Enzyme agent for manufacturing vegetable viscous food and drink

Abstract

Provided is a technique for enhancing the viscosity of a vegetable viscous food or beverage. The present technology provides an enzyme agent for producing a vegetable viscous food and drink, the enzyme agent containing an alpha-amylase and a cellulase. The enzyme agent for manufacturing vegetable viscous foods and drinks according to the present technology can be used as an enzyme agent for manufacturing vegetable fermented foods and drinks or as an enzyme agent for manufacturing vegetable lactic acid fermented foods and drinks. Also provided is a viscosity enhancer for a vegetable viscous food and drink, the viscosity enhancer containing an alpha-amylase and a cellulase. The present technology further provides a method for producing a vegetable viscous food or beverage, which comprises a step for causing an alpha-amylase and a cellulase to act on a vegetable raw material, and a method for enhancing the viscosity of a vegetable viscous food or beverage.

Description

Technical Field

[0001] The present technology relates to an enzyme for producing a plant-based viscous food or beverage. More specifically, the present technology relates to an enzyme for producing a plant-based viscous food or beverage that can increase the viscosity of the plant-based viscous food or beverage, a viscosity enhancer for the plant-based viscous food or beverage, a method for producing the plant-based viscous food or beverage, and a method for increasing the viscosity of the plant-based viscous food or beverage. Background Art

[0002] In recent years, the plant-based food and beverage market has been expanding as a dairy and meat substitute market, driven by sustainability and health concerns. Plant-based yogurt, for example, plays a key role in the dairy substitute market and continues to attract attention. The standard method for producing plant-based yogurt is to mix carbohydrate, protein, and lipid sources with plant-based milk, followed by lactic acid fermentation.

[0003] Against this backdrop, various technologies have been developed in recent years to improve the physical properties of plant-based foods and beverages. For example, Patent Document 1 discloses a method for producing plant-based yogurt by subjecting hydrolyzed cereal starch to lactic acid fermentation using enzymes produced by Aspergillus koji or an enzyme preparation derived from Aspergillus koji. Furthermore, Patent Document 2 discloses a method for producing an oat-based, non-dairy yogurt with enhanced viscosity by subjecting an oat base treated with α- or β-amylase to the action of transglutaminase. Patent Document 3 discloses a method for producing plant-based protein foods (particularly plant-based yogurt) with improved flavor and properties by subjecting a plant-based protein raw material to the action of protein deamidase and lipase. Non-Patent Document 1 discloses methods for improving the mouthfeel, sensory, nutritional, and functional properties of plant-based yogurt by using exopolysaccharide (EPS)-producing bacteria as a fermentation culture.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: International Publication No. 2022 / 037920

[0007] Patent Document 2: International Publication No. 2017 / 171601

[0008] Patent Document 3: International Publication No. 2022 / 014542

[0009] Non-patent literature

[0010] Non-patent document 1: Manufacture and characterization of a yogurt-likebeverage made with oat flakes fermented by selected lactic acid bacteria, International Journal of Microbiology 185 (2014) 17-26. Summary of the Invention

[0011] Plant-based foods and drinks differ from animal-based foods in their manufacturing methods. (1) They contain less protein, (2) have different coagulation properties, and (3) require the addition of structuring agents and emulsifiers. Consequently, these processes often require time and cost, and there is room for improvement to achieve satisfactory properties. In particular, when using plant-based ingredients to produce viscous foods and drinks that require viscosity, additives such as thickeners and emulsifiers used to increase viscosity do not conform to the concept of clean label products, and alternatives are sought.

[0012] Therefore, the main object of the present technology is to provide a new technology for increasing the viscosity of plant-based viscous food and beverages.

[0013] The inventors of this application have diligently studied and have succeeded in increasing the viscosity of viscous food and beverages produced using plant materials by allowing α-amylase and cellulase to act on the plant materials. This technology was developed through further research based on this finding.

[0014] That is, in the present technology, first, an enzyme agent for producing a plant-based viscous food or beverage is provided, the enzyme agent containing α-amylase and cellulase.

[0015] The enzyme agent for producing a plant-based viscous food or drink according to the present technology can be used to produce a plant-based fermented food or drink.

[0016] Furthermore, the enzyme agent for producing a plant-based viscous food or drink according to the present technology can be used to produce a plant-based lactic acid fermented food or drink.

[0017] Furthermore, the enzyme agent for producing a plant-based viscous food or drink according to the present technology can be used to produce plant-based yogurt.

[0018] The present technology further provides a viscosity enhancer for plant-based viscous food and beverages, comprising α-amylase and cellulase.

[0019] In the present technology, a method for manufacturing a plant-based viscous food and drink and a method for increasing the viscosity of a plant-based viscous food and drink are further provided. The method for manufacturing a plant-based viscous food and drink includes a process of allowing α-amylase and cellulase to act on a plant-based raw material, and the method for increasing the viscosity of a plant-based viscous food and drink includes a process of allowing α-amylase and cellulase to act on a plant-based raw material.

[0020] According to the present technology, the viscosity of plant-based viscous food and beverages can be increased even without using additives. DETAILED DESCRIPTION

[0021] Hereinafter, preferred modes for implementing the present technology will be described. In addition, the embodiment described below shows an example of a representative embodiment of the present technology, but the scope of the present technology should not be narrowly interpreted thereby.

[0022] 1. Enzymes for the production of plant-based viscous foods and beverages, and viscosity enhancers for plant-based viscous foods and beverages

[0023] The viscosity enhancer for the manufacture of vegetable viscous food and beverages involved in the present technology and the vegetable viscous food and beverages contains α-amylase and cellulase. In the present technology, as demonstrated in the examples described later, when manufacturing vegetable viscous food and beverages, by treating vegetable raw materials with α-amylase and cellulase, the viscosity of the vegetable viscous food and beverages manufactured was successfully and significantly enhanced. In addition, as demonstrated in the examples described later, when manufacturing vegetable viscous food and beverages, by treating vegetable raw materials with α-amylase and cellulase, the water retention of the vegetable viscous food and beverages manufactured was also successfully improved. Below, the components of the viscosity enhancer for the manufacture of vegetable viscous food and beverages involved in the present technology and the vegetable viscous food and beverages are described in detail.

[0024] (1) α-amylase

[0025] α-Amylase is an enzyme that acts on starch and mainly hydrolyzes α-1,4-glucosidic bonds. α-Amylases that can be used in the present technology may also have other functions as long as they have α-amylase activity.

[0026] The source of α-amylase that can be used in the present technology is not particularly limited as long as the action and effect of the present technology are not impaired. For example, filamentous fungi include Aspergillus aureus, Aspergillus foetidus, Aspergillus niger, and Aspergillus oryzae; actinomycetes include Saccharomonospora viridis, Streptomyces avermitilis, Streptomyces griseus, Streptomyces thermoviolaceus, Streptomyces violaceoruber, and Thermomonospora viridis; and bacteria include Alcaligenes latus, Arthrobacter (genus Arthrobacter), Bacillus amyloliquefaciens, Bacillus circulans, and Bacillus licheniformis. licheniformis), Bacillus stearothermophilus, Bacillus subtilis, Cellulosimicrobium cellulans, Microbacterium imperiale, Paenibacillus alginolyticus, and Sulfolobus solfataricus. Preferred are α-amylases derived from the genus Bacillus or α-amylases derived from Aspergillus, and more preferred are α-amylases derived from Bacillus amyloliquefaciens or α-amylases derived from Bacillus licheniformis.

[0027] In the present technology, the use of natural (wild-type) α-amylases is not limited to recombinant α-amylases. Commercially available α-amylases or α-amylase preparations can also be used. Examples of commercially available α-amylases or α-amylase preparations include α-amylase derived from Bacillus amyloliquefaciens manufactured by Amano Enzyme Co., Ltd.

[0028] The α-amylase used in this technology can be prepared from the culture broth of the microorganisms that serve as the source of the above-mentioned cellulase. Specific preparation methods include methods for recovering α-amylase from the culture broth or cells of the above-mentioned microorganisms. For example, when using microorganisms that secrete α-amylase, the enzyme can be isolated and / or purified after the cells are recovered from the culture broth by prefiltration, centrifugation, etc., as needed. Furthermore, when using microorganisms that do not secrete α-amylase, the enzyme can be isolated and / or purified after the cells are recovered from the culture broth in advance, then crushed by pressure treatment, ultrasonic treatment, etc., and the enzyme is extracted. The enzyme can be isolated and / or purified by any known protein separation and / or purification method, without particular limitation, including centrifugation, ultrafiltration (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 reduced-pressure drying. Suitable excipients and / or drying aids can also be used in this drying method to achieve powderization. Alternatively, the separated and / or purified enzyme may be made into a liquid by adding appropriate additives and sterilizing by filtration.

[0029] As long as the effects and effects of the present technology are not impaired, the content of α-amylase in the enzyme for producing plant-based viscous food and drink products and the viscosity enhancer for plant-based viscous food and drink products involved in the present technology is not particularly limited. The lower limit of the content of α-amylase can be set to, for example, 0.5U or more per 1g of plant raw material of the plant-based viscous food and drink product, preferably 1U or more, 5U or more, 10U or more, more preferably 20U or more, 30U or more, 40U or more, 50U or more, and further preferably 60U or more, 70U or more, 80U or more, 90U or more. In addition, the lower limit of the content of α-amylase can be set to, for example, 1U or more, preferably 2U or more, 5U or more, 10U or more, more preferably 20U or more, 40U or more, and further preferably 60U or more, 80U or more, 100U or more, 120U or more, of the carbohydrates (preferably starch) contained in the plant raw material of the plant-based viscous food and drink product per 1g.

[0030] On the other hand, the upper limit of the content of α-amylase can be set to, for example, less than 500,000 U, less than 100,000 U, less than 50,000 U, less than 10,000 U, or less than 5,000 U per 1 g of plant raw material of the plant-based viscous food and drink product, preferably less than 4,000 U, less than 2,000 U, or less than 1,000 U, and more preferably less than 800 U, less than 600 U, less than 400 U, less than 200 U, or less than 120 U. In addition, the upper limit of the content of α-amylase can be set to the carbohydrate (preferably starch) contained in the plant raw material per 1 g of the plant viscous food and beverage product, for example, 700,000 U or less, 150,000 U or less, 70,000 U or less, 15,000 U or less, 7,000 U or less, preferably 6,000 U or less, 3,000 U or less, 1,500 U or less, more preferably 1,000 U or less, 800 U or less, 600 U or less, 300 U or less, 200 U or less, 150 U or less.

[0031] In this technology, 1 unit (1 U) of α-amylase activity is defined as the amount of enzyme that reduces the color development of iodine in 1% potato starch by 10% within 1 minute when treated with 1% potato starch as a substrate at pH 5.0 and 37°C. In this technology, α-amylase activity is measured using the α-amylase activity assay described in the Examples below.

[0032] (2) Cellulase

[0033] Cellulase is an enzyme that hydrolyzes the glycosidic bonds of β-1,4-glucan. Cellulase that can be used in the present technology may be an enzyme that also has other functions as long as it has cellulase activity.

[0034] The source of the cellulase that can be used in the present technology is not particularly limited as long as the action and effect of the present technology are not impaired. Examples of cellulases include basidiomycetes such as Corticium, Irpex, and Pycnoporus coccineus; and filamentous fungi such as Acremonium cellulolyticus, Aspergillus aculeatus, Aspergillus awamori, Aspergillus niger, Humicola insolens, Penicillium funiculosum, Trichoderma harzianum, Trichoderma insolens, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, and Trichoderma viride. viride), actinomycetes include the genera Actinomyces and Streptomyces, and bacteria include Bacillus circulans and Bacillus subtilis. Preferred examples include cellulases derived from microorganisms of the genus Trichoderma. Examples of microorganisms of the genus Trichoderma include Trichoderma reesei and Trichoderma viride. Preferred examples include cellulases derived from Trichoderma viride.

[0035] In the present technology, the cellulases are not limited to natural (wild-type) cellulases; recombinant cellulases may also be used. Commercially available cellulases or cellulase preparations may also be used. Examples of commercially available cellulases or cellulase preparations include cellulases derived from Trichoderma microorganisms, such as cellulases derived from Trichoderma viride, manufactured by Amano Enzyme Products Co., Ltd., and cellulases derived from Aspergillus microorganisms, such as cellulases derived from Aspergillus niger, manufactured by Amano Enzyme Products Co., Ltd.

[0036] The cellulase used in the present technology can be prepared from the culture fluid of the microorganism that is the source of the above-mentioned cellulase. As a specific preparation method, a method of recovering the cellulase from the culture fluid or bacterial cells of the above-mentioned microorganisms can be cited. For example, when using a cellulase-secreting microorganism, the bacterial cells can be recovered from the culture fluid by prefiltration, centrifugation, etc. as needed, and the enzyme can be separated and / or purified. In addition, when using a cellulase-non-secreting microorganism, the bacterial cells can be recovered from the culture fluid in advance, and then the bacterial cells can be crushed by pressure treatment, ultrasonic treatment, etc. to extract the enzyme, and the enzyme can be separated and / or purified. As the enzyme separation and / or purification method, known protein separation and / or purification methods can be used without particular limitation, and examples include: centrifugation, UF concentration method, salting-out method, various chromatography methods using ion exchange resins, etc. The separated and / or purified enzyme can be powdered by drying methods such as freeze drying and reduced pressure drying. In addition, it can also be powdered using appropriate excipients and / or drying aids in the drying method. Alternatively, the separated and / or purified enzyme may be made into a liquid by adding appropriate additives and sterilizing by filtration.

[0037] As long as the effect of the present technology is not damaged, the content of the cellulase in the viscosity enhancer for the manufacture of the plant-based viscous food and beverage product involved in the present technology and the plant-based viscous food and beverage product is not particularly limited. The lower limit of the content of the cellulase can be set as the plant raw material of every 1g plant-based viscous food and beverage product, for example, more than 0.01U, preferably more than 0.1U, more than 0.5U, more than 1U, more preferably more than 2U, more than 5U, more preferably more than 10U, more than 15U, more than 20U. In addition, the lower limit of the content of the cellulase can be set as the dietary fiber included in the plant raw material of every 1g plant-based viscous food and beverage product, for example, more than 0.1U, preferably more than 1U, more than 5U, more than 10U, more preferably more than 20U, more than 50U, more preferably more than 100U, more than 150U, more than 180U.

[0038] On the other hand, the upper limit of the content of cellulase can be set as the plant raw material of every 1g plant viscosity food and drink article and for example be below 10000U, below 5000U, below 2000U, below 1000U, below 500U, preferably below 200U, below 100U, more preferably below 80U, below 60U, below 40U, below 30U, below 25U.In addition, the upper limit of the content of cellulase can be set as the dietary fiber comprised in the plant raw material of every 1g plant viscosity food and drink article and for example be below 100000U, below 50000U, below 20000U, below 10000U, below 5000U, preferably below 2000U, below 1000U, more preferably below 800U, below 600U, below 400U, below 300U, below 250U.

[0039] In this technology, 1 unit (1 U) of cellulase activity is defined as the amount of enzyme that produces reducing sugars equivalent to 1 μmole of glucose in 1 minute when treated with sodium carboxymethyl ether cellulose (also known as sodium carboxymethyl cellulose) as a substrate at pH 4.5 and 37°C. In this technology, cellulase activity is measured using the cellulase activity assay described in the Examples below.

[0040] As long as the effects and effects of the present technology are not impaired, the ratio of α-amylase to cellulase in the enzyme for producing plant-based viscous food and beverages and the viscosity enhancer for plant-based viscous food and beverages according to the present technology is not particularly limited. The ratio of α-amylase to cellulase in the enzyme for producing plant-based viscous food and beverages and the viscosity enhancer for plant-based viscous food and beverages according to the present technology can be set to, for example, α-amylase activity:cellulase activity=1:10-100:1, preferably 1:1-50:1, 2:1-20:1, 3:1-10:1, 4:1-6:1, 5:1-3:1, and more preferably 1:2-100:1, 1:1.5-50:1, 1:1.2-30:1, 1:1.2-20:1, 1:1.2-10:1.

[0041] (3) Plant-based raw materials

[0042] As plant-based raw materials for the production of the enzymes and viscosity enhancers for plant-based viscous foods and beverages according to the present technology, one or more plant-based raw materials that can be used in viscous foods and beverages can be freely selected and used, as long as the effects and benefits of the present technology are not impaired. Examples include: cereals such as wheat, barley, oats, rice, rye, buckwheat, millet, foxtail millet, and teff; beans such as soybeans, green peas, lentils, chickpeas, black beans, broad beans, mung beans, lupines, and kidney beans; nuts such as almonds, coconuts, peanuts, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, and pine nuts; and chia seeds, quinoa, amaranth seeds, canary grass seeds, and flax seeds. In the present technology, among these raw materials, preferably cereals and beans other than soybeans are used, and more preferably oats are used.

[0043] The content of starch contained in the vegetable raw material (based on the weight of the dried plant raw material) is not particularly limited, and the vegetable raw material containing a preferred amount of starch can be selected according to the type of the viscous food and beverage to be manufactured and the required physical properties. In the present technology, it is possible to use a vegetable raw material comprising, for example, more than 1% by weight, preferably more than 10% by weight, more preferably more than 30% by weight, and further preferably more than 60% by weight of starch. In addition, in the present technology, it is possible to use a vegetable raw material comprising, for example, less than 95% by weight, preferably less than 90% by weight, more preferably less than 85% by weight, further preferably less than 80% by weight, less than 70% by weight, less than 60% by weight, less than 50% by weight, less than 40% by weight, less than 30% by weight, and less than 20% by weight of starch.

[0044] In summary, as plant-based raw materials that can be used for the enzymes for producing plant-based viscous foods and beverages and the viscosity enhancers for plant-based viscous foods and beverages involved in this technology, it is preferred to use cereals and beans other than soybeans and / or grains whose edible parts have a starch content of more than 10% by weight.

[0045] As long as the effects of the present technology are not impaired, the properties of the plant raw materials in which the enzyme for producing plant-based viscous food and beverages and the viscosity enhancer for plant-based viscous food and beverages according to the present technology can be used are not particularly limited, and preferably, liquid (including suspension) and slurry can be used.

[0046] (4) Types of plant-based sticky foods and beverages

[0047] As long as the effects and benefits of the present technology are not impaired, the types of plant-based viscous food and beverages that can be manufactured using the plant-based viscous food and beverage manufacturing enzymes of the present technology and the types of plant-based viscous food and beverages that can be enhanced in viscosity using the viscosity enhancers for plant-based viscous food and beverages are not particularly limited. The plant-based viscous food and beverage manufacturing enzymes and the viscosity enhancers for plant-based viscous food and beverages of the present technology are particularly preferably used in plant-based fermented food and beverages.

[0048] In the present technology, "plant-based fermented food and drink" refers to food and drink obtained by fermenting plant-based food and drink materials. Specifically, examples include: plant-based fermented milk (plant-based yogurt), plant-based lactic acid bacteria beverages, plant-based yogurt sauce, plant-based cheese-like foods, alcoholic beverages (Japanese sake, beer, wine, shochu, etc.), various fermented condiments (soy sauce, miso, vinegar, etc.), pickles, kimchi, natto, kudzu cakes, etc. In addition, in the present technology, "plant-based fermented food and drink" also includes secondary processed food and drink using these plant-based fermented food and drink. Among them, in the present technology, it can be appropriately used for plant-based lactic acid fermented food and drink, and particularly can be appropriately used for plant-based yogurt.

[0049] (5) Others

[0050] As long as the effect and the effect of the present technology are not damaged, the viscosity enhancer for the manufacture of the plant-based viscous food and beverage product involved in the present technology and the plant-based viscous food and beverage product can be used in combination with other enzymes or other components. As other components, it is possible to use components such as excipients, pH regulators, coloring agents, flavoring agents, disintegrants, lubricants, stabilizers, etc. that are for example used for conventional formulations. In addition, it is also possible to suitably use a component with function disclosed or found in the future according to the purpose.

[0051] 2. Method for producing plant-based viscous food and beverages, and method for increasing the viscosity of plant-based viscous food and beverages

[0052] The method for producing a plant-based viscous food and beverage and the method for increasing the viscosity of a plant-based viscous food and beverage according to the present technology include a step of subjecting a plant-based raw material to the action of α-amylase and cellulase (hereinafter referred to as the "enzyme action step"). Furthermore, a fermentation step, a recovery step, etc. may also be performed as needed. Each step is described in detail below.

[0053] (1) Enzyme action process

[0054] The enzyme action step is a step in which α-amylase and cellulase are allowed to act on the plant-based raw material. In the enzyme action step, the step of allowing α-amylase to act on the plant-based raw material (hereinafter also referred to as the "α-amylase action step") and the step of allowing cellulase to act on the plant-based raw material (hereinafter also referred to as the "cellulase action step") can be performed simultaneously or sequentially.

[0055] (1-1) α-amylase action process

[0056] As long as the effect of the present technology is not damaged, the various conditions in the α-amylase action process can be freely set. For example, pH, temperature, action time, etc. can be set according to the physicochemical properties such as the optimal pH, stable pH range, optimal temperature, and temperature stability of the α-amylase used. pH can be set to, for example, pH 4.0 to 10.0, pH 4.5 to 9.0, preferably pH 5.0 to 8.0, and more preferably pH 5.5 to 7.5. Temperature can be set to, for example, 10°C to 80°C, preferably 25°C to 75°C, 35°C to 70°C, and more preferably 50°C to 70°C. Action time can be set to, for example, 5 minutes to 48 hours, preferably 30 minutes to 24 hours, 1 to 12 hours, 2 to 8 hours, and more preferably 3 hours to 6 hours. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0057] (1-2) Cellulase action process

[0058] As long as the effect of the present technology is not damaged, the various conditions in the cellulase action step can be freely set. For example, pH, temperature, action time, etc. can be set according to the physicochemical properties such as the optimal pH, stable pH range, optimal temperature, and temperature stability of the cellulase used. pH can be set to, for example, pH 3.0 to 8.5, pH 3.5 to 8.0, preferably set to pH 4.0 to 7.0, and more preferably set to pH 4.5 to 6.5. Temperature can be set to, for example, 20°C to 75°C, preferably set to 25°C to 70°C, 25°C to 70°C, and more preferably set to 35°C to 65°C. Action time can be set to, for example, 5 minutes to 48 hours, preferably set to 30 minutes to 24 hours, 1 to 12 hours, 2 to 8 hours, and more preferably set to 3 hours to 6 hours. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0059] (2) Fermentation process

[0060] In the case of plant-based fermented foods and beverages, the plant-based viscous foods and beverages manufactured using the method for manufacturing plant-based viscous foods and beverages according to the present technology or the plant-based viscous foods and beverages whose viscosity is increased using the method for increasing the viscosity of plant-based viscous foods and beverages according to the present technology may be subjected to the fermentation step.

[0061] The fermentation step is a step of fermenting a part or all of the plant-based raw material. The fermentation step may be performed after the enzyme action step or simultaneously with the enzyme action step.

[0062] In the fermentation process, fermentation can be carried out using one or more microorganisms selected from anaerobic microorganisms, aerobic microorganisms and facultative anaerobic microorganisms. As long as the effect of the present technology is not damaged, the microorganisms that can be used in the present technology can be freely selected from the microorganisms that can be used to make common fermented food and drink products according to the type of the plant-based fermented food and drink products manufactured, and more than one use can be used. As microorganisms, for example: lactic acid bacteria, yeast, filamentous fungi, Bacillus subtilis (Bacillus subtilis), acetic acid bacteria (Acetobacteraceae) etc. can be enumerated.

[0063] In the fermentation process of the present technology, as long as the effect of the present technology is not damaged, lactic acid fermentation can be carried out. The lactic acid bacteria used during lactic acid fermentation can be freely selected from the lactic acid bacteria that can be used to manufacture common lactic acid fermented food and beverages according to the type and purpose of the lactic acid fermented food and beverage to be manufactured. As lactic acid bacteria, for example: lactic acid cocci belonging to Lactococcus (Lactococcus), Streptococcus (Streptococcus), Pediococcus (Pediococcus), Leuconostoc (Leuconostoc), lactic acid bacteria belonging to Lactobacillus (Lactobacillus), Bifidobacterium (Bifidobacterium) etc., preferably Lactobacillus gasseri (Lactobacilus gasseri), Lactobacillus rhamnosus (Lactobacilus rhamnosus), Bifidobacterium infantis (Bifidobacterium infantis), Bifidobacterium adolescentis (Bifidobacterium adolescentis) etc. can be mentioned.

[0064] As long as the effect of the present technology is not impaired, the various conditions during the fermentation process can be freely set. For example, the fermentation conditions, heating conditions, etc. can be set according to the type of microorganism used. The fermentation temperature can be set to, for example, 20°C to 50°C, preferably 25°C to 45°C, and more preferably 30°C to 40°C. The fermentation time can be set to, for example, 1 to 30 hours, preferably 2 to 20 hours, and more preferably 4 to 10 hours. Heating conditions include, for example, high temperature short time sterilization (HTST), high temperature cooking (UHT), high pressure steam sterilization, etc., as long as the temperature of the food and beverage materials is above 90°C, preferably around 95°C. Examples include a method of treating the food and beverage materials at 90 to 100°C for 1 to 5 minutes or a method of treating the food and beverage materials at 90 to 95°C for 1 to 3 minutes.

[0065] (3) Recycling process

[0066] The recovery process refers to a process for recovering the vegetable sticky food and drink manufactured through an enzyme action process and a fermentation process as needed. The specific recovery method can be based on the type of vegetable sticky food and drink manufactured, using one or more recovery methods commonly used in the manufacture of vegetable sticky food and drink.

[0067] One embodiment of the method for producing a plant-based viscous food and drink and the method for increasing the viscosity of a plant-based viscous food and drink according to the present technology includes the following steps (1) and (2). In addition, an enzyme inactivation step may be added after step (2). In addition, the enzymes used in step (2) may be treated simultaneously or separately. The order in which the treatments are performed separately is not particularly limited. In addition, when the treatments are performed separately, an enzyme inactivation step may be added between each enzyme treatment as needed.

[0068] (1) Process of preparing plant-based raw materials

[0069] (2) Process of allowing α-amylase and cellulase to act on the prepared plant-based raw materials

[0070] One embodiment of the method for producing a plant-based viscous food and drink and the method for increasing the viscosity of a plant-based viscous food and drink according to the present technology includes the following steps (1) to (3). In addition, an enzyme inactivation step may be added after step (2). The enzymes used in step (2) may be treated simultaneously or separately. The order in which the treatments are performed separately is not particularly limited. In addition, when the treatments are performed separately, an enzyme inactivation step may be added between each enzyme treatment as needed.

[0071] (1) Process of preparing plant-based raw materials

[0072] (2) Process of allowing α-amylase and cellulase to act on the prepared plant-based raw materials

[0073] (3) Process for recycling plant-based sticky food and beverages

[0074] One embodiment of the method for producing a plant-based viscous food and drink and the method for increasing the viscosity of a plant-based viscous food and drink according to the present technology includes the following steps (1) to (4). In addition, steps (2) and (3) can be performed simultaneously. In addition, an enzyme inactivation step can be added after steps (2) and / or (3). The enzymes used in step (2) can be treated simultaneously or separately. The order in which the treatments are performed separately is not particularly limited. In addition, when the treatments are performed separately, an enzyme inactivation step can be added between each enzyme treatment as needed.

[0075] (1) Process of preparing plant-based raw materials

[0076] (2) Process of allowing α-amylase and cellulase to act on the prepared plant-based raw materials

[0077] (3) The process of fermenting part or all of the plant-based raw materials

[0078] (4) Process for recycling plant-based sticky food and beverages

[0079] Example

[0080] Hereinafter, the present invention will be described in more detail based on Examples. In addition, the Examples described below are representative examples of the present invention, but the scope of the present invention should not be narrowly interpreted thereby.

[0081] 1. Raw materials

[0082] The enzymes and plant-based raw materials used in the Examples are shown in Table 1 below.

[0083] [Table 1]

[0084]

[0085] 2. Enzyme Activity Assay

[0086] [α-Amylase activity determination method]

[0087] Accurately weigh 10 mL of 1% potato starch dissolved in 1M acetic acid-sodium acetate buffer (pH 5.0). After heating at 37°C for 10 minutes, accurately add 1 mL of a sample solution containing α-amylase and immediately shake to mix. After accurately standing at 37°C for 10 minutes, accurately weigh 1 mL of this solution and add 10 mL of a 0.1M hydrochloric acid reagent. Immediately shake to mix and stop the reaction. Next, accurately weigh 0.5 mL of this solution and add 10 mL of a 0.002M iodine reagent (Japan). After shaking and mixing, measure the absorbance at 660 nm using water as a control. Alternatively, accurately add 1 mL of water in place of the sample solution and repeat the same procedure to measure the absorbance. Under these conditions, the amount of enzyme that reduces the iodine-induced coloration of potato starch by 10% within 1 minute is considered one unit and calculated using the following formula.

[0088] α-amylase activity (U / g, U / mL) = {(AB-AT) / AB} × 1 / W

[0089] AT: absorbance of sample solution

[0090] AB: absorbance of blank solution

[0091] W: The amount of sample in 1 mL of sample solution (g or mL)

[0092] [Method for measuring cellulase activity]

[0093] The cellulase activity can be measured by a method based on the cellulase activity test method in the 9th edition of the Food Additive Standards.

[0094] Accurately weigh 4 mL of 0.625% sodium carboxymethyl ether cellulose (also known as sodium carboxymethyl cellulose) dissolved in 1M acetic acid-sodium acetate buffer (pH 4.5) and place it into a 50 mL Nessler colorimetric tube. After incubating at 37°C for at least 10 minutes, accurately weigh and add 1 mL of the sample solution containing cellulase and immediately shake to mix. This solution is incubated at 37°C for 30 minutes. Accurately add 2 mL of alkaline copper solution and shake to mix. Then, accurately add 3 mL of 0.5 M sodium hydroxide solution and shake to dissolve the precipitate. After incubating for 20 minutes, accurately add 13 mL of 1M acetic acid-sodium acetate buffer (pH 4.5). Accurately weigh 1 mL of this solution and add 9 mL of 1M acetic acid-sodium acetate buffer (pH 4.5) and shake thoroughly to mix. Measure the absorbance of this solution at 750 nm against water. Separately, accurately weigh 1 mL of the sample solution and place it in a 50 mL Nessler colorimetric tube. Accurately add 2 mL of alkaline copper test solution and shake to mix. Next, accurately weigh and add 4 mL of the substrate solution and shake to mix. Repeat the same procedure to measure the absorbance. Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 μmole of glucose in 1 minute is considered 1 unit and calculated using the following formula.

[0095] Cellulase activity (U / g, U / mL) = (A1-A2 / a) × (1 / 30) × (1 / 0.180) × n

[0096] A1: Absorbance of the sample solution

[0097] A2: Absorbance of blank solution

[0098] a: Slope of the glucose standard curve

[0099] 1 / 30: Conversion factor per minute

[0100] 1 / 0.180: 1 μmole of glucose = 0.180 mg

[0101] n: dilution factor per 1g or 1mL of sample

[0102] 3. Experimental Examples

[0103] <Experimental Example 1>

[0104] In Experimental Example 1, the effects of enzyme differences on the viscosity and water retention of plant-based foods were investigated. In Experimental Example 1, oatmeal powder (powder) was used as an example of a plant-based raw material, and plant-based yogurt was produced as an example of a plant-based viscous food or beverage.

[0105] (1) Production of plant-based yogurt

[0106] 7.5 g of oat flour was added to 42.5 g of purified water, and the mixture was mixed at 60 ° C for 30 minutes at a speed of 400 rpm using a stirrer with a heater. Then, the enzyme shown in Table 2 was added and stirred for another hour. After the reaction, 1.5 g of tapioca starch, 1.5 g of pea-derived protein, and 0.2 g of tricalcium phosphate were added, and the mixture was mixed at 60 ° C for 10 minutes at a speed of 400 rpm using a stirrer with a heater. Thereafter, the mixture was treated in a 95 ° C water bath for 20 minutes. Then, the mixture was cooled at room temperature, kept warm at 40 ° C for 30 minutes using a stirrer with a heater at a speed of 400 rpm, and 10 mg of yogurt starter was suspended in 100 μL of sterilized water. Thereafter, the mixture was allowed to stand and fermented in a constant temperature bath at 40 ° C. After 24 hours, the mixture was transferred to a refrigerator to stop the fermentation and produce plant-based yogurt.

[0107] (2) Measurement and evaluation

[0108] The viscosity and water retention of the produced plant-based yogurt were measured and evaluated by the following methods.

[0109] [Viscosity]

[0110] Transfer 35 ml of yogurt to a 50 ml volumetric tube and measure viscosity using a Brookfield Digital Viscometer (Model DV-E) according to the instructions in the Brookfield viscometer's instruction manual. Use a Model S61 spindle, set to 30 rpm, and maintain the sample temperature at approximately 20°C. Confirm that the value stabilizes 2 minutes after the spindle begins rotating. Use the average of the two measurements.

[0111] [Water retention]

[0112] Place 10g of yogurt in a 15mL tube and centrifuge at 600xg for 20 minutes. Collect the supernatant and measure its weight. Water retention is calculated using the following formula. An increase in water retention indicates an increase in the amount of water in the resulting plant-based yogurt (increased water retention).

[0113] Water retention (%) = (1-(weight of supernatant / weight of sample)) × 100

[0114] (3) Results

[0115] The results are shown in Table 2.

[0116] [Table 2]

[0117]

[0118] (4) Investigation

[0119] As shown in Table 2, while the viscosity of Comparative Example 2, which used hemicellulase in combination with pectinase, was slightly higher than that of Comparative Example 1, which used α-amylase alone, the water retention was comparable. Meanwhile, Example 1, which used α-amylase and cellulase in combination, exceeded the upper limit of the viscosity, so accurate values ​​could not be given. However, compared to Comparative Examples 1 to 3, the viscosity was significantly higher, and the water retention was also improved.

[0120] <Experimental Example 2>

[0121] In Experimental Example 2, the effects of varying the amount of cellulase added on the viscosity and water retention of a plant-based viscous food and drink were investigated. In Experimental Example 2, oatmeal flour was used as an example of a plant-based raw material, and plant-based yogurt was produced as an example of a plant-based viscous food and drink.

[0122] (1) Production of plant-based yogurt

[0123] Plant-based yogurt was produced by the same method as in Experimental Example 1 using the enzymes shown in Table 3 below.

[0124] (2) Measurement and evaluation

[0125] The viscosity and water retention of the produced plant-based yogurt were measured and evaluated by the following methods.

[0126] [Viscosity]

[0127] Since the viscosity upper limit was exceeded in Example 1 of Experimental Example 1, the rotation speed was set to 5 rpm in Experimental Example 2. The viscosity was measured by the same method as in Experimental Example 1 except for the rotation speed.

[0128] [Water retention]

[0129] The water retention was evaluated by the same method as in Experimental Example 1.

[0130] (3) Results

[0131] The results are shown in Table 3.

[0132] [Table 3]

[0133]

[0134] (4) Investigation

[0135] As shown in Table 3, viscosity was enhanced regardless of the amount of cellulase added. Furthermore, Example 6 exhibited a higher viscosity-enhancing effect than Comparative Examples 2 and 3 of Experimental Example 1. This indicates that even at an addition level of 1.1 U / g, the viscosity-enhancing effect can be fully exerted in oat flour.

[0136] It is found that the water retention improves as the amount of cellulase added increases. It is also found that in order to reliably exert the effect of water retention, it is preferable to use 10.5 U / g-oat flour or more of cellulase.

Claims

An enzyme preparation for producing a plant-based viscous food or beverage, comprising α-amylase and cellulase.

2. The enzyme agent for producing a plant-based viscous food or beverage according to claim 1, wherein The plant-based viscous food and beverage is a plant-based fermented food and beverage.

3. The enzyme agent for producing a plant-based viscous food or beverage according to claim 2, wherein The plant-based fermented food and beverage is a plant-based lactic acid fermented food and beverage.

4. The enzyme agent for producing a plant-based viscous food or beverage according to claim 3, wherein The plant-based lactic acid fermented food and beverage is plant-based yogurt.

5. A viscosity enhancer for plant-based viscous food and beverages, comprising α-amylase and cellulase.

6. A method for producing a plant-based viscous food or beverage, comprising the step of allowing α-amylase and cellulase to act on a plant-based raw material.

7. A method for increasing the viscosity of a plant-based viscous food or beverage, comprising the step of allowing α-amylase and cellulase to act on a plant-based raw material.

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