CN112055611A

CN112055611A - Microcapsules

Info

Publication number: CN112055611A
Application number: CN201980029521.0A
Authority: CN
Inventors: 立松绫子; 松藤久; 藤江望未
Original assignee: Tablemark Co Ltd
Current assignee: Tablemark Co Ltd
Priority date: 2018-05-01
Filing date: 2019-04-26
Publication date: 2020-12-08
Also published as: KR20210004970A; JP7341128B2; JPWO2019212050A1; WO2019212050A1

Abstract

The present invention relates to a microcapsule containing yeast cells remaining after separation of content components, a method for producing the same, and use thereof. The method for producing a microcapsule of the present invention comprises: the method for producing a yeast cell comprises the steps of treating a yeast cell with hot water to release the content component to the outside of the microorganism, thereby producing a yeast cell remaining after the content component is separated, and allowing the yeast cell remaining after the content component is separated to contain a second active component as a first active component, wherein the yeast cell remaining after the content component is separated is not subjected to an acid treatment.

Description

Microcapsules

Technical Field

The present invention relates to microcapsules containing yeast cells as an active ingredient, a method for producing the same, and use thereof.

Background

When considering the deliciousness of food, its flavor is an important factor. As physicochemical properties of flavor-related perfumes (flavors), molecular weights are generally as low as 300 or less, and reactive groups such as acetaldehyde, ketone, ester, and the like are present in the molecular structure. Therefore, the flavor is highly volatile and unstable to heat, light, and oxygen. In food processing, heat treatment is performed in the steps of drying, concentration, sterilization, and the like. In these processes, the flavor may be volatilized or oxidized, thereby causing deterioration in quality and flavor of the food. Therefore, it can be said that the stabilization of the seasoning is essential.

The flavor retention stabilization method comprises the following steps: an adsorption method in which the particles are adsorbed and held on activated carbon or zeolite, and a microencapsulation method in which the contents are protected by coating with a coating material. Microcapsules are fine particles in which a liquid, a solid, or a gas is enclosed and a thin film is uniformly coated around the microcapsules. Microcapsules in which medicines, agricultural chemicals, perfumes, food materials, and the like are coated have been industrially produced. By forming a thin film on the outside of a substance having a certain property, the property can be sealed therein together, and the substance covered therewith can be taken out when necessary. The food flavoring agent is expected to inhibit reaction and volatilization during cooking and processing, prevent decay caused by heat, light and oxidation, improve long-term storage property, and realize controlled release effect by microencapsulation technology. The method for producing microcapsules includes chemical methods such as an interface polymerization method, a liquid-in-solid skin membrane method, and a molecular encapsulation method using cyclodextrin, physical methods such as a spray drying method, a spray cooling method, and an in-air suspension skin membrane method, and bacterial methods such as a method in which a liquid substance is contained in bacterial cells of yeast (bacterial cell method).

Yeasts mainly used in the bacterial method have been essential for the production of alcoholic beverages by fermentation of sugars and the production of fermented foods such as bread, miso and pickles in food processing since ancient times. In recent years, due to rapid development of biotechnology and genetic engineering technology, amino acids, vitamins, nucleic acids, and the like have been used for producing various useful substances not only in the food field but also in the medical field. On the other hand, the method of using the skeletal part of yeast cells produced after extraction of useful substances is small, and the yeast cells are largely discarded as industrial waste. In order to effectively utilize cells containing the main component of the skeletal portion of the yeast cells (hereinafter referred to as yeast cells), new technical development is required.

In recent years, inclusion techniques focusing on the biofilm structure of yeast have been attracting attention, and flavor inclusion methods have been particularly attracting attention. The yeast microcapsules are capsules obtained by encapsulating yeast containing a flavor in its cell body. Yeast flavor powders are made by encapsulating flavors within yeast cells. Unlike spray-dried powders that do not use yeast cells, yeast flavor powders can maintain the shape of the powder in water and can be distinguished from spray-dried powders that do not use yeast cells that release flavor slowly immediately after water absorption, and yeast flavor powders are a flavor inclusion method, and applications thereof in fields other than dried foods are expected.

As a method for producing yeast microcapsules, a method for encapsulating yeast cells obtained by releasing yeast in vivo components to the outside of the cells has been disclosed.

Japanese patent application laid-open No. 8-243378 relates to a method for microencapsulating yeast cells that have been subjected to acid treatment after autodigestion. This is a method of reducing the electric repulsive force generated between the yeast cell wall surface and the contained substance and promoting the uptake by acid treatment (Japanese patent laid-open No. 8-243378, paragraph 0012). Jp 2009-268395 a relates to a yeast capsule in which the surface of a yeast microcapsule containing a flavor and a spice extract in the interior of a cell body is coated with an oil or fat. In this method, yeast cells are subjected to enzyme treatment, and then solid components are subjected to acid treatment, and a flavor is encapsulated in the yeast cells.

In the methods described in the above-mentioned Japanese patent application laid-open Nos. 8-243378 and 2009-268395, in order to coat more substances to be encapsulated, it is necessary to perform separation by enzyme treatment and then to perform treatment with a strong acid. Therefore, when encapsulation is performed, it is necessary to adjust the pH using an alkaline solution or the like, and therefore, there is a concern that the addition of an alkaline solution may change the components and affect the flavor.

Japanese patent laid-open No. 2016-514951 relates to a method for producing a microorganism having a separated cell wall, wherein a flavor or a fragrance is encapsulated in a composition comprising a homogeneous paste containing a microorganism such as a yeast having a separated cell wall, a flavor or a fragrance, and water at a predetermined ratio. After mixing and heating a mixture of plasmolyzed microorganisms with a carrier composed of a water-soluble emulsifier of a polymer and an optional carbohydrate, a molten mass is extruded using an extruder and cut into pellets to prepare glass particles or glass beads.

In addition, when yeast cells are used as a base for flavor powder, there is a possibility that waste can be sold as a high value-added product. It has been disclosed at present: the encapsulated flavor does not undergo a substantial release under oily conditions, swells in the case of wet cells and increases in cell wall pores, and the flavor undergoes a release, while in the case of dry yeast, the cell wall pores close and the release does not easily occur. However, little is known about the sustained release behavior of the included flavor and the stability of the included flavor.

International publication WO2016/080490 discloses: the peculiar off-flavors (bitterness, astringency, pungent flavor, etc.) and off-flavors can be reduced by subjecting yeast cells to enzyme treatment such as protease and cellulase and emulsifier addition treatment.

As described above, although the encapsulation method using yeast has been studied, a microcapsule having good flavor and excellent stability has not yet been obtained.

Documents of the prior art

Patent document

Patent document 1: japanese laid-open patent publication No. 8-243378

Patent document 2: japanese laid-open patent publication No. 2009-268395

Patent document 3: japanese patent laid-open publication No. 2016-514951

Patent document 4: international publication WO2016/080490

Patent document 5 WO2017/014253

Disclosure of Invention

Problems to be solved by the invention

The present inventors have conducted extensive studies to solve the above problems, and as a result, have found that microcapsules excellent in stability, sustained release properties, and flavor can be obtained by using yeast cells in which content components are released from the yeast cells and the content components are separated, and have arrived at the present invention.

The invention aims to provide a method for manufacturing microcapsules.

The present invention aims to provide a microcapsule containing a yeast cell from which a content component produced by the production method of the present invention has been separated, and a second active ingredient.

The present invention aims to provide a flavor enhancer for food or beverage, which comprises the microcapsule of the present invention.

The object of the present invention is to provide a food or beverage comprising the microcapsules or flavor enhancers of the present invention.

Means for solving the problems

The present invention includes, but is not limited to, the following schemes:

[ scheme 1] A production method which is a production method of a microcapsule, the production method comprising: the method for producing a yeast cell comprises the steps of treating a yeast cell with hot water to release the content component to the outside of the microorganism, thereby producing a yeast cell remaining after the content component is separated, and allowing the yeast cell remaining after the content component is separated to contain a second active component as a first active component, wherein the yeast cell remaining after the content component is separated is not subjected to an acid treatment.

[ claim 2] the method according to claim 1, which comprises a step of subjecting the yeast cells remaining after the separation of the content component to a protease and/or cellulase addition treatment.

[ solution 3] the method according to claim 2, which comprises a step of adding an emulsifier before, after or simultaneously with the step of subjecting the yeast cells remaining after the separation of the content component to the protease and/or cellulase addition treatment.

[ scheme 4] the method according to scheme 3, wherein the emulsifier is selected from the group consisting of glycerin fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, sucrose fatty acid ester, lecithin and saponin.

[ solution 5] the method according to any one of the aspects 1 to 4, wherein the second active ingredient is a fat-soluble substance selected from the group consisting of a perfume, a spice extract, a scented oil, an animal fat and oil, and a vegetable fat and oil.

[ solution 6] the method according to any one of the aspects 1 to 5, wherein the yeast cells remaining after the content components are separated are in a dry state.

[ claim 7] the method according to any one of claims 1 to 6, wherein the yeast cells remaining after the content component is separated contain a protein component in an amount of 45 to 70 wt%.

[ solution 8] the method according to any one of claims 1 to 7, wherein the step of including a second active ingredient in the yeast cells remaining after the separation of the content component, comprises: the yeast cells, the second active ingredient and the water remaining after the separation of the content ingredient are mixed and stirred.

[ solution 9] the method according to any one of claims 1 to 8, wherein the step of including a second active ingredient in the yeast cells remaining after the separation of the content component, comprises: the yeast cells, the second active ingredient, and the water remaining after the content ingredients are separated are mixed and stirred, and the obtained dispersion is dried.

[ solution 10] the method according to claim 8 or 9, wherein the mixing ratio of the yeast cells remaining after the content component is separated to the second active ingredient is in the range of 4:1 to 1: 1.

[ solution 11] the method according to any one of solutions 8 to 10, wherein the stirring time is 1 hour or more and 8 hours or less.

[ solution 12] the method according to any one of solutions 8 to 11, wherein the stirring temperature is 25 ℃ or higher and less than 50 ℃.

[ solution 13] the method according to any one of the solutions 1 to 12, wherein the inclusion efficiency of the second active ingredient is at least 40%.

[ solution 14] A microcapsule comprising yeast cells remaining after separation of the content component produced by the method of any one of solutions 1 to 13, and a second active ingredient.

[ claim 15] the microcapsule according to claim 14, which has an improved sustained-release property of the second active ingredient as compared with a microcapsule obtained using yeast cells which have been subjected to acid treatment and remain after the content ingredients have been separated.

[ solution 16] the microcapsule according to solution 15, which has oxidation stability equivalent to that of a microcapsule obtained using yeast cells which have been subjected to acid treatment and remain after separation of content components.

[ solution 17] the microcapsule according to any one of solutions 14 to 16, which contains 5% by weight or more of the second active ingredient.

[ solution 18] the microcapsule according to any one of claims 14 to 17, wherein the second active ingredient is a fat-soluble substance selected from the group consisting of perfumes, spice extracts, animal fats and oils, perfume oils, and vegetable fats and oils.

[ solution 19] the microcapsule according to any one of the solutions 14 to 18, which is in the form of a paste.

[ scheme 20] the microcapsule according to any one of scheme 19, which is capable of being stored under refrigeration.

[ scheme 21] A flavor enhancer for foods or beverages, which comprises the microcapsule according to any one of schemes 14 to 20.

[ solution 22] A food or beverage comprising the microcapsule according to any one of solutions 14 to 20 or the flavor enhancer according to solution 21.

[ claim 23] the food according to claim 22, which is a retort pouch food.

[ scheme 24] A composition for suppressing vegetable protein odor, comprising:

(1) a microcapsule containing the yeast cells remaining after the separation of the content component produced by the method according to any one of claims 1 to 13 and a second active ingredient; and

(2) a substance having a plant protein odor-suppressing effect.

[ solution 25] A food or drink comprising:

(2) a substance having a plant protein odor-suppressing effect.

Scheme 26 a method for suppressing vegetable protein odor comprising:

the following ingredients were added:

(2) a substance having a plant protein odor-suppressing effect.

ADVANTAGEOUS EFFECTS OF INVENTION

The microcapsule of the present invention can be produced by encapsulating yeast cells extracted with hot water, and thus can contain a larger amount of flavor, and can also produce a yeast microcapsule having an excellent sustained release property in which flavor is released over time and taste time. Further, since the microcapsules of the present invention are not subjected to acid treatment, they can contain a flavor such as vanilla flavor, which has not been used conventionally due to the influence of oxidative odor, and can be used not only for processed foods, seasonings, retort pouch foods, but also for various beverages and foods such as dairy products.

Drawings

FIG. 1 shows the relationship between the concentration of yeast dispersion (solid content concentration) (weight%) and the flavor content (mg/g powder microcapsules) of yeast cells (sample 2-D) (left vertical axis) and the water content (right vertical axis). The black dots indicate the inclusion ratio, and the white dots indicate the water content ratio.

Fig. 2 shows the relationship between the mixing ratio of flavor to yeast (sample 2) (flavor/yeast) and flavor inclusion rate (mg/powder microcapsules) (left vertical axis) and inclusion efficiency (%) (right vertical axis) in the microcapsules. Wherein squares indicate flavor inclusion rate and diamonds indicate inclusion efficiency. FIG. 2A shows the results of the yeast microcapsules (sample 2-S) using yeast cells obtained by Spray drying (Spray Dry) without enzyme or emulsifier treatment, and FIG. 2B shows the results of the yeast microcapsules using yeast cells (sample 2-D) obtained by heat drying (drum drying) without enzyme or emulsifier treatment.

FIG. 3 shows the relationship between the flavor to yeast mixing ratio (flavor/yeast) and the flavor inclusion rate (mg/powder microcapsules) in the microcapsules (left vertical axis). The black graph shows the results obtained using sample 1-D, and the shaded graph shows the results obtained using sample 2-D.

FIG. 4 shows the relationship between spray drying air inlet temperature and flavor inclusion rate (mg/powder microcapsules). Wherein, the circles represent the inclusion rates of the samples 1-D and D-limonene, the diamonds represent the inclusion rates of the samples 2-D and D-limonene, the squares represent the water content of the samples 1-D, and the triangles represent the water content of the samples 2-D.

Fig. 5 shows the relationship between the stirring time of the mixed solution of the cells, the second active ingredient, and water after extraction of the content ingredient and the flavor inclusion rate (mg/powder microcapsule). Where squares represent results for sample 1-D with D-limonene, circles represent results for sample 2-D with D-limonene, diamonds represent results for sample 1-D with ethyl hexanoate, and triangles represent results for sample 2-D with ethyl hexanoate.

Fig. 6 shows the relationship between the stirring temperature of the mixed solution of the cell (sample 1) from which the content component was extracted, the second active ingredient, and water, and the flavor inclusion rate (mg/powder microcapsule). The black plot on the right side of each temperature represents ethyl hexanoate, and the shaded plot on the left side represents the results for d-limonene.

Fig. 7 shows the relationship between the stirring temperature of the mixed solution of the cell (sample 2) from which the content component was extracted, the second active ingredient, and water, and the flavor inclusion rate (mg/powder microcapsule).

FIG. 8 shows the results of observation of the structure of a yeast microcapsule using yeast cells (sample 2-S) obtained by Spray drying (Spray Dry) without enzyme treatment or emulsifier treatment, using a scanning electron microscope. Wherein, the microcapsule is a control microcapsule without flavoring agent, a microcapsule containing ethyl caproate and a microcapsule containing d-limonene in sequence from the left side. The upper and middle pictures are 3000 times of photomicrographs, and the lower picture is 10000 times of photomicrographs.

FIG. 9 shows the results of observation of the structure of a yeast microcapsule using yeast cells (sample 2-D) obtained by drum drying without enzyme treatment or emulsifier treatment, using a scanning electron microscope. Wherein, the microcapsule is a control microcapsule without flavoring agent, a microcapsule containing ethyl caproate and a microcapsule containing d-limonene in sequence from the left side. The upper and middle pictures are 3000 times of photomicrographs, and the lower picture is 10000 times of photomicrographs.

FIG. 10 shows the results of observation of the structure of yeast microcapsules using spray-dried yeast cells (sample 1-D) treated with an enzyme or an emulsifier using a scanning electron microscope. The top panel shows the yeast cells (sample 1-D) before encapsulation, and the bottom panel shows the microcapsules containing D-limonene. The top picture is a 3000-fold micrograph, the bottom left picture is a 5000-fold micrograph, and the bottom right picture is a 1500-fold micrograph.

Fig. 11 shows the results of investigation of the sustained-release behavior of the microcapsules. In the figure, the horizontal axis represents the time after the microcapsules are manufactured and applied to dry or wet conditions, and the vertical axis represents the flavor residual rate. The slow release is in dry condition on the left and in wet condition (100% wt%) on the right. The top is the result for the microcapsules containing ethyl hexanoate and the bottom is the result for the microcapsules containing d-limonene.

Fig. 12 shows the results of the investigation of the oxidative stability of the microcapsules. Wherein the horizontal axis represents the storage time (days) in which the microcapsules were exposed to oxidizing conditions at 105 ℃. The vertical axes are the limonene oxide release rate (m g/powder microcapsules) (fig. 12A) and carvone release rate (mg/powder microcapsules) (fig. 12B). The circle indicates the substance obtained by encapsulating the sample 2-D, the square indicates the substance obtained by subjecting the sample 2-D to the acid treatment and encapsulating (comparative example 1), and the tangle-solidup is the result of the spray-dried product of dextrin.

In fig. 13, GC-MS measurements were performed on each of the experimental group containing only plant protein (experimental group 1), the experimental group containing lactic acid bacteria and yeast fermentation products (experimental group 2), the experimental group containing yeast microcapsules (experimental group 3), and the experimental group containing both lactic acid bacteria, yeast fermentation products, and yeast microcapsules (experimental group 4), and the total area of GC peaks corresponding to hexanal in each experimental group was shown (experimental groups 1 to 4 from above).

Detailed Description

1. Method for producing microcapsule

The present invention, in one embodiment, relates to a method of making a microcapsule.

Although not limited thereto, the method of the present invention comprises: a step of producing yeast cells remaining after separation of content components by subjecting yeast cells to hot water treatment to release the content components outside the cells; and a step of including the yeast cells remaining after the separation of the content component as a first active ingredient and a second active ingredient, wherein the yeast cells remaining after the separation of the content component are not subjected to an acid treatment.

(Yeast cell)

In the present invention, the kind of yeast cell is not particularly limited. In one embodiment, examples of the yeast cells include cells such as torula yeast, baker's yeast, brewer's yeast, and sake yeast. The yeast cells may be in various forms such as compressed yeast, dry yeast, active dry yeast, inactivated yeast, sterilized dry yeast, and the like. The yeast cell may be a yeast cell-derived material (for example, a crushed material or a powder of a yeast cell) having substantially the same composition as the yeast cell (bacterial cell).

The yeast used in the present invention is not particularly limited, and examples thereof include yeast belonging to the genus Saccharomyces cerevisiae (Saccharomyces) and yeast belonging to the genus Candida. For example, from the viewpoint of the rich experience in eating, it may be Saccharomyces cerevisiae (Saccharomyces cerevisiae) or Candida utilis (Candida utilis) from the viewpoint of the knowledge of many studies.

(Hot Water treatment)

The method of the present invention includes a step of producing yeast cells remaining after separation of content components by subjecting yeast cells to hot water treatment to release the content components outside the cells. As long as the solution has a function of releasing the content component to the outside of the bacteria, a solution having a high temperature other than water may be used. For example, a buffer or an emulsifier may be used.

The term "content component" refers to a component obtained by extracting yeast cells by decomposition. The main components of the feed comprise amino acids, nucleic acid-related substances, minerals and vitamins, and the feed can be used for seasonings, culture media for microbial culture, livestock feeds, health auxiliary foods and the like. The fermentation function and the fermentation product (metabolite) may be used for bread, alcoholic beverages, or the like.

The present invention uses the yeast cells remaining after the separation of the content component as the first active ingredient constituting the microcapsule. In the present specification, the yeast cell remaining after the content component is separated may be referred to as a "yeast cell", and a non-limiting example of the "yeast cell remaining after the content component is separated" may include Moistex STD (fuji food industry). The Moistex STD is obtained by treating yeast cells remaining after the separation of content components with an enzyme and an emulsifier. Other examples of the "yeast cells remaining after separation of the content" include so-called dry yeasts such as "DYP-SY-02" (Fuji food industry) "and" KR yeast "(KOHJIN Life science). These samples were obtained by killing yeast cells and drying them, either by extracting the content components or without extracting them, and were not subjected to the steps of enzyme treatment, emulsifier addition treatment, and the like, unlike the Moistex STD.

An example of a specific production process of these yeast cells is shown in table 1 below.

[ Table 1]

In the microcapsule of the present invention, the yeast cells remaining after the content component is separated may be in a dry state or may be in a paste state containing water. In one embodiment, the yeast cells remaining after the content components are separated are in a dry state.

In another embodiment, the yeast is a paste to which an appropriate amount of moisture is added. The amount of water to be added to the yeast is not particularly limited. Without limitation, the yeast may be made into a paste by adding water in an amount of 0.1 times or more the amount of the yeast.

In one embodiment of the present invention, after the heat treatment, or after the heat treatment and the enzyme treatment and/or the emulsifier addition treatment, the water-washed substrate is washed with water. The washing with water means washing by adding water to the yeast cells and stirring the mixture. Without limitation, it is preferable to remove content components and the like by performing solid-liquid separation using a centrifuge after washing with water. The washing with water is preferably carried out 2 or more times or 3 or more times. By repeating the washing with water, components causing odor and offensive odor can be removed, and the color tone thereof can be made close to white.

In one embodiment, the protein content of the yeast cells remaining after the content component is separated is preferably 70% by weight or less, 66% by weight or less, 60% by weight or less, 55% by weight or less, 53% by weight or less, or 51% by weight or less. The protein content of the yeast cells remaining after the separation of the content component is preferably 45% by weight or more, 48% by weight or more, or 50% by weight or more. The protein content of the yeast cells remaining after the separation of the content component is preferably 45% by weight or more and 70% by weight or less, 45% by weight or more and 66% by weight or less, 48% by weight or more and 55% by weight or less, and 48% by weight or more and 53% by weight or less.

(second active ingredient)

The production method of the present invention includes a step of including a yeast cell remaining after separation of a content component as a first active ingredient and a second active ingredient. In this step, a microcapsule in which the second active ingredient is contained in the yeast cells remaining after the separation of the content ingredient as the first active ingredient can be obtained.

The type of the second active ingredient is not particularly limited as long as it can be contained in the yeast cells remaining after the content component is separated.

The second active ingredient is, but not limited to, a fat-soluble substance selected from the group consisting of perfumes, spice extracts, perfume oils, animal fats and oils, and vegetable fats and oils. In one embodiment of the microcapsule of the present invention, it may be added to food, beverage, or the like. The second active ingredient is preferably an ingredient that is useful for improving the flavor, aroma, taste, and the like of foods, beverages, and the like.

Among the "spices (flavors)," include d-limonene, carvone, ethyl hexanoate, chili pepper, vanilla, barbecue, mustard, coffee, pepper, black pepper, mustard, curry, meat flavor, and the like, as one embodiment.

Examples of the second active ingredient include: simple lipids such as cyclic simple waxes represented by monoester-type chain simple waxes such as methyl palmitate and higher alcohols, sterol esters such as cholesterol esters and sitosterol esters and ergosterol esters, and esters of fat-soluble vitamins such as A, D, E, simple waxes containing nitrile lipids, monoglycerides and diglycerides or triolein such as glycerol monooleate and glycerol monostearate, soybean oil, corn oil, rice bran oil, safflower oil, cottonseed oil, olive oil, castor bean oil, cod oil, cuttlefish oil, sardine oil, lard, beef tallow, sheep oil, horse oil, triglycerides represented by other microbial oils, monoalkyl, dialkyl, monoalkyl monoacyl, monoalkyl diacyl, trialkyl-type alkyl glyceryl ether lipids such as squalane and batyl alcohol, and monoolefins, diolefins, fatty acids, Alkenyl glyceryl ether lipids such as monoolefin monoacyl, monoolefin diacyl, and triene type, and alcohols such as ceramides and isoamyl alcohol. In addition, there may be mentioned: long-chain hydrocarbons such as saturated and unsaturated straight-chain, single-branched and multi-branched chain hydrocarbons which are lipid-inducing, and oxidation-inducing long-chain alcohols such as octadecanol, long-chain amino alcohols such as sphinganine, sphingosine, phytosphingosine and dehydrosphingosine, citronellal, thymol, long-chain aldehydes such as insect pheromone, straight-chain ketones such as phylloquinone and ubiquinone, various fatty acids such as stearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, eicosapentaenoic acid, docosahexaenoic acid, isovaleric acid and isobutyric acid, long-chain acids such as hydroxy acid, keto acid and dicarboxylic acid, salts thereof, sesquiterpenes and limonene, monoterpenes such as menthol, citral and ionone, muscone, thymol, nerolidol, cyperone, and sesquiterpenes such as hinokitiol, campholesterone, sesquiterpenes, terpinene, limonene, terpinene, and limonene, Examples of the carotenoid include carotenoids such as phytol, hinokitiol, sugaol, abietic acid, chlorophyll, retinol, tocopherol, diterpenes such as diterpenes and triterpenes such as tetraterpene, menadione, and ubiquinone, cholesterol, sitosterol, ergosterol, bile acid, sex hormone, adrenocortical hormone, cardiotoxic ligand, steroids saponin, steroids such as solanine, and carotenoids such as phytoene, lycopene, carotene, lutein, citrulline, and capsorubin. In addition, there can be mentioned: lecithin such as soybean lecithin, glycerophospholipids such as phosphatidylethanolamine and phosphatidylserine, glycerophospholipids such as phosphatidylinositol, phosphatidylglycerol and cardiolipin, etherglycerolipids such as plasmalogen, ceramide phosphate, sphingomyelin and sphingomyelin, phospholipids such as sphingomyeline phosphonate (ceramide citrate), glycolipids such as other glyceroglycolipids and sphingoglycolipids, steroids such as saponin and solanine, glycolipids such as fatty acid sugar and lipopolysaccharide, phosphoglycolipid, sulfurized lipid and amino acid lipid.

In addition, liposoluble liquids such as fenitrothion and pyrazothion can be mentioned. Examples of the core material include emulsifiers typified by glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and polysorbates.

In one embodiment of the present invention, the "second active ingredient" may contain a non-fat-soluble substance. However, from the viewpoint of sustained release, in one embodiment, the "second active ingredient" may contain only a fat-soluble substance.

(mixing ratio)

The mixing ratio of the yeast cells remaining after the content component is separated and the second active ingredient can be appropriately selected. The mixing ratio of the yeast cells remaining after the separation of the content component and the second active component is preferably 6:1 to 1:2, 4:1 to 1:1, 3:1 to 1:1, 2:1 to 1:1, or about 2:1 in terms of weight ratio. Preferably, the yeast cells remaining after the content component is separated are mixed so as to account for a larger proportion of the second active component.

(step of including the second active ingredient)

The step of allowing the yeast cells remaining after the separation of the above-mentioned components to contain the second active ingredient is not particularly limited. In one embodiment, the step of allowing the yeast cells remaining after the separation of the content component to contain the second active ingredient comprises: mixing and stirring the yeast cells, the second active ingredient and the water remaining after the content ingredients are separated.

In addition to water, the yeast cells and the second active ingredient remaining after the separation of the content components can be mixed and dispersed in the same manner as water, and can be used as "water" in the method of the present invention. Examples thereof include buffer solutions, sugar solutions, saline solutions, and soup solutions.

The concentration of the yeast cells and the solid content of the second active ingredient remaining after the content components contained in the mixture are separated is not particularly limited. The solid content concentration is preferably, but not limited to, 10% or more, 15% or more, 20% or more, 23% or more, 25% or more, and 30% or more. In the examples of the present specification, the solid content is about 23% or more, and the content (inclusion rate) of the second active ingredient (flavor) is maximized.

In the present invention, the conditions of the stirring time and the stirring concentration are not particularly limited as long as the yeast cells remaining after the content component is separated, the second active ingredient, and the water are sufficiently mixed to obtain a dispersion. The stirring time is preferably, but not limited to, 1 minute or more, 10 minutes or more, 30 minutes or more, 1 hour or more, 2 hours or more, or 3 hours or more. The stirring time is preferably 10 hours or less, 8 hours or less, 6 hours or less, 5 hours or less, or 4 hours or less. The stirring time is preferably 30 minutes or more and 10 hours or less, 1 hour or more and 8 hours or less, 2 hours or more and 6 hours or less, 3 hours or more and 5 hours or less. In one embodiment, the stirring time is about 4 hours.

The stirring temperature is not particularly limited as long as it is a normal temperature and does not damage yeast cells, and the stirring may be performed at a normal temperature or a constant temperature. The stirring temperature is preferably 5 ℃ or more, 10 ℃ or more, 20 ℃ or more, 25 ℃ or more, 30 ℃ or more, or 35 ℃ or more. The stirring temperature is preferably less than 60 ℃, less than 50 ℃ and less than 45 ℃. The stirring temperature is preferably 20 ℃ or more and less than 60 ℃, 25 ℃ or more and less than 50 ℃, 30 ℃ or more and less than 45 ℃. In one embodiment, the stirring temperature is about 40 ℃.

For example, without limitation, a stirrer or homogenizer may be used, and the stirring is preferably carried out at a speed of 100 to 5000rpm, or 200 to 1000 rpm.

The step of bringing the second active ingredient into contact with the yeast cells remaining after the separation of the content ingredient is not particularly limited as long as the step is a step of bringing the second active ingredient into contact with the yeast cells remaining after the separation of the content ingredient, and the second active ingredient may be contained in the yeast cells remaining after the separation of the content ingredient by any method.

In one embodiment of the present invention, in the step of allowing the yeast cells remaining after the separation of the content component to contain the second active ingredient, the yeast cells remaining after the separation of the content component, the second active ingredient and water may be mixed and stirred, and the obtained dispersion may be used as microcapsules in a paste state as it is or dried.

In addition, as an embodiment, it is a paste in which a proper amount of water is added to yeast. The amount of water to be added to the yeast is not particularly limited. Without limitation, the yeast may be made into a paste by adding water in an amount of about 0.1 times or more the amount of the yeast.

In one embodiment of the present invention, the step of allowing the yeast cells remaining after the separation of the content component to contain the second active ingredient comprises: the yeast cells remaining after the content component is separated, the second active ingredient, and water are mixed and stirred, and the obtained dispersion is dried. By performing the drying, the microcapsule in a dry state is obtained. The drying may be carried out by, for example, spray drying, heat drying (for example, atmospheric heat drying), freeze drying, vacuum drying, or the like, but is not particularly limited.

Spray drying is a method in which a dispersion (emulsion) is sprayed as fine droplets by a sprayer and brought into contact with hot air at a high temperature to form a powder. The method can control particle size by sprayer rotation speed, and is used for spraying liquid food with high viscosity and slurry liquid containing crystal. Spray drying may be carried out using a Spray Dryer such as Mini Spray Dryer B290 Buchi.

The air inlet temperature of the spray dryer is not particularly limited. But not limited to, 140 ℃ or higher, 160 ℃ or higher, 180 ℃ or higher, and 200 ℃ or higher. But not limited to, 300 ℃ or lower, 280 ℃ or lower, 250 ℃ or lower, and 220 ℃ or lower. And in one embodiment, from 160 c to 220 c, and about 200 c. The temperature of the air outlet of the spray dryer is, but not limited to, 90 to 140 ℃, 100 to 125 ℃, and 105 to 120 ℃.

In one embodiment, the dispersion obtained by mixing and stirring the yeast cells remaining after the content component is separated, the second active component and water is preferably introduced into the spray dryer at a flow rate of 5ml/min to 15ml/min and at a flow rate of 8ml/min to 12ml/min, and more preferably at a flow rate of 10 ml/min.

In one embodiment, the rotation speed of the atomizer (the device for spraying the dispersion in the form of fine droplets) of the spray dryer is preferably 10000rpm to 50000rpm, 20000rpm to 40000rpm, 25000rpm to 35000rpm, or about 30000 rpm.

Further, as the heat drying, a cylinder dryer (cylinder drying), an oven or the like can be used and the heating can be carried out by a conventional method. The heat drying is preferably normal pressure heat drying. The vacuum drying can be performed by using a continuous vacuum drying apparatus (CVD) or the like. Freeze drying is a drying technique in which a substance containing moisture such as food is rapidly frozen at about-30 ℃ and evaporated under reduced pressure to dry the moisture. This technique is also known as Freeze Dry. For example, drying may be performed using a freeze dryer.

Without limitation, the inclusion efficiency of the second active ingredient is preferably at least 10%, 20%, 30%, 40%, 50%, 60%, 80%. The inclusion efficiency of the second active ingredient is a ratio of the second active ingredient contained by the microcapsule in the second active ingredient for mixing. The amount of the second active ingredient contained in the microcapsule can be measured by a quantitative gas chromatography analysis or the like such as a gas chromatography-flame ionization detector. The yeast microcapsules treated with the enzyme and the emulsifier have higher inclusion efficiency than the yeast microcapsules not treated with the enzyme and the emulsifier.

(without acid treatment)

In the method of the present invention, the yeast cells remaining after the separation of the content component are not subjected to acid treatment. The "acid treatment" is a treatment of heating and stirring an acidic aqueous solution containing yeast cells remaining after content components are separated for a certain period of time. The acidic aqueous solution includes hydrochloric acid, phosphoric acid, sulfuric acid, lactic acid, citric acid, acetic acid, ascorbic acid, and the like, but is not particularly limited. For example, Japanese patent application laid-open No. 8-243378 discloses a method for producing yeast microcapsules, which requires the following treatment with an acidic aqueous solution.

The pH of the acidic aqueous solution is preferably 2.0 or less, more preferably 0 to 1, and still more preferably 0 to 0.5. The yeast residue may be suspended in an acidic aqueous solution at a solid content concentration of 1 to 10%, preferably 2 to 5%. The heating temperature and time of the suspension are preferably set in accordance with the pH and ionic strength of the system, and for example, when the treatment is performed using an acidic aqueous solution having a pH of 0 to 0.5, the suspension may be heated at a temperature of 50 ℃ or more and 100 ℃ or less, preferably 85 ℃ or more and 100 ℃ or less for 5 minutes or more and 1 hour or less, and preferably 10 minutes or more and 30 minutes or less. In this case, if the treatment is carried out for a long time of 1 hour or more using an acidic aqueous solution having a pH of 0 to 0.5 or if the treatment is carried out using an acidic aqueous solution having too strong acidity having a pH of 0 or less, the strength of the cell wall of the yeast is reduced, and the yield of the acid-treated yeast may be reduced. When the treatment is carried out using the above acidic aqueous solution, various organic solvents, dispersants, and preservatives may be added as necessary. The organic solvent can be various alcohols such as methanol and ethanol, acetone and hexane, the dispersant can be sucrose ester and glyceride, the antiseptic can be benzoic acid, sorbic acid and salicylic acid, and they can be used alone or in combination.

One feature of the present invention is that the acid treatment as described above is not performed.

(enzyme treatment)

In a preferred embodiment of the present invention, the method may further comprise a step of subjecting the yeast cells remaining after the separation of the content component to an enzyme treatment. The enzyme treatment comprises: a treatment for allowing the cells from which the yeast extract has been extracted to more easily contain the second component, a treatment for improving the flavor-improving effect of the yeast as the first active ingredient, a treatment for improving the taste-improving effect of the yeast as the first active ingredient, a treatment for improving the oxidation stability of the yeast as the first active ingredient, a treatment for releasing the flavor of the second active ingredient by the cells from which the yeast extract has been extracted, and the like.

The enzyme may be used alone in 1 kind or in combination of 2 or more kinds to react with yeast cells. For example, multiple proteases may be combined or multiple cellulases may be combined. In one embodiment of the present invention, the enzyme used for the enzyme treatment is a protease, a cellulase, or a combination thereof, and in one embodiment of the present invention, "enzyme treatment" is a protease and/or cellulase addition treatment.

A protease (i)

Examples of the protease include serine protease, cysteine protease, aspartic protease, and metalloprotease, and examples thereof include protease derived from microorganisms, papain derived from plants, bromelain, and the like, trypsin, pepsin, and cathepsin derived from animals.

Examples of the microorganism include Aspergillus bacteria such as Aspergillus oryzae (Aspergillus oryzae) and Aspergillus melleus (Aspergillus melleus); rhizopus species such as Rhizopus niveus (Rhizopus niveus) and Rhizopus oryzae (Rhizopus oryzae); bacillus bacteria such as Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), Bacillus licheniformis (Bacillus licheniformis), and Bacillus stearothermophilus (Bacillus stearothermophilus).

The protease may be an endo-protease or an exo-protease. Preferably an endo-protease.

While not being bound by theory, it is understood that the flavor-improving effect of yeast cells can be enhanced by reacting yeast cells with protease to cleave proteins on the surface layer of yeast cell walls and remove lower alcohols, which are causative substances of foreign odor attached to the proteins, thereby removing foreign odor.

(ii) cellulase

The cellulase is not particularly limited as long as it is a cellulase capable of hydrolyzing the glycosidic bond of β -1, 4-glucan such as cellulose, and examples thereof include cellulases derived from the following microorganisms: trichoderma bacteria such as Trichoderma reesei (Trichoderma reesei) and Trichoderma viride (Trichoderma viride); aspergillus such as Aspergillus niger and Aspergillus niger; clostridium bacteria such as Clostridium thermocellum (Clostridium thermocellum) and Clostridium johnsonii (Clostridium johnsonii); cellulomonas faecalis (Cellulomonas fimi) and other genus Cellulomonas; acremonium species such as Acremonium cellulosum (Acremonium cellulosum) and the like; rake tooth genus such as Irpex lacteus (Irpex lacteus); humicola bacteria such as Humicola insolens; pyrococcus species such as Thermococcus (Pyrococcus horikoshii), and the like.

(iii) conditions of enzyme treatment

In this step, an enzyme is added to the yeast cells remaining after the separation of the content components to cause a reaction.

The amount of the enzyme to be added may be determined by the skilled person as appropriate depending on the type of the enzyme. For example, in the case of protease, the amount of protease added is preferably, but not limited to, 1 to 5000 units, more preferably 10 to 2000 units, and still more preferably 100 to 300 units per 1g of yeast cells (solid content). For example, in the case of cellulase, it is preferably 0.1 to 100 units, more preferably 0.5 to 50 units, and still more preferably 1 to 20 units per 1g of yeast cells (solid content).

The reaction temperature and time of the enzyme can be appropriately adjusted according to the enzyme to be used. The reaction temperature may be, for example, 10 to 80 ℃ or 25 to 60 ℃. Examples of the reaction time include 15 minutes to 48 hours, 30 minutes to 48 hours, and 2 hours to 12 hours.

(emulsifiers)

In one embodiment of the present invention, the method comprises a step of adding an emulsifier before, after, or simultaneously with the step of subjecting the yeast cells remaining after the separation of the content components to the enzyme treatment.

The step of adding the emulsifier may be performed before the enzyme reaction step, may be performed after the enzyme reaction step, or may be performed simultaneously with the enzyme reaction step. In the case where two enzymes are reacted, an emulsifier may be added between the two or more enzyme reactions after one enzyme reaction and before the second enzyme reaction.

The emulsifier preferably has an HLB value of 1 to 14. The HLB value of the emulsifier is more preferably 1 to 12, and still more preferably 1 to 7.

In one embodiment, the emulsifier is selected from the group consisting of glycerol fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, sucrose fatty acid esters, lecithin, and saponins. Further, 1 kind of emulsifier may be used alone or 2 or more kinds may be used in combination and added to the yeast cells remaining after the content components are separated.

Examples of the glycerin fatty acid ester include: a monoglyceride having a polymerization degree of glycerin of 1 and a carbon number of a fatty acid of 6 to 18; a polyglycerol fatty acid ester in which the polymerization degree of glycerol is 2 to 10 and the carbon number of the fatty acid is 6 to 18; organic acid monoglycerides, and the like.

Examples of the fatty acid constituting the glycerin fatty acid ester include caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, and the like.

Examples of the organic acid monoglyceride include monoglyceride caprylic acid succinate, monoglyceride stearate citric acid ester, monoglyceride stearate acetic acid ester, monoglyceride stearate succinic acid ester, monoglyceride stearate lactic acid ester, monoglyceride stearate diacetyl tartaric acid ester, and monoglyceride oleate citric acid ester.

The sorbitan fatty acid ester includes a fatty acid having 6 to 18 carbon atoms and sorbitan having 1 or more hydroxyl groups ester-bonded to each other. More specifically, examples thereof include: sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, and the like.

The propylene glycol fatty acid ester includes those obtained by ester-bonding a fatty acid having 6 to 18 carbon atoms with propylene glycol, and may be a monoester or diester. Examples of the fatty acid constituting the propylene glycol fatty acid ester include caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, and the like.

Examples of the sucrose fatty acid ester include those obtained by ester-bonding a fatty acid having 6 to 22 carbon atoms to 1 or more hydroxyl groups of sucrose, and examples thereof include sucrose laurate, sucrose myristate, sucrose palmitate, sucrose stearate, sucrose oleate, sucrose behenate, and sucrose erucate.

Examples of the lecithin include lecithin derived from plants such as soybean, corn, peanut, rapeseed, and barley; egg yolk, cattle, etc.; examples of various lecithins extracted from microorganisms such as Escherichia coli include: glycerolecithin such as phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylcholine, phosphatidylserine, bisphosphatidic acid, and diphosphatidylglycerol; sphingomyelin such as sphingomyelin. The lecithin may also be hydrogenated lecithin, enzymolyzed hydrogenated lecithin, hydroxy lecithin, etc.

Examples of the saponin include sophoricoside, quillajasaponins, refined soybean saponin, and yucca saponin.

The amount of the emulsifier added is preferably 0.01 to 1% by mass, more preferably 0.01 to 0.1% by mass, based on the yeast cells (wet mass) remaining after the separation of the content. In the present specification, the wet mass refers to the mass of yeast cells remaining after the content component containing the liquid (dispersion medium) is separated.

The temperature and time for the treatment with the emulsifier can be appropriately adjusted depending on the emulsifier selected. Examples of the temperature include, but are not particularly limited to, 50 ℃ to 95 ℃ and 70 ℃ to 95 ℃. The reaction time is not particularly limited, and examples thereof include 10 minutes to 5 hours and 20 hours to 3 hours.

While not being bound by theory, it is believed that the hydrophobic amino acids, which are substances responsible for the unpleasant tastes such as bitter taste, astringent taste, pungent taste derived from yeast cells, are easily washed away by enzyme treatment and/or addition of an emulsifier, thereby reducing these unpleasant tastes.

2. Microcapsules

The present invention relates in one embodiment to microcapsules.

The microcapsules of the present invention comprise: yeast cells remaining after the content component produced by the production method of the present invention is separated, and a second active ingredient. In the microcapsule of the present invention, the definition of the second active ingredient is the same as that described in "1. method for producing microcapsule". In one embodiment, the second active ingredient is a fat-soluble substance selected from the group consisting of flavors, spice extracts, flavor oils, animal fats and oils, and vegetable fats and oils.

The prepared microcapsule can be frozen, refrigerated or stored at normal temperature. The microcapsules prepared are preferably stored refrigerated. The freezing temperature is below-18 deg.C, the cold storage temperature is higher than-18 deg.C and below 10 deg.C, and the normal temperature is above 10 deg.C.

Although the dried yeast can be stored at room temperature for a long time, there is a possibility that the yeast cells constituting the microcapsule are destroyed in the drying step, and the inclusion state is lost. On the other hand, the yeast cells in the paste are not destroyed and can be contained for a certain period of time. The paste can be stored at room temperature, refrigerated or frozen. However, since yeast cells constituting microcapsules are decomposed in the case of storage at room temperature, they can be stored only for a shorter time than in the dry state, and in the case of cryopreservation, the yeast cells are easily damaged even in the freezing. In the case of cryopreservation, it is preferable because the degradation can be relatively suppressed without further destruction of the yeast cells.

In one embodiment of the invention, the yeast microcapsules are pasty yeast microcapsules that can be stored under refrigeration. The term "capable of being refrigerated" refers to a form suitable for refrigerated storage, and includes a form in which refrigerated storage is actually performed. By refrigerated it is meant, without limitation, higher than-18 ℃ and below 10 ℃. Without limitation, freezing means below-18 ℃ and ambient temperature means above 10 ℃. The yeast microcapsules in paste form are preferably capable of being stored under refrigeration for 1 week or more, 2 weeks or more, 1 month or more, 2 months or more, or 3 months or more, without limitation.

The microcapsule of the present invention has excellent sustained-release properties of the second active ingredient. In one embodiment, the microcapsule of the present invention exhibits excellent inclusion property under dry conditions (e.g., 80 ℃) and exhibits a residual ratio of the second active ingredient of about 80% or more even after 60 minutes from the production of the microcapsule. Depending on the kind of the second active ingredient, some of them show a residual ratio of 90% or more, for example. In one embodiment, the microcapsules of the invention show a sustained release also under wet conditions, for example a slow release of the second active principle after 10 minutes.

In one embodiment, the microcapsule of the present invention has an improved sustained release property of the second active ingredient as compared with a microcapsule obtained using yeast cells which have been subjected to acid treatment and remain after the content ingredients have been separated. The "acid treatment" in the "acid treatment performed" is the same as the acid treatment described in the "method for producing microcapsules" (without performing the acid treatment) "item 1.

By "the sustained release of the second active ingredient is improved" it is meant that more time is spent and/or more of the second active ingredient is released. For example, the microcapsule of the present invention releases the second active ingredient after a lapse of 1.5 times, 2 times or more, 3 times or more, and 5 times or more as long as the microcapsule obtained by using the yeast cells which are acid-treated and remain after the content ingredients are separated. The microcapsule of the present invention gradually releases the second active ingredient after 1 hour, 6 hours, 12 hours, 1 day, 3 days, 5 days, 8 days, 10 days, 15 days, and 20 days from the production of the microcapsule. In one embodiment, the microcapsule of the present invention has the ability to release the second active ingredient over 8 days or more, over 10 days, over 15 days, over 20 days from the production of the microcapsule. For example, the microcapsule of the present invention releases the second active ingredient in an amount of 1.5 times or more, 2 times or more, 3 times or more, and 5 times or more as compared with the microcapsule obtained using the yeast cells which are acid-treated and remain after the content ingredients are separated.

In one embodiment, the microcapsules of the present invention exhibit excellent oxidative stability. The term "oxidative stability" means that the properties of the microcapsule are not easily changed even under oxidative conditions, for example, even when heating is performed for a certain period of time or even when an oxidizing agent is added. For example, in one embodiment, the microcapsule has the ability to release the second active ingredient after 7 days or more, 10 days, 15 days, and 20 days under drying at a high temperature (e.g., 100 ℃ or more, e.g., 105 ℃). For example, in another embodiment, the microcapsules of the present invention remain uncolored and uncorrupted after drying at high temperatures (e.g., 100 ℃ or higher, e.g., 105 ℃) for 7 days or longer, 10 days, 15 days, and 20 days.

The yeast microcapsules treated with the enzyme and the emulsifier have higher oxidation stability of the second active ingredient contained therein and have better inclusion state of the second active ingredient than the yeast microcapsules not treated with these.

In one embodiment of the microcapsule of the present invention, the second active ingredient is contained in an amount of 5% by weight, preferably 10% by weight or more, preferably 15% by weight or more, and more preferably 20% by weight or more in the microcapsule in a dried state.

In one embodiment of the microcapsule, the yeast cells remaining after the content components are separated do not form monomers, but rather form aggregates. In one embodiment of the microcapsule, the surface is formed in a thin film shape as compared with a control group containing no second active ingredient.

3. Flavor enhancer

The present invention relates in one embodiment to a flavor enhancer for food or beverage comprising the above microcapsules.

The microcapsule contains, as a first active ingredient, yeast cells remaining after the content ingredient is separated. The yeast cells remaining after the content component is separated contain components derived from the yeast cells, which are useful for enhancing the flavor of foods or beverages. Meanwhile, the second active ingredient may further contain ingredients useful for enhancing the flavor of food or beverage, such as flavors, spice extracts, animal fats and oils, flavor oils, and vegetable fats and oils.

The flavor enhancer may contain other components as long as it contains microcapsules containing the yeast cells remaining after the content components are separated and the second active component. Examples of the component that can be further contained in the flavor enhancer include a seasoning and a pH adjuster.

The "containing as an active ingredient" is not particularly limited as long as the microcapsule exhibits a flavor-enhancing effect. That is, in one embodiment, the flavor enhancer contains the microcapsules in an amount of 30% by weight or more, 50% by weight or more, preferably 70% by weight or more, and more preferably 90% by weight or more. The flavor enhancer can be added to food or beverage. The flavor enhancer containing the microcapsule of the present invention may be in a dry state or may be in a paste state (paste).

The type of food or beverage to which the flavor enhancer of the present invention can be added is not particularly limited. The food product may be added directly to food materials including meat-based dishes (for example, dishes made with chicken, beef, pork, mutton, venison, wild pork, and the like, steaks, minced meat, and the like), aquatic animal-based dishes (dishes made with sea fish, river fish, shrimp, cuttlefish, shellfish, and the like), vegetable-based dishes (for example, dishes made with fruits and vegetables, leaf dishes, root dishes, and the like), soup, stew, and the like, or may be used as a by-product of fish production such as sauce, sugar coating, fish cake, and the like, which is added to the dishes. The meat and vegetable products can be used for, for example, a pickling solution for roasted chicken skewers, a seasoning for sausages, hams and the like, a seasoning for shaomai, dumplings and the like. It can also be added to materials for desserts (e.g., cookies, cakes), etc. The beverage can be added into water, fruit juice, tea (green tea, black tea), coffee, cocoa, milk, wine, milk beverage, etc. In addition, the microcapsules may be ingested as a nutritional product. In one embodiment, the food product to which the flavor enhancer of the present invention is added is a retort pouch food product. In one embodiment, the present invention comprises the use of a pasty flavor enhancer for retort pouch foods. The type of the retort pouch food is not particularly limited, and includes curry, stew, Lin's beef rice covered with soup, Chinese rice covered with soup, beef rice covered with soup, and the like.

In one embodiment of the present invention, the food to which the flavor enhancer of the present invention is added may be a chilled food or a frozen food, but is not particularly limited to this embodiment.

In one embodiment, the food or beverage comprises the flavor enhancer in a final concentration of preferably 0.01 to 99 wt.%, preferably 0.01 to 10 wt.%, preferably 0.03 to 5 wt.%, preferably 0.05 to 3 wt.%. In one embodiment, the final concentration of the food or beverage comprising microcapsules is preferably 0.01 to 99 wt%, preferably 0.01 to 10 wt%, preferably 0.03 to 5 wt%, preferably 0.05 to 3 wt%. In one embodiment, the food or beverage comprises the second active ingredient in a final concentration of preferably 0.01 to 99 wt.%, preferably 0.01 to 10 wt.%, preferably 0.03 to 5 wt.%, preferably 0.05 to 3 wt.%.

The microcapsule of the present invention can be used not only as a flavor enhancer but also to achieve a taste-improving effect. In one embodiment, the present invention relates to a mouthfeel improving agent for food, which comprises the above-described microcapsules. The agent comprising microcapsules of the present invention can function as a flavor enhancer and a taste modifier.

The microcapsule of the present invention may contain various ingredients as the second effective ingredient. The microcapsule can be used as aromatic, deodorant, deodorizer, and moisture remover, besides food.

4. Food or beverage comprising microcapsules or flavour enhancers

The present invention relates in one embodiment to a food or beverage comprising the above microcapsules or the above flavor enhancer. The contents of the beverage and the food are the same as those described in the section "3. flavor enhancer".

5. Composition for suppressing odor of vegetable protein

The present invention relates in one embodiment to a composition for suppressing vegetable protein odor comprising: (1) microcapsules, and (2) a substance having a plant protein odor-suppressing effect.

The "microcapsules" are the same as those described in the item "1. method for producing microcapsules" and "2. microcapsules".

The "substance having a plant protein odor-suppressing effect" is a substance having a plant protein odor-suppressing effect, and is not particularly limited as long as it is a substance other than the yeast microcapsule. Specifically, for example, a fermented product (lactic acid bacteria/yeast fermented product) of soybean milk or whey produced by lactic acid bacteria and yeast can be used, and as described in, for example, paragraphs [0017] to [0044] in WO2017/014253, the present invention is not limited thereto.

Examples of the raw material of the substance having the plant protein odor suppressing effect may be whole milk, whey or soybean milk derived from milk, and preferably whey or soybean milk derived from milk, but are not particularly limited. In the fermentation of whey or soybean milk, the order of lactic acid bacteria fermentation and yeast fermentation may be appropriately set, and in a preferred embodiment, lactic acid bacteria fermentation is first performed. The lactic acid bacteria used are not particularly limited as long as they are used for food production. The skilled person can suitably design the conditions for fermentation of lactic acid bacteria depending on the lactic acid bacteria used. The yeast used for yeast fermentation is not particularly limited as long as it is a yeast used for food production. The skilled person can appropriately design the conditions for yeast fermentation depending on the yeast used.

The substance having the plant protein odor suppressing effect may be in various forms. For example, the liquid material may be concentrated or dried as necessary, and made into paste, solid, powder, granule, or the like.

Examples of the substance having the plant protein odor-suppressing effect include CN-2 (manufactured by Fuji food industries Co., Ltd.), Masking shochu for "Jingbin" (manufactured by Takara Shuzo Co., Ltd.), and other cooking liquors.

Vegetable proteins, particularly soy proteins (including processed products made from such materials), have a unique oxidative odor (also referred to as "vegetable protein odor"). A composition comprising (1) microcapsules, and (2) a substance having a plant protein odor-suppressing effect can be used to suppress such plant protein odor. The "vegetable protein odor" refers to a unique oxidation odor of a vegetable protein, particularly a soybean protein (including a processed product using the same as a raw material).

WO2017/014253 describes: the lactobacillus and yeast fermented product can inhibit plant protein odor. However, as described in example 20 of the present specification, the use of only the lactic acid bacteria/yeast fermented product can reduce the vegetable protein odor, but it cannot completely eliminate the vegetable protein odor, and there is a problem that unpleasant odor derived from vegetable protein remains during chewing. The yeast microcapsule of the present invention has a plant protein odor-inhibiting effect and an effect of improving flavor. Further, by using the yeast microcapsule of the present invention in combination with lactic acid bacteria/yeast fermentation product, the plant protein odor-suppressing effect is maintained for a long period of time.

The amounts of the yeast microcapsules and the plant protein odor-suppressing substance contained in the composition are not particularly limited. The amount of the composition to be used is not particularly limited. In one embodiment, the composition is used in an amount of 0.5 wt% or more, 0.8 wt% or more, 1.0 wt% or more, 1.2 wt% or more, and 1.5 wt% or more of the yeast microcapsules based on the plant protein. The more the amount of the yeast microcapsule used, the more the flavor derived from the yeast microcapsule is sometimes felt (unpreferably). In one embodiment, the composition is used in an amount of 5.0 wt% or less, 3.0 wt% or less, or 2.0 wt% or less of the yeast microcapsules based on the plant protein.

The ratio of the yeast microcapsule to the substance having the plant protein odor suppressing effect is not particularly limited. In one embodiment, the ratio of the yeast microcapsule to the substance having the plant protein odor-suppressing effect is 2:1 to 1:4, or 1:1 to 1: 3. In one embodiment, the ratio of the content of yeast microcapsules to the content of the substance having vegetable protein odor-suppressing effect is about 1: 2.

The composition for suppressing vegetable protein odor of the present invention can preferably obtain a vegetable protein odor suppressing effect for a longer period of time than that obtained by using a substance having a vegetable protein odor suppressing effect alone, by using a yeast microcapsule in combination with a substance having a vegetable protein odor suppressing effect. While not being bound by theory, it is believed that in one embodiment, the vegetable protein odor-suppressing effect can be exhibited over a long period of time by including a substance having a vegetable protein odor-suppressing effect in the yeast capsule and slowly releasing the substance from the yeast capsule.

6. Food or drink

The present invention relates in one embodiment to a food or beverage comprising: (1) microcapsules, and (2) a substance having a plant protein odor-suppressing effect.

The "substance having a plant protein odor-suppressing effect" is the same as that described in "5. composition for suppressing plant protein odor".

The same contents as those in the section "3. flavor enhancer" are applied to beverages and foods. Examples of the food include foods preferably suppressing vegetable protein odor. In one embodiment, the food comprises processed food such as hamburger, fried meat pie, meat ball, baked meat roll, cabbage meat roll, sausage, etc. More preferably, the processed product is a processed product using soybean protein as a raw material.

7. Method for suppressing vegetable protein odor

The invention relates to a method for inhibiting plant protein odor, which comprises the following components: (1) microcapsules, and (2) a substance having a plant protein odor-suppressing effect.

The "vegetable protein smell" is the same as that described in "5. composition for suppressing vegetable protein smell".

The mode of adding the microcapsule (1) and the substance (2) having the plant protein odor-suppressing effect is not particularly limited, and both may be added simultaneously or may be added sequentially. Preferably at the same time. The microcapsule (1) and the substance (2) having the plant protein odor suppressing effect may be added as a single composition or may be added separately.

Examples

The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. The present invention can be easily modified, changed, and included in the technical scope of the present invention by the skilled person in the light of the description of the present specification.

In the examples of the present specification, the following materials and experimental instruments were used unless otherwise mentioned.

[ Material ]

(1) Flavouring agent

In the examples, as the flavor, d-limonene and ethyl hexanoate having the following structures were used.

[ chemical formula 1]

[ chemical formula 2]

Ethyl caproate is one of the esters, known primarily as an aroma component of pineapples and sake. d-limonene is a monoterpene and is known to be used mainly as a citrus aroma component such as lemon and orange. Examples of limonene oxide are limonene oxide, carvone.

(2) Sample for use

Unless otherwise indicated, the samples used in the examples are as follows:

d-limonene (Wako pure chemical industries, Ltd., Osaka, Japan);

ethyl caproate (Wako pure chemical industries, Osaka, Japan);

cyclohexanone (Wako pure chemical industries, Ltd., Osaka, Japan);

n-hexane (Wako pure chemical industries, Ltd., Osaka, Japan).

[ Experimental instruments ]

(1) Halogen moisture meter

The moisture content of the powder was measured using a halogen moisture meter (HB43, Mettler Toledo International inc., Greifensee, Switzerland). The weight of the sample at the start of measurement was measured, and then the sample was rapidly heated by a halogen heater module built in the body to evaporate the water. The sample weight was continuously measured during the drying process, and after the measurement was completed, the measurement was performed based on the temperature weight principle in which the water content was calculated from the amount of water reduction.

(2) Gas chromatography-flame ionization detector (GC-FID)

The flavor contained in the yeast containing the flavor was quantified using GC-FID (GC-2014, shimadzu, kyoto, japan). In the examples, a flame Ionization-Detector (FID) was used as a Detector in connection with gas chromatography. A capillary COLUMN was used in the COLUMN (ULBON HR-1COLUMN,

0.53mm i.d.. times.5 μm membrane, Kyoto chemical Co., Ltd., Kyoto, Japan) and packed column (PEG20M, Kyoto chemical Co., Ltd., Kyoto, Japan).

(3) Stirrer

The flavor mixture was stirred with a stirrer (Bio ShakerBR-13UM, TAITEC corporation, Saitama Yu, Japan) while being heated to obtain a flavor dispersion. The temperature and the stirring speed can be adjusted in this stirrer.

(4) Spray drier

The flavor dispersion was Spray-dried using a Spray Dryer (Mini Spray Dryer B290 Buchi).

Example 1 preparation of Yeast cells in which the content was released to the outside of the cells and the content was separated and left

In this example, yeast cells (sample 1) which were subjected to enzyme treatment and emulsifier treatment and yeast cells (sample 2) which were not subjected to these treatments were prepared.

(1) Preparation of enzyme-treated and emulsifier-treated Yeast cells (sample 1)

According to a predetermined method, the content components of the cultured baker's yeast (saccharomyces cerevisiae) are separated by hot water extraction, and the remaining yeast cells are used. Subsequently, 0.05 mass% of glycerin fatty acid ester based on yeast cells (wet mass) was added. Yeast cells were treated at 90 ℃ for 30 minutes and sterilized. Subsequently, after adjusting pH to 7.0 after cooling, the yeast cells were added with an endo-protease and reacted at 50 ℃ for 6 hours. Thereafter, the yeast cells were treated at 80 ℃ for 20 minutes to inactivate the enzyme. Subsequently, the yeast cells were cooled, washed with water (3 times), and dried by spray drying or drum drying to obtain powdered yeast cells. The spray dried sample was designated as sample 1-S and the cylinder dried sample was designated as sample 1-D.

(2) Preparation of yeast cells which were not subjected to enzyme treatment or emulsifier treatment (sample 2)

According to a predetermined method, the content components of the cultured baker's yeast (saccharomyces cerevisiae) are separated by hot water extraction, and the remaining yeast cells are used. Next, without adding particularly enzymes and emulsifiers thereto, the yeast cells were treated at 90 ℃ for 30 minutes and sterilized. Subsequently, the yeast cells were cooled, washed with water (3 times), and dried by spray drying or drum drying to obtain powdered yeast cells. The spray dried sample was designated as sample 2-S and the cylinder dried sample was designated as sample 2-D.

EXAMPLE 2 Yeast cell microencapsulation

In this example yeast cells were encapsulated and microcapsules were prepared. On this basis, the conditions of encapsulation were investigated.

(1) Encapsulation of sample 1

The flavor mixture was prepared by mixing the sample 1-D obtained in example 1 with 25 wt% of the solid content of the flavor (the weight ratio of yeast cells to the flavor in the solid content was 2:1) and 75 wt% of water, followed by shaking and stirring (250rpm, 4 hours, 40 ℃) with a stirrer (Bio ShakerBR-13UM, TAITEC corporation, Saitama jade, japan) to prepare a flavor dispersion, and encapsulating the yeast cells to obtain microcapsules.

Next, the flavor dispersion was introduced into a spray dryer at a flow rate of 10ml/min, and spray-dried at a nebulizer rotation speed of 30000 rpm. The inlet temperature of the spray dryer is 140-220 ℃, the outlet temperature is 64-108 ℃, and the air flow rate is 35m³H is used as the reference value. The spray-dried powder was recovered and used as a powdered yeast microcapsule (powdered yeast microcapsule of sample 1).

(2) Encapsulation of sample 2

The flavor mixture was prepared by mixing 30 wt% of the solid content of the flavor (the weight ratio of yeast cells to the flavor in the solid content was 2:1) obtained in example 1 with 70 wt% of water, shaking and stirring (250rpm, 4 hours, 40 ℃) with a stirrer (Bio ShakerBR-13UM, TAITEC K.K., Saitama Yu, Japan) to prepare a flavor dispersion, and encapsulating the yeast cells to obtain microcapsules.

Next, the flavor dispersion was introduced into a spray dryer at a flow rate of 10ml/min, and spray-dried at a nebulizer rotation speed of 30000 rpm. The inlet temperature of the spray dryer is 220 ℃, the outlet temperature is 110-118 ℃, and the airflow speed is 27m³H is used as the reference value. The spray-dried powder was recovered and used as a powdered yeast microcapsule (powdered yeast microcapsule of sample 2).

(3) Investigation of Yeast Dispersion concentration

In the same yeast microcapsule preparation as in example 2(1), the concentration of the yeast dispersion was examined by changing the solid content concentration of the yeast cells (sample 2) obtained in example 1.

An ethyl caproate yeast dispersion was prepared by stirring at a stirring temperature of 40 ℃ for 8 hours. The yeast dispersion was powdered by a spray dryer under the same spray drying conditions as in example 2(1), and microcapsules of powder were prepared. The amount of flavor contained in the yeast body in the prepared microcapsules was investigated using a gas chromatography-flame ionization detector (GC-FID).

Specifically, the yeast microcapsules were extracted using n-hexane containing 500ppm of cyclohexanone as an extraction solvent. After weighing 20mg of yeast microcapsules in a 12ml test tube (NR-10, MARUEM, osaka, japan), 300 μ l of distilled water was added, and after mixing for 10 minutes at room temperature with a Vortex mixer, 1700 μ l of an extraction solvent was added, and extraction was performed at a rate of 6 at room temperature using a Vortex mixer (Vortex genii us3, IKA (registered trademark) -Werke GmbH & co. The amount of surface flavour was determined by adding 1700 μ l of extraction solvent to 20mg of yeast microcapsules and washing the surface at room temperature using a 30 minute vortex mixer. After the extracted solution was centrifuged (3000rpm, 10 minutes), 1 μ l of an n-hexane layer was injected into GC-FID (split ratio 1: 10). The parameters of the GC-FID are shown in the following table.

[ Table 2]

Nitrogen was used as a carrier gas, and the total flow rate was set to 13.9 ml/min. Detection signals were collected by a chromatography data system (C DS-Lite ver 5.0, Co., Ltd., LA soft, Kyowa, Japan) with the injection port temperature set at 140 ℃, the column temperature set at 130 ℃ and the detector temperature set at 200 ℃. The flavor was quantified using the area of the detected peak. The flavor inclusion rate (mg/g powder microcapsules) was calculated using the following calculation.

[ chemical formula 3]

Flavor inclusion rate ═ C_E/C_M···(1)，

C_E: the amount of flavor (mg) included,

C_M: amount of microcapsule powder (g) for extraction

After the measurement, the flavor agent before and after heating was quantified, and the amount of the flavor agent evaporated was calculated, and then the net water content was calculated using the following calculation formula and was regarded as the water content.

[ chemical formula 4]

M_w+f＝M_b-M_a···(2)

M_w＝M_w+f-M_f···(3)

Water content (%) < M_w/M_b×100···(4)

M_b: amount of powder (g) (before heating),

M_a: amount of powder (g) (after heating),

M_w+f: the amount of flavor and moisture (g) evaporated,

M_w: the net water content (g),

M_f: amount of flavor evaporated (from GC-FID)

The results are shown in FIG. 1. When the solid content concentration was changed, the inclusion rate of ethyl caproate (flavor) was significantly increased to 23% of the solid content concentration. On the other hand, although the inclusion rate of 23% or more is improved, the increase rate is suppressed. In the examples hereinafter, the solid content concentration was 30% by weight (20% by weight of yeast, 10% by weight of ethyl hexanoate), unless otherwise specified.

(4) Study on blending ratio of flavor and Yeast

In the same yeast microcapsule preparation as in example 2(1), the untreated yeast cells (sample 2) obtained in example 1 were used to examine the content of ethyl hexanoate when the ratio of mixed ethyl hexanoate (flavor) to yeast cells was changed. The flavor inclusion rate was calculated in the same manner as in example 2 (3).

TABLE 3 mixing ratio of flavor agent to Yeast [ TABLE 3]

The results are shown in FIG. 2. The higher the proportion of flavor relative to the yeast cells, the more the amount of flavor contained tends to increase. However, if the blending ratio of the flavor and the yeast is not less than the value of 1/2, the flavor inclusion rate does not change so much, and as a result, the inclusion efficiency is lowered. In any of the yeast cells (samples 2-S and 2-D) obtained by the spray drying process or the drum drying process, when the yeast ratio is 2 times the amount of the flavor, the inclusion efficiency is high and the flavor inclusion rate is high, so that the most efficient encapsulation can be performed when the mixing ratio of the flavor and the yeast is 1/2.

Further, regarding the two kinds of cylinder-dried yeast cells (sample 1-D and sample 2-D) obtained in example 1, powder yeast microcapsules were prepared by changing the mixing ratio of the yeast cells and D-limonene, stirring and spray-drying the yeast cells, and the content of D-limonene was examined when the ratio of the yeast cells was changed. The amount of the inclusion rate of d-limonene was calculated in the same manner as in example 2 (3). The amount of the inclusion ratio when samples 1-D and 2-D were used is shown in FIG. 3.

As shown in FIG. 3, in the case of the yeast microcapsules of sample 1-D and the yeast microcapsules of sample 2-D, each again shows: the blending ratio of the flavor and the yeast cells was 1/2, which is more effective than the blending ratio of 1/4. Further, it was also shown that the yeast microcapsules of sample 1-D contained more efficiently than the yeast microcapsules of sample 2-D, indicating that treatments such as enzyme treatment and emulsifier treatment contribute to the improvement of the flavor-containing ratio of the yeast cells.

(5) Study of spray drying air inlet temperature

In the same yeast microcapsule preparation as in example 2, the influence of the dry inlet air temperature on the flavor (D-limonene) inclusion rate and the water content was examined using the yeast cells obtained in example 1 (samples 1-D and 2-D).

A flavor mixture was prepared by mixing 25 wt% yeast cells and flavor solids (the weight ratio of yeast cells to flavor in solids was 2:1) with 75 wt% water, followed by shaking and stirring (250rpm, 4 hours, 40 ℃ C.) to prepare a flavor dispersion, followed by spray drying in the same manner as in example 2 to obtain powder yeast microcapsules of sample 1 and powder yeast microcapsules of sample 2.

The flavor inclusion rate was calculated in the same manner as in example 2 (3). The water content was measured using a halogen moisture meter (HB43, Mettler Toledo International Inc., Greifensee, Switzerland). An aluminum pan for moisture determination (diameter 100mm, 1-5790-01, AS ONE Corporation) was used to take 1g of the wet yeast microcapsules for determination. The drying temperature was set to 160 ℃. After the measurement, the flavor agent before and after heating was quantified, the amount of the flavor agent evaporated was calculated, and then the net water content was calculated using the calculation formula of chemical formula 4 and used as the water content. The flavor inclusion rate and water content are shown in FIG. 4.

The yeast microcapsules of samples 1-D had higher flavor inclusion rates than the yeast microcapsules of samples 2-D, regardless of the dry inlet air temperature. In addition, the moisture content of the yeast microcapsules of sample 1-D was lower than that of the yeast microcapsules of sample 2-D regardless of the dry inlet air temperature. In particular, when the dry inlet air temperature was above 180 ℃, the moisture content of the yeast microcapsules of samples 1-D was significantly reduced, but the flavor inclusion rate was substantially unchanged. Particularly, the water content is as low as about 3% at 200 ℃.

Therefore, the yeast cell capsule of the present invention can reduce the water content while maintaining the inclusion rate of the flavor. In addition, the yeast microcapsules of samples 1-D had a higher flavor inclusion rate than the yeast microcapsules of samples 2-D.

From the above experiments, it can be considered that the optimum temperature at the inlet of the spray drying is 200 ℃.

(6) Study of stirring time

In the same yeast microcapsule preparation as in example 2(1), the influence of the stirring time of the flavor dispersion liquid on the inclusion rate of the flavor (d-limonene or ethyl hexanoate) was examined using the yeast cells (sample 1 and sample 2) obtained in example 1. The amount of flavor inclusion rate was calculated in the same manner as in example 2 (3).

The results are shown in FIG. 5. According to the results, when d-limonene or ethyl caproate was used, the inclusion rate increased with time in 0 to 4 hours in total. When the time was 4 hours or more, the inclusion ratio was almost unchanged.

In addition, the yeast cells showed a higher flavor-containing rate when sample 1-D was used than when sample 2-D was used.

On the other hand, when the time is 4 hours or more, the inclusion ratio is hardly changed. In the examples described below, the stirring time was set to 4 hours unless otherwise specified.

(7-1) examination of stirring temperature 1

In the same yeast microcapsule preparation as in example 2(1), using the sample 1-D obtained in example 1, the influence of the stirring temperature of the flavor dispersion liquid on the inclusion rate of the flavor (D-limonene or ethyl hexanoate) was examined. The amount of flavor inclusion rate was calculated in the same manner as in example 2 (3).

The results are shown in FIG. 6. The inclusion rate of d-limonene and ethyl hexanoate showed the highest values at 40 ℃.

(7-2) investigation of stirring temperature 2

In the same yeast microcapsule preparation as in example 2(1), the influence of the stirring temperature of the flavor dispersion liquid on the inclusion rate of the flavor agent (ethyl hexanoate) was examined using the yeast cells (sample 2-D) obtained in example 1. The amount of flavor inclusion rate was calculated in the same manner as in example 2 (3).

The results are shown in FIG. 7. The highest ethyl caproate inclusion rate is achieved at 40 ℃. In the examples described below, the stirring temperature is 40 ℃ unless otherwise specified.

Example 3 shape of Yeast microcapsules

In this example, the shape of the yeast microcapsules was examined.

Specifically, a yeast microcapsule was prepared by microencapsulating sample 2, which was subjected to spray drying or heat drying (drum drying), using ethyl caproate or d-limonene as a flavor agent, respectively. In addition, sample 1 precursor and sample 1 encapsulated yeast microcapsules were prepared separately. The structure of each substance obtained was observed using a scanning electron microscope (JSM-6060, Nippon electronics Co., Ltd., Tokyo, Japan). Specifically, will

The round carbon tape (NISSHIN EM K.K., Tokyo, Japan) was placed on a sample stand, and a small amount of the yeast microcapsules were put on the carbon tape with a doctor blade. The substrate was set in a magnetron sputtering apparatus (MPS-1S, VACUUM DEVICE, Kyowa, Japan) to adhere Pt-Pd electrons thereto. And placing the sample table into a sample frame, and installing the sample table to an electron microscope for observation.

The results are shown in fig. 8, 9 and 10. The bacterial cells of sample 1 and sample 2 both did not substantially exist as monomers, but formed polymers. It was found that the microcapsules formed a film on their surface compared to the control group which did not contain the flavor. In the case of encapsulating the spray-dried sample 2, almost no difference was observed between the two cases, as compared with the case of encapsulating the heat-dried sample 2. In addition, the original sample 1 and the microbial cells encapsulated in sample 1 both did not substantially exist as monomers, but formed polymers. The polymer surface has no significant unevenness, and is integrated into a slightly planar shape. This is considered to be because the excess protein is reacted in the step of enzyme treatment or addition of an emulsifier.

Comparative example 1

As a substance corresponding to the yeast cells subjected to the acid treatment in Japanese patent application laid-open No. 8-243378, a substance subjected to a treatment in which the yeast cells were left to stand with phosphoric acid at pH2.0 for 30 minutes (hereinafter referred to as acid treatment) was prepared and used in the following experiment. Hereinafter, comparative example 1 is assumed.

Example 4 analysis of nutrient composition of Yeast cells

In this example, while the mechanism of yeast microcapsules was examined, the nutrient analysis was performed on the three yeast cell precursors of samples 2 to S, 1 to S and comparative example 1. The contents of the respective components were analyzed by the following methods.

Moisture content: drying under atmospheric pressure (105 ℃, 3 hours)

Crude protein: kjeldahl method

Lipid: acid decomposition method

Ash content: direct ashing method (550 ℃, 10 hours)

Carbohydrate: the 4 components above are subtracted from the whole.

[ Table 4]

Components of Yeast cells

In the case of the samples 1-S subjected to the enzyme treatment and the emulsifier treatment, the amount of crude protein was smaller than that of the untreated sample 2-S, and the amount of the crude protein was smaller than that of the sample 1 subjected to the acid treatment. This indicates that these intracellular proteins are partially removed. Without being bound by theory, it is believed that the inclusion ability of the microcapsules can be further improved by removing the extra protein by enzyme treatment or treatment with an emulsifier. This suggests that the intracellular protein component can be further removed by the acid treatment in Japanese patent application laid-open No. 8-243378, and the inclusion ability of the capsule can be improved.

Example 5 sustained Release behavior of microcapsules under Dry and Wet conditions

By this example, the sustained-release behavior of the microcapsules under dry conditions and wet conditions was investigated.

Using the sample 2-S and the sample 2-D obtained in example 1, powder yeast microcapsules of the sample 2-S and the sample 2-D were prepared in the same manner as in example 2 (2). Flavor release behavior of yeast microcapsules under dry conditions of 80 ℃ and 100% wet conditions was observed. The amount of flavor released slowly was investigated according to the change in the amount of flavor in the yeast powder.

The results are shown in FIG. 11. In FIG. 11, the vertical axis represents the flavor residual rate and the horizontal axis represents time. The slow release rates of ethyl caproate and d-limonene were significantly faster in various yeasts under wet conditions, while they showed some flavor retention effect under dry conditions. The yeast microcapsules subjected to the drum drying treatment (2-D) released slightly more rapidly under the wet condition than the yeast microcapsules subjected to the spray drying treatment (2-S), but had almost the same sustained release ability.

Example 6 verification of oxidative stability and sustained Release Effect of Yeast microcapsules

In this example, a test of the oxidative stability of yeast microcapsules was performed.

Using sample 2-D obtained in example 1, powder yeast microcapsules of sample 2-D were prepared in the same manner as in example 2 (2). The flavour was measured using limonene and the rate of oxidation was determined by measuring the release of its oxides, limonene oxide and carvone. As a comparative example, after sample 2-D was treated with hydrochloric acid by the method of comparative example 1, a yeast microcapsule was prepared in the same manner as in example 2 (2). As a control, a capsule obtained by spray-drying dextrin was prepared. Next, by conducting an oxidation stability test under dry conditions of 105 ℃, the sustained-release behavior of each yeast microcapsule was observed separately.

The results are shown in FIG. 12. When the release of limonene oxide was verified at the beginning of the observation (about 1 to 10 days), the spray-dried product using dextrin released limonene oxide from the beginning of the experiment, and showed that oxidation occurred in a short time, whereas the release of limonene oxide was suppressed for each yeast microcapsule, and oxidation was suppressed when yeast cells were spray-dried together with limonene. In addition, in the case of the yeast cells subjected to the acid treatment, although the release of the oxides was suppressed, the oxides were not oxidized with time, whereas in the case of the yeast cells of the present invention, the release of the oxides was slightly more than that of the yeast cells subjected to the acid treatment, and it was found that the oxidation proceeded gradually with time.

In addition, the effect of releasing carvone, which is a decomposition product of limonene oxide, was also verified. In the case of spray-drying limonene together with dextrin, carvone was released immediately after the start of the experiment and then stabilized to a value around 1mg/g, whereas in the experimental group using microcapsules, the value was low at the beginning, but carvone gradually increased. In addition, the present invention showed higher values in the case of longer preservation time than the case of the acid treatment (comparative example 1), and it was found that limonene was gradually oxidized in the present invention, and particularly carvone was gradually released from the yeast microcapsule prepared from sample 2-D after 10 days. The yeast microcapsules of the present invention, which were not acid-treated, showed a unique oxidative stability, i.e., a short oxidation induction period, the oxidation was initially inhibited and then gradually started to be oxidized, and it was found that a flavor (limonene) could be included and kept stable, as compared with the case of the acid-treatment in comparative example 1.

Example 7 preparation of pasty Yeast cell microcapsules

Sample 1-D obtained in example 1 was mixed with a spice (black pepper extract) at a solid content weight ratio of 2:1, and then 3 times the amount of water was added, followed by stirring (250rpm) with a stirrer (Bio ShakerBR-13U M, TAITEC K.K., Saitama Jade, Japan) at 40 ℃ for 4 hours to obtain a paste-like yeast microcapsule (sample 1-P).

Example 8 experiments on limonene fragrance

In this example, sensory evaluation experiments were performed on the flavor of limonene in yeast microcapsules and measured using an odor sensor. It is shown in the sense that "aroma is strong-the amount contained is large", while it is shown in the odor sensor that "numerical value is high-the odor is strong".

A 3% aqueous solution of yeast microcapsules containing limonene was prepared for sensory evaluation and odor sensor measurements. Specifically, the yeast cells obtained in example 1 (sample 1-S and sample 2-S) and limonene/water were stirred at 40 ℃ for 1 hour to prepare a flavor dispersion, thereby obtaining paste-like yeast microcapsules (sample 1-P and sample 2-P). 3% of sample 1-P and sample 2-P were added to hot water at 80 ℃ and stirred for sensory evaluation of limonene flavor and yeast odor. Sensory evaluation was evaluated by 5 professional reviewers. After sensory evaluation, the odor sensor measured the maximum value at 1 minute as a result. The results are shown in the following table.

[ Table 5]

	Sample 2-P Yeast microcapsules	Sample 1-P Yeast microcapsules
			Sensory results (limonene fragrance)	△	○
Sensory results (Yeast smell)	△	○
			Odor sensor results	921	985

Limonene fragrance ≈ intensity Δ general

Yeast odor ≈ no Δ -

There was limonene fragrance and yeast odor in both sample 1-P and sample 2-P. The limonene fragrance in sample 1-P was stronger, with a higher value in the odor sensor and a higher amount of limonene contained.

Example 9 visual confirmation experiment of the inclusion status of Yeast microcapsule containing Capsici fructus flavor

This example was conducted to perform a visual confirmation experiment on the inclusion state of the yeast microcapsule containing the pepper flavor. Yeast microcapsules containing pepper flavour were prepared using sample 1 and sample 2. The chili FLAVOR used herein was chili sauce FLAVOR OS-64657 (manufactured by KOHKEN FOOD & FLAVOR Co., Ltd.). The yeast microcapsules of sample 1 were prepared in both a powder form (SD form) after spray-drying and a paste form (paste) without spray-drying.

The contained pepper flavors were each diluted to 0.2%, and their states were visually confirmed. If "less oil slick" is visually confirmed, it indicates that the cosmetic composition contains a flavor. The yeast microcapsules of sample 2 found oil slick immediately after dilution. Whereas in the yeast microcapsules (spray-dried, paste-like) of sample 1 almost no oil slick was found.

Example 10 sensory confirmation experiment of flavor inclusion status by adding yeast microcapsules containing pepper flavor to a fish cake

In this example, a sensory test for confirming the flavor-containing state was performed by adding yeast microcapsules containing a pepper flavor prepared in the same manner as in example 9 to a fish cake. Depending on the difference in the feeling of the flavor, "a light top note and a strong back note" means "included and encapsulated well". The ratio of the fish cake to the water is shown in the following table.

[ Table 6]

	Control group	Experimental group (1)	Experiment set 2	Experiment group III
					Minced fillet KA	100	100	100	100
Salt	3	3	3	3
					Ice water	40	40	40	40
Potato starch	5	5	5	5
					Granulated sugar	3	3	3	3
Glutamic acid sodium salt	1.8	1.8	1.8	1.8
					Chili flavoring agent	0.2	—	—	—
Original body	0.4	—	—	—
					Paste (sample 1-P)	—	2	—	—
SD product (sample 1-S)	—	—	0.6	—
					SD product (sample 2-S)	—	—	—	0.6

Sensory evaluation was evaluated by 5 professional reviewers. The results are shown in the following table.

[ Table 7]

	Control group	Experimental group (1)	Experiment set 2	Experiment group III
					Primary taste of pepper flavor	○	×	×	○
Aftertaste of pepper flavor	×	○	○	△

Strong Δ (normal X: weak)

Depending on the difference in the feeling of the flavor, "a light top note and a strong back note" means "the flavor is already contained and the encapsulation state is good". The yeast microcapsules of control and sample 2 experienced a hot pepper flavor immediately after tasting. In the case of the yeast microcapsules (spray-dried, paste-like) of sample 1, the taste of the capsicum flavor was suppressed and the taste of the minced fillet was perceived, and the capsicum flavor was gradually perceived with chewing. The yeast microcapsules of sample 2 were also able to feel an aftertaste of the pepper flavor, but were weaker than the yeast microcapsules of sample 1.

Example 11 organoleptic confirmation experiment of the oxidative stability by adding yeast microcapsules containing vanilla flavour in cookies

In this example, yeast microcapsules containing vanilla flavor (yeast microcapsules of samples 1-S and yeast microcapsules of samples 2-S) were prepared and an organoleptic confirmation experiment of oxidation stability when added to cookies was performed. The yeast microcapsules are spray dried. As the vanilla flavor, vanilla flavor MQ-9071 (manufactured by high-sand spice industries, Ltd.) was used. The vanilla flavour was kneaded into the cookie and baked so that the vanilla flavour in the yeast microcapsules containing vanilla flavour was 0.01% (added at 0.05% of the yeast microcapsules).

The cookies were stored at 45 deg.C for about one week and subjected to sensory evaluation. Sensory evaluation was evaluated by 5 professional reviewers. The results are shown in the following table.

[ Table 8]

Residual intensity of vanilla flavor:. general:. DELTA.weak: "extract

The yeast microcapsules had good oxidation stability because they had a vanilla flavor even after being stored for 1 week after the high-temperature baking treatment. The yeast microcapsules of samples 1-S are more fully retained with vanilla flavor, showing higher oxidative stability than the yeast microcapsules of samples 2-S.

Example 12 addition experiment to Worcester sauce

Using black pepper extract as a spice, Worcester sauce containing sample 1-P prepared by the method of example 7 was added to udon noodles and the taste-up property thereof was evaluated.

The yeast microcapsules were obtained by preparing a flavor dispersion in the same manner as in example 2(1) using sample 1 obtained in example, and encapsulating yeast cells (sample 1-P). Black PEPPER (O LEORESIN BLACK PEPER: Kalsec) was used as a flavor.

First, a preparation of worsted sauce (manufactured by KAGOME corporation) was prepared at the following mixing ratio.

[ Table 9]

TABLE 9 formulation ratio of Worcester sauce preparation

Subsequently, each Worcester sauce preparation was heated at 75 ℃ for 10 minutes. The prepared product of the Worcester sauce was poured into a large cup (15ml) on the udon noodle, and the mixture was stirred with udon noodle uniformly for sensory evaluation. The evaluation was classified into "strength of flavor", "strength of spicy taste", "flavor remaining after two days", and "speed of expressing flavor". Sensory evaluation was evaluated by 5 professional reviewers. The results are shown in the following table.

[ Table 10]

TABLE 10 sensory test results of Worcester sauce preparation

Feel strong Δ slightly feel x, no feel

The flavor derived from the worsted sauce was slightly perceived without adding spices, but the taste was relatively flat without pungent taste. By adding the spices, the overall taste is more uniform and the flavor is properly improved. In contrast, if the yeast microcapsules of samples 1-P were added, the flavor and pungency were strongly perceived. Further, if the yeast microcapsules were included, the flavor and the pungency could be retained even when the yeast microcapsules were stored for two days, and the storage stability was found to be high.

In addition, when the spice extract is developed, flavor and aroma are immediately perceived after tasting (flavor expression speed is high), whereas if the spice extract microencapsulated by yeast is added, the flavor and aroma perceived after tasting are weak, and the flavor and aroma are gradually perceived after a while, and finally, the flavor and aroma can be perceived stronger than the flavor and aroma obtained by adding the spice extract itself.

Example 13 experiment of addition effect in retort pouch treatment

Subsequently, flavor stability was verified when yeast microcapsules containing flavor were subjected to retort pouch processing. Flavor and yeast microcapsules the same paste-like black pepper extract as in example 12 was used.

First, experimental groups were prepared in the following ratios.

The control group contains black pepper extract 0.05%

Experimental group 0.5% of the pasty yeast microcapsule in example 12

After adding water to the retort pouch, each sample was added thereto in the above-mentioned amount. Subsequently, retort pouch heating and sterilization were performed. The heating condition of the cooking bag is 120 ℃ and 20 minutes.

After the retort pouch was heated, 5 professional panelists were allowed to evaluate "intensity of spicy taste" and "continuation of spicy taste" in a retort-heated state. The state of the retort pouch before heating was set to 5 minutes, and 5-stage evaluation of 1 to 5 minutes was performed. The higher the score, the state before the retort pouch was not changed. The average of the 5 panelists' evaluations is shown in the table below.

(intensity of pungent taste)

5 points of no change at all compared with the bag before the treatment

4 point: basically no change compared with before the retort pouch treatment

3 points that the strength was slightly suppressed compared to before retort pouch treatment

2 point no strength compared with the bag before the bag is steamed

1 point: no strength at all compared with before retort pouch treatment

(continuation of pungent taste)

5 min, compared with the product before the steaming bag treatment, the spicy taste is not changed at all, and the spicy taste is kept for a long time

4, compared with the product before the steaming bag treatment, the spicy taste is basically not changed, and the spicy taste is kept

3 min, the residual time of the peppery taste is not long compared with that before the steaming bag treatment

2 min, short duration of pungency compared with before retort pouch treatment

1 point of no strength at all and instant disappearance of the pungent taste compared with before retort pouch treatment

The results are shown in the following table.

[ Table 11]

TABLE 11 sensory evaluation results after retort pouch treatment

	Intensity of pungent taste	Continuation of pungent taste
			Control group	3.1	3.2
Experimental group	4.0	4.3

When yeast microencapsulated spice extract was added (experimental group), the decrease in pungency due to retort pouch treatment was suppressed as compared with the control group to which spice extract was directly added. In addition, the harshness in the control group did not remain for a long time and disappeared immediately, but if the yeast microcapsule of sample 1 was added, the harshness appeared at a later time and the harshness could be retained for a long time. This shows the behavior of gradually releasing aroma, since the flavor can be perceived for a long time.

Example 14 sensory confirmation experiment of flavor inclusion status by adding yeast microcapsules containing flavor to Curry sauce

This example is a sensory confirmation experiment for confirming the flavor-containing state by adding yeast microcapsules containing flavors to curry paste. Flavor and yeast microcapsules the same as in example 12 were used.

The curry base material was prepared in the following ratio.

[ Table 12]

	％
		Curry powder	11.62
High-gluten flour	40.40
		Salad oil	32.83
Salt-free butter	7.58
		Margarine	7.58
Total up to	100

Next, curry paste was prepared in the following ratio, and the retort pouch was heated and sterilized. The heating condition of the cooking bag is 120 ℃ and 20 minutes. The mixing ratio of the curry sauce is shown in the following table.

[ Table 13]

After the retort pouch was heated, 5 professional panelists were allowed to evaluate "strength of flavor", "strength of spicy taste" and "speed of expressing flavor". The results are shown in the following table.

[ Table 14]

Good flavor and strong hot taste (residue), and slightly strong flavor and hot taste (slightly residue)

In the yeast microcapsule of sample 1, the flavor was slightly strongly perceived when 0.5% was added, and the flavor was strongly perceived when 1% was added. In addition, the spicy taste was strongly felt both at 0.5% addition and at 1% addition. Regarding the speed of expression of flavor, if it is incorporated into yeast microcapsules, the speed of expression of flavor is reduced, and after a lapse of time after tasting, flavor and aroma are gradually perceived, and finally, flavor and aroma stronger than those by adding a spice extract itself are perceived. In addition, if only the spice extract is added, a chemical smell (unnatural taste) is felt, but if the yeast microcapsule is added, a natural flavor can be imparted thereto.

Example 15 flavor enhancement and juicy mouthfeel enhancement experiments by adding yeast microcapsules containing charcoal-roasted flavor to a roasted chicken meat string

In this experiment, yeast microcapsules containing charcoal-fired flavor (using samples 1-D) were prepared in the same manner as in example 7, and the flavor and mouthfeel when they were added to roast chicken skewers were examined. NG SMOKE FLAVOR NO50148-A (manufactured by KOHKEN FOOD & FLAVOR Co., Ltd.) was used as a charcoal-fired FLAVOR. The ratio of the pickling solution (the pickling solution for meat, fish, etc.) is as follows.

Blank group 1% salt solution

Control group, 1% salt water and 0.5 wt% flavoring agent for charcoal burn

Experimental group 1% salt Water + 0.5% by weight of pasty Yeast microcapsules containing charcoal-fired flavour (sample 1-P)

Adding 30 weight parts of the pickling solution into a bag filled with chicken, and rolling for 1 hour. Then, the mixture was left overnight and then roasted (12 minutes at 220 ℃). Thereafter, sensory examination was performed.

As a result, the control group slightly perceived the burnt flavor, but the burnt flavor adhered only to the surface and did not penetrate into the meat. A sharp unpleasant sensation characteristic of flavors remains in the mouth. In the experimental group, charcoal flavor (barbeque flavor) was perceived and very natural. The charcoal-burned flavor is felt from the meat, and therefore the feeling that it blends with the meat can be felt. As if charcoal fire grilling was actually performed. In addition, when the flavor is sensed, the meat juice can be sensed, and the taste becomes soft and juicy. The yeast microcapsule not only can realize the flavor enhancement effect, but also can realize the effect of improving the taste.

Example 16 preparation of Yeast microcapsules containing mustard oil (Green mustard) and storage stability test in retort pouch treatment

Pasty yeast microcapsules were prepared by the same method as in example 7 using mustard oil from green mustard. Subsequently, experimental groups were prepared with the following formulation.

Control group mustard oil from green mustard 0.2 wt%

Experimental group mushy yeast microcapsules containing 10 wt% mustard oil from green mustard 2%

After water was added to the retort pouch, each sample was added. Subsequently, retort pouch heating and sterilization were performed. The heating condition of the cooking bag is 110 ℃ for 10 minutes. After the retort pouch was heated, it was cooled and stored at 35 ℃ for sensory evaluation. As a result, the flavor and taste of the control group were significantly reduced compared to those before the retort pouch treatment, and the flavor and taste of mustard were not remained. On the other hand, in the experimental group, the flavor and taste of mustard remained remarkably. In addition, after a lapse of time, a stronger flavor was confirmed.

Example 17 Effect experiment of addition to Fried Chicken nuggets

Next, the aroma, flavor of yeast microcapsules containing mustard oil (mustard) prepared by the method of example 16 when added to fried chicken nuggets was verified.

The ratio of the pickling solution is as follows.

[ Table 15]

20g of chicken was dipped in the pickling solution and rolled for 1 hour under a vacuum of 0.08MPa and a rotation speed of 10 rpm. Then, the skin was covered, cooked at 170 ℃ for 4.5 minutes, and then frozen for storage. The frozen chicken was thawed and subjected to sensory evaluation for mustard flavor and taste, and as a result, the control group had almost no mustard flavor and taste. On the other hand, the aroma and flavor remained significantly in the experimental group. In addition, the taste was soft while retaining the flavor. The yeast microcapsule can not only realize the flavor improving effect, but also realize the effect of improving the taste.

EXAMPLE 18 quantitative analysis of aroma Components

The aroma components were quantitatively analyzed by flavor-containing yeast microcapsules. In this experiment, allyl isothiocyanate, which is a main pungent ingredient of yellow mustard, green mustard, and the like, was used as a fragrance ingredient. In addition, the same microcapsules as in example 7 (sample 1-P) were used as the yeast microcapsules.

First, the following respective samples were prepared.

Control group containing mustard oil 0.2% water solution

[ Experimental method ]

2ml of each sample was taken in a vial and adsorbed by Solid Phase Microextraction (SPME) at 50 ℃ for 18 hours. The adsorbed SPME was determined by GC-MS. The residual rate of allyl isothiocyanate was determined by measuring the amount of allyl isothiocyanate in each sample of the experimental group and the control group immediately after the preparation and after the storage at 45 ℃ for 3 days.

[ aroma component: GC-MS method ]

Device VariancP-3800

Chromatographic column of Agilent DB-WAX60m X0.25 mm, I.D.0.25 μm

Heating to 50-240 deg.c at 5 deg.c/min

(results)

The measurement results are shown below.

[ Table 16]

	Control group	Experimental group
			Residual rate	39.27％	65.96％

By including mustard oil in sample 1 and microencapsulating the mustard oil, the decomposition of allyl isothiocyanate can be suppressed.

Example 19 storage stability of pasty Yeast microcapsules

In this example, the storage stability of the pasty yeast microcapsules was verified.

[ sample preparation ]

The same flavor dispersion as in example 2(1) was prepared in the same manner as in example 12 using sample 1 obtained in example 1, and yeast cells were encapsulated to prepare a yeast microcapsule in a paste form (sample 1-P). Black PEPPER (OLEORESIN BLACK PEPPER: Kalsec corporation) was used as the flavor.

The resulting pasty yeast microcapsules were stored for one month under conditions of freezing (-20 ℃ C.), cold storage (4 ℃ C.), 25 ℃ C., 45 ℃ C. The yeast microcapsules as prepared were used as a control.

A pickling solution containing the preserved pasty yeast microcapsules was prepared. A1% salt solution containing 2.5% of paste was used as a pickling solution.

Adding 30 wt% of the pickling solution into the chicken which is peeled and bagged in advance, and rolling for 1 hour. After further standing overnight, the mixture was baked (220 ℃ C., 12 minutes) and subjected to sensory examination.

(results)

The results of the sensory examination are shown in the following table. Based on the case where the yeast microcapsules immediately after preparation were used. Sensory evaluation was evaluated by 3 professional reviewers.

[ Table 17]

Feeling strong

A slight feeling

No feel of X

The flavor of black pepper can be strongly sensed in refrigerated products. There was little change in the yeast microcapsules whether they were used after storage or just after preparation. The flavor of black pepper was only slightly perceived when stored at 25 ℃. When the product is stored at 35 ℃ and 45 ℃, the flavor of black pepper is hardly perceived.

Example 20 inhibitory Effect of Yeast microcapsules on plant-derived protein odor component

In this example, the effect of yeast microcapsules on inhibiting plant-derived protein odor components was examined.

(1) Sensory evaluation test

In this example, sensory evaluation experiments were conducted on the effect of yeast microcapsules containing black pepper extract on the suppression of vegetable protein odor by using the same.

[ sample preparation ]

[ preparation of Experimental group ]

Experimental group 1 vegetable proteins

Experimental group 2 CN-2 (manufactured by Fuji food industries Co., Ltd.)

CN-2 is a compound fermented seasoning obtained by fermenting soybean milk with lactic acid bacteria and yeast.

Experimental group 3 sample 1-P

Experimental group 4 Simultaneous use of CN-2 and sample 1-P

As a raw material of plant protein, APPEX (manufactured by shin-sho oil corporation) was used.

[ Experimental method ]

Each experimental group was prepared according to the following ratio.

[ Table 18]

Sample 1-P containing vegetable protein in an amount of 2 wt%

Adding CN-2 in an amount of 4 wt% to vegetable protein

All raw materials (average 75g for each experimental group) were weighed and placed in a cooking bag for mixing. After standing for about 30 minutes (water reconstitution), each cooking bag is put into boiling water for cooking for 10 minutes. Then, the mixture was cooled to room temperature and stored in a refrigerator for 1 day and night. Samples of each experimental group were cooked and used for sensory evaluation experiments.

[ sensory evaluation test ]

Each experimental group was compared with the blank group (experimental group 1) for "smell" and "flavor" and subjected to sensory evaluation by 3 professional reviewers.

[ Table 19]

…. good. no unpleasant odor, O. unpleasant odor was suppressed

…. excellent flavor of vegetable protein, no unpleasant flavor, and

(results)

From the results of the experimental group 2, it was found that the addition of the fermentation liquid (CN-2) to the plant protein suppressed the smell of the plant protein, but the smell of the plant protein remained when the plant protein was chewed, as compared with the experimental group 1.

In addition, according to the results of experimental group 3, if the yeast microcapsule containing black pepper extract of the present invention (sample 1-P) was added to vegetable proteins, vegetable protein odor was suppressed. However, it was found that a faint odor remained and the odor increased with time. No flavor from vegetable proteins was perceived.

Further, according to the results of experimental group 4, if the fermentation broth and the yeast microcapsule containing black pepper extract of the present invention (sample 1-P) were added to the plant protein, the physical protein odor disappeared and the state could be maintained for a long time. At the same time, it was found that the flavor of vegetable proteins was not perceived.

The flavor and the yeast microcapsule of the present invention containing yeast cells can suppress the odor of plant proteins and the unpleasant flavor derived from plant proteins. In addition, by using the yeast microcapsule of the present invention and the fermentation liquid together, the effect of suppressing the odor of vegetable proteins can be maintained for a longer period of time.

(2) GC-MS analysis

In this example, GC-MS analysis was performed on the samples of each experimental group prepared in (1).

The test samples of each experimental group, which were kept in the refrigerator for 1 day, were taken out of the refrigerator the next day and returned to room temperature. 2ml of each experimental group sample was collected from a vial and adsorbed by Solid Phase Micro Extraction (SPME) at 45 ℃ for 2 hours. The adsorbed SPME was determined by GC-MS.

Device VariancP-3800

Chromatographic column of Agilent DB-WAX60m × 0.25mm, I.D., 0.25 μm

The temperature is increased to 45-240 ℃ and 5 ℃/min

The total area of the GC peaks corresponding to hexanal in each experimental group is shown in FIG. 13 according to the GC-MS measurement results.

In the experimental group 3 and the experimental group 4 to which the yeast microcapsule (sample 1-P) containing the black pepper extract of the present invention was added, the amount of hexanal was reduced in the total area of the GC peak as compared with the experimental group 1.

In the experimental group 2 to which the fermentation broth (CN-2) was added, the behavior of hexanal was not changed.

(3) Investigation of

Based on the results of (1) sensory evaluation experiments and (2) GC-MS analysis, the following investigation can be made.

The addition of the fermentation broth (CN-2) has an effect of suppressing the odor of vegetable proteins.

The test group 3 to which the yeast microcapsules (samples 1 to P) were added had the effect of suppressing the odor of vegetable proteins and the effect of improving the flavor. But in terms of odor, vegetable protein odor increases over time. In the other method, the effect of suppressing the odor of vegetable proteins in the experimental group 4 using both the fermentation broth (CN-2) and the yeast microcapsules (sample 1-P) can be sustained for a long period of time.

From the results of the GC-MS analysis in (2), in the experimental group 3 to which the yeast microcapsules were added, the amount of hexanal, which is the main component of vegetable protein odor, was reduced compared to the experimental group 1. From the results of the experimental group 2, the amount of hexanal was not decreased in the case of adding the fermentation broth (CN-2).

From the results of experimental group 3, it was found that the black pepper in the capsule was slowly released by adding the yeast microcapsule, thereby improving the smell and flavor. Further, as in experimental group 4, by using both the fermentation liquid (CN-2) and the yeast microcapsules (sample 1-P), the fermentation liquid enters the capsules and is slowly released from the yeast microcapsules, so that the effect of suppressing the odor of the plant protein can be exhibited over a long period of time.

(4) Study of the amount of Yeast microcapsule added

In this example, the amount of yeast microcapsules (sample 1-P) added was examined.

As seen from the results of the test group 3 of the sensory evaluation test (1), when 2% of the sample 1-P was added to vegetable protein, the odor-suppressing effect of vegetable protein was exhibited. Therefore, in order to confirm the optimum amount of addition, the amount of addition of sample 1-P was reduced and the sensory evaluation test was again conducted.

The experiment was conducted so that the amount of sample 1-P added was 1.6 wt%, 1.0 wt%, 0.8 wt% with respect to the vegetable protein. The results are shown in the following table.

[ Table 20]

Flavor · · · · · · · · · · · · · · · · · · · · · · · o that unpleasant odor was hardly perceived, and Δ · · · flavor of residual vegetable protein

As shown in table 20, when sample 1-P was added in an amount of 1.0 wt% or more based on the plant protein, unpleasant odor was suppressed. When sample 1-P was added in an amount of 1.0 to 2.0 wt% based on the plant protein, the flavor of the yeast microcapsules (sample 1-P) was not perceived.

Claims

1. A production method of a microcapsule, comprising:

a step of producing yeast cells remaining after separation of the content components by subjecting the yeast cells to a hot water treatment to release the content components outside the cells,

a step of including the yeast cells remaining after the separation of the content component as a first active ingredient and a second active ingredient,

the yeast cells remaining after the content components are separated are not subjected to acid treatment.

2. The method according to claim 1, comprising the step of subjecting the yeast cells remaining after the separation of the content component to a protease and/or cellulase addition treatment.

3. The method according to claim 2, which comprises a step of adding an emulsifier before, after or simultaneously with the step of adding a protease and/or a cellulase to the yeast cells remaining after the separation of the content component.

4. The method of claim 3, wherein,

the emulsifier is selected from glycerin fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, sucrose fatty acid ester, lecithin and saponin.

5. The method according to any one of claims 1 to 4,

the second effective component is liposoluble substance selected from perfume, spicery extract, perfume oil, animal fat and vegetable fat.

6. The method according to any one of claims 1 to 5,

the yeast cells remaining after the content components are separated are in a dry state.

7. The method according to any one of claims 1 to 6,

the yeast cells remaining after the content component is separated contain a protein component in an amount of 45 to 70 wt%.

8. The method according to any one of claims 1 to 7,

the step of allowing the yeast cells remaining after the separation of the content component to contain the second active ingredient includes: the yeast cells, the second active ingredient and the water remaining after the separation of the content ingredient are mixed and stirred.

9. The method according to any one of claims 1 to 8,

the step of allowing the yeast cells remaining after the separation of the content component to contain the second active ingredient includes: the yeast cells, the second active ingredient, and the water remaining after the content ingredients are separated are mixed and stirred, and the obtained dispersion is dried.

10. The method of claim 8 or 9,

the mixing ratio of the yeast cells remaining after the content component is separated to the second active component is in the range of 4:1 to 1: 1.

11. The method according to any one of claims 8 to 10,

the stirring time is 1 hour or more and 8 hours or less.

12. The method according to any one of claims 8 to 11,

the stirring temperature is above 25 ℃ and below 50 ℃.

13. The method according to any one of claims 1 to 12,

the second active ingredient has an inclusion efficiency of at least 40%.

14. A microcapsule comprising the yeast cells remaining after the content component produced by the method according to any one of claims 1 to 13 is separated, and a second active ingredient.

15. The microcapsule according to claim 14, wherein the sustained-release property of the second active ingredient is improved as compared with a microcapsule obtained using yeast cells which are acid-treated and remain after the content ingredients are separated.

16. The microcapsule according to claim 15, which has oxidation stability equivalent to that of a microcapsule obtained using the yeast cells which are subjected to the acid treatment and remain after the content components are separated.

17. A microcapsule according to any one of claims 14 to 16, which comprises 5% by weight or more of the second active ingredient.

18. The microcapsule according to any one of claims 14 to 17,

the second effective component is liposoluble substance selected from perfume, spicery extract, animal oil and fat, perfume oil and vegetable oil and fat.

19. A microcapsule according to any one of claims 14 to 18 which is paste-like.

20. The microcapsule of any one of claims 19, which is capable of being stored under refrigeration.

21. A flavor enhancer for food or beverage comprising the microcapsule according to any one of claims 14 to 20.

22. A food or beverage comprising a microcapsule according to any one of claims 14 to 20 or a flavour enhancer according to claim 21.

23. The food product of claim 22, which is a retort pouch food product.

24. A composition for suppressing vegetable protein odor comprising:

(1) a microcapsule containing the yeast cells remaining after the separation of the content component produced by the method according to any one of claims 1 to 13, and a second active ingredient; and

(2) a substance having a plant protein odor-suppressing effect.

25. A food or beverage comprising:

(2) a substance having a plant protein odor-suppressing effect.

26. A method for suppressing vegetable protein odor, the method comprising:

the following ingredients were added:

(2) a substance having a plant protein odor-suppressing effect.