step, a pulverization step, and an emulsification step.

[0137] -Sterilization process- The sterilization step is a step of sterilizing the yeast cell walls that serve as the raw material before the enzyme treatment step. The method for sterilizing the yeast cell walls in the sterilization step is not particularly limited and can be appropriately selected from known methods, such as a method of heating under pressurized conditions and an ultrahigh temperature short time (UHT) method. In the method of heating under pressurized conditions, the pressure, temperature, and time are not particularly limited as long as they are capable of sterilizing the yeast cell walls, and can be appropriately selected depending on the purpose.

[0138] -Neutralization process- The neutralization step is a step of neutralizing the enzyme-treated product obtained in the enzyme treatment step or the heat-treated product obtained in the heat treatment step. Depending on the intended use of the enzyme-treated product and the heat-treated product, they may be neutralized in the neutralization step. The neutralization method is not particularly limited and can be appropriately selected from known methods, and examples thereof include a method of appropriately adding an alkali such as sodium hydroxide, potassium hydroxide, sodium citrate, sodium acetate, or sodium phosphate, or an acid such as hydrochloric acid, sulfuric acid, citric acid, or formic acid to the enzyme-treated product or the heat-treated product. The pH of the enzyme-treated product or the heat-treated product after neutralization is preferably 4 to 8, and more preferably 5 to 7.

[0139] -Enzyme deactivation process- The enzyme deactivation step is a step of deactivating the enzyme in the enzyme-treated product obtained in the enzyme treatment step. As described in the heat treatment step, the enzymes used in the enzyme treatment step and remaining in the enzyme-treated product are inactivated by the heat treatment step. However, in the method for producing a composition containing a yeast cell wall-derived degradation product, the enzyme treatment step and the heat treatment step do not have to be performed consecutively. In such a case, the enzyme treatment step may be followed by the enzyme inactivation step to inactivate the enzymes in the enzyme-treated product. The method for inactivating the enzyme in the enzyme-treated product is not particularly limited and can be selected appropriately depending on the purpose. For example, the pH of the enzyme-treated product can be adjusted to a pH that inactivates the enzyme. The timing of the enzyme deactivation step is not particularly limited as long as it is after the enzyme treatment step, and can be selected appropriately depending on the purpose. However, in order to obtain the desired enzyme-treated product, it is preferable to perform the enzyme deactivation step immediately after the enzyme treatment step in order to prevent the enzyme reaction from proceeding too far.

[0140] -Cooling process- The cooling step is a step of cooling the heat-treated product obtained in the heat treatment step to room temperature (15° C. to 25° C.) or a temperature lower than room temperature. The cooling method is not particularly limited and can be appropriately selected from among publicly available methods, and examples thereof include standing to cool and water cooling.

[0141] -Centrifugation or filtration process- The centrifugation or filtration step is a step of removing impurities by centrifuging or filtering the enzyme-treated product obtained in the enzyme treatment step or the product obtained in the heat treatment step. The enzyme-treated products and heat-treated products have significantly better solubility and dispersion stability than conventional products, but when clarity is required for the use of the enzyme-treated products and heat-treated products (for example, food and beverages such as beer, beer-flavored beverages, non-alcoholic beverages, RTDs (ready-to-drink: e.g., low-alcohol beverages), spirits, whiskey, etc.), impurities can be removed by the centrifugation or filtration process, resulting in an enzyme-treated product or heat-treated product with higher clarity.

[0142] - Concentration or drying process - The concentrating or drying step is a step of concentrating or drying the enzyme-treated product obtained in the enzyme treatment step or the heat-treated product obtained in the heat treatment step. The concentration or drying method is not particularly limited and can be appropriately selected from among publicly available methods, such as spray drying, air drying, drying using a dry drum, freeze drying, and vacuum drying. The concentration rate of the enzyme-treated product or the heat-treated product is not particularly limited and can be selected appropriately depending on the purpose. However, since the enzyme-treated product or the heat-treated product has an improved solubilization rate compared to the yeast cell wall, even if the enzyme-treated product or the heat-treated product is concentrated to a high concentration of about 40% to 50% by mass, it can be made into a liquid or paste with good fluidity, which is advantageous in terms of good work efficiency and production efficiency.

[0143] -Crushing process- The pulverization step is a step of pulverizing the enzyme-treated product obtained in the enzyme treatment step or the heat-treated product obtained in the heat treatment step into powder. The pulverization step is preferably performed on the concentrated or dried product of the enzyme-treated product obtained in the concentration or drying step, or the concentrated or dried product of the heat-treated product. The pulverization method is not particularly limited and can be appropriately selected from conventional methods, such as shredding, cutting, milling, etc. These methods may be used alone or in combination of two or more. The means used for the pulverization is not particularly limited and can be appropriately selected from known pulverizers, such as a food processor.

[0144] -Emulsification process- The emulsification step is a step of emulsifying the enzyme-treated product obtained in the enzyme treatment step or the heat-treated product obtained in the heat treatment step. The raw material used to emulsify the enzyme-treated product or the heat-treated product is not particularly limited and can be appropriately selected from common oil and fat raw materials, and examples thereof include vegetable oils and fats such as sunflower oil, soybean oil, and rapeseed oil; and animal oils and fats such as lard, lard, beef tallow, and chicken oil. The amount of the fat or oil raw material to be added to the enzyme-treated product or the heat-treated product is not particularly limited and can be appropriately selected depending on the purpose. The method for emulsifying the enzyme-treated product or the heat-treated product is not particularly limited and can be appropriately selected from known emulsification methods, such as a method of vigorously stirring the mixture using a general mixer, or a method using a homogenizer or a high-pressure emulsifier.

[0145] (Composition for imparting beer-like aroma) The composition for imparting a beer-like aroma of the present invention contains at least the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) of the present invention, and may further contain other components as necessary.

[0146] <Composition containing yeast cell wall-derived decomposition products> The composition containing a degradation product derived from a yeast cell wall is the composition containing a degradation product derived from a yeast cell wall (enzyme-treated product) of the present invention, and is preferably a composition containing a degradation product derived from a yeast cell wall of brewer's yeast. The present inventors have unexpectedly found that the composition containing a decomposition product derived from the yeast cell walls of brewer's yeast has a very strong beer-like aroma despite having little off-flavor. This beer-like aroma is thought to be due to at least one compound selected from ethyl caproate, ethyl caprylate, ethyl caprate, phenylethyl alcohol, and capric acid in the composition.

[0147] In the present invention, the term "beer-like aroma" refers to an aroma similar to the aroma (also referred to as "brewed aroma") derived from an alcoholic beverage fermented with brewer's yeast using malt, hops, and water as the main ingredients, with other ingredients such as rice, corn, and starch used as secondary ingredients, if necessary, within a certain range (preferably less than half the mass of the malt). Although capric acid is not usually a compound directly related to brewing aroma, it is known to be an important factor in imparting thickness and body to beers, beer-flavored beverages, non-alcoholic beverages, RTDs (e.g., low-alcohol beverages), spirits, whiskey, etc. Therefore, capric acid is a very important and essential substance in the composition for imparting beer-like aroma.

[0148] In the present invention, the term "off-flavor" refers to a smell that is perceived as an "unpleasant smell" or a negative indicator when it is judged to deviate from a "normal smell" due to a deterioration smell, an oxidation smell, or the like.

[0149] The method for producing the composition containing a yeast cell wall-derived degradation product is not particularly limited and can be appropriately selected depending on the purpose, but it is preferable that the composition containing a cell wall-derived degradation product be produced by a method similar to that used when brewer's yeast is selected as the type of yeast used as the raw material for the yeast cell walls. In this case, the composition containing a yeast cell wall-derived degradation product of brewer's yeast has properties equivalent to those of the enzyme-treated product (the enzyme-treated product (1) or the enzyme-treated product (2)), and preferred embodiments are also similar.

[0150] The brewer's yeast is not particularly limited and can be appropriately selected depending on the purpose. Surplus yeast generated during beer brewing or its by-products may be used. The surplus yeast generated during beer brewing or its by-products has a unique aroma when used as is, and therefore its applications have been limited. Furthermore, yeast cell walls generated from brewer's yeast obtained by autolysis have a putrid odor and an oxidized hop odor, further limiting their applications. On the other hand, the composition for imparting a beer-like aroma of the present invention is advantageous in that it contains a composition containing a degradation product derived from the yeast cell walls of brewer's yeast, resulting in reduced off-flavors and a good beer-like aroma.

[0151] The content of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) in the composition for imparting a beer-like aroma is not particularly limited and can be appropriately selected depending on the purpose. The composition for imparting a beer-like aroma may be the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) itself.

[0152] <Other ingredients> The other components in the composition for imparting a beer-like aroma are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include solvents, pH adjusters, antiseptics such as salts, glycerin, and propylene glycol. These may be used alone or in combination of two or more. The composition for imparting a beer-like aroma may also contain enzymes such as exoglucanases, proteases, and exopeptidases derived from the composition containing a yeast cell wall-derived degradation product. The content of the other components in the composition for imparting a beer-like aroma is not particularly limited and can be appropriately selected depending on the purpose.

[0153] The use of the composition for imparting a beer-like aroma is not particularly limited and can be appropriately selected depending on the purpose. However, since the composition has excellent solubility and dispersion stability, little off-flavor, and a good beer-like aroma, it can be used for at least one of enhancing a beer-like aroma and imparting a beer-like aroma in foods and beverages, pet foods, palatability improvers for pet foods, etc. The composition for imparting a beer-like aroma can also be used as an additive or raw material for flavorings, flavoring raw materials, food ingredients, extract raw materials, etc., for at least one of enhancing a beer-like aroma and imparting a beer-like aroma. For example, the composition for imparting a beer-like aroma can be added when brewing, blending, or mixing beverages such as beers, beer-flavored beverages, non-alcoholic beverages, RTDs, spirits, and whiskey. Furthermore, it is known that such beer-like aroma and yeast odor also have the effect of attracting insects and mites, and therefore the composition for imparting a beer-like aroma can also be used as an insecticide or a mite repellent.

[0154] (Composition for imparting roasted flavor and composition for imparting roasted color) The composition for imparting roast flavor of the present invention contains at least the yeast cell wall-derived degradation product-containing composition (heat-treated product) of the present invention, and further contains other components as necessary. The roasted color-imparting composition of the present invention contains at least the yeast cell wall-derived degradation product-containing composition (heat-treated product) of the present invention, and may further contain other components as necessary.

[0155] <Composition containing yeast cell wall-derived decomposition products> The yeast cell wall-derived decomposition product-containing composition in the composition for imparting a roast flavor or the composition for imparting a roast color is the yeast cell wall-derived decomposition product-containing composition (heat-treated product) of the present invention, and therefore preferred aspects, etc. are as described in the section "--Yeast cell wall-derived decomposition product--" above. Therefore, the method for producing the yeast cell wall-derived decomposition product-containing composition (heat-treated product) is not particularly limited and can be selected appropriately depending on the purpose, but it is preferable that it be produced by a method similar to the method for producing the cell wall-derived decomposition product-containing composition.

[0156] The content of the yeast cell wall-derived degradation product-containing composition (heat-treated product) in the roast flavor-imparting composition or the roast color-imparting composition is not particularly limited and can be appropriately selected depending on the purpose. The roast flavor-imparting composition or the roast color-imparting composition may be the yeast cell wall-derived degradation product-containing composition (heat-treated product) itself.

[0157] <Other ingredients> The other components in the roasted flavor-imparting composition or the roasted color-imparting composition are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include solvents, pH adjusters, antifungals such as salts, glycerin, propylene glycol, etc. These may be used alone or in combination of two or more. The content of the other components in the roasted flavor-imparting composition or the roasted color-imparting composition is not particularly limited and can be appropriately selected depending on the purpose.

[0158] The use of the roasted flavor-imparting composition or the roasted color-imparting composition is not particularly limited and can be appropriately selected depending on the purpose. However, since the composition has excellent solubility and dispersion stability and has at least one of a good roasted flavor, oily feel, and roasted color, it can be used for at least one of enhancing roasted flavor, imparting roasted flavor, enhancing roasted color, imparting roasted color, enhancing oily feel, and imparting oily feel in foods and beverages, pet foods, palatability improvers for pet foods, and the like. The roast flavor-imparting composition or the roast color-imparting composition can also be used as an additive or a raw material for a flavoring or flavoring raw material for at least one of enhancing roast flavor, imparting roast flavor, enhancing roast color, imparting roast color, enhancing oily and fatty feel, and imparting oily and fatty feel.

[0159] (food and drink) The food and beverage of the present invention contains at least one of the yeast cell wall-derived decomposition product-containing composition of the present invention (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition of the present invention (heat-treated product), and, if necessary, further contains other ingredients.

[0160] The food and beverages referred to above refer to those which are unlikely to be harmful to human health and which are taken orally or by administration through the digestive tract in normal social life, and are not limited to administrative categories such as food and beverages, medicines, and quasi-drugs, but rather broadly include, for example, general foods, health foods, health functional foods, beauty foods, quasi-drugs, medicines, and the like which are taken orally.

[0161] The food and drink products are not particularly limited and can be appropriately selected depending on the purpose. Examples of the food and drink products include beers, beer-flavored drinks, non-alcoholic drinks, RTDs, spirits, whiskey, soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (including concentrates and powders for preparing these drinks); frozen desserts such as ice cream, ice sorbet, and shaved ice; noodles such as soba, udon, harusame, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; candies, candies, gum, chocolate, tablet sweets, snacks, biscuits, jelly, jam, cream, peanut cream, baked goods, and bread, or confectionery ingredients therefor; crab, salmon, clams, tuna, sardines, shrimp, bonito, mackerel, whale, oysters, saury, squid, ark shells, scallops, and abalone. Seafood such as sea urchin, salmon roe, and tokobushi sea bream; processed seafood and livestock foods such as kamaboko, ham, and sausage; dairy products such as processed milk and fermented milk; oils and fats and oil-based foods such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressing; condiments such as sugar, salt, vinegar, soy sauce, miso, sauces, consommé bases, and bouillon; retort pouch foods such as curry, stew, oyakodon, porridge, rice porridge, Chinese rice bowl, katsudon, tempura bowl, unadon, hayashi rice, oden, mapo dolphin, beef bowl, meat sauce, egg soup, omelet rice, gyoza, shumai, hamburger steak, and meatballs; side dishes such as salads and pickles; health, beauty, and nutritional supplements in various forms; pharmaceuticals and quasi-drugs such as tablets, granules, capsules, drinks, and lozenges. The food and drink products are not limited to those exemplified above.

[0162] The content of at least one of the yeast cell wall-derived decomposition product-containing composition (heat-treated product) and the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) in the food or beverage is not particularly limited and can be selected appropriately depending on the purpose.

[0163] The other ingredients include auxiliary raw materials and additives that are commonly used in producing foods and beverages. The auxiliary raw materials and additives are not particularly limited and can be selected appropriately depending on the purpose. Examples include glucose, fructose, xylose, ribose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, glutathione, calcium salts, colorants, flavorings, and preservatives. The content of the other ingredients in the food or drink is not particularly limited and can be appropriately selected depending on the purpose.

[0164] The method for producing the food and beverage product is not particularly limited, as long as it contains at least one of the yeast cell wall-derived decomposition product-containing composition (heat-treated product) and the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product), and any known method for producing food and beverage products can be selected as appropriate.

[0165] Since the food and beverage of the present invention contains at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product), these roast flavors, beer-like aromas, or roast color can be utilized to create food and beverages that have the original flavor or roast color of the food and beverage itself, and that further impart or enhance the roast flavor, beer-like aroma, or roast color. Furthermore, at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product) is derived from the yeast cell wall and can impart or enhance roast flavor, beer-like aroma, or roast color without incorporating other additives, which is advantageous in that it can be labeled as clean label and natural in the EU.

[0166] (Pet food palatability enhancer) The pet food palatability improver of the present invention contains at least one of the yeast cell wall-derived decomposition product-containing composition of the present invention (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition of the present invention (heat-treated product), and, if necessary, further contains other ingredients.

[0167] In the present invention, the term "pet food palatability enhancer" refers to a pet food ingredient that has the effect of improving palatability (e.g., increasing appetite and food intake by pets) when added in small amounts to pet food. Typically, yeast materials or fish meal made from ingredients such as dry yeast, Maillard substances, amino acids, or sugars are added to pet food in amounts of 1% to 2% by mass. Other known pet food palatability improvers include those made from yeast, yeast extract, or yeast cell walls. However, conventional pet food palatability improvers are yeast materials such as yeast, yeast extract, or yeast cell walls, or are the yeast materials to which amino acids (lysine, cysteine, methionine, etc.) or sugars (reducing sugars, such as xylose) are added to cause a Maillard reaction. When these materials are added to pet food, they are labeled as additives. On the other hand, the pet food palatability improver of the present invention contains at least one of the yeast cell wall-derived decomposition product-containing composition of the present invention (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition of the present invention (heat-treated product), and therefore the pet food palatability improver and pet food containing it can be supplied without additives, and are advantageous in that they can be labeled as clean label and natural. Such pet food palatability improvers have not previously existed.

[0168] <Composition containing yeast cell wall-derived decomposition products> The yeast cell wall-derived degradation product-containing composition is at least one of the yeast cell wall-derived degradation product-containing composition (enzyme-treated product) of the present invention and the yeast cell wall-derived degradation product-containing composition (heat-treated product) of the present invention. Therefore, preferred embodiments are as described in the above sections "--Yeast cell wall-derived degradation product-containing composition (enzyme-treated product)-" and "--Yeast cell wall-derived degradation product-containing composition (heat-treated product)-". Among these, the pet food palatability improver preferably contains the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) containing at least one compound selected from ethyl caproate, ethyl caprylate, ethyl caprate, phenylethyl alcohol, and capric acid, or the yeast cell wall-derived decomposition product-containing composition (heat-treated product) containing at least one compound selected from pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol, and more preferably contains the yeast cell wall-derived decomposition product-containing composition (heat-treated product) containing at least one compound selected from pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol. The pet food palatability improver is preferable in that it contains at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product) that contain these compounds (aroma components), thereby stimulating the pet's sense of smell and increasing attraction and food intake, i.e., improving palatability.

[0169] The methods for producing the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product) are not particularly limited and can be selected appropriately depending on the purpose, but it is preferable that they be produced by a method similar to the method for producing the cell wall-derived decomposition product-containing composition.

[0170] The content of at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product) in the pet food palatability improver is not particularly limited and can be appropriately selected depending on the purpose. The pet food palatability improver may be at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product) themselves.

[0171] <Other ingredients> Examples of the other components include raw materials, auxiliary raw materials, and additives that are commonly used when producing a palatability improver for pet food. The content of the other components in the pet food palatability improver is not particularly limited and can be appropriately selected depending on the purpose.

[0172] The method for producing the pet food palatability improver is not particularly limited, as long as it contains at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product), and any known method for producing a pet food palatability improver can be appropriately selected.

[0173] The form of the pet food palatability improver is not particularly limited and can be appropriately selected depending on the purpose. Examples include a form in which the agent is mixed during pet food production (e.g., kneaded into pet food pellets), and a form in which the agent is sprinkled on or coated onto finished pet food.

[0174] The amount of the pet food palatability improver added to the pet food is not particularly limited and can be appropriately selected depending on the purpose, but the upper limit is preferably less than 1% by mass, more preferably 0.5% by mass or less, and particularly preferably 0.3% by mass or less, based on the total amount of the pet food. The lower limit of the amount of the pet food palatability improver added to the pet food is preferably 0.1% by mass or more, and more preferably 0.2% by mass or more. The upper and lower limits of the amount of the pet food palatability improver to be added to the pet food can be appropriately combined, but are preferably 0.1% by mass or more and less than 1% by mass, more preferably 0.1% by mass or more and 0.5% by mass or less, and particularly preferably 0.2% by mass or more and 0.3% by mass or less. When a concentration of about 1% by mass to 2% by mass is desired for the mixing process, the agent may be diluted with other palatability improvers, dispersants, excipients, etc. to achieve the above-mentioned concentrations.

[0175] The pets to be treated with the pet food palatability improver are not particularly limited and can be appropriately selected depending on the purpose. Examples include non-human animals such as monkeys, pigs, cows, sheep, goats, dogs, cats, mice, rats, hamsters, birds, and fish.

[0176] The method for confirming the palatability of the pet is not particularly limited and can be appropriately selected from known methods. For example, a typical method involves simultaneously giving a pet food to which the pet food palatability improver has been added and a control pet food (for example, pet food to which the pet food palatability improver has not been added, or pet food to which a commercially available pet food palatability improver has been added), and evaluating the pet food based on the above two items using a two-point evaluation method, in which the pet food that the pet puts in its mouth first (hereinafter, sometimes referred to as "First Choice (FC)" or "First Bite") and the pet food that is eaten by the greatest amount are judged to be the pet food with the highest palatability.

[0177] The pet food palatability improver contains at least one of the yeast cell wall-derived decomposition product-containing composition (enzyme-treated product) and the yeast cell wall-derived decomposition product-containing composition (heat-treated product), and is therefore advantageous in that it can utilize the flavor, i.e., at least one of a beer-like aroma, a roast flavor (e.g., meat flavor), an oily feel, and a roast color, to impart or enhance at least one of a beer-like aroma, a roast flavor, an oily feel, and a roast color to the pet food in addition to the flavor or color of the original pet food itself. [Example]

[0178] The present invention will be specifically explained below with reference to preparation examples and test examples, but the present invention is not limited to these preparation examples and test examples.

[0179] (Preparation Example 1-1: Autolyzed yeast cell wall slurry) Saccharomyces ( Saccharomyces A slurry of yeast cell walls (autolytic yeast cell walls) derived from brewer's yeast belonging to the genus Bacillus was prepared so that the concentration of the slurry was 10% by mass. Specifically, 100 g of yeast cell walls (autolytic yeast cell walls) (product name: Yeast Cell Wall, manufactured by Asahi Group Foods Co., Ltd.) was suspended in 900 g of water to prepare an "autolytic yeast cell wall slurry." The "yeast cell wall" (manufactured by Asahi Group Foods Co., Ltd.) used as the yeast cell wall (autolytic yeast cell wall) was a spray-dried powder.

[0180] (Preparation Example 1-2: Autoclaved product of autolyzed yeast cell wall (1)) The autolyzed yeast cell wall slurry (slurry concentration: 10% by mass) prepared in Preparation Example 1-1 was heat-treated in an autoclave at 120°C for 15 minutes to obtain "autoclaved autolyzed yeast cell wall product (1)."

[0181] (Preparation Example 1-3: Autoclaved product of autolyzed yeast cell wall (2)) The autolyzed yeast cell wall slurry (slurry concentration: 10% by mass) prepared in Preparation Example 1-1 was heat-treated in an autoclave at 120°C for 30 minutes to obtain "autoclaved autolyzed yeast cell wall product (2)."

[0182] (Preparation Example 2-1: Hot Water Extraction Type Yeast Cell Wall Slurry) Saccharomyces ( SaccharomycesA slurry of yeast cell walls (hot water-extracted yeast cell walls) derived from baker's yeast belonging to the genus Bacillus was prepared so that the concentration of the slurry was 16% by mass. Specifically, 160 g of yeast cell walls (hot water-extracted yeast cell walls) (product name: HG-YCW, manufactured by Asahi Group Foods Co., Ltd.) was suspended in 840 g of water to prepare a "hot water-extracted yeast cell wall slurry." The yeast cell wall (hot water-extractable yeast cell wall) used, "HG-YCW" (manufactured by Asahi Group Foods Co., Ltd.), was in the form of a spray-dried powder.

[0183] (Preparation Example 2-2: Autoclaved hot water-extracted yeast cell wall (1)) The hot water-extracted yeast cell wall slurry of Preparation Example 2-1 (slurry concentration: 16% by mass) was heat-treated in an autoclave at 120°C for 15 minutes to obtain "autoclaved hot water-extracted yeast cell wall product (1)."

[0184] (Preparation Example 2-3: Autoclaved hot water-extracted yeast cell wall (2)) The hot water-extracted yeast cell wall slurry of Preparation Example 2-1 (slurry concentration: 16% by mass) was heat-treated in an autoclave at 120°C for 30 minutes to obtain "autoclaved hot water-extracted yeast cell wall product (2)."

[0185] (Preparation Example 3-1: Composition containing autolyzed yeast cell wall-derived decomposition product (1E)) 1 kg of the autoclaved product (1) of autolyzed yeast cell walls obtained in Preparation Example 1-2 was placed in a 2 L jar, and the pH was adjusted to 5.7 using sodium hydroxide. Talaromyces glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd., Bacillus A protease derived from the genus Alcalase (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.), and AspergillusThree enzymes, including an exopeptidase derived from the genus Pseudomonas aeruginosa (Flavorzyme® 1000L, manufactured by Novozymes Japan Co., Ltd.), were simultaneously added in an amount of 0.5% by mass based on the solid mass of the autoclaved autolyzed yeast cell wall product (1), and the autolyzed yeast cell wall product was treated with the enzymes for 24 hours while stirring at 50°C and 200 rpm to obtain "composition (1E) containing a degradation product derived from an autolyzed yeast cell wall." The pH of the composition (1E) containing a degradation product derived from an autolyzed yeast cell wall after the 24-hour enzyme treatment was 5.5. The pH was measured using a pH meter (model: F-52 / glass electrode type, manufactured by Horiba, Ltd.) In the following preparation examples, the pH was measured in the same manner.

[0186] (Preparation Example 3-2: Composition containing autolyzed yeast cell wall-derived decomposition product (2E)) In Preparation Example 3-1, when performing the enzyme treatment, Aspergillus No exopeptidases derived from the genus are used. Talaromyces Glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd., and Bacillus A composition containing a degradation product derived from an autolyzed yeast cell wall (2E) was obtained in the same manner as in Preparation Example 3-1, except that only two enzymes, a protease derived from the genus Alcalase (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.), were added simultaneously in an amount of 0.5% by mass based on the mass of the solid content of the autoclaved autolyzed yeast cell wall (1). The pH of the composition containing a degradation product derived from an autolyzed yeast cell wall (2E) after 24 hours of enzyme treatment was 5.5.

[0187] (Preparation Example 3-3: Composition containing autolyzed yeast cell wall-derived decomposition product (3E)) In Preparation Example 3-3, protease, glucanase, and exopeptidase were separately treated with the enzyme in two steps by the following method.

[0188] In the first step, 1 kg of the autoclaved product (1) of autolyzed yeast cell walls obtained in Preparation Example 1-2 was placed in a 2 L jar, and the pH was adjusted to 8.0 using sodium hydroxide. Bacillus A protease derived from the genus Alcalase (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.) was added in an amount of 0.5% by mass based on the mass of the solid content of the autoclaved autolyzed yeast cell wall product (1), and the autoclaved autolyzed yeast cell wall product (1) was subjected to enzymatic treatment for 6 hours while stirring at 65°C and 200 rpm. The pH of the autoclaved autolyzed yeast cell wall product (1) after the protease treatment was 7.3.

[0189] Next, in the second step, the autoclaved product (1) of autolyzed yeast cell walls treated with protease was adjusted to pH 5.5 using hydrochloric acid. Talaromyces a glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd.; Aspergillus An exopeptidase derived from the genus Pseudomonas aeruginosa (Flavourzyme® 1000L, manufactured by Novozymes Japan Co., Ltd.) was added in an amount of 0.5% by mass based on the solid mass of the autoclaved autolyzed yeast cell wall product (1), and the autolyzed yeast cell wall product was treated with enzymes at 50°C and 200 rpm for 18 hours while stirring to obtain "composition (3E) containing a degradation product derived from an autolyzed yeast cell wall." The pH of composition (3E) containing a degradation product derived from an autolyzed yeast cell wall after enzymatic treatment with glucanase and exopeptidase was 5.5.

[0190] (Preparation Example 3-4: Composition containing autolyzed yeast cell wall-derived decomposition product (4E)) 1 kg of the autoclaved product (1) of autolyzed yeast cell walls obtained in Preparation Example 1-2 was placed in a 2 L jar, and the pH was adjusted to 5.5 using sodium hydroxide. TalaromycesA glucanase derived from the genus Pseudomonas aeruginosa (FILTRASE® BRX, manufactured by DSM Japan Co., Ltd.) was simultaneously added in an amount of 0.5% by mass relative to the solid mass of the autoclaved autolyzed yeast cell wall product (1), and the autolyzed yeast cell wall product was subjected to an enzymatic treatment for 24 hours while stirring at 50°C and 200 rpm to obtain "a composition containing a decomposition product derived from an autolyzed yeast cell wall (4E)." The pH of the composition containing a decomposition product derived from an autolyzed yeast cell wall (4E) after the 24-hour enzymatic treatment was 5.5.

[0191] (Preparation Example 4-1: Composition containing hot water extractable yeast cell wall-derived decomposition product (1E)) 1 kg of the autoclaved hot water-extracted yeast cell wall product (1) obtained in Preparation Example 2-2 was placed in a 2 L jar, and the pH was adjusted to 5.7 using sodium hydroxide. Talaromyces glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd., Bacillus A protease derived from the genus Alcalase (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.), and Aspergillus Three enzymes, including an exopeptidase derived from the genus Pseudomonas aeruginosa (Flavourzyme® 1000L, manufactured by Novozymes Japan Co., Ltd.), were simultaneously added in an amount of 0.5% by mass based on the solid mass of the autoclaved hot-water-extracted yeast cell wall product (1), and the mixture was subjected to enzyme treatment for 24 hours with stirring at 50°C and 200 rpm to obtain "Hot-water-extracted yeast cell wall-derived degradation product-containing composition (1E)." The pH of the hot-water-extracted yeast cell wall-derived degradation product-containing composition (1E) after the 24-hour enzyme treatment was 5.5.

[0192] (Preparation Example 4-2: Hot water extractable yeast cell wall-derived decomposition product-containing composition (2E)) In Preparation Example 4-1, during the enzyme treatment, Aspergillus No exopeptidase derived from the genus (Flavourzyme® 1000L, manufactured by Novozymes Japan Co., Ltd.) was used. Talaromyces Glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd. BacillusA "hot-water-extracted yeast cell wall-derived degradation product-containing composition (2E)" was obtained in the same manner as in Preparation Example 4-1, except that only two enzymes, a protease derived from the genus Pseudomonas aeruginosa (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.) and a lactic acid bacteria-derived protease, were added simultaneously in an amount of 0.5% by mass relative to the solid mass of the hot-water-extracted yeast cell wall autoclaved product (1). The pH of the hot-water-extracted yeast cell wall-derived degradation product-containing composition (2E) after 24 hours of enzyme treatment was 5.5.

[0193] (Preparation Example 4-3: Composition containing hot water extractable yeast cell wall-derived decomposition product (3E)) In Preparation Example 4-3, protease, glucanase, and exopeptidase were separately treated with the enzyme in two steps by the following method.

[0194] In the first step, 1 kg of the autoclaved hot water-extractable yeast cell wall product (1) obtained in Preparation Example 2-2 was placed in a 2 L jar, and the pH was adjusted to 8.0 using sodium hydroxide. Bacillus A protease derived from the genus Alcalase (Alcalase® 2.4 L FG, manufactured by Novozymes Japan Co., Ltd.) was added in an amount of 0.5% by mass based on the solid mass of the autoclaved hot-water-extracted yeast cell wall product (1), and the autoclaved hot-water-extracted yeast cell wall product (1) was subjected to enzymatic treatment for 6 hours while stirring at 65°C and 200 rpm. The pH of the autoclaved hot-water-extracted yeast cell wall product (1) after the enzymatic treatment with the protease was 7.0.

[0195] Next, in the second step, the autoclaved product (1) of hot water-extracted yeast cell walls treated with protease was adjusted to pH 5.5 using hydrochloric acid. a glucanase derived from the genus FILTRASE (registered trademark) BRX, manufactured by DSM Japan Co., Ltd.; TalaromycesThe autoclaved hot-water-extracted yeast cell wall product (1) was treated with an exopeptidase derived from the yeast genus (Flavourzyme® 1000L, Novozymes Japan Co., Ltd.) in an amount of 0.5% by mass based on the solid content, and the mixture was stirred at 50°C and 200 rpm for 18 hours to obtain "Composition (3E) containing a degradation product derived from a hot-water-extracted yeast cell wall." The pH of the composition (3E) containing a degradation product derived from a hot-water-extracted yeast cell wall after the enzymatic treatment with glucanase and exopeptidase was 5.5.

[0196] (Preparation Example 4-4: Composition containing hot water extractable yeast cell wall-derived decomposition product (4E)) 1 kg of the autoclaved hot water-extracted yeast cell wall product (1) obtained in Preparation Example 2-1 was placed in a 2 L jar, and the pH was adjusted to 5.5 using sodium hydroxide. Aspergillus Talaromyces A glucanase derived from the genus Pseudomonas aeruginosa (FILTRASE® BRX, manufactured by DSM Japan Co., Ltd.) was simultaneously added in an amount of 0.5% by mass based on the solid mass of the autoclaved hot water-extracted yeast cell wall product (1), and the mixture was subjected to an enzymatic treatment for 24 hours with stirring at 50°C and 200 rpm to obtain "a composition containing a hot water-extracted yeast cell wall-derived degradation product (4E)." The pH of the composition containing a hot water-extracted yeast cell wall-derived degradation product (4E) after the 24-hour enzymatic treatment was 5.5.

[0197] The conditions for Preparation Examples 3-1 to 4-4 are summarized in Table 1 below.

[0198] [Table 1]

[0199] (Preparation Example 5-1: Composition containing autolyzed yeast cell wall-derived decomposition product (1M)) The autolytic yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 3-1 was adjusted to pH 8.0 using sodium hydroxide. The composition was then heat-treated at 120°C for 30 minutes using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 6.4. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain "an autolytic yeast cell wall-derived decomposition product-containing composition (1M)."

[0200] (Preparation Example 5-2: Composition containing autolyzed yeast cell wall-derived decomposition product (2M)) The autolytic yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 3-1 was adjusted to pH 8.0 using sodium hydroxide. The composition was then heat-treated at 90°C for 3 hours using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 6.5. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "composition (2M) containing autolytic yeast cell wall-derived decomposition product."

[0201] (Preparation Example 5-3: Composition containing autolyzed yeast cell wall-derived decomposition product (3M)) The autolytic yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 3-1 was adjusted to pH 10.0 using sodium hydroxide. The composition was then heat-treated at 120°C for 30 minutes using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 7.3. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "composition (3M) containing an autolytic yeast cell wall-derived decomposition product."

[0202] (Preparation Example 5-4: Composition containing autolyzed yeast cell wall-derived decomposition product (4M)) The autolytic yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 3-1 was adjusted to pH 10.0 using sodium hydroxide. The composition was then heat-treated at 90°C for 3 hours using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 7.6. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "composition (4M) containing autolytic yeast cell wall-derived decomposition product."

[0203] (Preparation Example 6-1: Composition containing hot water extractable yeast cell wall-derived decomposition product (1M)) The hot water-extracted yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 4-1 was adjusted to pH 8.0 using sodium hydroxide. The composition was then heat-treated at 120°C for 30 minutes using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 6.7. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "hot water-extracted yeast cell wall-derived decomposition product-containing composition (1M)."

[0204] (Preparation Example 6-2: Composition containing hot water extractable yeast cell wall-derived decomposition product (2M)) The hot water-extracted yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 4-1 was adjusted to pH 8.0 using sodium hydroxide. The composition was then heat-treated at 90°C for 3 hours using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 6.8. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "hot water-extracted yeast cell wall-derived decomposition product-containing composition (2M)."

[0205] (Preparation Example 6-3: Composition containing hot water extractable yeast cell wall-derived decomposition product (3M)) The hot water-extracted yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 4-1 was adjusted to pH 10.0 using sodium hydroxide. The composition was then heat-treated at 120°C for 30 minutes using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 7.9. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "hot water-extracted yeast cell wall-derived decomposition product-containing composition (3M)."

[0206] (Preparation Example 6-4: Composition containing hot water extractable yeast cell wall-derived decomposition product (4M)) The hot water-extracted yeast cell wall-derived decomposition product-containing composition (1E) obtained in Preparation Example 4-1 was adjusted to pH 10.0 using sodium hydroxide. The composition was then heat-treated at 90°C for 3 hours using an autoclave (LSX-300, manufactured by Tomy Seiko Co., Ltd.). The pH of the heat-treated product was 8.0. The heat-treated product was adjusted to pH 6.0 using hydrochloric acid to obtain a "hot water-extracted yeast cell wall-derived decomposition product-containing composition (4M)."

[0207] The conditions for Preparation Examples 5-1 to 6-4 are summarized in Table 2 below.

[0208] [Table 2]

[0209] <Test Example 1: Solubilization rate> The solubilization rates were measured by the following method using the test samples obtained in the preparation examples shown in Tables 3-1 and 3-2 below. The mass of the dried product obtained by drying 50 g of the test sample at 105° C. for 5 hours was defined as the “solid mass B” (g) of the test sample. In addition, 50 g of the test sample of yeast cell wall slurry was centrifuged at 5,000 G for 5 minutes, and the resulting supernatant was dried at 105°C for 5 hours. The mass of the dried matter obtained was defined as the "solid mass A" (g) solubilized in the test sample. The solubilization rate of the solid content in the test sample was calculated based on the solid content mass A and the solid content mass B according to the following formula (1). The results are shown in Tables 3-1 and 3-2 below. Solubilization rate (%)=solid mass A / solid mass B×100...Formula (1)

[0210] [Table 3-1]

[0211] [Table 3-2]

[0212] It was found that the solubilization rate of yeast cell walls did not change simply by autoclaving them, but that the solubilization rate of yeast cell walls was improved by enzymatically treating them with exoglucanase alone or with exoglucanase and other enzymes simultaneously. However, when the exoglucanase and protease were treated separately in two stages, the improvement in solubilization rate was slightly less than when these enzymes were treated simultaneously, even though the optimal pH for each enzyme was achieved. Furthermore, the volume of the precipitated residue was larger when the enzymes were treated separately in two stages than when they were treated simultaneously. It was found that the solubilization rate of yeast cell walls was further improved by heat-treating the enzyme-treated yeast cell walls.

[0213] <Test Example 2: Color Tone> The test samples obtained in the preparation examples shown in Tables 4-1 and 4-2 below were used to measure the color tone by the following method. In accordance with JIS Z-8722, the color tone of each test sample was measured using a color difference meter under the following measurement conditions, and the L * value, a * value, and b * The results are shown in Tables 4-1 and 4-2 below. [Color measurement conditions] Color difference meter: ZE 6000 (manufactured by Nippon Denshoku Industries Co., Ltd.) Lighting conditions: Reflection, conforms to JIS Z-8722 Measurement method: Double beam method Light source: Halogen lamp 12V 20W (NA55919) · Color system: L * a * b *

[0214] [Table 4-1]

[0215] [Table 4-2]

[0216] By the heat treatment process, * value and b * The value (chromaticity) did not change significantly, but the L * The value (brightness) of the meat dropped significantly, indicating browning (blackening), which suggests that a strong Maillard reaction was occurring.

[0217] <Test Example 3: Roasted Aroma> The test samples obtained in the preparation examples shown in Tables 5-1 and 5-2 below were evaluated for roasted aroma by the following method. Five expert panelists evaluated the roasted aroma of the test samples based on the following criteria, using the aroma of the yeast cell wall slurry used as a raw material in each preparation example as a control (0 points). (That is, for samples obtained using autolyzed yeast cell walls as a raw material, the aroma of the autolyzed yeast cell wall slurry was used as a control, and for samples obtained using hot water-extracted yeast cell walls as a raw material, the aroma of the hot water-extracted yeast cell wall slurry was used as a control.) The evaluation results are shown as the average score of the five expert panelists. The results are shown in Tables 5-1 and 5-2 below. [Evaluation criteria] -3 points: Very weak roasted aroma -2 points: Weak roasted aroma -1 point: slightly weak roasted aroma 0 points: Control +1 point: Slightly stronger roasted aroma +2 points: Strong roasted aroma +3 points: Very strong roasted aroma

[0218] [Table 5-1]

[0219] [Table 5-2]

[0220] The results showed that the roasted aroma did not change when yeast cell walls were autoclaved or treated with enzymes alone, but that the roasted aroma was obtained by heat treatment after the enzyme treatment. Furthermore, the stronger the roasted aroma, the more meat-like the flavor tended to be. From the results of Test Examples 2 and 3, it was inferred that the autolysis-type yeast cell wall-derived decomposition product-containing compositions (1M) to (4M) and the hot water extraction-type yeast cell wall-derived decomposition product-containing compositions (1M) to (4M) were Maillard reaction products.

[0221] <Test Example 4: Beer-like aroma> The products obtained in the preparation examples shown in Table 6 below were used as test samples and evaluated for beer-like aroma by the following method. Five expert panelists evaluated the strength of the beer-like aroma of the test samples based on the following evaluation criteria, using the aroma of the yeast cell wall slurry used as a raw material in each preparation example as a control (0 points) (i.e., for samples obtained using autolyzed yeast cell walls as a raw material, the aroma of the autolyzed yeast cell wall slurry was used as a control, and for samples obtained using hot water-extracted yeast cell walls as a raw material, the aroma of the hot water-extracted yeast cell wall slurry was used as a control). The evaluation results are shown as the average score of the five expert panelists' evaluation criteria. The results are shown in Table 6 below. [Evaluation criteria] -3 points: Very weak beer-like aroma -2 points: Weak beer-like aroma -1 point: Beer-like aroma is slightly weak 0 points: Control +1 point: Slightly strong beer-like aroma +2 points: Strong beer-like aroma +3 points: Very strong beer-like aroma

[0222] [Table 6]

[0223] The brewer's yeast cell wall slurry had off-flavors such as a unique microbial putrid odor, other putrid odors, and the oxidized odor of hops. In contrast, enzyme treatment of the brewer's yeast cell wall slurry in the enzyme treatment step not only reduced the off-flavors but also unexpectedly imparted a strong beer-like aroma. Furthermore, the composition (1E) (Preparation Example 3-1) containing a decomposition product derived from an autolyzed yeast cell wall had a complex and rich beer-like aroma. Furthermore, the composition (4E) (Preparation Example 3-4) containing a decomposition product derived from an autolyzed yeast cell wall had a sharp beer-like aroma, demonstrating that it can impart a superior beer-like aroma. On the other hand, when baker's yeast cell walls were used as the raw material, no beer-like aroma was observed. Furthermore, when the enzyme-treated brewer's yeast cell walls were heat-treated, the beer-like aroma weakened, and a weak green aroma and a strong roasted aroma were observed. Previously, beer yeast cell walls alone could not be used as a flavor ingredient due to the aforementioned off-flavors and the absence of substances derived from brewing and fermentation. However, it is believed that by solubilizing the yeast through low molecular weight conversion and further by monomerization, the solubility rate is increased and the yeast dissolves, allowing the aroma components derived from brewing and fermentation that were encapsulated in the cell wall at the molecular level to be released. Furthermore, as shown in Test Example 1, although some insoluble matter remains in the yeast cell walls even after the enzyme treatment process, it is assumed that these aromas were obtained for the first time because, unlike conventional methods, they are in a dispersed state with little precipitation, close to solubilization.

[0224] <Test Example 5: Reducing Monosaccharide Content and Total Free Amino Acid Content> Using the test samples obtained in the preparation examples shown in Table 7 below, the glucose content as a reducing monosaccharide and the total free amino acid content in each test sample were measured by the following methods. The results are shown in Table 7 below. For the products obtained in the preparation examples shown in Table 8 below, the results of the content of each free amino acid are shown in Table 8 below.

[0225] -Measurement of solids concentration in test samples- The test samples were dried at 105° C. for 5 hours, and the mass of the dried product (solid mass) was measured. The solid concentration (T) of each test sample was calculated based on the following formula (3). Solid content concentration (T) (mass%) = solid content mass / test sample mass × 100 Equation (3)

[0226] -Measurement of glucose content- The glucose concentration (G) in the test sample was measured by an enzyme-catalyzed reaction and hydrogen peroxide electrode detection method using a multifunctional biosensor (BF-7D, manufactured by Oji Scientific Instruments Co., Ltd.) using a glucose electrode and a sucrose electrode at 30°C using a dedicated buffer solution. In Table 7 below, "Glucose content" indicates the glucose content relative to the solid mass of the test sample, calculated based on the measured glucose concentration (G) and the solid content (T) in each test sample according to the following formula (4). Glucose content (mass%) = glucose concentration (G) / solid concentration (T) Equation (4)

[0227] -Measurement of total free amino acid content- The free amino acids in the test samples were derivatized using the AccQ-Tag Ultra Derivatization kit (Waters) according to the product protocol. The concentration (F) of each free amino acid in each test sample containing the derivatized free amino acids was measured by ultra-high performance high-resolution liquid chromatography under the following measurement conditions. The "free amino acid content" in Table 8 below indicates the content of each free amino acid relative to the solid mass of the test sample, calculated based on the measured free amino acid concentration (F) and the solid concentration (T) in each test sample according to the following formula (5). In addition, the "total free amino acid content" in Table 7 below indicates the total content of each free amino acid relative to the solid mass of the test sample, calculated based on the measured free amino acid concentration (F) and the solid concentration (T) in each test sample according to the following formula (5). Free amino acid content (mass %) = free amino acid concentration (F) / solid content concentration (T)...Equation (5) [Free amino acid measurement conditions] Analytical equipment: Ultra Performance LC (UPLC (registered trademark)) (Waters) Detector: Photodiode array (PDA) (Waters) Column: AccQ-Tag Ultra RP Column (130 Å, 1.7 μm, 2.1 mm × 100 mm, manufactured by Waters) Column temperature: 55℃ Sample temperature: 10℃ Mobile phase: AccQ-Tag Ultra Eluent A (Waters) as solution A, and AccQ-Tag Ultra Eluent B (Waters) as solution B

[0228] [Table 7]

[0229] [Table 8]

[0230] It was found that the glucose content increased by enzymatic treatment with exoglucanase alone or exoglucanase and other enzymes simultaneously. Therefore, it was presumed that the improvement in the solubilization rate of yeast cell walls by enzymatic treatment as shown in Test Example 1 was due to the decomposition of the yeast cell walls by the enzymatic treatment. On the other hand, the content of total free amino acids did not increase with exoglucanase alone, but increased with simultaneous enzymatic treatment with exoglucanase and other enzymes. When the enzyme treatment was divided into two stages and performed under optimal conditions, first with protease and then with exoglucanase (Preparation Example 3-4), the glucose and total free amino acid contents were expected to increase further because the protein was decomposed with protease and then with glucanase, and the sugars were decomposed with glucanase. However, the glucose content was lower than when the enzyme treatment was performed simultaneously with protease and exoglucanase. Furthermore, the total free amino acid content was slightly higher than when the enzyme treatment was performed simultaneously with protease and exoglucanase, but was lower than when the enzyme treatment was performed simultaneously with exoglucanase, protease, and exopeptidase. Furthermore, after the heat treatment, both the glucose content and the total free amino acid content were significantly reduced, and in particular, the total free amino acid content was reduced to the same level as that of the autoclaved autolyzed yeast cell wall product (1) and the autoclaved hot water-extracted yeast cell wall product (1) before the enzyme treatment. From this, it was inferred that the autolyzed yeast cell wall-derived decomposition product-containing composition (1M) and the hot water-extracted yeast cell wall-derived decomposition product-containing composition (1M) are Maillard reaction products.

[0231] <Test Example 6: Oil and fat content of composition containing yeast cell wall-derived decomposition product> The test samples were the products obtained in the preparation examples shown in Table 9 below, as well as an autolyzed yeast extract (S-Ps, manufactured by Asahi Group Foods Co., Ltd.) and a hot water-extracted yeast extract (HG-Ps, manufactured by Asahi Group Foods Co., Ltd.), and the oil and fat content of these test samples was analyzed by the Japan Food Research Laboratories, Inc. The reported results are shown in Table 9 below.

[0232] [Table 9]

[0233] <Test Example 7-1: Aroma component 1 of the composition containing a yeast cell wall-derived decomposition product> Using yeast cell walls (autolyzed yeast cell walls) (product name: yeast cell walls, manufactured by Asahi Group Foods Co., Ltd.), the composition containing an autolyzed yeast cell wall-derived decomposition product (1E) obtained in Preparation Example 3-1, and the composition containing an autolyzed yeast cell wall-derived decomposition product (1M) obtained in Preparation Example 5-1 as test samples, the aroma components of each test sample were analyzed by solid-phase microextraction (SPME)-gas chromatography-mass spectrometry under the following analytical conditions.

[0234] [Analysis conditions] --Pretreatment conditions for test samples-- The autolytic yeast cell wall-derived decomposition product-containing composition (1E) and the autolytic yeast cell wall-derived decomposition product-containing composition (1M) were each spray-dried, and the powdered yeast cell wall-derived decomposition product-containing compositions were used as test samples.

[0235] --Solid-phase microextraction conditions-- Solid-phase microextraction (SPME) fiber: PDMS / DVB (film thickness: 65 μm, coating phase: divinylbenzene-dispersed polydimethylsiloxane, manufactured by SUPELCO) Volatile component extraction device: MPS GC-MS multifunction autosampler (manufactured by Gestell Corporation) Extraction conditions: 2.0 g of the test sample was weighed into a 20 mL SPME vial, and the volatile components were extracted at 60°C for 30 minutes, allowing the volatile components to be adsorbed onto the SPME fiber.

[0236] --Gas chromatograph mass spectrometry conditions-- Measurement equipment: Agilent 6890GC-5975MSD (manufactured by Allegient Technologies, Inc.) Column: DB-WAX (60m x 0.250mm ID x 0.25μm F.T., manufactured by Allegient Technologies, Inc.) Temperature conditions: 38°C (10 minutes) → Heat to 230°C at 3°C / minute → Hold at 230°C (20 minutes) → Heat to 245°C at 5°C / minute → Hold at 245°C (6 minutes) Carrier: He gas, gas flow rate 1.0 mL / min Inlet: Split / splitless inlet · Inlet temperature: 230℃ · Injection volume: 1μL Injection method: Pulsed splitless (pulse pressure 250KPa, up to 1.01m) Split ratio: 5:1 Liner: Straight, SPME taper, Ultra Inert liner Transfer line temperature: 230℃ Ionization method: EI (ionization voltage 70 eV) Ion source type: Extractor ion source Ion source temperature: 230℃ · Quadrupole temperature: 150℃ Measurement mode: Scan Scan mass: m / z 29.0~550.0

[0237] The results of solid phase microextraction (SPME)-gas chromatography mass spectrometry of the test sample are shown in Figures 1 to 7. In the composition (1E) containing the decomposition product derived from autolyzed yeast cell walls, new peaks were observed for ethyl caproate (hexanoic acid, ethyl ester), ethyl caprylate (octanoic acid, ethyl ester), ethyl caprate (decanoic acid, ethyl ester), phenylethyl alcohol, and capric acid (n-decanoic acid), which were not observed in the autolyzed yeast cell walls (untreated) (see Figures 1, 2, and 4). Furthermore, in the composition (1M) containing a decomposition product derived from an autolyzed yeast cell wall, new peaks were observed for pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,3-dimethylpyrazine, and furaneol, which were not observed in the composition (1E) containing a decomposition product derived from an autolyzed yeast cell wall (untreated) (see Figures 1 to 7). The retention time, peak height, and peak area of ​​the peaks detected in each test sample are shown in Table 10-1 below.

[0238] [Table 10-1]

[0239] The peaks of the compounds listed in the composition (1E) containing the autolytic cell wall-derived decomposition product in Table 10-1 are peaks that were not detected when the yeast cell walls of the raw material brewer's yeast (autolytic yeast cell walls) were measured under the same conditions, and are presumed to be peaks of aroma components that produce a beer-like aroma. Furthermore, the peaks of the compounds listed in the composition (1M) containing an autolyzed cell wall-derived decomposition product in Table 10-1 are peaks that were not detected when the yeast cell walls of the raw material brewer's yeast (autolyzed yeast cell walls) and the composition (1E) containing an autolyzed cell wall-derived decomposition product obtained by enzymatically treating them were measured under the same conditions, and are presumed to be peaks of aroma components that produce a roasted flavor.

[0240] <Test Example 7-2: Aroma component 2 of the composition containing a yeast cell wall-derived decomposition product> The composition containing the autolyzed yeast cell wall-derived decomposition product (1E) obtained in Preparation Example 3-1 and the composition containing the autolyzed yeast cell wall-derived decomposition product (4E) obtained in Preparation Example 3-4 were used as test samples, and the aroma components of each test sample were analyzed by solid-phase microextraction (SPME)-gas chromatography mass spectrometry under the following analytical conditions.

[0241] [Analysis conditions] --Pretreatment conditions for test samples-- The autolytic yeast cell wall-derived decomposition product-containing composition (1E) and the autolytic yeast cell wall-derived decomposition product-containing composition (4E) were each spray-dried, and the powdered yeast cell wall-derived decomposition product-containing compositions were used as test samples.

[0242] --Solid-phase microextraction conditions-- Solid-phase microextraction (SPME) fiber: PDMS / DVB (film thickness: 65 μm, coating phase: divinylbenzene-dispersed polydimethylsiloxane, manufactured by SUPELCO) Volatile component extraction device: MPS GC-MS multifunction autosampler (manufactured by Gestell Corporation) Extraction conditions: 2.0 g of the test sample was weighed into a 20 mL SPME vial, and the volatile components were extracted at 60°C for 30 minutes, allowing the volatile components to be adsorbed onto the SPME fiber.

[0243] --Gas chromatograph mass spectrometry conditions-- Measurement equipment: Agilent 8890B / 5977B (single quadrupole) (manufactured by Allegient Technologies, Inc.) Column: DB-WAX UI (30 m x 0.250 mm ID x 0.25 μm F.T., manufactured by Allegient Technologies, Inc.) Temperature conditions: 40°C (3 minutes) → Heat to 250°C at 10°C / minute → Hold at 250°C (6 minutes) Carrier: He gas, gas flow rate 1.1 mL / min Inlet: Split / splitless inlet · Inlet temperature: 250℃ · Injection volume: 1μL Injection method: Pulsed splitless (pulse pressure 20 psi, 3 minutes) Split ratio: 5:1 Liner: Straight, SPME taper, Ultra Inert liner Transfer line temperature: 230℃ Ionization method: EI (ionization voltage 70 eV) Ion source type: Extractor ion source Ion source temperature: 250℃ · Quadrupole temperature: 150℃ Measurement mode: Scan Scan mass: m / z 29.0~550.0

[0244] The results of solid phase microextraction (SPME)-gas chromatography mass spectrometry of the test sample are shown in FIG. The retention time, peak height, and peak area of ​​the peak detected in each test sample are as shown in Table 10-2 below. The peak areas shown in Table 10-2 below are graphed in Figure 9.

[0245] [Table 10-2]

[0246] It was found that the composition containing autolyzed yeast cell wall-derived decomposition products treated with one type of enzyme (4E) had an increased amount of aroma components that produce a beer-like aroma compared to the composition containing autolyzed cell wall-derived decomposition products treated with three types of enzymes (1E).

[0247] <Test Example 8: Particle size distribution of the insoluble fraction of the composition containing a yeast cell wall-derived decomposition product> The autolyzed yeast cell wall slurry obtained in Preparation Example 1-1 and the composition (1E) containing the autolyzed yeast cell wall-derived decomposition product obtained in Preparation Example 3-1 were used as test samples, and the particle size distribution (d10, d50, and d90), volume average particle size (MV), number average particle size (MN), area average particle size (MA), and standard deviation (SD; a measure of the distribution width of the measured particle size distribution) of the insoluble fraction in the test samples were measured by the following method. The results are shown in Table 11 below and Figure 10.

[0248] [Particle size distribution measurement conditions] Measuring equipment: Microtrac particle size distribution meter (MT3300EX, manufactured by Nikkiso Co., Ltd.) Wet method: Uses water as a solvent · Transparency: Transparent Refractive index: 1.333 Measurement time: 10 seconds Measurement count: 3 times Method: Laser diffraction scattering method

[0249] [Table 11]

[0250] Test Example 9: Dispersion stability of composition containing yeast cell wall-derived decomposition product The autoclaved product of autolyzed yeast cell walls (1) obtained in Preparation Example 1-2, the composition containing a decomposition product derived from autolyzed yeast cell walls (1E) obtained in Preparation Example 3-1, and the composition containing a decomposition product derived from autolyzed yeast cell walls (4E) obtained in Preparation Example 3-4 were used as test samples to evaluate the dispersion stability by the following method. The results are shown in Table 12 below.

[0251] The solid content of each test sample was calculated by the method described in Test Example 5, and the solid content of each test sample was adjusted to 10% by mass. 100 mL of a test sample with a solid content of 10% by mass was placed in a 100 mL glass measuring cylinder (manufactured by Shibata Scientific Co., Ltd.) and allowed to stand for 48 hours under normal pressure. The volume of the deposit was then measured using the graduations on the measuring cylinder.

[0252] [Table 12]

[0253] It was found that by treating with exoglucanase alone or with exoglucanase and other enzymes simultaneously, not only the solubilization rate of yeast cell walls was improved but also the dispersion stability of the solid content was improved.

[0254] <Test Example 10: Dog Preference Test 1> -Test item I- A pet food (hereinafter sometimes referred to as "test product I") was prepared by sprinkling 0.3% by mass of a composition (4M) containing an autolyzed yeast cell wall decomposition product having a roasted aroma prepared in Preparation Example 5-4 onto dog food for experimental animals (no added palatable agents, manufactured by Kitayama Labes Co., Ltd.) and mixing it uniformly.

[0255] -Control 1- Only dog ​​food for experimental animals (without added palatability agent, manufactured by Kitayama Labes Co., Ltd.) was used as "Control 1."

[0256] -Two-point comparison test- The average first-choice rate and average intake rate were tested by the following pairwise comparison using Test Product I and Control 1. In order to avoid statistical differences, this pairwise comparison test was performed by switching the positions of Test Product I and Control 1 between Test 1 and Test 2, as shown below. The beagle dogs used in the following two-point comparison test had passed discrimination training for the preference test and were undergoing regular training.

[0257] [First test] Test 1 was conducted 24 hours after the last feeding of Control 1. In the first test, Test Product I and Control 1 were fed in separate containers (250 g each, FA) with Test Product I placed on the left and Control 1 placed on the right. These were then fed to beagle dogs (n=20; 2-11 years old; mixed group of female (F), male (M), or neutered (XM); Kitayama Labes Co., Ltd.). Water was available ad libitum. In this case, the first item placed in the mouth (Test Product I or Control 1) was designated as the first choice (FC), and it was confirmed whether each beagle dog chose Test Product I or Control 1 as its first choice. In Table 13-1, "〇" indicates the first-choice item. In the first test, the first-choice rate was calculated from the number of dogs out of 20 that chose Test Product I or Control 1 as their first choice (hereinafter, sometimes referred to as "FC1"). The results are shown in Table 13-1.

[0258] Next, 20 minutes after the start of feeding, the amount of Test Product I and Control 1 left uneaten (hereinafter sometimes referred to as "remaining food amount") (g) for each Beagle dog was used to calculate the first feeding intake (FI; Feeding Intake) for each Beagle dog (hereinafter sometimes referred to as "FI1"). The first feeding intake was calculated based on the amount of Test Product I and the amount of Control 1 eaten. The results are shown in Table 13-2.

[0259] [Second exam] 24 hours after the first feeding, in the second test, 250g of each of Test Product I and Control 1 was placed in separate containers, with Control 1 placed on the left and Test Product I placed on the right, and given to each beagle dog in the same manner as in the first test. Water was available ad libitum. In this case, as in the first test, it was confirmed whether each beagle dog chose Test Product I or Control 1 as its first choice. In Table 13-1, "〇" indicates the first choice. In the second test, the first choice rate was calculated from the number of dogs out of 20 that chose Test Product I or Control 1 as their first choice (hereinafter, sometimes referred to as "FC2"). The results are shown in Table 13-1.

[0260] Next, as in the first test, 20 minutes after the start of feeding, the second food intake (FI) of each beagle dog was calculated from the remaining food amounts (g) of Test Food I and Control Food 1 (hereinafter sometimes referred to as "FI2"). The results are shown in Table 13-3.

[0261] Next, the average first choice rate (average FC rate) (%) of test product I or control 1 was calculated using the following formula (6). The results are shown in Table 13-5 below. Average FC rate (%)=(FC1+FC2) / 2 ··· Formula (6) In formula (6), "FC1" represents the first choice rate of test product I or control 1 in the first test, and "FC2" represents the first choice rate of test product I or control 1 in the second test.

[0262] The intake rate (%) of Test Article I or Control 1 for each beagle dog was calculated using the following formula (7). The results are shown in Table 13-4. The average intake rate (%) of Test Article I or Control 1 for the 20 beagle dogs was calculated from the intake rate (%) of Test Article I or Control 1 for each beagle dog. The results are shown in Table 13-5 below. Feeding rate (%) = (FI1-X + FI2-X) / (FI1-Y + FI2-Y) × 100 Equation (7) In formula (7), "FI1-X" indicates the amount (g) of intake of test item I or control 1 in the first test, "FI2-X" indicates the amount (g) of intake of test item I or control 1 in the second test, "FI1-Y" indicates the total amount (g) of intake of test item I and control 1 in the first test, and "FI2-Y" indicates the total amount (g) of intake of test item I and control 1 in the second test. In formula (7), when calculating the intake rate (%) of test item I, "FI1-X" and "FI2-X" simultaneously indicate the amount (g) of intake of test item I, and when calculating the intake rate (%) of control 1, "FI1-X" and "FI2-X" simultaneously indicate the amount (g) of intake of control 1.

[0263] [Table 13-1]

[0264] [Table 13-2]

[0265] [Table 13-3]

[0266] [Table 13-4]

[0267] [Table 13-5] The results in Table 13-5 show that compared to Control 1, Test Product I had significantly higher average first choice rates and average intake rates, and also had significantly lower amounts of leftover food, demonstrating its high palatability to dogs.

[0268] <Test Example 11: Dog Preference Test 2> -Control 2- 0.3% by mass of the powdered yeast cell wall (autolytic yeast cell wall) (product name: yeast cell wall, manufactured by Asahi Group Foods Co., Ltd.) used in Preparation Example 1-1 was sprinkled onto dog food for experimental animals (no added palatable agent, manufactured by Kitayama Labes Co., Ltd.) and mixed uniformly to prepare a pet food (hereinafter sometimes referred to as "Control 2").

[0269] -Two-point comparison test- A pairwise comparison test was conducted in the same manner as in Test Example 10, except that Control 1 was changed to Control 2 in the pairwise comparison test of Test Example 10, and the average first choice rate and average intake rate were tested. The results are shown in Table 14.

[0270] [Table 14] The results in Table 14 show that compared to Control 2, Test Product I had significantly higher average first choice rates and average intake rates, and also had significantly lower amounts of leftover food, demonstrating its high palatability to dogs.

[0271] <Test Example 12: Dog Preference Test 3> -Preparation of Test Sample II- A pet food (hereinafter sometimes referred to as "test product II") was prepared by sprinkling 0.3% by mass of the composition (1E) containing the autolyzed yeast cell wall decomposition product having a beer-like aroma prepared in Preparation Example 3-1 onto dog food for experimental animals (no added palatable agent, manufactured by Kitayama Labes Co., Ltd.) and mixing it uniformly.

[0272] -Two-point comparison test- In the paired comparison test of Test Example 11, except that Test Product I was changed to Test Product II, a paired comparison test was conducted in the same manner as in Test Example 11, and the average first choice rate and average intake rate were tested. The results are shown in Table 15.

[0273] [Table 15] The results in Table 15 show that compared to Control 2, Test Product II had significantly higher average first choice rates and average intake rates, and also had significantly lower amounts of leftover food, demonstrating its high palatability to dogs.

[0274] <Test Example 13: Dog Preference Test 4> -Preparation of Test Sample III- A pet food (hereinafter sometimes referred to as "test product III") was prepared by sprinkling 0.3% by mass of a hot water-extracted yeast cell wall-derived decomposition product-containing composition (2M) prepared in Preparation Example 6-2, which has a roasted flavor similar to meat flavor, onto dog food for experimental animals (no added palatable agents, manufactured by Kitayama Labes Co., Ltd.).

[0275] -Control 3- 0.3% by mass of the powdered yeast cell wall (hot water-extracted yeast cell wall) (product name: HG-YCW, manufactured by Asahi Group Foods Co., Ltd.) used in Preparation Example 2-1 was sprinkled onto dog food for experimental animals (no added palatable agents, manufactured by Kitayama Labes Co., Ltd.) and mixed uniformly to prepare a pet food (hereinafter sometimes referred to as "Control 3").

[0276] -Two-point comparison test- A pairwise comparison test was conducted in the same manner as in Test Example 10, except that Test Product I was changed to Test Product III and Control 1 was changed to Control 3, and the average first choice rate and average intake rate were tested. The results are shown in Table 16.

[0277] [Table 16] The results in Table 16 show that compared with Control 3, Test Product III had significantly higher average first choice rate and average intake rate, and also had significantly lower residual food amount, demonstrating its high palatability to dogs.

[0278] <Test Example 14: Cat Preference Test 1> -Test item IV- A pet food (hereinafter sometimes referred to as "test product IV") was prepared by sprinkling 0.3% by mass of a composition (4M) containing autolyzed yeast cell wall-derived decomposition product having a roasted aroma, prepared in Preparation Example 5-4, onto cat food for experimental animals (without added palatable agents, manufactured by Kitayama Labes Co., Ltd.) and mixing it uniformly.

[0279] -Control 4- Only cat food for experimental animals (without added palatability agent, manufactured by Kitayama Labes Co., Ltd.) was used as "Control 4."

[0280] -Two-point comparison test- In the pairwise comparison test of Test Example 10, Test Product I was replaced with Test Product IV, Control 1 was replaced with Control 4, and Beagle dogs were replaced with cats (n=20, 2-17 years old, mixed group of female (F), male (M), or neutered (XF), Kitayama Labes Co., Ltd.), and the feeding amount in Test 1 and Test 2 was changed from 250 g to 70 g. The pairwise comparison test was conducted in the same manner as in Test Example 10, and the average first-choice rate and average intake rate were tested. The results are shown in Tables 17-1 to 17-5. In Table 17-1, "〇" indicates the first-choice item. The cats used in the two-point comparison test were cats that had passed discrimination training for the preference test and were undergoing regular training.

[0281] [Table 17-1]

[0282] [Table 17-2]

[0283] [Table 17-3]

[0284] [Table 17-4]

[0285] [Table 17-5] The results in Table 17-5 show that compared to Control 4, Test Product IV had significantly higher average first choice rates and average intake rates, and also had significantly lower amounts of leftover food, demonstrating its high palatability to cats.

[0286] <Test Example 15: Cat Preference Test 2> -Control 5- 0.3% by mass of the powdered yeast cell wall (autolytic yeast cell wall) (product name: yeast cell wall, manufactured by Asahi Group Foods Co., Ltd.) used in Preparation Example 1-1 was sprinkled onto cat food for experimental animals (no added palatable agent, manufactured by Kitayama Labes Co., Ltd.) and mixed uniformly to prepare a pet food (hereinafter sometimes referred to as "Control 5").

[0287] -Two-point comparison test- A pairwise comparison test was conducted in the same manner as in Test Example 14, except that Control 4 was changed to Control 5, and the average first choice rate and average intake rate were tested. The results are shown in Table 18.

[0288] [Table 18] The results in Table 18 show that compared to Control 5, Test Product IV had significantly higher average first choice rate and average intake rate, and also had significantly less residual food, demonstrating its high palatability to cats.

[0289] <Test Example 16: Cat Preference Test 3> -Preparation of test sample V- A pet food (hereinafter sometimes referred to as "test product V") was prepared by sprinkling 0.3% by mass of the composition (1E) containing the autolyzed yeast cell wall decomposition product having a beer-like aroma prepared in Preparation Example 3-1 onto cat food for experimental animals (without the addition of palatable agents, manufactured by Kitayama Labes Co., Ltd.) and mixing it uniformly.

[0290] -Two-point comparison test- In the pairwise comparison test of Test Example 15, a pairwise comparison test was conducted in the same manner as in Test Example 15, except that Test Product IV was changed to Test Product V, and the average first choice rate and average intake rate were tested. The results are shown in Table 19.

[0291] [Table 19] The results in Table 19 show that compared to control 5, test product V had significantly higher average first choice rate and average intake rate, and also had significantly less residual food, demonstrating its high palatability to cats.

[0292] <Test Example 17: Cat Preference Test 4> -Preparation of test sample VI- A hot water-extracted yeast cell wall-derived decomposition product-containing composition (2M) with a meat-like roasted flavor prepared in Preparation Example 6-2 was sprinkled on cat food for experimental animals (without added palatable agents, manufactured by Kitayama Labes Co., Ltd.) at a concentration of 0.3% by mass, and the mixture was mixed uniformly to prepare a pet food (hereinafter referred to as "test product VI").

[0293] -Control 6- "Control 6" was prepared by sprinkling 0.3% by mass of the powdered yeast cell wall (hot water-extracted yeast cell wall) (product name: HG-YCW, manufactured by Asahi Group Foods Co., Ltd.) used in Preparation Example 2-1 onto cat food for experimental animals (no added palatable agents, manufactured by Kitayama Labes Co., Ltd.).

[0294] -Two-point comparison test- In the paired comparison test of Test Example 14, a paired comparison test was conducted in the same manner as in Test Example 14, except that Test Product IV was changed to Test Product VI and Control 4 was changed to Control 6, and the average first choice rate and average intake rate were tested. The results are shown in Table 20.

[0295] [Table 20] The results in Table 20 show that compared to Control 6, Test Product VI had significantly higher average first choice rate and average intake rate, and also had significantly less residual food, demonstrating its high palatability to cats.

[0296] In Test Examples 10 to 17, when comparing pet foods containing no additives (Control 1 or Control 4) with pet foods containing yeast cell walls (Control 2, Control 3, Control 5, or Control 6), adding yeast cell walls to the pet food slightly improved the animal's palatability. This is thought to be because the cell walls of brewer's yeast contain a certain amount of yeast odor, etc. However, when the pet food was added with a composition containing a yeast cell wall-derived decomposition product (enzyme-treated product) or a composition containing a yeast cell wall-derived decomposition product (heat-treated product), the palatability was not only significantly improved compared to regular pet food, but also significantly improved compared to the pet food containing the yeast cell walls. In particular, it was found that the composition containing a yeast cell wall-derived decomposition product (heat-treated product) was able to favorably improve the palatability of cats. [Industrial Applicability]

[0297] The method for producing a composition containing a yeast cell wall-derived degradation product of the present invention can improve the solubilization rate and dispersion stability of yeast cell walls, does not require the separation and removal of the insoluble fraction of yeast cell walls, has high work efficiency and production efficiency, and is low cost, so it can be suitably used as a method for effectively utilizing yeast cell walls, which have traditionally been discarded as insoluble matter after extracting yeast extract. The cell wall-derived degradation product-containing composition of the present invention has excellent solubility and dispersion stability and is therefore suitable for use in various fields such as the food, bio, and cosmetics fields, and is particularly suitable for use in foods and beverages, alcoholic beverages, pet food ingredients, pet food palatability improvers, and culture media. The yeast cell wall-derived degradation product-containing composition, roast flavor-imparting composition, and roast color-imparting composition of the present invention have excellent solubility and dispersion stability and exhibit good roast flavor, oily feel, and roast color, and therefore can be suitably used for at least one of enhancing roast flavor, imparting roast flavor, enhancing roast color, imparting roast color, enhancing oily feel, and imparting oily feel in foods and beverages, pet foods, pet food palatability improvers, etc. They can also be suitably used as additives or raw materials for flavorings and flavoring ingredients for at least one of enhancing roast flavor, imparting roast flavor, enhancing roast color, imparting roast color, enhancing oily feel, and imparting oily feel. The composition for imparting a beer-like aroma of the present invention has excellent solubility and dispersion stability, little off-flavor, and a favorable beer-like aroma, and is therefore suitable for at least one of enhancing a beer-like aroma in foods and beverages and imparting a beer-like aroma to them. It can also be suitably used as an additive or a raw material for flavors, flavoring ingredients, food ingredients, extract ingredients, and the like, for at least one of enhancing a beer-like aroma and imparting a beer-like aroma to them. The food and drink of the present invention contains the yeast cell wall-derived decomposition product-containing composition of the present invention, and therefore can be suitably used as a food and drink having a beer-like aroma, roasted flavor, oily texture, roasted color, and the like. The pet food palatability improver of the present invention contains the yeast cell wall-derived decomposition product-containing composition of the present invention, and therefore can be suitably used as an additive for pet food or as a raw material therefor.

Claims

1. A brewer's yeast cell wall-derived decomposition product derived from the cell wall of brewer's yeast (excluding those containing brewer's yeast extract), Contains ethyl caproate and ethyl caprylate, further containing at least one compound selected from ethyl caprate, phenylethyl alcohol, and capric acid; A decomposition product derived from brewer's yeast cell walls, characterized in that the volume of the sediment when measured under the following measurement conditions is 50 mL or less. <Measurement conditions> Water was added to the brewer's yeast cell wall-derived decomposition product to adjust the solid content to 10% by mass, and then 100 mL of the brewer's yeast cell wall-derived decomposition product to which water had been added was placed in a 100 mL glass measuring cylinder and left to stand at 25°C and normal pressure for 48 hours, measuring the volume of the sediment.

2. A composition for imparting a beer-like aroma, a palatability improver for pet food, or a food or drink, comprising the brewer's yeast cell wall-derived decomposition product according to claim 1.

3. A method for producing a brewer's yeast cell wall-derived degradation product, The method includes an enzyme treatment step of treating the cell walls of brewer's yeast (excluding those containing brewer's yeast extract) with exoglucanase, The brewer's yeast cell wall-derived decomposition product contains ethyl caproate and ethyl caprylate, The method for producing a decomposition product derived from brewer's yeast cell walls, characterized in that the decomposition product derived from brewer's yeast cell walls further contains at least one compound selected from ethyl caprate, phenylethyl alcohol, and capric acid.

4. The method according to claim 3 , wherein the enzyme treatment step further comprises treating the cell walls of the brewer's yeast with a protease.

5. The method according to claim 4, wherein the enzyme treatment step further comprises exopeptidase treatment of the cell wall of the brewer's yeast.

6. The method for producing a brewer's yeast cell wall-derived decomposition product according to any one of claims 3 to 5, wherein the brewer's yeast cell wall-derived decomposition product has a sediment volume of 50 mL or less when measured under the following measurement conditions. <Measurement conditions> Water was added to the brewer's yeast cell wall-derived decomposition product to adjust the solid content to 10% by mass, and then 100 mL of the brewer's yeast cell wall-derived decomposition product to which water had been added was placed in a 100 mL glass measuring cylinder and left to stand at 25°C and normal pressure for 48 hours, measuring the volume of the sediment.

7. A method for producing a composition for imparting a beer-like aroma, a palatability improver for pet food, or a food or beverage, which comprises producing a composition for imparting a beer-like aroma, a palatability improver for pet food, or a food or beverage, containing a brewer's yeast cell wall-derived hydrolysate produced by the method for producing a brewer's yeast cell wall-derived hydrolysate according to any one of claims 3 to 6.

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