JP2023104264A - Cheese flavor imparting agent - Google Patents

Abstract

To provide a production method for a flavor imparting agent that imparts the flavor of mature cheese.SOLUTION: A production method for a cheese flavor imparting agent includes a step to process raw material with protease and a step to fermentation-process the raw material with Limosilactobacillus reuteri.SELECTED DRAWING: None

Description

The present disclosure relates to methods for making cheese flavoring agents.

Cheese is made by adding lactic acid bacteria to fresh milk, adding an enzyme (rennet) that solidifies the milk to make it solid, and removing excess water and whey from it. In Japan, natural cheese, processed cheese, and cheese food are defined as cheese (Ministerial Ordinance Concerning Ingredient Standards for Milk and Dairy Products (December 27, 1951 Ordinance No. 52 of the Ministry of Health and Welfare, hereinafter Ministerial Ordinance for Milk, etc.)) . Natural cheeses are fresh or aged products obtained by coagulating milk, cream, buttermilk or a mixture thereof and then removing the whey. Processed cheese is obtained by pulverizing, mixing, heating and emulsifying natural cheese. Cheese food is obtained by pulverizing natural cheese or processed cheese, mixing, heating and melting, and emulsifying, and contains 51% or more of cheese content in the product. Non-dairy fats, proteins or carbohydrates should not exceed 10% of the final product weight. Cheese-like foods refer to foods that are not originally cheese but have the same texture, flavor, physical properties, etc. as cheese, or foods that are processed by blending cheese as a component.

The above natural cheese is roughly classified into non-ripened type and aged type depending on the presence or absence of the aging process. Unaged types include cream cheese, mascarpone, etc., and aged types include cheddar, gouda, edam, emmental, parmesan, camembert, blue, and the like. In ripened cheese, the flavor is primarily formed by enzymatic reactions during ripening. The following enzymes are involved in this enzymatic reaction.

(1) Enzymes derived from raw milk (enzymes derived from heat-resistant bacteria that remain even after sterilizing raw milk)
(2) Enzymes derived from lactic acid bacteria (peptidase, aminopeptidase, etc.)
(3) Enzymes derived from rennet (4) Enzymes derived from microorganisms other than lactic acid bacteria such as mold (in the case of cheese using mold etc.)
Here, in the case of cheese using mold, etc., the enzyme derived from microorganisms such as mold in (4) above most affects the generation of flavor, but in the case of other cheeses, usually the above (2) ) greatly influences the production of flavor during aging.

Ripening of natural cheese proceeds mainly by enzymes produced by starter lactic acid bacteria. Proteolytic enzymes (proteases) are involved in umami and texture. Proteases are classified into endoproteases and exoproteases according to their mechanism of action. Endoproteases cleave peptide bonds located within the protein molecule. Exoproteases act from the N-terminal side or C-terminal side of low-molecular-weight peptides to cleave them into amino acids or dipeptides. When lactic acid bacteria survive, they release enzymes outside the cells (extracellular enzymes, mainly endoproteases), and when they die during ripening, they release intracellular enzymes (mainly exoproteases) by bacteriolysis. It is mainly exoproteases that produce umami (amino acids).

In general, the longer the ripening period, the more the milk-derived proteins, fats, carbohydrates, and the like are decomposed, and the more intense the characteristic flavor becomes. In particular, it is thought that the generation of free amino acids and dipeptides exhibiting various flavors during the aging process enhances the various flavors and richness peculiar to aged cheese.

Demand for cheese has increased in Japan in recent years. In particular, aged cheese contains amino acids, etc. generated in the aging process as described above, and has a variety of flavors and unique richness. Demand is also increasing for processed cheese flavored as On the other hand, the production of matured cheese takes a long period of time (many days), and installation costs and operating costs for storage facilities (aging warehouses) are required. Therefore, a method for imparting the flavor of aged cheese in a short period of time and at low cost is desired.

In addition, a raw material called Enzyme Modified Cheese (EMC) is generally used as a cheese flavor imparting agent. In general, enzyme-treated cheese is obtained by subjecting raw cheese to protease treatment or lipase treatment, and is used for the purpose of imparting the flavor of natural cheese at a relatively low cost. However, in the ripening process of cheese, in addition to protease and lipase, other enzymes possessed by microorganisms function to produce various components. For this reason, it is considered difficult to impart various flavors and richness peculiar to matured cheese only with ordinary enzyme-treated cheese.

By the way, it is known that there are two types of amino acids, D-type (D-amino acid) and L-type (L-amino acid). D-type was not the subject of research because the technology for analyzing L-type and D-type with high accuracy had not been established. However, recent advances in analytical techniques have revealed the presence of some D-amino acids in vivo, and it has been reported that they play important physiological functions in the brain, hormone secretion, and the like.

D-amino acids have also attracted attention in terms of taste, and it has been reported that free amino acids exhibit greatly different flavors depending on the D-type or L-type. For example, D-aspartic acid suppresses acidity and bitterness, D-glutamic acid suppresses salty taste, and D-proline maintains sweetness and umami. It is clear. Fermented foods such as soy sauce, miso, and cheese are rich in D-amino acids, and are believed to play a role in the flavor of fermented foods (Non-Patent Document 1). In addition, traditional sake (kimoto-zukuri) tends to contain a high concentration of D-amino acids, and these D-amino acids enhance the umami and overall evaluation of sake (Non-Patent Document 2).

Among fermented foods, D-amino acids are contained in large amounts in foods containing a lactic acid fermentation step. For example, D-alanine, D-glutamic acid, and D-aspartic acid are found to be produced by a wide range of lactic acid bacteria species, and D-alanine in particular tends to be produced at high concentrations by a relatively wide range of bacterial species, while D-glutamic acid and It has been clarified that the ability to produce D-aspartic acid tends to vary greatly depending on the bacterial species (Non-Patent Document 3).

Based on the above information, the present inventors considered that not only L-amino acids but also D-amino acids are involved in cheese ripening, and conducted studies. As a result, even in the same kind of cheese, the longer the aging period, the higher the concentration of D-amino acid is contained, and the addition of the fermented product of lactic acid bacteria containing D-amino acid can impart the aged flavor of cheese. .

JP 2020-188724 Patent No. 6449418 Patent No. 6576611 Special table 2005-510254

Trace Nutrients Research 31, 59-65 (2014) Trace Nutrients Research 29, 1-6 (2012) Springer Plus 2(1), 691 (2013)

There is a demand for a method capable of imparting a mature cheese flavor that can be implemented in a short period of time at low cost.

First, the present inventors verified the flavor of aged cheese. Specifically, the same kind of cheese by aging period (Gouda: aged for 1 month and aged for 500 days, Mimolette: aged for 3 months and aged for 6 months, Manchego: aged for 3 months and aged for 12 months) was sensory evaluated and the results were compared. By doing so, we narrowed down the flavor that strengthens with aging. As a result, it was confirmed that all of the three types of cheese, after ripening, have a stronger flavor, salty taste, bitterness, and milk fat feeling, and that each of the above flavors is strongly felt, resulting in a greatly enhanced complexity. Based on this evaluation result, the contents described in the prior literature were verified in detail. Patent Literature 1 describes a method for producing a fermented milk raw material in which milk protein is treated with protease and then fermented with lactic acid bacteria. However, it only describes its use as a milk flavor imparting agent, and even when it is actually prepared and imparted to cheese, the flavor is weak and the effect of imparting complexity seen in aged cheese was not observed. Patent document 2 shows that a mixture of D-alanine, D-aspartic acid, D-glutamic acid and D-proline improves the richness and persistence. The present inventors actually verified the effect of imparting flavor by adding a mixture of D-alanine, D-aspartic acid, D-glutamic acid, and D-proline. It did not become strong and did not lead to the impartation of aged cheese-like flavor. Patent Document 3 describes a method for improving umami persistence by coexisting one or more of D-proline, D-glutamic acid and D-aspartic acid with L-glutamic acid. However, it was thought that this content improved the umami lasting, but did not lead to enhancement of other flavors (saltiness, bitterness, etc.) found in aged cheese. Patent Document 4 describes a method for producing a cheese flavor component in which protein is treated with protease and then fermented with bacteria belonging to the genera Enterococcus, Staphylococcus and Pseudomonas. However, this method is said to impart a flavor "having an intense smeared, salty, sulphurous character described as the odor of dirty socks", but not to aged flavors of cheese. It was thought that there is no umami, saltiness, bitterness, milk fat feeling, and complexity that can be felt after aging.

In order to solve the above problems, the inventors of the present invention have made further extensive studies. As a result, it was found that a high concentration of D-amino acid can be produced by fermenting the raw material with a specific lactic acid bacteria species after protease treatment, and the addition of this fermented liquid can impart a mature cheese flavor. , completed this disclosure.

That is, means for solving the above problems are as follows.
[1] A method for producing a cheese flavor imparting agent, comprising the steps of subjecting a protein-containing raw material to protease treatment and performing fermentation treatment with Remosilactobacillus reuteri.
[2] The method for producing the cheese flavor-imparting agent according to [1], wherein the flavor to be imparted is the aged flavor of cheese.
[3] The method for producing a cheese flavor-imparting agent according to [1] or [2], wherein yeast extract is contained in the raw material.
[4] The method for producing a cheese flavor-imparting agent according to any one of [1] to [3], wherein the raw material containing protein is a raw material containing milk protein, soybean protein, or yeast protein.
[5] The cheese flavor imparting agent according to any one of [1] to [4], wherein the total concentration of D-glutamic acid and D-aspartic acid that increases in the fermentation process is 500 ppm or more. Production method.
[6] A method for producing a cheese flavored food, which comprises adding the cheese flavor imparting agent obtained by the method according to any one of [1] to [5] to the food.

INDUSTRIAL APPLICABILITY According to the present invention, a matured cheese flavoring agent containing a high concentration of D-amino acid can be efficiently produced.

The present invention will be described in detail below.

The present disclosure relates to methods for producing cheese flavoring agents containing D-amino acids.

It is preferable that the lactic acid bacteria used in the fermentation treatment according to the present disclosure have a high ability to produce D-glutamic acid and D-aspartic acid. In the fermentation of lactic acid bacteria, D-glutamic acid, D-aspartic acid and D-alanine are produced in a wide range of bacterial species. It is believed that D-glutamic acid is converted from L-glutamic acid, D-aspartic acid from L-aspartic acid, and D-alanine from L-alanine. Among these, D-alanine can be used as a food additive as DL-alanine, and can be added at a relatively low cost and easily at a high concentration. On the other hand, D-glutamic acid and D-aspartic acid have not been approved for use as food additives, making it difficult to blend them in high concentrations compared to D-alanine. For this reason. A food material containing D-glutamic acid or D-aspartic acid at a higher concentration than D-alanine is more preferable. A person skilled in the art can select and use lactic acid bacteria with high productivity by performing fermentation tests or the like as necessary.

The lactic acid bacteria used in the fermentation treatment according to the present disclosure are not particularly limited as long as they have the ability to produce D-glutamic acid and D-aspartic acid. Panis, Lactobacillus delbrueckii subspecies bulgaricus, Lactobacillus acidophilus, Levilactobacillus brevis, Lactobacillus paracasei subspecies paracasei, Lactobacillus gasseri, Lactobacillus plantarum subspecies Plantarum, Remosilactobacillus fermentum, Lactococcus lactis subspecies lactis, Lactobacillus helveticus, Streptococcus thermophilus, and the like, with Remosilactobacillus reuteri being particularly preferred. A person skilled in the art can use one or more selected from these.

Examples of strains of the species include Remocilactobacillus reuteri including strains such as Remocilactobacillus reuteri JCM1112, Remocilactobacillus reuteri JCM1081, and Remocilactobacillus reuteri JCM1084. Examples of Remosilactobacillus panis include Remosilactobacillus panis JCM11053. Examples of Lactobacillus delbrueckii subspecies bulgaricus include Lactobacillus delbrueckii subspecies bulgaricus JCM1002, and examples of Lactobacillus acidophilus include strains such as Lactobacillus acidophilus JCM1034. Examples of Levilactobacillus brevis include strains such as Levilactobacillus brevis JCM1061, and examples of Lacticaseibacillus paracasei subspecies paracasei include strains such as Lacticaseibacillus paracasei subspecies paracasei JCM1109. Examples of Lactobacillus gasseri include strains such as Lactobacillus gasseri JCM1131, and examples of Lactobacillus plantarum subspecies plantarum include strains such as Lactobacillus plantarum subspecies plantarum JCM1149. mentioned. Examples of Limocilactobacillus fermentum include strains such as Limocilactobacillus fermentum JCM1173, and examples of Lactococcus lactis subspecies lactis include Lactococcus lactis subspecies lactis JCM5805. be done. Lactobacillus helveticus includes Lactobacillus helveticus FL-65 strain and the like, and Streptococcus thermophilus includes Streptococcus thermophilus FL-176 strain and the like. The above strains can be obtained from domestic and overseas public microorganism storage institutions such as the RIKEN BioResource Center and the National Institute of Technology and Evaluation Patent Microorganisms Depositary Center.

The raw material used in the present disclosure should contain at least protein, and its type and shape are not particularly limited. Other ingredients may be included, and commercially available products such as animal protein and vegetable protein can be used.

The medium for lactic acid fermentation used in the present disclosure may be the raw material itself or may contain only the raw material, and in addition to the raw material, yeast extract, sugar, pH adjuster, flavor, and any one or more of other nutrients required for growth of lactic acid bacteria, or may contain other components in addition to these. Alternatively, the raw material containing protein may be treated with protease and then mixed with the aforementioned yeast extract, sugar, pH adjuster, etc. to form a culture medium. Alternatively, a medium that has been treated with protease may be used.

Yeast extract is preferably used in the medium for lactic acid fermentation used in the present disclosure. Although the type of yeast extract is not particularly specified, it preferably contains a large amount of L-glutamic acid and L-aspartic acid, which are substrates for D-amino acids, and can sufficiently promote the fermentation of lactic acid bacteria. Examples of such yeast extracts include Ajitop (Mitsubishi Corporation Life Sciences Co., Ltd.), yeast extract SL-W (Mitsubishi Corporation Life Sciences Co., Ltd.), Hypermest HG-Pd D20 (Asahi Food and Healthcare Co., Ltd.). ), SK yeast extract Hi-KC (T) (Nippon Paper Industries Co., Ltd.), YP-21CM (Fuji Food Industry Co., Ltd.) and the like. The amount of yeast extract to be blended can be 0.5 to 12%, preferably 1 to 10%, more preferably 2 to 8%, or any number therebetween. Specific amounts of yeast extract are, for example, about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 12%.

Examples of protein-containing raw materials used in the medium for lactic acid fermentation used in the present disclosure include raw materials containing milk protein, soybean protein, yeast protein, wheat protein, and pea protein. Raw materials containing protein may contain either undegraded protein or partially degraded protein (peptone, peptide). In particular, raw materials containing milk protein are preferable because the cheese itself contains milk protein and is considered to have good flavor compatibility. For milk allergen-free cheese-flavored cheese-like foods made from soymilk or the like, it is preferable to use proteins other than milk proteins.

Sources containing milk protein include, for example, animal-derived liquid milk such as cow's milk, goat's milk, and sheep's milk, skimmed milk powder, whole milk powder or powdered milk, and milk reconstituted from concentrated milk. In particular, skim milk or powdered skim milk is preferable because it is easy to treat after lactic acid fermentation and easy to manage.
The amount of the raw material containing milk is not specified, but the non-fat milk solid content is 3.0 to 22.0%, preferably 5.0 to 20.0%, more preferably 8.0 to 18.0%. , or any number therebetween, but is not limited to these. Illustrative non-fat milk solids are, for example, about , and 22%.

Raw materials containing soy protein include, for example, soy milk, powdered soy protein, soy peptide and the like. The amount of the raw material containing soy protein is not particularly specified, but the protein is 0.5 to 10.0%, preferably 1.0 to 8.0%, more preferably 1.5% to 6.0%, or Any number between these can be blended, but is not limited to these. Specific soy protein amounts are, for example, about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10%.

Raw materials containing yeast proteins include, for example, yeast peptone and yeast cells. The amount of the raw material containing the yeast protein is not particularly specified, but the protein is 0.5 to 10.0%, preferably 1.0 to 8.0%, more preferably 1.5% to 6.0%, or Any number between these can be blended, but is not limited to these. Specific yeast protein amounts are, for example, about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10%.

Sugar is preferably added to the medium for lactic acid fermentation used in the present disclosure. The type and amount of sugar are not particularly limited, and a mixture may be used, and glucose, fructose-glucose liquid sugar, fructose, maltose, lactose, etc. can be added. Moreover, when sugar is contained in raw materials containing proteins, it can be assimilated by lactic acid bacteria. For example, skimmed milk contains lactose, so when lactic acid bacteria capable of assimilating lactose are used, it can be fermented with sugar derived from skimmed milk.

The medium for lactic acid fermentation used in the present disclosure can be pH-adjusted, flavored, and the like. The range of pH adjustment and the substances used are not particularly limited, and inorganic salts such as sodium hydroxide and potassium carbonate, trisodium citrate, and organic acids such as lactic acid, citric acid, malic acid, and acetic acid are used. , can be adjusted to a pH range suitable for lactic acid fermentation. The type and amount of flavoring agent to be used are also not particularly limited, and can be adjusted according to the required flavor.

The medium can also be supplemented with nutrients such as L-cysteine, manganese, and oleic acid that are required for the growth of some lactic acid bacteria. These can be used as they are or can be added to foods containing them.

In the present disclosure, proteolytic treatment with a protease is preferably performed prior to the fermentation treatment. In cheese, protein degradation progresses as it matures, so it is possible to reproduce that change. In addition, by degrading proteins, L-amino acids that serve as substrates for D-amino acids are supplied, making it possible to further increase the concentration of D-amino acids. Although the protease used in the present disclosure is not particularly specified, those having exoprotease activity with high L-amino acid supplying effect are preferred. As such a protease, for example, Sumiteam FP-G (Shin Nippon Chemical Industry Co., Ltd.), Protease M "Amano" SD (Amano Enzyme Co., Ltd.), Peptidase R (Amano Enzyme Co., Ltd.), and the like are preferable. Moreover, if necessary, an enzyme other than protease can be added, for example, lactase or lipase can be used in combination.

The enzyme treatment time in the present disclosure can be appropriately selected depending on the raw material, the enzyme used, the treatment temperature, the desired flavor, etc., and is usually about 0.5 to about 12 hours, preferably about 1 to about 6 hours. , more preferably from about 1.5 hours to about 4 hours, or any number therebetween, but is not limited to these times. Specific enzyme treatment times are, for example, about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7, 8 , 9, 10, 11, and 12 hours.

The enzyme treatment temperature in the present disclosure can be appropriately selected depending on raw materials, enzymes used, treatment time, desired flavor, etc., and is usually about 20 to about 65°C, preferably about 25 to about 60°C, more preferably. is from about 30° C. to about 55° C. or any number therebetween, but is not limited to these temperatures. Specific enzyme treatment times are, for example, about 20, 25, 30, 35, 40, 45, 50, 55, 60, and 65°C.

The pH at the start of enzyme treatment in the present disclosure can be appropriately selected depending on the raw material, the enzyme used, the treatment temperature, the desired flavor, etc., and is usually about pH 3-9, preferably about pH 4-8, more preferably about pH 4-8. About pH 5-7 or any number therebetween, but is not limited to these pH's. Specific pHs are, for example, about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8 .0, 8.5, and 9.0.

It is preferable that the enzyme-treated product prepared by the enzyme treatment be inactivated by heat treatment. Heat treatment conditions can be appropriately selected depending on the enzyme used, the desired flavor, and the like. The temperature of the heat treatment is usually about 70 to 150°C, preferably about 75 to 145°C, more preferably about 80 to 140°C, or any number therebetween, but is not limited to these temperatures. Specific temperatures are, for example, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, and 150°C. Similarly, the heat treatment time can be appropriately selected depending on the heating temperature, the enzyme used, the desired flavor, and the like. seconds or any number therebetween, but is not limited to these times.

A starter can be prepared as appropriate by those skilled in the art prior to the fermentation process in the present disclosure. The culture time for producing the starter can be appropriately selected depending on the raw materials, the microorganisms used, the fermentation temperature, the desired flavor, etc., and is usually about 8 hours to about 108 hours, preferably about 12 hours to about 96 hours. More preferably from about 16 hours to about 72 hours or any number therebetween, but not limited to these times. Specific fermentation times are, for example, 96, and 108 hours.

Fermentation treatment time in the present disclosure can be appropriately selected depending on raw materials, microorganisms used, fermentation temperature, desired flavor, etc., and is usually about 8 hours to about 108 hours, preferably about 12 hours to about 96 hours, More preferably from about 16 hours to about 72 hours or any number therebetween, but not limited to these times. Specific fermentation times are, for example, 96, and 108 hours.

The fermentation treatment temperature in the present disclosure can be appropriately selected depending on the raw materials, microorganisms used, fermentation time, desired flavor, etc., and is usually about 20 to about 45°C, preferably about 25 to about 43°C, more preferably. is from about 30 to about 40° C. or any number therebetween, but is not limited to these temperatures. Specific fermentation times are, for example, about 20, 25, 30, 35, 37, 40, 43, and 45°C.

The pH at the start of the fermentation treatment in the present disclosure can be appropriately selected depending on the raw material, the microorganism used, the fermentation temperature, the desired flavor, etc., and is usually about pH 3 to 9, preferably about pH 4 to 8, more preferably about pH 4 to 8. About pH 5-7 or any number therebetween, but is not limited to these pH's. Specific pHs are, for example, about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8 .0, 8.5, and 9.0.

The pH at the end of the fermentation treatment in the present disclosure can be appropriately selected depending on the raw materials, microorganisms used, fermentation temperature, fermentation time, desired flavor, etc., and is usually about pH 2.5 to 6, preferably about pH 3 to 3. 5.5, more preferably about pH 3.5 to 5 or any number therebetween, but is not limited to these pHs. Specific pHs are, for example, about 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, and 6.0.

The fermented product in the present disclosure can be subjected to pH adjustment, flavoring, and the like, if necessary. The range of pH adjustment and the substance used are not particularly limited. For example, organic acids such as lactic acid, citric acid, malic acid, and acetic acid can be used to adjust the pH to an acidic range, sodium hydroxide, potassium carbonate, etc. The pH can also be adjusted to a neutral range with an inorganic salt of The type and amount of flavoring agent to be used are also not particularly limited, and can be adjusted according to the required flavor.

The fermented product in the present disclosure can be stabilized by adding a stabilizer such as pectin or sugar before sterilization, if necessary. The types and amounts of stabilizers and sugars to be added are not particularly limited, and can be adjusted as necessary. As the stabilizer, it is preferable to use HM pectin, soybean polysaccharides, or the like, which are generally used for stabilizing acidic milk beverages. As sugar, it is preferable to use sugar (sucrose) that is generally used for stabilizing acidic beverages.

The fermented product produced by the fermentation treatment can be brought to a state in which the lactic acid bacteria are inactivated by heat treatment. Heat treatment conditions can be appropriately selected depending on the microorganisms used, the desired flavor, and the like. The temperature of the heat treatment is usually about 70 to 150°C, preferably about 75 to 145°C, more preferably about 80 to 140°C, or any number therebetween, but is not limited to these temperatures. Specific temperatures are, for example, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, and 150°C. Similarly, the heat treatment time can be appropriately selected depending on the heating temperature, the fermentation microorganism used, the desired flavor, etc. For example, it is usually 10 to 1200 seconds, preferably 15 to 900 seconds, more preferably 20 to 20 seconds. 600 seconds or any number therebetween, but is not limited to these times.

The fermented product produced by the fermentation treatment can be stored in a container in a cooled state in a refrigerator or the like. Alternatively, it can be stored in a frozen state in a freezer or the like.

The fermented milk of the present disclosure may or may not contain organic acids such as lactic acid, acetic acid, malic acid and citric acid, and aroma components such as diacetyl, acetoin, acetaldehyde, ethanol and 2-methyl alcohol.

In the production of the cheese flavor imparting agent of the present disclosure, in addition to the fermentation treatment described above, those skilled in the art may appropriately perform additional steps alone or in combination. Examples of such additional steps include addition of other ingredients, homogenization, filtration, dialysis, heating, cooling, pressurization, and decompression.

The cheese flavor imparting agent of the present disclosure can be used by adding to foods. In the present disclosure, food refers to all foods and drinks, including but not limited to solid, semi-solid, or liquid foods, and cheese-flavored foods are particularly preferred.

The food and drink mentioned above may be in liquid form, paste form, capsule form, jelly form, solid gel form, powder form, etc. In addition to tablets, liquid foods, etc., for example, breads, noodles, cake mixes, etc. Flour products such as fried chicken powder, bread crumbs; instant noodles, cup noodles, retort and cooked foods, cooked canned foods, microwave oven foods, instant soups and stews, instant miso soups and soups, canned soups, freeze-dried foods, other instant foods Ready-to-eat foods such as: canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, cereals (processed grains) and other processed agricultural products; canned marine products, fish hams and sausages, fish paste products, marine products Processed marine products such as delicacies and tsukudani; Processed livestock products such as canned livestock products, pastes, meat hams and sausages; Milk and dairy products such as cream and other dairy products; Oils and fats such as butter, margarine, and vegetable oils; Basic seasonings such as soy sauce, miso, sauces, processed tomato seasonings, mirin, and vinegar; Complex seasonings and foods such as curry ingredients, sauces, dressings, noodle soups, spices, and other complex seasonings; Ingredients Frozen foods such as frozen foods, half-cooked frozen foods, and cooked frozen foods; caramel , candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, jelly, and other confectionery; carbonated drinks, natural fruit juices, fruit juice drinks, fruit juices Preference for soft drinks, fruit drinks, fruit drinks with fruit, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic drinks, and other beverages Beverages, baby food, furikake, ochazuke seaweed and other commercially available foods; infant formula; enteral nutrition; functional foods (food for specified health use, food with nutrient function claims);

The cheese flavoring agent of the present disclosure can be concentrated or dried, such as by vacuum concentration, membrane concentration, drum drying, air drying, spray drying, vacuum drying or freeze drying, or combinations thereof.

As used herein, "about" means a range of ±10%, preferably ±5% of the stated numerical value.

Examples according to the present disclosure will be described below, but the technical scope of the present disclosure is not limited to this description.

<Method for analyzing free amino acids>
The contents of free amino acids in the following commercially available cheeses, examples, and comparative examples are amino acids using the following ortho-phthalaldehyde/N-acetyl-L-cysteine chiral derivatization method (OPA-NAC chiral derivatization method) Measured by quantitative analysis. The commercially available cheeses subjected to analysis are as follows. Gouda aged for 1 month (country of origin: Holland), Gouda aged for 500 days (country of origin: Netherlands), Mimolette aged for 3 months (country of origin: France), Mimolette aged for 6 months (country of origin: France), Manchego aged for 3 months (country of origin: France) Country: Spain), Manchego aged for 12 months (country of origin: Spain). In addition, each amino acid was described with the following abbreviations.
D-aspartic acid: D-Asp
L-aspartic acid: L-Asp
D-glutamic acid: D-Glu
L-glutamic acid: L-Glu
D-alanine: D-Ala
L-alanine: L-Ala
L-serine: L-Ser
L-histidine: L-His
L-arginine: L-Arg
L-tyrosine: L-Tyr
L-valine: L-Val
L-methionine: L-Met
L-phenylalanine: L-Phe
L-Isoleucine: L-Ile
L-Leucine: L-Leu
L-Lysine: L-Lys

Two volumes of methanol were added to the liquid fermented product according to each example, stirred, and then centrifuged. . After crushing commercial cheese finely, add 10 times the amount of water, stir, homogenize with a homogenizer, centrifuge, and dilute the obtained supernatant 3 times with distilled water to make a chiral derivative. It was used as a sample for A sample for chiral derivatization was prepared by directly diluting the supernatant by 2 to 100 times with distilled water depending on the amount of amino acid contained.

<Chiral derivatization procedure>
To 60 μl of the sample for chiral derivatization, 40 μl of 1% sodium tetraborate aqueous solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 20 μl of 1% N-acetyl-L-cysteine aqueous solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 20 μl of 1.6% ortho-phthalaldehyde methanol solution (manufactured by Sigma-Aldrich) was added, and filtered through a 0.45 μm cellulose acetate membrane filter (manufactured by Advantec Toyo Co., Ltd.) to obtain a chiral derivatization treatment solution. Amino acid analysis was performed by high performance liquid chromatography (HPLC, manufactured by Shimadzu Corporation, limit of detection: 0.5 ppm) using the chiral derivatization treatment solution as an analysis sample.

Further, the conditions shown in Table 1 below were selected as the analysis conditions for the amino acid analysis by HPLC using the chiral derivatization treatment liquid as the analysis sample.

<Analysis of pH>
The pH of the following examples, comparative examples, medium samples before protease treatment, and medium samples after protease treatment were analyzed using a desktop pH meter LAQUA (Horiba, Ltd.).

<Preparation example of lactic acid bacteria starter>
A culture medium consisting of 5.53 parts by weight of MRS (de Man Rogosa Sharpe) medium (Difco® product, Nippon Becton Dickinson) was prepared, sterilized at 121°C for 15 minutes, and then cooled to 37°C. Then, after cooling, the culture medium was inoculated with 0.02 parts by weight of Remosilactobacillus reuteri JCM1112 strain to obtain a viable cell count of about 1.0×10 ⁵ to 1.0×10 ⁷ cfu/ml. cultured for hours. Thereafter, the fermented product was centrifuged to collect the cells, suspended in the same amount of sterilized physiological saline, and centrifuged again to wash the cells. The finally obtained cells were resuspended in sterilized physiological saline and adjusted to a viable cell count of about 1.0×10 ⁸ to 2.0×10 ⁹ cfu/ml, which was used as JCM1112 starter.

A JCM1081 starter was prepared by replacing the lactic acid bacteria to be inoculated with Remocilactobacillus reuteri JCM1081 from the JCM1112 starter preparation example and otherwise taking the same steps.

A JCM1084 starter was prepared by replacing the lactic acid bacteria to be inoculated with Remocilactobacillus reuteri JCM1084 from the JCM1112 starter preparation example and otherwise taking the same steps.

A JCM1002 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactobacillus delbrueckii subsp.

A JCM1034 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactobacillus acidophilus JCM1034 from the above JCM1112 starter preparation example and otherwise taking the same steps.

A JCM1061 starter was prepared by replacing the lactic acid bacteria to be inoculated with Levilactobacillus brevis JCM1061 from the above JCM1112 starter preparation example, setting the culture temperature to 30 ° C., and otherwise taking the same steps.

A JCM1109 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lacticaceae Bacillus paracasei subsp.

A JCM1131 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactobacillus gasseri JCM1131 from the above JCM1112 starter preparation example and otherwise taking the same steps.

JCM1149 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactobacillus plantarum subsp.

A JCM1173 starter was prepared by replacing the lactic acid bacteria to be inoculated with Remosilactobacillus fermentum JCM1173 from the JCM1112 starter preparation example and otherwise taking the same steps.

A JCM11053 starter was prepared by replacing the lactic acid bacteria to be inoculated with Remocilactobacillus panis JCM11053 from the JCM1112 starter preparation example, and otherwise taking the same steps.

A JCM5805 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactococcus lactis subsp.

FL-65 starter was prepared by replacing the lactic acid bacteria to be inoculated with Lactobacillus helveticus FL-65 from the above JCM1112 starter preparation example and otherwise taking the same steps.

FL-176 starter was prepared by replacing the lactic acid bacteria to be inoculated with Streptococcus thermophilus FL-176 from the above JCM1112 starter preparation example and otherwise taking the same steps.

<Example 1-1: Preparation example of fermented milk>
10 parts by weight of powdered skim milk (Morinaga Milk Industry Co., Ltd.), 5 parts by weight of Ajitop (Mitsubishi Corporation Life Sciences Co., Ltd.), and 83.5 parts by weight of water were mixed to collect a sample for analysis (referred to as Comparative Example 1-1 ). After that, the temperature is adjusted to 50° C. in a hot water bath, 0.5 parts by weight of Sumiteam FP-G (Shin Nippon Chemical Industry Co., Ltd.) is added, and the mixture is stirred with a stirrer and held at 50° C. for 2 hours to perform protease treatment. rice field. After the protease treatment, it was sterilized in a hot water bath at 90° C. for 20 minutes, cooled to 37° C., and a sample for analysis was aseptically recovered (referred to as Comparative Example 1-2). After sterilization, the medium was inoculated with 1 part by weight of the JCM1112 starter prepared above and fermented at 37° C. for 24 hours. Fermentation was stopped by heating at 80° C. for 1 minute, and the resulting fermented product was designated as Example 1-1.

<Example 1-2: Preparation example of fermented milk>
Example 1-2 was prepared by replacing the starter JCM1112 with the starter JCM1081 in Example 1-1, and otherwise taking the same steps.

<Example 1-3: Preparation example of fermented milk>
Example 1-3 was prepared by replacing the starter JCM1112 with the starter JCM1084 in Example 1-1 and otherwise taking the same steps.

<Comparative Example 1-3: Preparation example of fermented milk>
Comparative Example 1-3 was prepared by replacing Sumizyme FP-G with water, omitting the protease treatment step, and taking other steps similar to those of Example 1-1.

<Comparative Example 1-4: Preparation example of fermented milk>
Comparative Example 1-4 was prepared by replacing the starter JCM1112 with the starter JCM1002 in Example 1-1, and otherwise taking the same steps.

<Comparative Example 1-5: Preparation example of fermented milk>
Comparative Example 1-5 was prepared by replacing the starter JCM1112 with the starter JCM1034 in Example 1-1 and otherwise taking the same steps.

<Comparative Example 1-6: Preparation example of fermented milk>
Comparative Example 1-6 was prepared by replacing the starter JCM1112 with the starter JCM1061 in Example 1-1, setting the culture temperature to 30° C., and otherwise performing the same steps.

<Comparative Example 1-7: Preparation example of fermented milk>
Comparative Example 1-7 was prepared by replacing the starter JCM1112 with the starter JCM1109 in Example 1-1 and otherwise taking the same steps.

<Comparative Example 1-8: Preparation example of fermented milk>
Comparative Example 1-8 was prepared by replacing the starter JCM1112 with the starter JCM1131 in Example 1-1, and otherwise taking the same steps.

<Comparative Example 1-9: Preparation example of fermented milk>
Comparative Example 1-9 was prepared by replacing the starter JCM1112 with the starter JCM1149 in Example 1-1, setting the culture temperature to 30° C., and otherwise performing the same steps.

<Comparative Example 1-10: Preparation example of fermented milk>
Comparative Example 1-10 was prepared by replacing the starter JCM1112 with the starter JCM1173 in Example 1-1 and otherwise taking the same steps.
<Comparative Example 1-11: Preparation example of fermented milk>
Comparative Example 1-11 was prepared by replacing the starter JCM1112 with the starter JCM11053 in Example 1-1, and otherwise taking the same steps.

<Comparative Example 1-12: Preparation example of fermented milk>
Comparative Example 1-12 was prepared by replacing the starter JCM1112 with the starter JCM5805 in Example 1-1, setting the culture temperature to 30° C., and otherwise performing the same steps.

<Comparative Example 1-13: Preparation example of fermented milk>
In Example 1-1, 1 part by weight of the starter JCM1112 was replaced with 0.5 parts by weight of the starter FL-65 and 0.5 parts by weight of the starter FL-176. 13 was prepared.

<Comparative Examples 1-14 to 1-17: Preparation examples of fermented milk for comparison>
10 parts by weight of powdered skim milk and 88.5 parts by weight of water were mixed, and a sample for analysis was collected (comparative example 1-14). After that, the temperature is adjusted to 50° C. in a hot water bath, 0.5 parts by weight of Sumiteam FP-G (Shin Nippon Chemical Industry Co., Ltd.) is added, and the mixture is stirred with a stirrer and held at 50° C. for 2 hours to perform protease treatment. rice field. After protease treatment, it was sterilized in a hot water bath at 90° C. for 20 minutes, cooled to 37° C., and a sample for analysis was aseptically recovered (comparative example 1-15). After sterilization, the medium was inoculated with 1 part by weight of the JCM1112 starter prepared above and fermented at 37° C. for 24 hours. Fermentation was stopped by heating at 80° C. for 1 minute, and the resulting fermented product was designated as Comparative Example 1-17. On the other hand, Comparative Example 1-15 was prepared by replacing Sumizyme FP-G with water, omitting the protease treatment step, and taking other steps similar to those of Comparative Example 1-17.

<Example 2: Preparation example of soy protein fermented liquid>
5 parts by weight of Prolina HD101R (Fuji Oil Co., Ltd.), 5 parts by weight of Ajitop, 5 parts by weight of whole sugar glucose glue final (Shimizuko Sugar Refining Co., Ltd.), and 83.5 parts by weight of water were mixed to obtain a sample. It was recovered (referred to as Comparative Example 2-1). After that, the temperature was adjusted to 50° C. in a hot water bath, 0.5 parts by weight of Sumizyme FP-G was added, and the mixture was stirred with a stirrer and held at 50° C. for 2 hours for protease treatment. After the protease treatment, it was sterilized in a hot water bath at 90° C. for 20 minutes, cooled to 37° C., and the sample was aseptically recovered (referred to as Comparative Example 2-2). After sterilization, the medium was inoculated with 1 part by weight of the JCM1112 starter prepared above and fermented at 37° C. for 24 hours. Fermentation was stopped by heating at 80° C. for 1 minute, and the resulting fermented product was used as Example 2.

<Comparative Example 2-3: Preparation example of soy protein fermented liquid>
Comparative Example 2-3 was prepared by replacing Sumizyme FP-G with water, omitting the protease treatment step, and taking the same steps as in Example 2 above.

<Example 3: Preparation example of yeast peptone fermentation liquid>
5 parts by weight of yeast peptone HYP-A581 (Oriental Yeast Co., Ltd.), 5 parts by weight of Ajitop, 5 parts by weight of total sugar glucose glue final, and 83.5 parts by weight of water were mixed, and a sample was collected (comparative example 3-1). After that, the temperature was adjusted to 50° C. in a hot water bath, 0.5 parts by weight of Sumizyme FP-G was added, and the mixture was stirred with a stirrer and held at 50° C. for 2 hours for protease treatment. After the protease treatment, it was sterilized in a hot water bath at 90° C. for 20 minutes, cooled to 37° C., and the sample was aseptically recovered (referred to as Comparative Example 3-2). After sterilization, the medium was inoculated with 1 part by weight of the JCM1112 starter prepared above and fermented at 37° C. for 24 hours. Fermentation was terminated by heating at 80° C. for 1 minute, and the resulting fermented product was designated as Example 3.

<Comparative Example 3-3: Preparation example of yeast peptone fermentation liquid>
Comparative Example 3-3 was prepared by replacing Sumizyme FP-G with water, omitting the protease treatment step, and taking the same steps as in Example 3 above.

Table 2 shows the free amino acid analysis results for the commercially available cheese.

Comparing the concentrations of free amino acids for each aging period of Gouda, Mimolette, and Manchego in Table 2, it was found that the overall free amino acid concentration tended to be higher in samples with longer aging periods for all three types of cheese. Furthermore, it was shown that not only the L-form, but also three D-form (D-Asp, D-Glu, D-Ala) free amino acids increased in concentration in samples with a long aging period. From this result, it is confirmed that both free L-amino acids and free D-amino acids are increased in matured cheese, and by supplementing these concentrations, the free amino acid composition can be brought closer to matured cheese, and derived from amino acids It was thought that the flavor could also be brought closer.

Table 3 shows the analysis results of free amino acids in Examples 1-1 to 1-3 and Comparative Examples 1-1 to 1-13.

Comparing Comparative Examples 1-1 and 1-2 in Table 3, it was shown that the concentrations of all free amino acids were improved. This was probably because proteins in the medium were degraded by protease treatment, resulting in an increase in free amino acids. In Table 2, it was confirmed that the concentration of all free amino acids in cheese increases with ripening, so it was shown that protease treatment of the medium can cause an increase in free amino acid concentration in the same manner as cheese ripening.

Comparing Example 1-3 and Comparative Examples 1-1 to 1-3, the concentrations of two D-amino acids (D-Asp, D-Glu) are clearly higher in Examples 1-1 to 1-3. It was shown that In Examples 1-1 to 1-3, the protease-treated medium was fermented with Remosilactobacillus reuteri. On the other hand, Comparative Examples 1-1 and 1-2 were not fermented with lactic acid bacteria and contained almost no D-amino acids. Further, in Comparative Example 1-3, the medium without protease treatment was fermented with Remocilactobacillus reuteri JCM1112, but compared with Examples 1-1 to 1-3 in which fermentation was performed after protease treatment, D -Asp, D-Glu concentrations were low. From this, it was shown that the concentration of D-Asp and D-Glu produced by fermentation can be improved by performing lactic acid fermentation after protease treatment.

Comparing Examples 1-1 to 1-3 with Comparative Examples 1-4 to 1-13, two types of D-amino acids (D-Asp, D-Glu ) was shown to be higher. In Examples 1-1 to 1-3, Remosilactobacillus reuteri was used, and in Comparative Examples 1-4 to 1-13, lactic acid bacteria species other than Remosilactobacillus reuteri were used. Other conditions such as protease treatment and fermentation time were similar. From this result, it was shown that significantly higher concentrations of D-Asp and D-Glu can be obtained by fermentation with Remosilactobacillus reuteri after protease treatment than when other lactic acid bacteria are used.

Table 4 shows the free amino acid analysis results of Comparative Examples 1-14 to 1-17.

Comparing Comparative Examples 1-14 to 1-17 with Comparative Examples 1-1 to 1-3 and Example 1-1 shown in Table 3, Comparative Examples 1-1 to 1-3 and Example 1-1 , all amino acids tended to have high concentrations. Furthermore, when comparing Comparative Examples 1-16 and 1-17 with Comparative Examples 1-3 and Example 1-1, Comparative Example 1-3 and Example 1-1 clearly show three types of D-amino acid (D-Asp, D-Glu, D-Ala) concentrations were shown to be elevated. Comparative Examples 1-14 to 1-17, Comparative Examples 1-1 to 1-3, and Example 1-1 differ only in the addition of yeast extract to the medium. For this reason, it was considered that the addition of the yeast extract promoted the fermentation of Remosilactobacillus reuteri and promoted the conversion of D-amino acids.

Table 5 shows the analysis results of free amino acids in Example 2 and Comparative Examples 2-1 to 2-3.

In Example 2 and Comparative Examples 2-1 to 2-3 shown in Table 5, soybean protein, yeast extract, and glucose were used as medium raw materials instead of skimmed milk powder and yeast extract. Comparing Comparative Examples 2-1 and 2-2, the concentrations of all free amino acids are improved, and it was shown that even when soy protein is used, the free amino acid concentration can be increased by protease treatment in the same manner as milk. . In addition, in Comparative Example 2-3 and Example 2, the D-Asp and D-Glu concentrations were increased, and in particular, in Example 2 in which fermentation was performed with Remosilactobacillus reuteri after protease treatment, higher concentrations were obtained. D-Asp, D-Glu were detected. These results indicate that even when soybean protein is used as a medium, significantly high concentrations of D-Asp and D-Glu can be obtained by fermentation with Remocilactobacillus reuteri after protease treatment.

Table 5 shows the analysis results of free amino acids in Example 3 and Comparative Examples 3-1 to 3-3.

In Example 3 and Comparative Examples 3-1 to 3-3 shown in Table 6, yeast peptone, yeast extract, and glucose were used as medium raw materials instead of skimmed milk powder and yeast extract. Comparing Comparative Examples 3-1 and 3-2, the concentrations of all free amino acids are improved, indicating that even when yeast peptone is used, protease treatment can increase the concentration of free amino acids in the same manner as milk. . In addition, in Comparative Example 3-3 and Example 3, the D-Asp and D-Glu concentrations are increased, and in Example 3 in which fermentation was performed with Remosilactobacillus reuteri after protease treatment, a higher concentration of D -Asp, D-Glu detected. These results indicate that high concentrations of D-Asp and D-Glu can be obtained by fermentation with Remocilactobacillus reuteri after protease treatment even when yeast peptone is used as a medium.

As described above, a cheese flavor improving agent containing high concentrations of D-Asp and D-Glu could be prepared using various raw materials.

<Taste sensory test of food and drink to which cheese flavor improver is added>

Regarding the taste of food and drink to which the cheese flavor improving agents according to Examples 1-1 to 1-3, Example 2, and Example 3 were added, Comparative Examples 1-1 to 1-17 and Comparative Example 2- A sensory test was conducted while comparing the taste of food and drink to which 1 to 2-3 and Comparative Examples 3-1 to 3-3 were added.

Tables 7 to 10 show a list of formulations in evaluation test plots subjected to sensory evaluation. In Tables 7 to 10, Examples 1-1 to 1-3, Example 2, and Example 3, and Comparative Examples 1-1 to 1-17, with respect to 100 parts by weight of blank food It was prepared by adding 0.5 parts by weight of any of Comparative Examples 2-1 to 2-3 and Comparative Examples 3-1 to 3-3.

Each evaluation test plot was subjected to a sensory evaluation test. Specifically, 5 panelists (n=5) who are well trained and who evaluate food and drink on a daily basis evaluated the food and drink. From the evaluation results of commercially available cheeses by ripening period, the longer the ripening period, the stronger the umami, saltiness, bitterness, milk fat feeling, and complex taste (variety of flavors that can be felt). It was judged that the strength of the whole flavor became stronger and more complex. Therefore, the aged cheese feeling was evaluated by the following items for each evaluation test section. That is, for umami, saltiness, bitterness, and milk fat, the intensity of each flavor was scored, and the average score given by 10 people was used as the evaluation. Complex taste was evaluated based on the variety of perceived flavors (for example, the degree to which various flavors are mixed together rather than a strong single taste such as umami or bitterness). In addition, the average of the above five items was used as a comprehensive evaluation. According to this evaluation method, it was determined that when the overall evaluation was high, the aged cheese-like flavor was strong. In addition, the evaluation points are uniformly set to 2.0 for each item of the target food and drink itself (blanks 1 to 3), and based on this 2.0 point, each item in each comparative example and example A score of "1, 2, 3, 4, or 5" was assigned to give a large score if the variety of taste and flavor was strong.

In Tables 11 and 12, processed cheese (baby cheese (plain) manufactured by Rokko Butter Co., Ltd.) was used as a cheese blank (hereinafter referred to as blank 1). The evaluation item was set to 2.0. In the case, the fermented milk according to Examples 1-1 to 1-3 was added to Blank 1, and Examples 4-1 to 4-3 were prepared according to the recipes in Table 7, and to Blank 1 The sensory evaluation results of Comparative Examples 4-1 to 4-17 prepared according to the recipes in Table 7 and Table 8 with the addition of the fermented milk according to Comparative Examples 1-1 to 1-17 are shown. In addition, each fermented milk was added by heating and dissolving the blank 1 in a microwave oven, followed by thorough mixing.

From the results of Table 11, Examples 4-1 to 4-3 exceeded the evaluation point of 2.0 of Blank 1, which was used as the evaluation criteria, in any evaluation item, and Comparative Examples 4-1 to 4-13 or higher. got the result. Examples 4-1 to 4-3 all use Remosilactobacillus reuteri in fermented milk, and Comparative Examples 4-4 to 4-13 use other lactic acid bacteria, so that Remosilacto It was found that the addition of fermented milk using Bacillus reuteri strongly improves the aged flavor of cheese. Further, when Example 4-1 and Comparative Example 4-3 were compared, Example 4-1 exceeded Comparative Example 4-3 in all evaluation items. Since Example 4-1 and Comparative Example 4-3 differ only in the presence or absence of protease treatment of the fermented milk, by using Remosilactobacillus reuteri after performing the protease treatment of the raw material, either It was shown that a stronger cheese ripening flavor can be imparted than when only is performed.

From the results in Table 12, Comparative Examples 4-14 to 4-17 were lower than Comparative Examples 4-1 to 4-3 and Example 4-1 in Table 11 in all evaluation items. Between Comparative Examples 4-14 to 4-17, Comparative Examples 4-1 to 4-3, and Example 4-1, all differ only in the presence or absence of addition of yeast extract to the raw material of fermented milk. All use the same process and strain. Therefore, by adding yeast extract to the raw material, the fermentation of Remosilactobacillus reuteri is promoted, the amount of D-amino acids is greatly increased, and the effect is greatly improved in imparting cheese ripening flavor at the time of addition. was shown. Furthermore, when comparing Comparative Example 4-2 in which only protease treatment was performed without fermentation with Remosilactobacillus reuteri, and Comparative Example 4-15, Comparative Example 4- in which yeast extract was added to the raw material 2, it exceeded Comparative Examples 4-15 in which yeast extract was not added in all evaluation items. Therefore, it was suggested that protease treatment of the yeast extract itself, in addition to promoting the fermentation of Remosilactobacillus reuteri, contributes to enhancing the cheese ripening flavor.

In Table 13, ice cream (excellent mascarpone manufactured by Morinaga Milk Industry Co., Ltd.) is used as a blank related to frozen dessert (hereinafter referred to as blank 2). Example 5 was prepared by adding the fermented milk according to Example 1-1 according to the recipe in Table 9, and blank 2 was added with fermented milk according to Comparative Examples 1-1 to 1-3. were prepared according to the recipes in Table 9 and were used as Comparative Examples 5-1 to 5-3, and the sensory evaluation results are shown.

From the results in Table 13, Example 5 exceeded the evaluation score of 2.0 for blank 2, which was used as the evaluation criteria, in all evaluation items, and obtained results equal to or higher than those of Comparative Examples 5-1 to 5-3. Here, when Example 5 and Comparative Example 5-3 were compared, Example 5 exceeded Comparative Example 5-3 in all evaluation items. Since Example 5 and Comparative Example 5-3 differ only in the presence or absence of protease treatment of the fermented milk, cheese-flavored frozen desserts can be obtained by using Limosilactobacillus reuteri after protease treatment of the raw materials. It was also shown that it is possible to impart a strong cheese ripening flavor to the cheese.

Table 14 shows the blank 3 when the evaluation item of the cheese-like food (Soy milk shred manufactured by Marsanai Co., Ltd.) is used as a blank that does not contain milk (hereinafter referred to as blank 3) is 2.0. The soy protein fermented liquid according to Example 2 and the yeast peptone fermented liquid according to Example 3 were added to Examples 6-1 to 6-3 prepared according to the recipe in Table 10, and blank 3 To the soy protein fermented liquid according to Comparative Examples 2-1 to 2-3 and the yeast peptone fermented liquid according to Comparative Examples 3-1 to 3-3 were added according to the recipe in Table 10, and Comparative Example 6-1 The results of the sensory evaluation are shown for those with ˜6-9.

From the results of Table 14, Example 6-1 and Example 6-2 exceeded the evaluation score of 2.0 of Blank 3, which was used as the evaluation criterion, in any evaluation item, and Comparative Examples 6-1 to 6- 6 or more results were obtained. The results in Table 14 show that even when soybean protein and yeast peptone are used, a strong cheese ripening flavor can be imparted by combining protease treatment and fermentation with Remosilactobacillus reuteri. Since the soybean protein fermented liquid and the yeast peptone fermented liquid do not contain dairy materials, it is thought that they can be used for the purpose of imparting a matured flavor to dairy allergen-free cheese-flavored foods.

In the present disclosure, the aged cheese has increased free L-amino acids and D-amino acids, and the addition of a cheese flavor improver with increased L-amino acids and D-amino acids also improves the aged flavor of cheese. It turned out to be reproducible. L-amino acids are known to exhibit umami, bitterness, etc., depending on the type. Similarly, simple D-amino acids are also known to mainly exhibit sweetness, and it has been shown that D-amino acids also affect perception of other flavors. From these, by bringing the composition of L-amino acids and D-amino acids closer to matured cheese, it is possible to enhance the various flavors derived from each amino acid itself while also enhancing the influence of D-amino acids on the flavor, resulting in ripening. It was considered that a state in which various flavors were felt strongly like cheese was reproduced.

By adding the cheese flavor improving agent according to the present disclosure, umami, salty taste, bitterness, milk fat feeling, and complex taste can be enhanced, and mature cheese flavor can be imparted to a wide range of food and drink.

By adding the flavor improving agent according to the present invention, it is possible to improve the body feeling and sharpness of the aftertaste to a wide range of food and drink, and to improve the flavor balance as a whole.

Claims (6)

A method for producing a cheese flavor imparting agent, comprising a step of treating a protein-containing raw material with protease, and a step of performing fermentation treatment with Remocilactobacillus reuteri. 2. The method for producing a cheese flavor-imparting agent according to claim 1, wherein the flavor to be imparted is the aged flavor of cheese. 3. The method for producing a cheese flavor-imparting agent according to claim 1 or 2, wherein yeast extract is contained in the raw material. 4. The method for producing a cheese flavor imparting agent according to any one of claims 1 to 3, wherein the raw material containing protein is a raw material containing milk protein, soybean protein, or yeast protein. 5. The method for producing a cheese flavor-imparting agent according to any one of claims 1 to 4, wherein the total concentration of D-glutamic acid and D-aspartic acid, which increases in the fermentation process, is 500 ppm or more. A method for producing a cheese-flavored food, which comprises adding the cheese flavor-imparting agent obtained by the method according to any one of claims 1 to 5 to the food.