Enzymatic agent, processed coffee-containing beverage, and use thereof

Abstract

The present invention addresses the problem of improving an effect for suppressing the coagulation that occurs when coffee is mixed with an animal-based milk or a plant-based milk. The present invention relates to an enzymatic agent that contains an oxidase and that is for suppressing coagulation that occurs when coffee and milk are mixed. The present invention also relates to: a processed coffee obtained by causing a reaction between an enzymatic agent and coffee; a processed coffee-containing beverage that contains milk and the processed coffee obtained by causing a reaction between an enzymatic agent and coffee; and methods for producing the processed coffee and for producing the processed coffee-containing beverage.

Description

Enzyme preparations, processed coffee-containing beverages and their applications

[0001] The present invention relates to an enzyme preparation, processed coffee, a processed coffee-containing beverage, a method for producing processed coffee, a method for producing a processed coffee-containing beverage, and a method for inhibiting aggregation.

[0002] Beverages rich in nutrients such as protein have long been popular because they provide a convenient way to ingest nutrients. In recent years, in addition to animal-based milks such as cow's milk, plant-based milks made from plant materials have also become popular. Known plant-based milks include those made from soybeans, oats, almonds, and other plant-based materials.

[0003] It is known that key components of coffee bind to the proteomes of animal and plant-based milks (Non-Patent Document 1). Therefore, animal and plant-based milks may aggregate in heat and / or low pH environments. For example, aggregation occurs when animal and plant-based milks are added to hot, acidic beverages such as coffee and tea.

[0004] In order to suppress aggregation when coffee is mixed with animal milk or plant milk, it has been investigated to treat the milk raw material with an enzyme. For example, Patent Literature 1 discloses a method of treating plant milk such as pea, oat, hemp, almond, peanut, cashew nut, pistachio, or hazelnut with a protein deamidating enzyme, which shows that this method has the effect of suppressing aggregation when coffee is mixed with the milk.

[0005] International Publication No. 2020 / 171106

[0006] Identify the interactions between phytochemicals and proteins in the complicated food matrix. Food chemistry, 356, 129641.

[0007] However, conventional techniques have not been effective in suppressing aggregation when coffee is mixed with animal milk or plant milk, and further improvements are needed.

[0008] Therefore, an object of the present invention is to improve the aggregation suppression effect when coffee is mixed with animal milk or plant milk.

[0009] Examples of specific embodiments of the present invention are given below.

[0010] [1] An enzymatic preparation containing an oxidase for inhibiting aggregation when coffee and milk are mixed. [2] The enzymatic preparation according to [1], wherein the oxidase is a copper oxidase. [3] The enzymatic preparation according to [1] or [2], wherein the oxidase is a multicopper oxidase. [4] The enzymatic preparation according to any one of [1] to [3], wherein the oxidase is a laccase. [5] The enzymatic preparation according to any one of [1] to [4], wherein the oxidase is derived from the genus Trametes. [6] Processed coffee obtained by allowing the enzymatic preparation according to any one of [1] to [5] to act on coffee. [7] A processed coffee-containing beverage containing processed coffee obtained by allowing the enzymatic preparation according to any one of [1] to [5] to act on coffee, and milk. [8] The processed coffee-containing beverage according to [7], wherein the milk is plant-based milk. [9] The processed coffee-containing beverage according to [8], wherein the vegetable milk raw material is at least one selected from the group consisting of soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, kidney beans, wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, potatoes, almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia seeds, amaranth, canary seeds, linseed, and algae.

[10] A method for producing processed coffee, comprising allowing the enzyme preparation according to any one of [1] to [5] to act on coffee.

[11] A method for producing a processed coffee-containing beverage, comprising allowing the enzyme preparation according to any one of [1] to [5] to act on coffee, and then mixing with milk.

[12] The method for producing a processed coffee-containing beverage according to

[11] , wherein the milk is plant-based milk.

[13] The method for producing a processed coffee-containing beverage according to

[12] , wherein the raw material for the plant-based milk is at least one selected from the group consisting of soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, kidney beans, wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, potatoes, almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia, amaranth, canary seeds, linseed, and algae.

[14] A method for suppressing coagulation when coffee and milk are mixed, comprising applying the enzyme preparation according to any one of [1] to [5] to coffee.

[0011] [A] Use of an enzymatic preparation containing an oxidase to suppress aggregation when coffee and milk are mixed. [B] Use of an enzymatic preparation containing the oxidase according to [A], wherein the oxidase is a copper oxidase. [C] Use of an enzymatic preparation containing the oxidase according to [A] or [B], wherein the oxidase is a multicopper oxidase. [D] Use of an enzymatic preparation containing the oxidase according to any of [A] to [C], wherein the oxidase is a laccase. [E] Use of an enzymatic preparation containing the oxidase according to any of [A] to [D], wherein the oxidase is derived from the genus Trametes.

[0012] According to the present invention, it is possible to improve the aggregation suppression effect when coffee is mixed with animal milk or plant milk. Furthermore, according to the present invention, a coffee-containing beverage in which the formation of aggregates is suppressed is provided.

[0013] The present invention will be described in detail below. The following description may be based on representative embodiments or specific examples, but the present invention is not limited to such embodiments. In this specification, a numerical range expressed using "to" means a range that includes the numerical values ​​before and after "to" as the lower and upper limits.

[0014] (Enzyme Preparation) This embodiment relates to an enzyme preparation containing oxidase that suppresses aggregation when coffee and milk are mixed. In this embodiment, by allowing oxidase, an oxidizing enzyme, to act on coffee, aggregation that occurs when coffee and milk are mixed can be suppressed. This provides a coffee-containing beverage in which the generation of aggregates is suppressed.

[0015] In this specification, milk includes both animal-based milk and plant-based milk. In particular, since plant-based milk tends to cause aggregation, the milk is preferably plant-based milk. In other words, this embodiment may relate to an enzyme agent for suppressing aggregation when coffee and plant-based milk are mixed.

[0016] The enzyme preparation may be any preparation containing an oxidase, or may consist of an oxidase. The enzyme preparation may be in any form, such as powder, solid, gel, or liquid.

[0017] <Oxidase> Oxidase is an enzyme that electron-reduces oxygen to produce water molecules. Examples of oxidases include glutathione peroxidase, ascorbate peroxidase, heme peroxidase, lactoperoxidase, myeloperoxidase, horseradish peroxidase, copper oxidase, iron oxidase, lignin peroxidase, manganese peroxidase, and P450. Among these, copper oxidase is preferred.

[0018] The copper oxidase may be a copper-containing monooxidase or a copper-containing multicopper oxidase (hereinafter also referred to as a multicopper oxidase). Examples of copper-containing monooxidases include dopamine-β-monooxygenase, peptidylglycine α-hydroxylating monooxygenase, tyrosinase, and methane monooxygenase.

[0019] The copper oxidase is preferably a multicopper oxidase. A multicopper oxidase is an enzyme containing 2 to 8 copper atoms in the molecule, which are necessary for enzymatic activity, and which generates water molecules by electron reduction of oxygen. The multicopper oxidase is preferably at least one selected from phenol oxidase and bilirubin oxidase, more preferably phenol oxidase, and even more preferably polyphenol oxidase. Examples of multicopper oxidases include laccase, bilirubin oxidase, ascorbic acid oxidase, ceruloplasmin, Fet3p, CueO, CotA, stellacyanin, tyrosinase, catechol oxidase, and nitrite reductase. Among these, the multicopper oxidase is preferably at least one selected from the group consisting of laccase, bilirubin oxidase, ascorbic acid oxidase, and tyrosinase, and particularly preferably laccase.

[0020] Laccase is an enzyme with phenol oxidase activity (EC 1.10.3.2) and can be used in any application that utilizes the oxidation of a substrate and / or various accompanying chemical reactions resulting from the radical species of the reaction intermediates generated by the oxidation. Examples of such oxidation reactions or accompanying chemical reactions include the oxidation of phenolic compounds such as o- and p-diphenols, urushiol, and laccol; the oxidation of aromatic amines such as p-phenylenediamine; the degradation of lignin; and the cross-linking of proteins with easily oxidizable functional groups such as tyrosine side chains (phenolic hydroxyl groups), cysteine ​​side chains (sulfhydryl groups), lysine side chains (ε-amino groups), and histidine side chains (imidazole groups). Chemically altering a substrate through the oxidation reaction and accompanying chemical reactions of laccase can also be referred to as "oxidative modification."

[0021] Laccases are found in plants, fungi, bacteria, animals, etc. Among these, the laccase used in the present invention is preferably derived from fungi or bacteria. Specific examples include laccases derived from the genera Aspergillus, Neurospora, Podospora, Botrytis, Collybia, Formes, Lentinus, Pleurotus, Pycnoporus, Pyricularia, Trametes, Rhizoctonia, Rigidoporus, Coprinus, Psatyrella, Myceliophtera, Schtalidium, Polyporus, Phlebia, Coriolus, Bacillus, etc. Among these, laccases derived from the genus Trametes are particularly preferred. By using laccase derived from the genus Trametes, the effect of inhibiting aggregation when coffee and milk are mixed can be further improved.

[0022] Laccase can be prepared from the culture medium of the microorganism from which the laccase is derived. Specific preparation methods include recovering laccase from the culture medium or cells of the microorganism. For example, when a laccase-secreting microorganism is used, the cells are recovered from the culture medium, if necessary, by filtration, centrifugation, or the like, and the enzyme is then isolated and / or purified. When a laccase-nonsecreting microorganism is used, the cells are recovered from the culture medium, if necessary, and then disrupted by pressure treatment, ultrasonic treatment, or the like to expose the enzyme, after which the enzyme is isolated and / or purified. The enzyme isolation and / or purification method is not particularly limited, and known protein isolation and / or purification methods can be employed, including centrifugation, UF concentration, salting out, and various chromatography methods using ion exchange resins. The isolated and / or purified enzyme can be powdered by drying methods such as freeze-drying and vacuum drying, or by using appropriate excipients and / or drying aids in the drying methods. The isolated and / or purified enzyme can also be liquefied by adding an appropriate additive and sterilizing by filtration.

[0023] Commercially available laccases can also be used, and a preferred example of a commercially available laccase is laccase derived from Trametes sp., manufactured by Amano Enzyme Inc.

[0024] Bilirubin oxidase is an enzyme having bilirubin oxidase activity (EC 1.3.3.5). Bilirubin oxidase derived from microorganisms belonging to the genera Myrothecium, Coprinus, Penicillium, and Bacillus can be used, for example. Examples of Myrothecium bacteria include preserved strains such as Myrothecium verrucaria MT-1 and FERM-BP 653 (see Agricultural and Biological Chemistry, Vol. 45, pp. 2383-2384 (1981)), Myrothecium verrucaria IFO 6113, Myrothecium verrucaria IFO 6133, Myrothecium verrucaria IFO 6351, Myrothecium verrucaria IFO 9056, Myrothecium tinctum IFO 9950, and Myrothecium verrucaria IFO 9531. Examples of Coprinus bacteria include preserved strains such as Coprinus cinereus IFO 8371 and Coprinus lagopides IFO 30120. Examples of Penicillium bacteria include Penicillium jansinerum (see JP-A-63-309187). Examples of Bacillus bacteria include Bacillus licheniformis (see JP-A-61-209587). These strains are cultured in liquid or solid form by conventional methods, and the culture solution is subjected to extraction, salting out, dialysis, ion exchange, gel filtration, and the like to obtain purified preparations of the enzyme bilirubin oxidase. Meanwhile, bilirubin oxidases derived from Schizophyllum commune (see JP-A-59-135886), Asteraceae plants (see JP-A-62-285782), and alfalfa (see JP-A-6-319536), as well as recombinant forms (see JP-A-5-199882), are also known, and these bilirubin oxidases can also be used.

[0025] Commercially available bilirubin oxidases can also be used, and preferred examples of commercially available products include those available from Amano Enzyme Inc., Takara Bio Inc., Asahi Kasei Corporation, Sigma-Aldrich Corporation, etc.

[0026] When an enzyme preparation is allowed to act on coffee, the amount of oxidase to be applied is not particularly limited, but it is preferable to add it so that the enzymatic activity per 1 g of coffee is 1 U or more. From the viewpoint of further enhancing the aggregation-inhibiting effect when coffee is mixed with milk, it is preferable to add oxidase so that the enzymatic activity per 1 g of coffee is preferably 10 U or more, more preferably 50 U or more, even more preferably 100 U or more, even more preferably 200 U or more, even more preferably 400 U or more, even more preferably 600 U or more, and particularly preferably 800 U or more. The upper limit of the amount of oxidase to be added is not particularly limited, but it is preferable to add it so that the enzymatic activity per 1 g of coffee is preferably 80,000 U or less, more preferably 60,000 U or less, even more preferably 40,000 U or less, even more preferably 20,000 U or less, even more preferably 10,000 U or less, and even more preferably 9,000 U or less.

[0027] Among these, the oxidase is preferably a multicopper oxidase, and the amount of the multicopper oxidase to be reacted is preferably within the above range.

[0028] The enzymatic activity of multicopper oxidase is measured as follows. First, a substrate solution is prepared by mixing 0.25 mol / L phenol test solution, 0.009 mol / L 4-aminoantipyrine test solution, and 1 mol / L acetic acid / sodium acetate buffer (pH 4.5) in a volume ratio of 1:1:0.5. 2.5 ml of this substrate solution is placed in a cuvette and preheated to 30°C. 0.5 ml of enzyme solution is added, stirred, and incubated at 30°C. The absorbance at 505 nm is measured after 10 and 40 seconds. The amount of enzyme contained in 1 mL of reaction solution is defined as 1 unit (U) when the absorbance increases by 0.1 per minute.

[0029] <Coffee> The coffee is liquid or semi-liquid coffee. The coffee is preferably a coffee extract, and examples thereof include an extract obtained by extracting roasted coffee beans or their crushed material with water (including hot water or steam), and a concentrate obtained by concentrating the extract. Note that the coffee may also be instant coffee obtained by drying the extract or concentrate and dissolving it in water (including hot water or steam).

[0030] The pH of coffee is, for example, 3.0 to 7.0, preferably 3.5 to 6.5, more preferably 4.0 to 6.0, and even more preferably 4.2 to 5.0.

[0031] In this embodiment, liquid foods and beverages other than coffee (drinks, liquid foods) can also be used. Examples of liquid foods and beverages other than coffee include tea (black tea, green tea, oolong tea, etc.), tea drinks (flavored tea, milk tea, tea drinks containing fruit juice, etc.), fruit juice, fruit juice drinks, sports drinks, nutritional supplement drinks (protein drinks, nutritional drinks for care, etc.), soups (bouillon soups, stews, chowder, borscht, vegetable soups (e.g., tomato soup, corn soup, potage, pumpkin soup), miso soup, curry, cocoa, chocolate drinks, etc. Among these, it is preferable that the liquid foods and beverages other than coffee are polyphenol-containing beverages. Note that liquid foods and beverages include those obtained by processing (e.g., concentrating, freeze-drying) extracts and then reconstituting them. These liquid foods and beverages can be used in combination with coffee or can be used in place of coffee.

[0032] <Milk> In this specification, milk is a general term for animal milk and plant milk. Animal milk is preferably mammalian milk, and examples include cow's milk, goat's milk, sheep's milk, and horse's milk. Plant milk is preferably a liquid obtained by extracting components containing protein and fat from plant-derived raw materials such as beans, nuts, and grains with water. The raw material for the plant-based milk is preferably at least one selected from the group consisting of pulses such as soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, and kidney beans; cereals such as wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, and potatoes; nuts and seeds such as almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia, amaranth, canary seeds, and linseed; and algae.

[0033] The pH of the milk is, for example, 5.0 to 8.0, preferably 5.5 to 7.5, more preferably 5.5 to 7.0, and even more preferably 6.8 to 7.0.

[0034] The protein concentration in milk is preferably 0.01 to 10.0% by mass, more preferably 0.1 to 7.0% by mass, and even more preferably 0.3 to 6.0% by mass.

[0035] (Processed Coffee) This embodiment relates to processed coffee in which the above-described enzyme agent is applied to coffee. In this specification, coffee refers to liquid or semi-liquid coffee. For example, coffee includes an extract obtained by extracting coffee ingredients (green coffee beans, roasted coffee beans, or ground coffee beans) with water (including hot water or steam), and a concentrate obtained by concentrating the extract. The concentrate may also be a semi-liquid substance such as a paste obtained by concentrating the coffee extract. Furthermore, coffee may be instant coffee obtained by dissolving a dried product (such as instant coffee powder) obtained by drying the above-described extract or concentrate in water (including hot water or steam). Furthermore, coffee also includes a slurry in which coffee ingredients are dispersed and a solution in which the coffee ingredients have settled.

[0036] Processed coffee is obtained by treating coffee with an enzyme, and contains enzyme-treated products of some components of coffee.

[0037] In this specification, processed coffee also includes a slurry in which coffee ingredients are dispersed, a solution in which coffee ingredients have settled, and a solid form of dried processed coffee.

[0038] (Processed Coffee-Containing Beverage) This embodiment relates to a processed coffee-containing beverage containing processed coffee obtained by allowing the enzyme agent described above to act on coffee, and milk. Because the processed coffee-containing beverage contains processed coffee and milk, it is sometimes called coffee milk.

[0039] Examples of milk include the milks mentioned above. Among them, the milk is preferably plant-based milk, and the raw material of the plant-based milk is preferably obtained from at least one selected from the group consisting of pulses such as soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupine beans, and kidney beans; cereals such as wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, and potato; nuts and seeds such as almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia seeds, amaranth, canary seeds, and linseed; and algae.

[0040] The mixing ratio of processed coffee to milk in the processed coffee-containing beverage is, for example, preferably 1:9 to 30:1, more preferably 1:9 to 9:1, and even more preferably 2:8 to 8:2.

[0041] <Optional Ingredients> The processed coffee-containing beverage may contain optional ingredients, such as flavorings, sweeteners, acidulants, bittering agents, coloring agents, antioxidants, pH adjusters, vitamins, amino acids, minerals, and antifoaming agents.

[0042] When a flavoring is contained in the processed coffee-containing beverage, examples of the flavoring include black tea flavor, bergamot flavor, matcha flavor, chocolate flavor, white chocolate flavor, vanilla flavor, caramel flavor, cocoa flavor, coffee flavor, strawberry flavor, whiskey flavor, etc. Of these, the flavoring is preferably coffee flavor.

[0043] Furthermore, because the occurrence of aggregation is suppressed in the processed coffee-containing beverage of this embodiment, it may be substantially free of, for example, emulsifiers (glycerin fatty acid esters, sucrose fatty acid esters, lecithin, saponin, etc.), thickening polysaccharides (pectin, carboxymethylcellulose, etc.), and salts (table salt, calcium salts, phosphates, etc.). In this specification, "substantially free" means that the amount of these added is 0.1% by mass or less relative to the total mass of the processed coffee-containing beverage.

[0044] (Method for Producing Processed Coffee) This embodiment relates to a method for producing processed coffee, which includes allowing the enzyme agent described above to act on coffee.

[0045] The reaction time, temperature, and pH of the reaction solution for allowing the enzyme agent to act on coffee are not particularly limited. The reaction temperature is, for example, 1 to 80°C, preferably 5 to 70°C, and more preferably 40 to 60°C. The pH of the reaction solution is, for example, 3 to 8, preferably 3.5 to 7, and more preferably 4 to 6. The reaction time is, for example, 30 seconds to 72 hours, preferably 1 minute to 48 hours, and more preferably 30 minutes to 24 hours. The above reaction conditions make it easier to obtain processed coffee that can exhibit an aggregation-inhibiting effect when mixed with milk. These reaction conditions are selected appropriately depending on the type of coffee used, etc. The optimal reaction conditions can be determined through preliminary experiments.

[0046] One aspect of the method for producing processed coffee according to the present embodiment includes the following steps (1) and (2). Note that an enzyme deactivation step may be added after step (2): (1) Preparing coffee (2) Treating the prepared coffee with an enzyme agent By using the above production method, it is possible to produce processed coffee that can exhibit an aggregation-inhibiting effect when mixed with milk.

[0047] (Method for producing a processed coffee-containing beverage) This embodiment relates to a method for producing a processed coffee-containing beverage, which includes allowing the enzyme agent to act on coffee and then mixing milk. In this embodiment, milk is mixed with the processed coffee obtained by allowing the enzyme agent to act on coffee.

[0048] One aspect of the method for producing a processed coffee-containing beverage of this embodiment includes the following steps. Note that an enzyme deactivation step may be added after step (A): (A) A step of treating coffee with an enzyme agent to obtain processed coffee (B) A step of mixing the processed coffee with milk. Note that the mixing conditions with milk in step (B) are the same as those described above.

[0049] The temperature of the processed coffee mixed in step (B) is preferably 30 to 90°C, more preferably 40 to 80°C, and even more preferably 50 to 70°C. The pH of the processed coffee mixed in step (B) is preferably 3.0 to 7.0, more preferably 3.5 to 6.5, even more preferably 4.0 to 6.0, and even more preferably 4.2 to 5.0. The temperature of the milk mixed in step (B) is preferably 0 to 70°C, more preferably 0.1 to 50°C, and even more preferably 0.5 to 30°C.

[0050] Examples of milk used in the method for producing a processed coffee-containing beverage of this embodiment include the milks described above. Among these, the milk is preferably plant-based milk, and the raw material for the plant-based milk is preferably at least one selected from the group consisting of pulses such as soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, and kidney beans; cereals such as wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, and potato; nuts and seeds such as almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, amaranth, canary seeds, and linseed; and algae.

[0051] In the step of mixing the processed coffee with milk, optional ingredients may be added as needed. Examples of optional ingredients include the optional ingredients described above. For example, in the step of mixing the processed coffee with milk, a flavoring may be added, and in this case, it is preferable to add a coffee flavor.

[0052] (Aggregation Inhibition Method) This embodiment relates to a method for inhibiting aggregation when coffee and milk are mixed, which includes allowing the enzyme agent described above to act on coffee. In the aggregation inhibition method of this embodiment, aggregation when coffee and milk are mixed can be inhibited by allowing the enzyme agent described above to act on coffee. In conventional technology, it has been considered to enhance the aggregation inhibition effect by subjecting milk to enzyme treatment, but this embodiment includes allowing oxidase to act on coffee. This makes it possible to significantly improve the aggregation inhibition effect when coffee and milk are mixed.

[0053] Examples of milk used in the aggregation-inhibiting method of this embodiment include the milks described above. Among them, the milk is preferably plant-based milk, and the raw material of the plant-based milk is preferably obtained from at least one selected from the group consisting of pulses such as soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, and kidney beans; cereals such as wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, and potato; nuts and seeds such as almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, amaranth, canary seeds, and linseed; and algae.

[0054] This embodiment may relate to the use of an enzyme preparation containing oxidase for suppressing aggregation when coffee and milk are mixed. The enzyme preparation of this embodiment is used to suppress aggregation that occurs when coffee and milk are mixed by allowing the oxidase to act on coffee.

[0055] The features of the present invention will be explained in more detail below with reference to examples and comparative examples. The materials, amounts used, ratios, treatment contents, treatment procedures, etc. shown in the following examples can be changed as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention should not be construed as being limited by the specific examples shown below.

[0056] (Materials used)

[0057] (Method for Measuring Enzyme Activity) Laccase Activity Measurement The reagent used for measuring laccase activity was prepared based on the polyphenol oxidase activity measurement test method in the 9th edition of the Japanese Standards for Food Additives. Specifically, a substrate solution was prepared by mixing 0.25 mol / L phenol solution, 0.009 mol / L 4-aminoantipyrine solution, and 1 mol / L acetic acid / sodium acetate buffer (pH 4.5) in a volume ratio of 1:1:0.5. 2.5 ml of this substrate solution was placed in a cuvette and preheated to 30°C. 0.5 ml of enzyme solution diluted to a specified concentration was added, stirred, and incubated at 30°C. The absorbance at 505 nm was measured after 10 and 40 seconds. Under these conditions, an increase in absorbance of 0.1 per minute was defined as one unit (U) of enzyme content in 1 mL of reaction solution.

[0058] Test Example 1: 140 mL of water and 2 g of instant coffee powder were mixed, and laccase was added at concentrations of 840 U, ​​4200 U, and 8400 U per 1 g of instant coffee powder. The mixture was then incubated overnight (12-16 hours) at 50°C. Coffee flavor was then added to a final concentration of 0.01% by mass. After the reaction was completed, the coffee beverage was boiled at 100°C to inactivate the enzyme. Processed coffee was thus obtained. 40 mL of processed coffee heated to 80-100°C and 10 mL of oat drink stored in the cold were mixed in a 100 mL beaker. The flocculation ability was evaluated by measuring the turbidity (absorbance 660 nm) at the center of the liquid for 60 minutes immediately after mixing. A higher relative turbidity indicates a lower flocculation ability of the coffee. The relative turbidity after each time point was calculated as follows: The control was coffee to which no laccase was added, and the relative turbidity at the start of the reaction (storage time 0 minutes) for both the coffee (control) and the processed coffee (enzyme-treated coffee) was set to 100%. The control was also produced under the same conditions except that no enzyme was added. Relative turbidity (%) = (absorbance at 660 nm after each time elapsed after mixing with coffee) / (absorbance at 660 nm 0 minutes after mixing with coffee) x 100

[0059]

[0060] In the control coffee that had not been treated with laccase, aggregates began to settle five minutes after mixing with the oat drink, whereas in the enzyme-treated coffee, no aggregates were observed even after 60 minutes.

[0061] As shown in Table 3, there was no significant difference in pH between the coffee not treated with laccase (control) and the processed coffee treated with laccase.

[0062]

[0063] (Test Example 2) A test was also conducted on an almond drink using the same procedure as in Test Example 1. 140 mL of water and 2 g of instant coffee powder were mixed, and 840 U of laccase per 1 g of instant coffee powder was added. The mixture was incubated overnight (12 to 16 hours) at 50°C. After the reaction was completed, the coffee drink was boiled at 100°C to inactivate the enzyme. In this way, processed coffee was obtained. 40 mL of processed coffee heated to 80 to 100°C was mixed with 10 mL of almond drink (the paste was diluted with tap water to a protein concentration of 0.5%) that had been stored in the cold. The flocculation property was evaluated by measuring the turbidity (absorbance at 660 nm) for 60 minutes immediately after mixing.

[0064]

[0065] In the case of the non-laccase-treated coffee (control), aggregates began to settle 5 minutes after mixing with the almond milk, whereas in the case of the processed coffee (enzyme-treated coffee), no aggregates were observed even after 60 minutes.

[0066] (Test Example 3) A test was also conducted on soy milk using the same procedure as Test Example 1. 140 mL of water and 2 g of instant coffee powder were mixed, and 840 U of laccase per 1 g of instant coffee powder was added, followed by incubation at 50°C overnight (12 to 16 hours). After the reaction was completed, the coffee beverage was boiled at 100°C to inactivate the enzyme. In this way, processed coffee was obtained. 40 mL of processed coffee heated to 80 to 100°C was mixed with 10 mL of soy milk (Meiraku Unadjusted Organic Soy Milk, manufactured by Sujata) that had been stored in the cold. The flocculation property was evaluated by measuring the turbidity (absorbance at 660 nm) for 60 minutes immediately after mixing.

[0067]

[0068] In the case of the non-laccase-treated coffee (control), aggregates began to settle 15 minutes after mixing with soy milk, whereas in the case of the processed coffee (enzyme-treated coffee), no aggregates were observed even after 60 minutes.

[0069] As described above, according to the present technology, coffee in which the occurrence of aggregation when coffee is mixed with animal milk or plant milk is suppressed (anti-curdling coffee) can be prepared.

Claims

1. An enzyme preparation containing oxidase to inhibit coagulation when coffee and milk are mixed.

2. The enzyme preparation according to claim 1, wherein the oxidase is a copper oxidase.

3. The enzyme preparation according to claim 1, wherein the oxidase is a multicopper oxidase.

4. The enzyme preparation according to claim 1, wherein the oxidase is laccase.

5. The enzyme preparation according to claim 1, wherein the oxidase is derived from the genus Trametes.

6. Processed coffee obtained by allowing the enzyme preparation according to claim 1 to act on coffee.

7. A processed coffee-containing beverage comprising processed coffee obtained by reacting the enzyme preparation according to claim 1 with coffee, and milk.

8. The processed coffee-containing beverage of claim 7, wherein the milk is a vegetable milk.

9. The processed coffee-containing beverage according to claim 8, wherein the raw material for the plant-based milk is at least one selected from the group consisting of soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, kidney beans, wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, potatoes, almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia, amaranth, canary seeds, linseeds and algae.

10. A method for producing processed coffee, which comprises allowing the enzyme preparation according to claim 1 to act on coffee.

11. A method for producing a processed coffee-containing beverage, which comprises allowing the enzyme preparation according to claim 1 to act on coffee and then mixing with milk.

12. The method of producing a processed coffee-containing beverage according to claim 11, wherein the milk is a vegetable milk.

13. The method for producing a processed coffee-containing beverage according to claim 12, wherein the raw material for the vegetable milk is at least one selected from the group consisting of soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, kidney beans, wheat, barley, oats, sorghum, rice, rye, buckwheat, barnyard millet, foxtail millet, teff, quinoa, corn, potatoes, almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, pine nuts, hemp seeds (industrial hemp), chia seeds, chia, amaranth, canary seeds, linseeds and algae.

14. A method for inhibiting coagulation when coffee and milk are mixed, comprising allowing the enzyme preparation according to claim 1 to act on coffee.

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