Method for producing vegetable food and drink

Abstract

Provided is a technique for increasing the cyclodextrin content in a vegetable food or beverage produced by sufficiently performing liquefaction during the production of the vegetable food or beverage. The present technology provides a method for producing a vegetable food or beverage, the method comprising: a cyclodextrin-producing enzyme action step for causing a cyclodextrin-producing enzyme to act on a vegetable raw material; and a cyclodextrin-producing enzyme and alpha-amylase action step in which, after the cyclodextrin-producing enzyme action step, a cyclodextrin-producing enzyme and an alpha-amylase act on the plant raw material.

Description

Technical Field

[0001] The present technology relates to a method for producing a plant-based food and drink. More specifically, the present technology relates to a method for producing a plant-based food and drink, a method for increasing the cyclodextrin content of a plant-based food and drink, an agent for increasing the cyclodextrin content of a plant-based food and drink, and a plant-based food and drink. Background Art

[0002] In recent years, due to the rise of health awareness, plant-based food and beverages have attracted much attention. For example, oat beverages are prepared by suspending oat materials in an aqueous medium and using one or more amylases to decompose the starch of the oat materials. The amylase used in this case is β-amylase, or a mixture of α-amylase and β-amylase. The oat beverage prepared by this method produces maltose through the action of β-amylase, thereby presenting a suitable sweet taste (Patent Document 1).

[0003] Here, the enzyme used in the present technology is described. Cyclodextrin-producing enzyme is an enzyme that acts on starch to produce α-, β-, and γ-cyclodextrin. In addition, it is known that cyclodextrin-producing enzyme acts on starch to produce cyclodextrin without liquefying starch granules (Patent Document 2).

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: Japanese Patent Application Publication No. 2018-075029

[0007] Patent Document 2: Japanese Patent Application Laid-Open No. 09-262091 Summary of the invention

[0008] As mentioned above, in recent years, the plant-based food and drink products that receive attention have the local flavor from plant raw materials, and it is desired to reduce unpreferable local flavor. In order to reduce unpreferable local flavor, it is effective to make the plant-based food and drink products contain the cyclodextrin with masking effect. This cyclodextrin can be generated by starch through cyclodextrin generating enzyme.

[0009] On the other hand, when manufacturing plant-based food and drink, by making α-amylase act on plant-based raw material, the liquefaction of plant-based raw material can be fully promoted. But, as shown in the embodiment described later, it is clear that if α-amylase and cyclodextrin generating enzyme are acted on plant-based raw material, then compared with the situation that cyclodextrin generating enzyme is acted on separately, the content of cyclodextrin in the plant-based food and drink is reduced.

[0010] Therefore, the main object of the present technology is to provide a technology for sufficiently performing liquefaction during production of plant-based foods and drinks and increasing the cyclodextrin content in the produced plant-based foods and drinks.

[0011] The inventors of the present application have conducted in-depth research on the technology of increasing the cyclodextrin content in plant-based foods and beverages, and found that after allowing cyclodextrin-producing enzyme to act on plant raw materials that serve as raw materials for plant-based foods and beverages, α-amylase is allowed to act on the plant raw materials while continuing the treatment with the cyclodextrin-producing enzyme. This allows the liquefaction during the manufacture of plant-based foods and beverages to be fully carried out while suppressing the reduction in the cyclodextrin content in the plant-based foods and beverages caused by α-amylase. As a result, the cyclodextrin content in the plant-based foods and beverages is successfully increased, thereby completing the present technology.

[0012] That is, in the present technology, firstly, a method for producing a plant-based food and beverage is provided, which comprises:

[0013] a cyclodextrin-generating enzyme action step of allowing the cyclodextrin-generating enzyme to act on the plant-based raw material; and

[0014] After the cyclodextrin-forming enzyme action step, a cyclodextrin-forming enzyme and α-amylase action step is performed in which the cyclodextrin-forming enzyme and α-amylase are allowed to act on the plant-based raw material.

[0015] In the present technology, in addition, a method for increasing the cyclodextrin content of a plant-based food product is provided, which comprises:

[0016] a cyclodextrin-generating enzyme action step of allowing the cyclodextrin-generating enzyme to act on the plant-based raw material; and

[0017] After the cyclodextrin-forming enzyme action step, a cyclodextrin-forming enzyme and α-amylase action step is performed in which the cyclodextrin-forming enzyme and α-amylase are allowed to act on the plant-based raw material.

[0018] In the present technology, there is also provided a cyclodextrin content increasing agent for a plant-based food and beverage, comprising:

[0019] A first dose containing a cyclodextrin-forming enzyme; and

[0020] The second dose contains alpha-amylase.

[0021] The present technology also provides a plant-based food and drink using the cyclodextrin content-increasing agent according to the present technology.

[0022] According to the present technology, liquefaction can be sufficiently performed during production of plant-based food and drinks, and the cyclodextrin content in the produced plant-based food and drinks can be increased. DETAILED DESCRIPTION

[0023] The preferred mode for implementing the present technology will be described below. 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.

[0024] 1. Method for producing plant-based food and beverage, method for increasing cyclodextrin content in plant-based food and beverage

[0025] The manufacturing method of the plant-based food and drink involved in the present technology and the method for increasing the cyclodextrin content are methods including a process of causing a cyclodextrin generating enzyme to act on a plant-based raw material (hereinafter also referred to as a "cyclodextrin generating enzyme action process") and a process of causing a cyclodextrin generating enzyme and an α-amylase to act on a plant-based raw material (hereinafter also referred to as a "cyclodextrin generating enzyme and α-amylase action process"). In addition, depending on the type of the plant-based food and drink, a general food manufacturing process can also be performed before and after each process or simultaneously with each process within the range that does not damage the effect of the present technology. In addition, in the manufacturing method of the plant-based food and drink and the method for increasing the cyclodextrin content, a recovery process of recovering the manufactured plant-based food and drink and the plant-based food and drink with increased cyclodextrin content can also be further implemented.

[0026] In the manufacturing method of the plant-based food and drink product involved in the present technology and the method for increasing the cyclodextrin content, before the cyclodextrin generating enzyme action step, it is preferred to perform a step (hereinafter also referred to as "α-amylase action step") of causing α-amylase to act on the plant-based raw material. In addition, as long as it does not affect the production amount of cyclodextrin, the α-amylase action step can be performed before the cyclodextrin generating enzyme action step. Hereinafter, the raw materials and each step used are described in detail.

[0027] (1) Plant-based raw materials

[0028] As long as the plant raw materials that can be used in this technology do not damage the effect of this technology, the origin and type of the plant raw materials are not particularly limited, and can be freely selected according to the target plant food and drink. For example, beans such as soybeans, peas, lentils, chickpeas, black beans, broad beans, mung beans, lupines, kidney beans, etc.; cereals such as wheat, barley, oats, rice, rye, buckwheat, barnyard grass, millet, and teff; nuts such as almonds, coconuts, peanuts, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, biri nuts, chestnuts, sesame seeds, and pine nuts; hemp seeds (industrial hemp), chia seeds, quinoa, Amaranthus, canary seeds, flax seeds, etc. In this technology, they can be used alone or in combination of two or more. Among these raw materials, cereals are preferred, and oats are more preferred.

[0029] In the present technology, the properties of the plant raw material when subjected to various enzyme treatments are not particularly limited unless the effects of the present technology are impaired, but liquid, slurry, and paste are preferred.

[0030] The plant raw material that can be used in the present technology contains sugars. The content of the plant sugar contained in the plant raw material (based on the weight of the plant raw material in a dry state) is not particularly limited, and for example, 20% by weight or more can be cited. Preferably, 30% by weight or more can be cited, more preferably 40% by weight or more, and further preferably 50% by weight or more. The upper limit of the content range is not particularly limited, and for example, 90% by weight or less, 80% by weight or less, and 70% by weight or less can be cited.

[0031] In the present technology, the vegetable raw material when carrying out the cyclodextrin generating enzyme action step described later preferably uses the vegetable raw material that has not been treated with α-amylase. When the starch contained in the vegetable raw material is liquefied, by treating with cyclodextrin generating enzyme before α-amylase treatment, then treating with α-amylase and cyclodextrin generating enzyme, the cyclodextrin in the plant food and drink after the liquefaction can be increased.

[0032] (2) Cyclodextrin production enzyme action process

[0033] The cyclodextrin generating enzyme action step is the step of making the cyclodextrin generating enzyme act on the vegetable raw material. In the cyclodextrin generating enzyme action step, preferably the cyclodextrin generating enzyme is acted on the vegetable raw material alone, but as long as in the scope of the production amount that does not affect cyclodextrin, just can select and use one or more other enzymes that can be used for the manufacture of vegetable food and drink products.

[0034] The cyclodextrin-producing enzyme that can be used in the present technology is an enzyme that has the activity of producing cyclodextrin composed of a degree of polymerization of 6 to 8 using a polysaccharide such as starch having a structure in which glucose is bonded in α-1,4 bonds as a substrate. As the cyclodextrin-producing enzyme that can be used in the present technology, as long as the activity of producing cyclodextrin is the main activity, it may also be an enzyme that has other functions.

[0035] In the present technology, by allowing a cyclodextrin-producing enzyme to act on a plant-based raw material, the starch in the plant-based raw material can be liquefied, and the cyclodextrin in the produced plant-based food and drink can be increased. The cyclodextrin in the produced plant-based food and drink can mask foreign smells.

[0036] The source of the cyclodextrin-producing enzyme that can be used in the present technology is not particularly limited, and examples thereof include: enzymes from the genus Bacillus, such as Bacillus stearothermophilus, Bacillus megaterium, Bacillus circulans, Bacillus macerans, Bacillus ohbensis, and Bacillus clarkii; from the genus Geobacillus, such as Geobacillus stearothermophilus; from the genus Paenibacillus, such as Paenibacillus macerans; from the genus Klebsiella, such as Klebsiella pneumoniae; pneumoniae); cyclodextrin-producing enzymes of the genus Thremoanaerobactor; and the genus Brevibacterium. These cyclodextrin-producing enzymes may be used alone or in combination of two or more. Among them, from the viewpoint of improving the flavor of the plant raw material, the cyclodextrin-producing enzyme is preferably a cyclodextrin glucosyltransferase derived from Paenibacillus and / or a cyclodextrin glucosyltransferase derived from Anoxybacillus caldiproteolyticus (including a substance formerly also known as Geobacillus stearothermophilus), and more preferably a cyclodextrin glucosyltransferase derived from Paenibacillus macerans and / or a cyclodextrin glucosyltransferase derived from Anoxybacillus caldiproteolyticus (including a substance formerly also known as Geobacillus stearothermophilus).

[0037] The “cyclodextrin-producing enzyme derived from Paenibacillus macerans” and / or the “cyclodextrin-producing enzyme derived from Bacillus thermolyticus (including a substance previously known as Geobacillus stearothermophilus)” herein refers to a cyclodextrin-producing enzyme produced by a microorganism (which may be a wild strain or a mutant strain) classified as Paenibacillus macerans and / or Bacillus thermolyticus (including a substance previously known as Geobacillus stearothermophilus), or a cyclodextrin-producing enzyme obtained by a genetic engineering method using a cyclodextrin-producing enzyme gene. Therefore, a recombinant produced by a host microorganism into which a cyclodextrin-producing enzyme gene (or a gene in which the gene is modified) obtained from Paenibacillus macerans and / or Bacillus thermolyticus (including a substance formerly known as Geobacillus stearothermophilus) is introduced also corresponds to “cyclodextrin-producing enzyme from Paenibacillus macerans” and / or “cyclodextrin-producing enzyme from Bacillus thermolyticus (including a substance formerly known as Geobacillus stearothermophilus)”.

[0038] The cyclodextrin generating enzyme used in the present technology can be prepared from the culture solution of the microorganism that is the source of the above-mentioned cyclodextrin generating enzyme. As a specific preparation method, a method for recovering the cyclodextrin generating enzyme from the culture solution or the bacterial cell of the above-mentioned microorganism can be cited. For example, in the case of using a cyclodextrin generating enzyme secretory microorganism, as needed, after recovering the bacterial cell from the culture solution by prefiltration, centrifugation, etc., the enzyme can be separated and / or purified. In addition, in the case of using a cyclodextrin generating enzyme non-secreting microorganism, as needed, after recovering the bacterial cell from the culture solution in advance, the bacterial cell can be crushed by pressure treatment, ultrasonic treatment, etc., the enzyme is extracted, and then the enzyme is separated and / or purified. As an enzyme separation and / or purification method, a known protein separation and / or purification method can be used without particular limitation, and examples thereof include: centrifugal separation method, 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, and in addition, a suitable excipient and / or drying aid can be used in the drying method to be powdered. Alternatively, the separated and / or purified enzyme may be made into a liquid by adding appropriate additives and sterilizing by filtration.

[0039] In the present technology, commercially available products can also be used as cyclodextrin-producing enzymes. Examples of preferred commercially available products include cyclodextrin glycosyltransferase from Paenibacillus macerans manufactured by Amano Enzyme Co., Ltd., cyclodextrin glycosyltransferase from Anoxybacillus caldiproteolyticus (formerly also classified as Geobacillus stearothermophilus) manufactured by Amano Enzyme Co., Ltd., and the like.

[0040] As long as the effect of the present technology is not damaged, the addition of the cyclodextrin generating enzyme in the cyclodextrin generating enzyme action step can be freely set. The lower limit of the addition of the cyclodextrin generating enzyme can be set to more than 0.01U relative to 1g of plant raw material, for example, from the viewpoint of further improving the cyclodextrin content in the plant-based food and drink, it can be set to preferably more than 0.03U, more preferably more than 0.1U, further preferably more than 0.3U, and further preferably more than 1U.

[0041] In addition, the lower limit of the amount of cyclodextrin generating enzyme added can be set to, for example, 0.02U or more relative to 1g of polysaccharide (preferably starch) used for the manufacture of plant-based food and beverages. From the viewpoint of further increasing the cyclodextrin content in the plant-based food and beverage, it can be set to preferably 0.05U or more, more preferably 0.5U or more, further preferably 2U or more, and further preferably 5U or more.

[0042] Unless the effects of the present technology are impaired, the upper limit of the amount of cyclodextrin-producing enzyme added is not particularly limited, and can be set to, for example, 200 U or less, 100 U or less, 50 U or less, 20 U or less, or 10 U or less per 1 g of the plant raw material.

[0043] The upper limit of the amount of cyclodextrin generating enzyme added can be set to, for example, 500 U or less, 300 U or less, 150 U or less, 80 U or less, 60 U or less, or 40 U or less per 1 g of polysaccharide (preferably starch) used for producing plant-based food and beverage products.

[0044] In the present technology, the activity of the cyclodextrin-producing enzyme refers to a value measured by the measurement method described in the Examples described later.

[0045] As long as the effect of the present technology is not damaged, the various conditions of the cyclodextrin generating enzyme 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 cyclodextrin generating enzyme used. pH can be set to, for example, pH5.0-8.0, preferably pH5.5-7.5, and more preferably pH5.5-7.0. Temperature can be set to, for example, 30°C-90°C, preferably 40°C-85°C, and more preferably 50°C-80°C. As the reaction time of Total, the action time can be set to, for example, 10 minutes-24 hours, preferably 20 minutes-12 hours, and more preferably 30 minutes-240 minutes. As the reaction time of the cyclodextrin generating enzyme alone, it can be set to, for example, 5 minutes-12 hours, preferably 10 minutes-6 hours, and more preferably 20 minutes-150 minutes. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0046] (3) Cyclodextrin-generating enzyme and α-amylase action process

[0047] The cyclodextrin generating enzyme and α-amylase action step are steps of making the cyclodextrin generating enzyme and α-amylase act on the vegetable raw material through the cyclodextrin generating enzyme action step. In the present technology, by making the cyclodextrin generating enzyme and α-amylase act on the vegetable raw material that has carried out the cyclodextrin generating enzyme treatment, the cyclodextrin content in the manufactured vegetable food and drink can be increased.

[0048] In addition, in the present technology, after the cyclodextrin generating enzyme action step, cyclodextrin generating enzyme and α-amylase may be added to perform the cyclodextrin generating enzyme and α-amylase action steps, but it is preferred that after the cyclodextrin generating enzyme action step, α-amylase is added without inactivating the cyclodextrin generating enzyme, thereby performing the cyclodextrin generating enzyme and α-amylase action steps. By adding α-amylase while continuing the cyclodextrin generating enzyme treatment of the plant raw material, the addition of each enzyme is completed at one time, so that the production process can be simplified and the workability can be improved.

[0049] The α-amylase that can be used in the present technology is an enzyme that acts on starch and mainly hydrolyzes α-1,4 glycosidic bonds. The α-amylase that can be used in the present technology may also have other actions as long as the α-amylase activity is the main activity.

[0050] The source of the α-amylase that can be used in the present technology is not particularly limited, and examples thereof include α-amylases derived from organisms of the genus Aspergillus (e.g., Aspergillus oryzae, Aspergillus niger, etc.), the genus Bacillus (e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.), preferably α-amylases derived from the genus Bacillus or α-amylases from Aspergillus, more preferably α-amylases derived from Bacillus amyloliquefaciens or α-amylases derived from Bacillus licheniformis.

[0051] The α-amylase that can be used in the present technology can be prepared by the culture solution of the microorganism that is the source of the above-mentioned α-amylase. As a specific preparation method, it can be easily prepared by culturing α-amylase-producing bacteria and isolating α-amylase using public means, and by using genetic recombination technology.

[0052] In the present technique, commercially available α-amylase may be used, and preferred examples of commercially available α-amylases include α-amylase (derived from Bacillus amyloliquefaciens) manufactured by Amano Enzyme Products Co., Ltd.

[0053] As long as the effect of the present technology is not impaired, the amount of α-amylase added in the cyclodextrin generating enzyme and α-amylase action step can be freely set. The lower limit of the specific amount of α-amylase added in the present technology can be set to, for example, 0.05U or more relative to 1g of the plant raw material, and from the viewpoint of further improving the effect of increasing the cyclodextrin content, it can be set to preferably 0.1U or more, more preferably 0.5U or more.

[0054] In addition, the lower limit of the amount of α-amylase added can be set to, for example, 0.1 U or more relative to 1 g of polysaccharides (preferably starch) used in the manufacture of plant-based food and beverages, and from the viewpoint of further improving the effect of increasing the cyclodextrin content, it can be set to preferably 0.2 U or more, more preferably 1 U or more.

[0055] As long as the effect of the present technology is not impaired, the upper limit of the addition amount of α-amylase is not particularly limited, and can be set, for example, to 1000U or less, 200U or less, 100U or less, preferably 50U or less, more preferably 10U or less, further preferably 5U or less, 3U or less, or 1U or less, relative to 1g of plant raw material.

[0056] In addition, the upper limit of the content of α-amylase can be set to, for example, 2000U or less, 400U or less, 200U or less, preferably 100U or less, more preferably 20U or less, further preferably 10U or less, 4U or less, relative to 1g of polysaccharides (preferably starch) used for the manufacture of plant-based food and beverage products.

[0057] In addition, in the present technology, the activity of α-amylase refers to a value measured by the measurement method described in the Examples described later.

[0058] In the step of acting on the cyclodextrin generating enzyme and α-amylase, the amount of the cyclodextrin generating enzyme added when it is added is preferably the specific amount of the cyclodextrin generating enzyme described in the description of the cyclodextrin generating enzyme action step, the sum of the amount added in the cyclodextrin generating enzyme action step and the amount added in the cyclodextrin generating enzyme and α-amylase action step.

[0059] As long as the effect of the present technology is not damaged, the various conditions of the cyclodextrin generating enzyme and the alpha-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 cyclodextrin generating enzyme and the alpha-amylase used. pH can be set to, for example, pH 5.0 to 8.0, preferably pH 5.5 to 7.5, and more preferably pH 6.0 to 7.0. The temperature can be set to, for example, 30°C to 90°C, preferably 40°C to 85°C, and more preferably 50°C to 80°C. The action time can be set to, for example, 5 minutes to 12 hours, preferably 10 minutes to 6 hours, and more preferably 20 minutes to 150 minutes. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0060] (4) Recycling process

[0061] The recovery process is a process for recovering the plant food and drink manufactured through the cyclodextrin generating enzyme action process and the cyclodextrin generating enzyme and α-amylase action process. The specific recovery method can be used according to the type of the manufactured plant food and drink, using one or a free combination of two or more general recovery methods in the manufacture of plant food and drink.

[0062] The plant-based food and drink manufactured through the cyclodextrin generating enzyme action step and the cyclodextrin generating enzyme and alpha-amylase action step is characterized in that the content of cyclodextrin increases. Specifically, for example, the content of cyclodextrin in the plant-based food and drink manufactured through the cyclodextrin generating enzyme action step and the cyclodextrin generating enzyme and alpha-amylase action step is more than 3.0mg relative to every 1mL plant-based food and drink, preferably more than 3.5mg, more preferably more than 4.0mg. That is, in the present technology, the content of cyclodextrin can be recovered relative to every 1mL plant-based food and drink for more than 3.0mg, preferably more than 3.5mg, more preferably more than 4.0mg, and more preferably the recovery process of the plant-based food and drink more than 5.0mg.

[0063] One embodiment of the method for producing a plant-based food and beverage and a method for increasing the cyclodextrin content of a plant-based food and beverage according to the present technology comprises the following steps (1) to (3). In addition, after step (3), a step (4) of inactivating an enzyme and / or a step (5) of recovering the plant-based food and beverage with an increased cyclodextrin content may be added.

[0064] (1) Process of preparing raw materials containing plant sugars

[0065] (2) Process of treating the prepared raw materials with cyclodextrin-generating enzyme

[0066] (3) A step of treating the raw material treated with cyclodextrin-generating enzyme with cyclodextrin-generating enzyme and α-amylase

[0067] One embodiment of the method for producing a plant-based food and beverage and a method for increasing the cyclodextrin content of a plant-based food and beverage according to the present technology comprises the following steps (1) to (3). In addition, after step (3), a step (4) of inactivating an enzyme and / or a step (5) of recovering the plant-based food and beverage with an increased cyclodextrin content may be added.

[0068] (1) Process of preparing raw materials containing plant sugars

[0069] (2) Process of treating the prepared raw materials with cyclodextrin-generating enzyme

[0070] (3) A step of adding α-amylase to the raw material being treated with the cyclodextrin-forming enzyme while continuing the step (2) to treat the raw material with the cyclodextrin-forming enzyme and the α-amylase.

[0071] In the step (5), a plant-based food or beverage having a cyclodextrin content of 3.0 mg or more per 1 mL of the plant-based food or beverage may be recovered.

[0072] 2. Cyclodextrin content increasing agent for plant-based food and beverage

[0073] The cyclodextrin content increasing agent of the plant-based food and beverage involved in the present technology includes a first agent containing a cyclodextrin generating enzyme and a second agent containing an alpha-amylase. For example, after adding a first agent to the plant-based raw material used for the manufacture of plant-based food and beverage for treatment based on a cyclodextrin generating enzyme, the second agent is added without inactivating the cyclodextrin generating enzyme, thereby enabling the cyclodextrin generating enzyme and alpha-amylase action process to be performed, as described above, the cyclodextrin content in the manufactured plant-based food and beverage can be increased. Below, the composition of the cyclodextrin content increasing agent is described in detail.

[0074] (1) First dose

[0075] The first dose contains a cyclodextrin generating enzyme. The first dose may also contain the cyclodextrin generating enzyme alone. That is, the first dose may also be the cyclodextrin generating enzyme itself. In addition, in the first dose, as long as it does not affect the production amount of cyclodextrin, it is also possible to select to contain one or more other enzymes that can be used for the manufacture of plant-based food and drink products. In addition, the details of the cyclodextrin generating enzyme are the same as the details of the cyclodextrin generating enzyme described in the manufacturing method of the above-mentioned plant-based food and drink products and the method for increasing the cyclodextrin content of the plant-based food and drink products, so the description is omitted here.

[0076] The content of the cyclodextrin generating enzyme in the first agent can be freely set as long as the effect of the present technology is not impaired. The content of the cyclodextrin generating enzyme is the same as the amount of the cyclodextrin generating enzyme added in the above-mentioned cyclodextrin generating enzyme action step, so the description thereof is omitted here.

[0077] (2) Second dose

[0078] The second dose contains α-amylase. The second dose may also contain α-amylase alone. That is, the second dose may also be α-amylase itself. In addition, in addition to α-amylase, the second dose may also contain a cyclodextrin generating enzyme. Furthermore, in the second dose, as long as it does not affect the production amount of cyclodextrin, it is also possible to select to contain one or more other enzymes that can be used for the manufacture of plant-based food and drink products. In addition, the details of α-amylase are the same as the details of α-amylase described in the above-mentioned method for manufacturing plant-based food and drink products and the method for increasing the cyclodextrin content of plant-based food and drink products, so the description is omitted here.

[0079] The content of α-amylase in the second agent can be freely set as long as the effect of the present technology is not impaired. The content of α-amylase is the same as the amount of α-amylase added in the above-mentioned cyclodextrin generating enzyme and α-amylase action step, so the description is omitted here.

[0080] (3) Others

[0081] As long as the effect of the present technology is not damaged, it is also possible to freely select to contain one or more other components in the first dose and the second dose of the cyclodextrin content increaser of the plant-based diet product involved in the present technology. As other components, it is possible to use components such as excipients, pH adjusting agents, coloring agents, flavoring agents, disintegrants, lubricants, stabilizers, etc., which are used for conventional formulations. And then, it is also possible to appropriately and use the components with functions disclosed or found in the future according to the purpose.

[0082] In addition, as long as it does not affect the amount of cyclodextrin produced, the cyclodextrin content increasing agent for the plant-based food and beverage involved in the present technology may also contain a third agent, a fourth agent, etc. that are different from the first and second agents and contain one or more other enzymes that can be used in the manufacture of plant-based food and beverages, or functional ingredients that are publicly known or discovered in the future.

[0083] 3. Plant-based food

[0084] The plant-based food and beverages involved in the present technology are plant-based food and beverages manufactured using the above-mentioned cyclodextrin content increasing agent. As a specific example of a plant-based beverage, oat beverages can be cited, and more specifically, oat milk (also known as "oat milk") can be cited. As a specific example of a plant-based food, plant-based yogurt can be cited.

[0085] The amount of cyclodextrin in the plant-based food and drink involved in the present technology increases, and by the increase of cyclodextrin, it is possible to mask peculiar smell. The amount of cyclodextrin contained in the plant-based food and drink involved in the present technology is not particularly limited either. As the amount of cyclodextrin contained in the plant-based food and drink, in terms of the total amount of α-cyclodextrin, relative to each 1mL of the plant-based food and drink, for example, 3.0 mg or more, preferably 3.5 mg or more, more preferably 4.0 mg or more, and further preferably 4.5 mg or more.

[0086] Example

[0087] Hereinafter, the present invention will be described in more detail based on examples. In addition, the examples described below are examples of representative embodiments of the present invention, but the scope of the present invention is not to be narrowly interpreted thereby.

[0088] 1. Use of enzymes

[0089] Cyclodextrin-generating enzyme (CGT): Amano Enzyme Co., Ltd. Cyclodextrin glycosyltransferase from Anoxybacillus caldiproteolyticus (formerly classified as Geobacillus stearothermophilus)

[0090] α-Amylase (AMY): α-amylase from Bacillus amyloliquefaciens manufactured by Amano Enzyme Co., Ltd.

[0091] 2. Enzyme activity assay

[0092] [Method for measuring the activity of cyclodextrin-generating enzyme]

[0093] The activity of the cyclodextrin-forming enzyme can be measured by a method based on the cyclodextrin-forming enzyme activity test method in the 9th edition of the Food Additive Code.

[0094] Add 20 mL of water to 1.0 g of potato starch, and slowly add 5 mL of sodium hydroxide test solution (1 mol / L) while stirring to make a paste. After heating in a boiling water bath for 3 minutes while stirring, add 25 mL of water, cool with running water, adjust the pH to 5.5 with acetic acid test solution (1 mol / L), and add water to make a 100 mL solution as the substrate solution. Measure 10 mL of the substrate solution, heat at 40°C for 10 minutes, add 1 mL of the enzyme solution and mix. After incubating at 40°C for 10 minutes, add 10 mL of hydrochloric acid test solution (0.1 mol / L) to stop the reaction. Add 10 mL of iodine-potassium iodide test solution (0.4 mmol / L) to 1 mL of the reaction solution and mix to make a test solution. In addition, the iodine-potassium iodide test solution is prepared by dissolving 10.0 g of potassium iodide and 1.0 g of iodine in water, and diluting 200 times with water after making 100 mL. Water was used as a comparative solution instead of the reaction solution, and the absorbance of the test solution and the comparative solution at 660 nm was measured. The amount of enzyme that reduced the blue iodine color of starch by 1% in 1 minute was defined as 1 unit (U).

[0095] [Method for determining the activity of α-amylase]

[0096] After heating 10 mL of 1% potato starch matrix solution (0.1 mol / L acetic acid (pH 5.0)) at 37°C for 10 minutes, add 1 mL of the sample solution containing α-amylase and immediately shake and mix. After the liquid is placed at 37°C for 10 minutes, 1 mL of the liquid is added to 10 mL of 0.1 mol / L hydrochloric acid test solution and immediately shake and mix. Next, 0.5 mL of the liquid is measured, 10 mL of 0.0002 mol / L iodine test solution (Japanese Bureau) is added, and after shaking and mixing, the absorbance (AT) at a wavelength of 660 nm is measured with water as a control. In addition, 1 mL of water is added instead of the sample solution, and the same operation is performed to measure the absorbance (AB). In addition, 0.0002 mol / L iodine test solution (Japanese Bureau) is prepared as follows: 10 mL of water is added to 12.7 g of iodine and 25 g of potassium iodide and mixed thoroughly, water is added to make 100 mL, and then diluted 2500 times with water. The α-amylase activity was calculated using the following mathematical formula: 1 unit (1 U) was defined as the amount of enzyme that reduced the color development of iodine based on potato starch by 10% within 1 minute.

[0097] α-amylase activity (U / g, U / mL) = {(AB-AT) / AB} × 1 / W

[0098] AT: absorbance of the reaction solution

[0099] AB: absorbance of blank solution

[0100] W: The amount of sample in 1 mL of sample solution (g or mL)

[0101] 3. Experimental Examples

[0102] (1) Experimental methods

[0103] 100 mL of water was added to 12.5 g of oat flour (PREMIUM OAT FLOUR, sugar content 67.7 g / 100 g, manufactured by Slow Food Kitchen) and suspended, and cyclodextrin generating enzyme (CGT) was added thereto in an amount corresponding to the amount listed in Table 1 relative to the raw materials. After the enzyme was added, the enzyme was allowed to act for 0 to 60 minutes while stirring in a 60°C water bath. Next, α-amylase (AMY) was added to the cyclodextrin generating enzyme reaction solution in an amount corresponding to the amount listed in Table 1 relative to the raw materials, and the enzyme was allowed to react for 30 to 120 minutes while stirring in a 60°C water bath. After the reaction, the enzyme was inactivated by treating at 95°C for 10 minutes. After the enzyme was inactivated for the quantitative determination of α-cyclodextrin, 1% of glucoamylase was added, the enzyme was allowed to act at 60°C for 2 hours, and the enzyme was inactivated by treating at 95°C for 10 minutes. 190 μL of 1 mol / L trichloroacetic acid was added to 1.5 mL of the enzyme-inactivated oatmeal solution and allowed to stand at room temperature for 10 minutes. After 10 minutes, centrifugation was performed (12,000 rpm, 10 minutes) to recover the supernatant. An equal amount of diethyl ether was added to the recovered supernatant and suspended. Centrifugation was performed (12,000 rpm, 10 minutes), the organic solvent layer was discarded, and the water layer was recovered. This operation was repeated twice. An equal amount of 99.5% ethanol (for high performance liquid chromatography, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added to the recovered water layer and stirred, and the substance filtered with a membrane filter was provided for analysis. High performance liquid chromatography was used for the analysis, and the chromatographic column used was MCL GEL CK04S manufactured by Mitsubishi Chemical Corporation. The amount of α-cyclodextrin produced was quantified based on the peak area. In addition, for the viscosity of plant-based food and beverages, the case where the viscosity was significantly higher than that of water when stirred with a scraper was evaluated as "+", and the case where the viscosity was close to that of water was evaluated as "-".

[0104] (2) Results

[0105] The results are shown in Table 1 below.

[0106] [Table 1]

[0107]

[0108] (3) Investigation

[0109] As shown in samples 1-1 to 1-3, 2-1 and 2-2 of Table 1, by using cyclodextrin generating enzyme and α-amylase in combination, liquefaction can be fully performed and viscosity can be reduced, but the content of α-cyclodextrin in the plant-based food and drink is reduced in proportion to the amount of α-amylase added. On the other hand, as shown in samples 3-1, 3-2, 4-1 and 4-2 of Table 1, before the treatment of cyclodextrin generating enzyme and α-amylase, only the cyclodextrin generating enzyme is used to treat the plant-based raw material, thereby significantly suppressing the reduction of the content of α-cyclodextrin, and although liquefaction is fully performed, it is possible to confirm the same α-cyclodextrin content increase effect as that without the addition of α-amylase.

Claims

1. A method for producing a plant-based food and beverage, comprising: A cyclodextrin-generating enzyme action step of allowing the cyclodextrin-generating enzyme to act on the plant raw material; as well as After the cyclodextrin-forming enzyme action step, a cyclodextrin-forming enzyme and α-amylase action step is performed in which the cyclodextrin-forming enzyme and α-amylase are allowed to act on the plant-based raw material.

2. A method for increasing the cyclodextrin content of a plant-based food product, comprising: A cyclodextrin-generating enzyme action step of allowing the cyclodextrin-generating enzyme to act on the plant raw material; as well as After the cyclodextrin-forming enzyme action step, a cyclodextrin-forming enzyme and α-amylase action step is performed in which the cyclodextrin-forming enzyme and α-amylase are allowed to act on the plant-based raw material.

3. A cyclodextrin content increasing agent for a plant-based food or beverage, comprising: A first dose containing a cyclodextrin-generating enzyme; and The second dose contains alpha-amylase.

4. A plant-based food or beverage using the cyclodextrin content-increasing agent according to claim 3.

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