Mold-ripened cheese flavoring agent and method for producing mold-ripened cheese flavoring agent

Abstract

The present invention addresses the problem of providing: a flavoring agent that imparts, to foods and beverages, a good flavor like a natural cheese, particularly a mold-ripened cheese; and a food or beverage in which the flavoring agent is used.　Provided is a mold-ripened cheese flavoring agent that satisfies all of requirements (1) to (3).　Requirements: (1) a mold fermentation product is contained; (2) molds cannot be analyzed in PCR sequence analysis; and (3) the mass ratio of the contained free glutamic acid to the contained methyl ketones, which is expressed as (free glutamic acid) / (methyl ketones), is 0.1 to 40 inclusive. The expression "molds cannot be analyzed" means that the amount of mold DNA contained in a PCR amplification product is at or below the detection limit due to the absence of molds in the product.

Description

Mold-type cheese flavoring agent and method for producing mold-type cheese flavoring agent

[0001] RELATED ART This application claims the benefit of priority from Japanese Patent Office Application No. 2025-050046 filed on March 25, 2025 (Reiwa 7). The entire content of the priority basic application is incorporated herein by reference.

[0002] The present invention relates to a mold-type cheese flavoring agent and a method for producing a mold-type cheese flavoring agent.

[0003] Cheese is a type of dairy product produced through processes such as coagulation and fermentation using milk collected from cows and other animals as a raw material. It is eaten not only as it is but also in combination with other foods. Among cheeses, mold-type cheeses such as blue cheese and Camembert cheese that utilize mold fermentation have unique aromas and aged flavors, and are used in various dishes. In recent years, initiatives aimed at achieving the Sustainable Development Goals (SDGs) have been progressing globally. In particular, due to global population growth, social awareness of crisis regarding global issues such as "food resource shortage" and "environment" has been increasing. Under such market circumstances, cheese flavoring agents and cheese flavor-like lactic acid fermented products have been studied as materials that can impart cheese flavor without using cheese itself (Patent Documents 1 and 2). In addition, it has been conventionally known to add cheese flavoring to foods and drinks in order to impart cheese flavor.

[0004] Japanese Unexamined Patent Application Publication No. 2021-108562, Japanese Unexamined Patent Application Publication No. 2018-68182

[0005] The inventor has studied flavoring agents that impart cheese flavor, particularly mold-type cheese flavor, to foods and drinks. As means for imparting mold-type cheese flavor, it is conceivable to blend mold-type cheese itself or add a flavoring agent. However, the flavor titer of mold-type cheese itself is insufficient, and there have been cases where sufficient flavor cannot be imparted to the target food or drink. In addition, flavoring agents may impart artificial aromas and flavors, and there have been cases where it is difficult to impart a good mold-type cheese-like flavor to foods and drinks.

[0006] Furthermore, as shown in Patent Document 1, simply adding specific amino acids sometimes made it difficult to achieve the aroma and aged flavor of moldy cheese. Similarly, as shown in Patent Document 2, even cheese-flavored lactic acid fermented products sometimes made it difficult to achieve the aroma and aged flavor of moldy cheese. Therefore, the object of the present invention is to provide a flavoring agent that imparts a particularly good flavor reminiscent of moldy cheese to food and beverages, and to provide food and beverages using this flavoring agent.

[0007] As a result of diligent research into the above-mentioned problems, the inventors have discovered that the above-mentioned problems can be solved when a product contains fungal fermentation products, when the fungi cannot be analyzed by PCR sequencing, and when the mass ratio of free glutamic acid to methyl ketones, as represented by free glutamic acid / methyl ketones, is within a specific range. This led to the completion of the present invention.

[0008] In other words, the present invention encompasses the following inventions: [1] A mold-based cheese flavoring agent that satisfies all of the following requirements 1) to 3): 1) Contains mold fermentation products; 2) Molds are unanalyzable in PCR sequencing analysis; 3) The mass ratio of free glutamic acid to methyl ketones, as expressed by free glutamic acid / methyl ketones, is 0.1 or more and 40 or less, provided that "molds are unanalyzable" means that the amount of mold DNA contained in the PCR amplified product is below the detection limit because no molds are included. [2] The mold-based cheese flavoring agent according to [1] having a blue cheese flavor. [3] The mold-based cheese flavoring agent according to [1] or [2] containing 0.3% by mass or more of free glutamic acid and 0.05% by mass or more of methyl ketones per solid content. [4] The mold-based cheese flavoring agent according to [1] or [2] further satisfying 4) below. 4) Contains free fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms in an amount of 0.4% by mass or more and 50% by mass or less per solid content. [5] Furthermore, the mold-based cheese flavoring agent described in [3] satisfies the following 4). 4) Contains free fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms in an amount of 0.4% by mass or more and 50% by mass or less per solid content. [6] A cheese-like food containing the mold-based cheese flavoring agent described in [1] or [2]. [7] A cheese-like food containing the mold-based cheese flavoring agent described in [3]. [8] A cheese-like food containing the mold-based cheese flavoring agent described in [4]. [9] A cheese-like food containing the mold-based cheese flavoring agent described in [5].

[10] Food and beverages containing the cheese-like food described in [6].

[11] Food and beverages containing the cheese-like food described in [7].

[12] Food and beverages containing the cheese-like food described in [8].

[13] Food and beverages containing the cheese-like food described in [9].

[14] A method for producing a mold-based cheese flavoring agent, comprising the following steps (A) and (B): (A) a step of extracting a mold fermentation product to obtain a mold fermentation product; (B) a step of mixing the mold fermentation product obtained in step (A) with a lactic acid bacteria fermentation product and / or a lipase enzyme reaction product of oil and fat.

[15] A method for imparting a mold-based cheese flavor to food and beverages, comprising incorporating a composition that satisfies all of the following requirements 1) to 3).1) Contains fungal fermentation products. 2) Fungi are undetectable by PCR sequencing analysis. 3) The mass ratio of free glutamate to methyl ketones, expressed as free glutamate / methyl ketones, is 0.1 or higher and 40 or lower. However, "fungi are undetectable" means that the amount of fungal DNA contained in the PCR amplified product is below the detection limit because no fungi are present.

[0009] The present invention provides a flavoring agent that imparts a particularly good flavor to food and beverages, similar to that of moldy cheeses, among other types of natural cheeses. Furthermore, it enables the provision of food and beverages using this flavoring agent.

[0010] In this specification, moldy cheeses include all types of cheese in which mold is grown inside or on the surface of natural cheese. Blue cheeses refer to moldy cheeses that primarily utilize blue mold, and examples include Roquefort, Gorgonzola, Blue, Stilton, Fourme d'Ambert, Dana Blue, Cabrales, Queso de Valdeón, Cambozola, and Bresse Blue.

[0011] The mold-based cheese flavoring agent of the present invention is a flavoring agent that can impart the aforementioned mold-based cheese flavor to various foods and beverages. The mold-based cheese flavoring agent can be used in forms such as extracting mold fermentation products from oils and fats, sterilizing mold fermentation products with a filter, or removing mold precipitates from mold fermentation products by centrifugation. The mold-based cheese flavoring agent of the present invention may be concentrated or diluted as needed to adjust its flavor potency. For example, the flavor potency can be adjusted from a flavor intensity that exhibits a clear effect when added at 0.01% to 9% by mass, to a flavor intensity that exhibits a clear effect when added at 10% to 90% by mass. The amount of mold-based cheese flavoring agent added to foods and beverages can be adjusted according to the desired flavor.

[0012] In this specification, the characteristic aroma of blue cheese refers to a blue cheese-like aroma, which constitutes the blue cheese flavor. In this specification, the aged flavor refers to the umami, richness, or both that are perceived when eating, which constitutes the blue cheese flavor. A particularly natural and authentic blue cheese flavor can be perceived when all of these components are perceived in a balanced manner.

[0013] The mold-based cheese flavoring agent of the present invention contains mold fermentation products. Mold fermentation products refer to components produced from molds through fermentation. By including these mold fermentation products in the mold-based cheese flavoring agent, a good mold-based cheese flavor that cannot be reproduced with fragrances alone can be imparted. Typical and non-limiting production examples of mold fermentation products used in the mold-based cheese flavoring agent are given below, but those skilled in the art can appropriately design the fermentation method of the molds and the production conditions of the mold fermentation products. The mold fermentation products to be included in the mold-based cheese flavoring agent of the present invention can be obtained by fermentation by filamentous fungi, which are a type of mold. Examples of such fungi include those belonging to the genera Aspergillus and Penicillium. Examples of molds belonging to the genus Aspergillus include Aspergillus oryzae, Aspergillus tamari, Aspergillus parasiticus, Aspergillus awamori, Aspergillus kawachii, Aspergillus niger, or Aspergillus pheonicis. Aspergillus oryzae, Aspergillus awamori, and Aspergillus kawachii are commercially available as starter cultures and can also be used. Examples of fungi belonging to the genus Penicillium include Pen. roqueforti, Pen. camemberti, Pen. casei, or Pen. janthinellum.

[0014] In one embodiment, the mold-based cheese flavoring agent of the present invention preferably contains 0.2% by mass or more and 45% by mass or less of mold fermentation products. More preferably, the lower limit of the mold fermentation products contained in the mold-based cheese flavoring agent is 0.3% by mass, 0.4% by mass, and even more preferably 0.5% by mass. Furthermore, the upper limit of the mold fermentation products contained in the mold-based cheese flavoring agent is more preferably 40% by mass, 35% by mass, and even more preferably 30% by mass. If the amount of mold fermentation products contained in the mold-based cheese flavoring agent is appropriate, the mold-based cheese flavoring agent can impart a good flavor to food and beverages that particularly gives a natural and authentic blue cheese flavor.

[0015] The mold-based cheese flavoring agent of the present invention contains mold fermentation products, but molds are not detected in PCR sequencing analysis, making analysis impossible. In this specification, "molds are not analyzable" means that, in PCR sequencing analysis, the amount of mold DNA contained in the PCR amplified product is below the detection limit because no molds are present. Below the detection limit means that the amount is below the minimum value at which molds can be detected when PCR sequencing analysis is performed using a PCR amplified product prepared with primers specific to the gene region of molds. Even with PCR sequencing equipment, molds cannot be detected, and analysis is impossible. Because molds are not detected and analysis is impossible in PCR sequencing analysis, mold cells are removed from the mold fermentation product. The method for removing mold cells from the mold fermentation product is not particularly limited, but examples include oil extraction methods such as a method of recovering the oil fraction after contacting the mold fermentation product with oils and fats, or a method of recovering the oil fraction contained in the mold fermentation product. Furthermore, examples include methods of removing mold cells by centrifuging the mold fermentation product and collecting only the supernatant, or by filtering the mold fermentation product with a filter to remove mold cells. Preferred methods include oil extraction or filtering the mold fermentation product with a filter to remove mold cells, and more preferably, oil extraction. By removing mold cells, it is possible to prepare a highly potent flavoring agent with a natural and authentic blue cheese flavor without suppressing the expression of the aroma and flavor characteristic of mold cheeses. On the other hand, if molds are detected in sequencing PCR analysis of the flavoring agent, it may contain a large amount of mold cells, which may suppress the expression of the aroma and flavor characteristic of blue cheeses, resulting in a decrease in the potency of the flavoring agent or the perception of an undesirable flavor derived from the mold cells.

[0016] In this specification, PCR sequencing analysis can be performed by extracting DNA from a sample, purifying the obtained DNA, and then sequencing the PCR amplified product. While there are no particular limitations on the method for extracting and purifying DNA from a sample, the FastDNA SPIN Kit for Soil (manufactured by MP Biomedicals) can be used. Following the kit's protocol, the reagents included in the kit are mixed with the mold-based cheese flavoring agent, and the DNA is purified by adsorbing it onto a silica matrix from tissue disrupted using a FastPrep Instrument. The purified DNA can then be PCR amplified using an Applied Biosystems SeqStudio Genetic Analyzer, followed by sequencing analysis. In PCR sequencing analysis, no molds are detected in the mold-based cheese flavoring agent of the present invention, making analysis impossible.

[0017] The oils and fats contained in the mold-based cheese flavoring agent of the present invention and the oils and fats used in the manufacturing process are not particularly limited, but examples include vegetable oils such as soybean oil, sunflower seed oil, high oleic sunflower oil, cottonseed oil, rapeseed oil, high erucic acid rapeseed oil, peanut oil, rice oil, rice bran oil, corn oil, olive oil, kapok oil, sesame oil, safflower oil, evening primrose oil, linseed oil, palm oil, palm kernel oil, coconut oil, shea butter, sal fat, etc., as well as animal oils such as milk fat, beef tallow, lard, fish oil, algae oil, oils and fats derived from microbial fermentation, medium-chain triglyceride (MCT), and processed oils and fats such as hydrogenated oils, fractionated oils, hydrogenated fractionated oils, fractionated hydrogenated oils, transesterified oils, etc., and mixed oils and fats thereof.

[0018] In one embodiment, the mold-based cheese flavoring agent of the present invention is particularly characterized by the amount of free glutamic acid contained in the solid content. By appropriately adjusting the amount of free glutamic acid contained in the solid content of the mold-based cheese flavoring agent, a good aged mold cheese flavor can be imparted. Preferably, the amount of free glutamic acid contained in the solid content is 0.3% by mass or more. More preferably, it is 0.35% by mass or more, 0.4% by mass or more, and even more preferably, 0.45% by mass or more, 0.5% by mass or more, or 0.55% by mass or more. By adjusting the amount of free glutamic acid contained in the solid content of the mold-based cheese flavoring agent to an appropriate range, a good aged mold cheese flavor can be imparted. Note that the solid content of the mold-based cheese flavoring agent refers to the amount of solid content excluding water contained in the mold-based cheese flavoring agent.

[0019] In this specification, the free glutamic acid content in mold-based cheese flavorings can be quantified using the post-columnar hydrin method. Specifically, various flavorings can be dispersed in a 5% by mass trichloroacetic acid solution so that the protein content is 0.4% by mass, the peptide components can be precipitated and removed, and the soluble components can be quantified using a Hitachi L-8500 high-speed amino acid analyzer.

[0020] In this specification, methyl ketones refer to 2-heptanone, 2-nonanone, and 2-undecanone. 2-heptanone is also known as amyl methyl ketone, n-amyl methyl ketone, heptan-2-one, methylpentyl ketone, pentyl methyl ketone, etc. 2-nonanone is also known as heptyl methyl ketone, nonan-2-one, methylheptyl ketone, etc. 2-undecanone is also known as nonyl methyl ketone, n-nonyl methyl ketone, undecane-2-one, methylnonyl ketone, etc. The methyl ketone content of the mold-based cheese flavoring agent of the present invention refers to the total content of 2-heptanone, 2-nonanone, and 2-undecanone contained in the mold-based cheese flavoring agent. In one embodiment, the mold-based cheese flavoring agent of the present invention is characterized by the amount of methyl ketones contained in the solid content. By appropriately adjusting the amount of methyl ketones contained in the solid content of the mold-type cheese flavoring agent, a good aged flavor of mold-type cheese can be imparted. The amount of methyl ketones contained in the solid content of the mold-type cheese flavoring agent of the present invention is preferably 0.05% by mass or more. More preferably, it is 0.06% by mass or more, 0.07% by mass or more, and even more preferably, 0.08% by mass or more, 0.09% by mass or more, and 0.1% by mass or more. By adjusting the amount of methyl ketones contained in the solid content of the mold-type cheese flavoring agent to an appropriate range, the aroma of mold-type cheese, especially the aroma characteristic of blue cheese, can be imparted.

[0021] The methyl ketone content in the mold-based cheese flavoring agent of the present invention is quantitatively analyzed by gas chromatography. Specifically, a sample pretreated according to a conventional method can be measured under the conditions shown below, and a calibration curve can be created from 2-heptanone, 2-nonanone, and 2-undecanone standard solutions (manufactured by Tokyo Chemical Industry Co., Ltd.) to quantify the content. Measurement conditions: Instrument: Shimadzu GC-2010Plus Detector: FID Column: UA+-1 (HT) (100% dimethyl polysiloxane) 15m, film thickness 0.15μm, 0.53mmφ Injection volume: 1μL (split 30:1) Column temperature: 35℃ (no hold) -4℃ / min -140℃ -25℃ / min -350℃ (15min hold) Injection port temperature: 350℃ Detector temperature: 375℃ Carrier gas: Helium

[0022] The mold-type cheese flavoring agent of the present invention has a mass ratio of 0.1 to 40, preferably 0.2 to 38, and more preferably 0.3 to 36, of which the amount of free glutamic acid relative to the methyl ketones contained (free glutamine / methyl ketones) is appropriate. If this mass ratio is appropriate, a mold-type cheese flavoring agent with a good mold-type cheese flavor can be prepared. In particular, a flavoring agent having a blue cheese flavor with a good balance of the characteristic aroma of mold-type cheese and the aged flavor can be prepared.

[0023] Typical and non-limiting production examples of free glutamic acid are given below, but those skilled in the art can appropriately design the production conditions for free glutamic acid. The free glutamic acid contained in the mold-based cheese flavoring agent of the present invention can be a seasoning containing sodium glutamate, a protein hydrolysate, an enzyme-treated product of a protein-containing food and / or a fermented product, etc. Among these, the use of a lactic acid bacteria ferment is preferred. In one embodiment, the mold-based cheese flavoring agent of the present invention contains a lactic acid bacteria ferment containing free glutamic acid. As an example of the production of the lactic acid bacteria ferment, a lactic acid bacteria ferment is produced by adding lactic acid bacteria to a mixture containing dairy products and / or other raw materials and fermenting it. Other raw materials can include, for example, a carbon source, a nitrogen source, minerals, vitamins, etc. as nutrients for the lactic acid bacteria. Also, for example, a molten salt can be added for the purpose of melting milk protein or cheese. Also, for example, starch, thickening polysaccharides, emulsifiers, egg products, etc. can be added for the purpose of improving the stability of the lactic acid bacteria ferment. Furthermore, for example, enzymes, edible lipid sources, yeast extracts, seasonings, protein hydrolysates, flavorings, etc., can be added to improve the flavor potency or flavor quality of the lactic acid bacteria fermented product. The fermentation method of the lactic acid bacteria fermented product is not particularly limited, but the fermentation temperature is preferably 8°C to 50°C, more preferably 15°C to 47°C. The fermentation period is 1 day to 40 days, more preferably 1 day to 30 days, and more preferably 1 day to 20 days. It is also preferable that the fermentation be carried out under stirring conditions. During the lactic acid bacteria fermentation process, the pH can be adjusted by adding a pH adjuster as appropriate to 4.5 to 6.5. The lactic acid bacteria fermented product can be concentrated as needed to improve its flavor potency. It can also be heat-sterilized as needed.

[0024] A typical and non-limiting example of the production of methyl ketones is given below, but those skilled in the art can appropriately design the production conditions for methyl ketones. Oils and fats containing saturated or unsaturated fatty acids with 4 to 14 carbon atoms as constituent fatty acids are added to a liquid culture medium, a culture of molds is inoculated, and fermentation is carried out for 6 to 144 hours. During the fermentation process, the pH can be adjusted by adding an alkali such as sodium hydroxide. After the fermentation process is completed, the oil phase in the fermentation liquid can be separated by a centrifuge or the like to extract the oils and fats. The fermentation liquid may be concentrated or diluted as needed to adjust the concentration of methyl ketones to a predetermined concentration. The oils and fats are not particularly limited, but it is preferable that they include one or more oils and fats selected from the group consisting of palm kernel oil, coconut oil, and medium-chain fatty acid triglycerides (MCTs), as well as hydrogenated oils, fractionated oils, hydrogenated fractionated oils, fractionated hydrogenated oils, processed oils and fats that have undergone transesterification, and mixed oils and fats thereof. More preferably, the product includes one or more oils and fats selected from the group consisting of palm kernel oil and coconut oil, as well as processed oils and fats obtained by hydrogenating, fractionating, hydrogenating fractionating, fractionating hydrogenating, transesterification, etc., and further mixed oils and fats thereof. Even more preferably, it includes one or more oils and fats selected from the group consisting of palm kernel oil and coconut oil.

[0025] In one embodiment, the mold-based cheese flavoring agent of the present invention is characterized by the total amount of free fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms contained in the solid content. By appropriately adjusting the amount of free fatty acids contained in the solid content of the mold-based cheese flavoring agent, a good aged flavor can be imparted. In this specification, fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms refer to caproic acid, caprylic acid, and capric acid, respectively. Caproic acid is also called n-caproic acid, hexanoic acid, n-hexanoic acid, hexylic acid, hexonic acid, n-hexanoic acid, etc. Caprylic acid is also called n-caprylic acid, octanoic acid, n-octanoic acid, octicic acid, etc. Capric acid is also called n-capric acid, decanoic acid, n-decanoic acid, etc. The free fatty acid content of the mold-based cheese flavoring agent of the present invention refers to the total content contained in the mold-based cheese flavoring agent. The total amount of free fatty acids with 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms contained in the solid content is preferably 0.4% by mass or more. More preferably 0.5% by mass or more, 0.6% by mass or more, and even more preferably 0.7% by mass or more, 0.8% by mass or more, and 0.9% by mass or more. Furthermore, the amount of free fatty acids contained in the solid content is preferably 50% by mass or less. More preferably 48% by mass or less, 46% by mass or less, and even more preferably 44% by mass or less, 42% by mass or less, and 40% by mass or less. By adjusting the total amount of free fatty acids with 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms contained in the solid content of the mold-type cheese flavoring agent to an appropriate range, a more natural and authentic blue cheese flavor can be imparted.

[0026] The total amount of free fatty acids with 6, 8, and 10 carbon atoms contained in the mold-based cheese flavoring agent of the present invention can be quantitatively analyzed by derivatizing the sample with nitrophenylhydrazine and using a high-performance liquid chromatograph mass spectrometer. More specifically, by combining a pretreatment method including a derivatization step with 3-nitrophenylhydrazine (3-NPH) (LC / MS / MS Method Package Short-Chain Fatty Acids (Shimadzu Corporation)) with a high-performance liquid chromatograph mass spectrometer (LCMS-8050, Shimadzu Corporation), the amount of free fatty acids with 6, 8, and 10 carbon atoms can be measured, and a calibration curve can be created from standard solutions of caproic acid, caprylic acid, and capric acid (Tokyo Chemical Industries, Ltd.) to quantify the content. Detailed measurement conditions using a high-performance liquid chromatograph mass spectrometer are shown below. Column: Inertsil™ ODS-4HP (3μm 2.1×150mm *2 columns linked) Mobile phase: A CO2, B 0.1% (w / v) ammonium acetate in methanol Gradient: B conc. 2% (0 min) - 30% (3 min) - 30% (3-5 min) - 2% (5.1-8 min) Flow rate: 1.0 mL / min Column temperature: 40℃ BPR: 10 MPa Detector: LCMS-8050 (ESI, SIM mode, MRM mode) Makeup solvent: 0.1% (w / v) ammonium acetate in methanol Makeup solvent flow rate: 0.05 mL / min Injection volume: 1 μL

[0027] Typical and non-limiting production examples of free fatty acids having 6, 8, and 10 carbon atoms that can be used in the mold-based cheese flavoring agent of the present invention are given below, but those skilled in the art can appropriately design the production conditions for the free fatty acids. Lipase is added to a composition containing oil and water and an enzymatic reaction is carried out. The method of the enzymatic reaction is not particularly limited, but the reaction temperature is preferably 8°C to 70°C, more preferably 15°C to 65°C, the reaction period is 1 to 48 hours, more preferably 2 to 36 hours, and even more preferably 3 to 24 hours, and the reaction is preferably carried out under stirring conditions. During the enzymatic reaction, the pH can be adjusted by adding a pH adjuster as appropriate to 4 to 7. The enzymatic reaction product can be concentrated as needed to improve its flavor potency. Furthermore, after the enzymatic reaction, it is preferably heat-treated at a temperature of 75°C or higher. The aforementioned oils and fats are not particularly limited, but preferably include one or more oils and fats selected from the group consisting of palm kernel oil, coconut oil, medium-chain triglycerides (MCT), and milk fat, as well as processed oils and fats that have undergone hydrogenation, fractionation, hydrogenated fractionation, fractionated hydrogenated oil, transesterification, etc., and further mixed oils and fats thereof. More preferably, it includes one or more oils and fats selected from the group consisting of palm kernel oil, medium-chain triglycerides (MCT), and milk fat, as well as processed oils and fats that have undergone hydrogenation, fractionation, hydrogenated fractionation, fractionated hydrogenated oil, transesterification, etc., and further mixed oils and fats thereof. Even more preferably, it is one or more oils and fats selected from the group consisting of palm kernel oil, medium-chain triglycerides (MCT), and milk fat.

[0028] In this specification, "cheese-like food" refers to food and beverages such as natural cheese, processed cheese made primarily from natural cheese, and processed products prepared by combining oils and fats, proteins, starches, etc. Furthermore, analog cheeses that have been prepared to have a cheese-like flavor without incorporating natural or processed cheese, and processed products prepared by combining such analog cheeses with oils and fats, etc., are also included in "cheese-like food." The mold-based cheese flavoring agent of the present invention can be used by incorporating it into cheese-like food. The resulting cheese-like food will have the characteristic aroma and aged flavor of blue cheese. Therefore, this aroma and flavor will allow consumers to experience a flavor particularly characteristic of blue cheese among natural cheeses. One embodiment of the present invention, the cheese-like food, can be used in combination with other food and beverages as a cheese filling, cheese sauce, cheese food, processed cheese, shredded cheese, powdered cheese, block cheese, diced cheese, cheese whip, cheese cream, etc. A composite food formed by combining this cheese-like food with other food and beverages will have a more natural and authentic blue cheese flavor.

[0029] The mold-based cheese flavoring agent of the present invention can be appropriately incorporated into various foods and beverages. The amount incorporated can be appropriately adjusted depending on the desired intensity of the mold-based cheese flavor to be imparted to the food or beverage. Furthermore, cheese-like foods containing the mold-based cheese flavoring agent can be incorporated into various foods and beverages, or a composite food having a mold-based cheese flavor can be prepared by combining such cheese-like foods with food or beverages.

[0030] In other words, the present invention can also be described as a method for imparting a moldy cheese flavor to food and beverages using a composition that contains a moldy fermentation product, in which the molds are unanalyzable in PCR sequencing analysis, and in which the mass ratio of free glutamic acid to methyl ketones, represented by free glutamic acid / methyl ketones, is 0.1 or more and 40 or less.

[0031] The mold-based cheese flavoring agent of the present invention is substantially mold-free, as PCR sequencing analysis reveals that molds are unanalyzable. Therefore, compared to incorporating cheese-like foods containing molds into food and beverages, it is expected that the deterioration of food shelf life due to molds can be suppressed. The use of the mold-based cheese flavoring agent of the present invention also suggests a new industrial benefit: the reduction of food waste.

[0032] Embodiments of the present invention will be described in more detail below with reference to examples and other relevant information. Hereinafter, "%" and "parts" refer to "mass percent" and "parts by mass," respectively, unless otherwise specified.

[0033] ● Preparation of mold fermentation products: Refined palm kernel oil (manufactured by Fuji Oil Co., Ltd.), soybean peptone (manufactured by Fuji Oil Co., Ltd.), glucose, and water were mixed and heat-sterilized at 121°C for 20 minutes. After the heat-sterilized mixture cooled, the mold Aspergillus oryzae was inoculated into the mixture, and aerobic fermentation was carried out at 30°C for 3 to 5 days. After the fermentation process was completed, the fermentation liquid was heat-sterilized at 90°C for 20 minutes, and the fermentation liquid was concentrated as appropriate to prepare mold fermentation products. Samples obtained by filtering the mold fermentation product or by centrifuging the mold fermentation product and recovering the oil phase were prepared, and the microbial cells were removed from each sample. These were designated as mold fermentation product A and mold fermentation product B, respectively.

[0034] ●Preparation of Lactic Acid Bacteria Fermented Product 50 parts by mass of mozzarella cheese, 1 part by mass of sodium polyphosphate, and 49 parts by mass of water were mixed and heat-sterilized at 95°C for 5 minutes, then cooled. To the cooled mixture, 0.01 parts by mass of protease derived from Aspergillus oryzae, and Streptococcus thermophilus, Lactococcus lactis ssp. lactis, Lactobacillus brevis, and Lactobacillus helveticus were added, and fermentation was carried out at 30°C for 10 days with stirring. During the fermentation process, sodium hydroxide solution or hydrochloric acid solution was added as needed to adjust the pH to a range of 4.5 to 6.5. After the completion of the fermentation process, the fermentation liquid was heat-sterilized at 80°C for 1 minute to prepare the lactic acid bacteria fermented product.

[0035] ●MCT Enzyme-treated product: 20 parts by mass of medium-chain fatty acid-bound triglycerides (MCT), 10 parts by mass of skim milk powder, 70 parts by mass of water, and 0.1 parts by mass of lipase derived from Candida cylindracea were mixed, and an enzymatic reaction was carried out at 50°C for 5 hours with stirring. After the enzymatic reaction step, the enzyme reaction solution was heat-sterilized at 90°C for 20 minutes to prepare the MCT enzyme-treated product.

[0036] ● Preparation of mold-type cheese flavorings: Various mold-type cheese flavorings were prepared by mixing the raw materials according to the formulations described in Table 1 or Table 2 and then heat-sterilizing them at 80°C for 1 minute. For comparison with the mold-type cheese flavorings, a blue cheese-like cheese sauce was also prepared with added flavorings containing monosodium glutamate and methyl ketones.

[0037] ●PCR Sequencing Analysis: DNA was extracted and purified from samples of mold-based cheese flavoring agents prepared according to the formulations described in Table 1 or Table 2. The FastDNA SPIN Kit for Soil (MP Biomedicals) was used for DNA extraction and purification. The purified DNA was amplified using an Applied Biosystems SeqStudio Genetic Analyzer, and sequencing analysis of the fungi was performed.

[0038] ●Preparation of Cheese Sauce Base and Blue Cheese-like Cheese Sauce ・Cheese Sauce Base A cheese sauce base was prepared by mixing 40 parts by mass of cottage cheese, 34 parts by mass of sour cream, 15 parts by mass of mayonnaise, 5 parts by mass of milk, and 1 part by mass of lemon juice. ・Blue Cheese-like Cheese Sauce A blue cheese-like cheese sauce was prepared by mixing 5 parts by mass of various mold-based cheese flavoring agents into the cheese sauce base. The blue cheese-like cheese sauce with added mold-based cheese flavoring agents was compared in flavor with the cheese sauce base without added mold-based cheese flavoring agents.

[0039] ● Sensory Evaluation The criteria for sensory evaluation were set at 5 points each for the blue cheese-like aroma, aged flavor, and overall blue cheese flavor of the blue cheese-like cheese sauce prepared by mixing 30 parts by mass of blue cheese into the cheese sauce base. Similarly, the blue cheese-like aroma, aged flavor, and overall blue cheese flavor of the blue cheese-like cheese sauce prepared by mixing 5 parts by mass of blue cheese were each set at 2 points. Furthermore, the sensory evaluation in the examples referred to the above criteria, and comparative evaluations were conducted for each food and beverage with and without the addition of various mold-type cheese flavoring agents. The evaluation items were confirmed for the blue cheese-like aroma and aged flavor, and the effect of imparting blue cheese flavor was evaluated. Specifically, a score of 1 was given to food and beverages with various mold-type cheese flavoring agents added compared to food and beverages without mold-type cheese flavoring agents, indicating no effect whatsoever on imparting blue cheese-like aroma or aged flavor, and a score of 5 was used, with higher scores indicating a greater effect. Sensory evaluation was conducted by 10 trained panelists, using a 5-point scale from 1 to 5, and the average score was calculated. Furthermore, the overall blue cheese-like sauce was evaluated to determine whether it had a natural and authentic blue cheese flavor. The overall evaluation was based on the blue cheese flavor of the entire blue cheese sauce. The scores were recorded as follows: 1 point for an average of 1.0-1.4, 2 points for 1.5-2.4, 3 points for 2.5-3.4, 4 points for 3.5-4.4, and 5 points for 4.5-5.0, with a passing score of 3 or higher. Preferably, a passing score of 4 or higher was used. The results of the PCR sequencing analysis, sensory evaluation, and various component values ​​of the mold-type cheese flavoring agent are shown in Tables 1 and 2.

[0040]

[0041]

[0042] The blue cheese-like cheese sauce using the mold-based cheese flavoring agent in the example exhibited a strong blue cheese-like aroma and mature flavor, and the entire cheese sauce had a natural and authentic blue cheese flavor.

[0043] From the above, it has been clarified that any mold-type cheese flavoring agent satisfying all of the following requirements 1) to 3) can impart a favorable mold-type cheese-like flavor to foods and drinks. 1) comprising a fermentation product of molds 2) the molds are undetectable in PCR sequence analysis 3) the mass ratio of the amount of free glutamic acid to the methyl ketones contained therein, represented by free glutamic acid / methyl ketones, is 0.1 or more and 40 or less

[0044] According to the present invention, there can be provided a flavoring agent capable of imparting a favorable mold-type cheese-like flavor to foods and drinks, and there can be provided foods and drinks using said flavoring agent.

Claims

1. A mold-based cheese flavoring agent that satisfies all of the following requirements 1) to 3): 1) Contains mold fermentation products; 2) Molds are undetectable in PCR sequencing analysis; 3) The mass ratio of free glutamate to methyl ketones, as expressed by free glutamate / methyl ketones, is 0.1 or higher and 40 or lower. However, "molds are undetectable" means that the amount of mold DNA contained in the PCR amplified product is below the detection limit because no molds are included.

2. The mold-type cheese flavoring agent according to claim 1, having a blue cheese flavor.

3. A mold-based cheese flavoring agent according to claim 1 or 2, comprising 0.3% by mass or more of free glutamic acid and 0.05% by mass or more of methyl ketones based on solid content.

4. A mold-based cheese flavoring agent according to claim 1 or 2, further satisfying the following 4): 4) Contains 0.4% by mass or more and 50% by mass or less of free fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms based on solid content.

5. The mold-based cheese flavoring agent according to claim 3, further satisfying the following 4): 4) Contains 0.4% by mass or more and 50% by mass or less of free fatty acids having 6 carbon atoms, 8 carbon atoms, and 10 carbon atoms based on solid content.

6. A cheese-like food comprising the mold-based cheese flavoring agent described in claim 1 or 2.

7. A cheese-like food containing the mold-based cheese flavoring agent described in claim 3.

8. A cheese-like food comprising the mold-based cheese flavoring agent described in claim 4.

9. A cheese-like food comprising the mold-based cheese flavoring agent described in claim 5.

10. Food and beverage containing the cheese-like food described in claim 6.

11. Food and beverages comprising the cheese-like food described in claim 7.

12. Food and beverage containing the cheese-like food described in claim 8.

13. Food and beverage containing the cheese-like food described in claim 9.

14. A method for producing a mold-based cheese flavoring agent, comprising the following steps (A) and (B): (A) A step of extracting a mold fermentation product to obtain a mold fermentation product. (B) A step of mixing the mold fermentation product obtained in step (A) with a lactic acid bacteria fermentation product and / or a lipase enzyme reaction product of oil and fat.

15. A method for imparting moldy cheese flavor to food and beverages by incorporating a composition that satisfies all of the following requirements 1) to 3): 1) Contains mold fermentation products; 2) Molds are undetectable in PCR sequencing analysis; 3) The mass ratio of free glutamic acid to methyl ketones, as expressed by free glutamic acid / methyl ketones, is 0.1 or higher and 40 or lower. However, "molds are undetectable" means that the amount of mold DNA contained in the PCR amplified product is below the detection limit because no molds are present.

Images