Method for improving foamability of plant-based foods and beverages

Abstract

To provide a technique for improving the foamability of plant-based foods and beverages.　The present technology provides a method for improving foamability of plant-based foods and beverages, the method comprising: a cyclodextrin-producing enzymatic action step in which a cyclodextrin-producing enzyme is caused to act on a plant-based raw material; and a protein deamidation step in which a protein deamidase is applied.

Description

Method for improving foaming properties of plant-based foods and drinks

[0001] The present technology relates to a method for improving the foaming property of a plant-based food or drink, more specifically, the present technology relates to a method for improving the foaming property of a plant-based food or drink, a foaming property enhancer for a plant-based food or drink, and a plant-based food or drink.

[0002] In recent years, growing health consciousness has led to increased attention being paid to plant-based foods and beverages. For example, oat beverages are prepared by suspending oat material in an aqueous medium and hydrolyzing the starch in the oat material using one or more amylases. The amylase used here is β-amylase or a mixture of α-amylase and β-amylase. Oat beverages prepared by this method exhibit a moderate sweetness due to the production of maltose by the action of β-amylase (Patent Document 1). Furthermore, because plant-based milks such as soy milk have inferior foaming stability compared to cow's milk, techniques using various additives have been investigated (Patent Document 2).

[0003] Here, the enzymes used in this technology will be described. Cyclodextrin-forming enzymes act on starch to produce α-, β-, and γ-cyclodextrins. It is also known that cyclodextrin-forming enzymes act on starch to produce cyclodextrins without liquefying starch granules (Patent Document 3).

[0004] JP 2018-075029 A JP 2020-054379 A JP 09-262091 A

[0005] As mentioned above, plant-based foods and beverages that have been attracting attention in recent years have inferior foaming properties compared to milk, and there is a demand for improved foaming properties.

[0006] Therefore, the main object of this technology is to provide a technology for improving the foaming properties of plant-based foods and beverages.

[0007] As a result of intensive research into technologies for improving the foaming properties of plant-based foods and beverages, the inventors of the present application succeeded in improving the foaming properties of plant-based foods and beverages by applying cyclodextrin-forming enzymes and protein deamidating enzymes to the plant-based raw materials that are the raw materials for plant-based foods and beverages, and thus completed this technology.

[0008] That is, the present technology first provides a method for improving the foaming properties of a plant-based food or beverage, which includes a cyclodextrin-forming enzyme treatment step in which a cyclodextrin-forming enzyme is allowed to act on a composition containing a plant-based raw material, and a protein deamidating enzyme treatment step in which a protein deamidating enzyme is allowed to act on the composition. The method for improving the foaming properties of a plant-based food or beverage according to the present technology may also include an α-amylase treatment step in which an α-amylase is allowed to act on the composition containing the plant-based raw material. The method for improving the foaming properties of a plant-based food or beverage according to the present technology may also include an oil / fat addition step in which an oil / fat is added.

[0009] The present technology further provides a foaming enhancer for plant-based foods and beverages, which comprises a cyclodextrin-forming enzyme and a protein deamidating enzyme.

[0010] Additionally, the present technology provides plant-based foods and beverages that use the foaming enhancer according to the present technology.

[0011] According to the present technology, the foaming properties of plant-based foods and beverages can be improved.

[0012] Preferred embodiments for carrying out the present technology will be described below. Note that the embodiments described below are examples of typical embodiments of the present technology, and the scope of the present technology should not be construed as being narrow.

[0013] 1. Method for Improving the Foaming Properties of Plant-Based Foods and Beverages The method for improving the foaming properties of plant-based foods and beverages according to the present technology includes a step of treating a plant-based raw material with a cyclodextrin-forming enzyme (hereinafter also referred to as the "cyclodextrin-forming enzyme treatment step") and a step of treating a plant-based raw material with a protein deamidating enzyme (hereinafter also referred to as the "protein deamidating enzyme treatment step"). The method for improving the foaming properties of plant-based foods and beverages according to the present technology preferably further includes a step of treating a plant-based raw material with α-amylase (hereinafter also referred to as the "α-amylase treatment step"). Furthermore, the method for improving the foaming properties of plant-based foods and beverages according to the present technology preferably further includes an oil / fat addition step of adding an oil / fat. Depending on the type of plant-based food or beverage, general food manufacturing processes can be performed before, after, or simultaneously with each step, as long as the effects of the present technology are not impaired. Furthermore, the method for improving the foaming properties of plant-based foods and beverages can further include a recovery step of recovering the plant-based food or beverage with improved foaming properties. The raw materials used and each step are described in detail below.

[0014] (1) Plant-based raw materials The plant-based raw materials that can be used in the present technology are not particularly limited in terms of origin or type, and can be freely selected depending on the intended plant-based food or beverage, as long as they do not impair the effects of the present technology. Examples include beans such as soybeans, green peas, lentils, chickpeas, black beans, broad beans, mung beans, lupin beans, and kidney beans; grains such as wheat, barley, oats, rice, rye, buckwheat, barnyard millet, foxtail millet, and teff; nuts such as almonds, coconuts, peanuts, cashew nuts, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, and pine nuts; hemp seeds (industrial hemp), chia seeds, quinoa, amaranth, canary seeds, and flaxseed. In the present technology, these may be used alone or in combination of two or more. Among these raw materials, preferred are cereals, and more preferred are oats.

[0015] In the present technology, the state of the plant raw material when subjected to various enzyme treatments is not particularly limited as long as it does not impair the effects of the present technology, but preferably, the plant raw material is in a liquid or slurry state.

[0016] The plant-derived raw material that can be used in the present technology contains carbohydrates. The content of the plant-derived carbohydrates contained in the plant-derived raw material (based on the weight of the plant-derived raw material in a dry state) is not particularly limited, but may be, for example, 20% by weight or more. Preferably, it is 30% by weight or more, more preferably 40% by weight or more, and even more preferably 50% by weight or more. The upper limit of the content range is not particularly limited, but may be, for example, 90% by weight or less, 80% by weight or less, or 70% by weight or less.

[0017] (2) Cyclodextrin-forming enzyme action step The cyclodextrin-forming enzyme action step is a step in which a cyclodextrin-forming enzyme is allowed to act on a plant-based raw material. In the cyclodextrin-forming enzyme action step, it is preferable to allow the cyclodextrin-forming enzyme to act alone on the plant-based raw material, but one or more other enzymes that can be used in the production of plant-based foods and beverages can be selected and used in combination, as long as the amount of cyclodextrin produced is not affected.

[0018] The cyclodextrin-forming enzyme that can be used in the present technology is an enzyme that has the activity of producing cyclodextrin with a degree of polymerization of 6 to 8 using a polysaccharide having an α-1,4-linked glucose structure, such as starch, as a substrate. An example of the cyclodextrin-forming enzyme is cyclodextrin glucanotransferase (EC 2.4.1.19). The cyclodextrin-forming enzyme that can be used in the present technology may also be an enzyme that has other functions as long as it has the activity of producing cyclodextrin. It is preferable that the activity of producing cyclodextrin is the main activity.

[0019] In this technology, the foaming properties of plant-derived raw materials can be improved by allowing cyclodextrin-forming enzymes to act on the plant-derived raw materials.

[0020] The origin of the cyclodextrin-forming enzyme that can be used in the present technology is not particularly limited, and examples thereof include the genus Bacillus, such as Bacillus stearothermophilus, Bacillus megaterium, Bacillus circulans, Bacillus macerans, Bacillus ohbensis, and Bacillus clarkii; the genus Geobacillus, such as Geobacillus stearothermophilus; stearothermophilus; the genus Paenibacillus, for example Paenibacillus macerans; the genus Klebsiella, for example Klebsiella pneumoniae; the genus Anoxybacillus, for example Anoxybacillus caldiproteolyticus, Anoxybacillus pashukinoensis, Examples of cyclodextrin-producing enzymes include those derived from the genera Thermoanaerobacter, Brevibacterium, Bacillus pushchinoensis, Anoxybacillus flavisthermus, Thermoanaerobacter, and Brevibacterium. These cyclodextrin-producing enzymes may be used singly or in combination.Among these, the cyclodextrin-forming enzyme is preferably cyclodextrin glucanotransferase derived from a microorganism of the genus Paenibacillus and / or cyclodextrin glucanotransferase derived from a microorganism of the genus Anoxybacillus, and more preferably cyclodextrin glucanotransferase derived from Paenibacillus macerans and / or cyclodextrin glucanotransferase derived from Anoxybacillus caldiproteolyticus (including the strain formerly known as Geobacillus stearothermophilus).

[0021] Here, "cyclodextrin-producing enzyme derived from Paenibacillus macerans" and / or "cyclodextrin-producing enzyme derived from Anoxybacillus caldiproteolyticus" means a cyclodextrin-producing enzyme produced by a microorganism (whether a wild-type strain or a mutant strain) classified as Paenibacillus macerans and / or Geobacillus stearothermophilus, or a cyclodextrin-producing enzyme obtained by genetic engineering techniques using a cyclodextrin-producing enzyme gene. Therefore, recombinants produced by a host microorganism into which a cyclodextrin-producing enzyme gene obtained from Paenibacillus macerans and / or Anoxybacillus caldiproteolyticus (or a modified version of said gene) has been introduced also fall under the category of "cyclodextrin-producing enzyme derived from Paenibacillus macerans" and / or "cyclodextrin-producing enzyme derived from Anoxybacillus caldiproteolyticus."

[0022] The cyclodextrin-forming enzyme used in the present technology can be prepared from the culture broth of the microorganism from which the cyclodextrin-forming enzyme is derived. Specific preparation methods include methods in which the cyclodextrin-forming enzyme is recovered from the culture broth or cells of the microorganism. For example, when a cyclodextrin-forming enzyme-secreting microorganism is used, the cells can be recovered from the culture broth, if necessary, by filtration, centrifugation, or the like, and the enzyme can then be separated and / or purified. When a cyclodextrin-forming enzyme-nonsecreting microorganism is used, the cells can be recovered from the culture broth, if necessary, and then disrupted by pressure treatment, ultrasonic treatment, or the like to extract the enzyme, which can then be separated and / or purified. The enzyme can be separated and / or purified by any known protein separation and / or purification method, without any particular limitation, and examples thereof include centrifugation, UF concentration, salting out, and various chromatography methods using ion exchange resins, etc. The isolated and / or purified enzyme can be powdered by a drying method such as freeze-drying or vacuum drying, or can be powdered using an appropriate excipient and / or drying aid in the drying method. The isolated and / or purified enzyme can also be liquefied by adding an appropriate additive and sterilizing by filtration.

[0023] In the present technology, commercially available cyclodextrin-producing enzymes can also be used, and preferred examples of commercially available products include cyclodextrin glucanotransferase derived from Paenibacillus macerans, manufactured by Amano Enzyme Inc., and cyclodextrin glucanotransferase derived from Anoxybacillus caldiproteolyticus (Geobacillus stearothermophilus), also manufactured by Amano Enzyme Inc.

[0024] The amount of cyclodextrin-forming enzyme added in the cyclodextrin-forming enzyme reaction step can be freely set as long as it does not impair the effects of the present technology. The lower limit of the amount of cyclodextrin-forming enzyme added can be set to, for example, 0.01 U or more per 1 g of plant-based raw material, and from the viewpoint of further enhancing the foaming properties of the plant-based food or beverage, it can be set to preferably 0.03 U or more, more preferably 0.1 U or more, even more preferably 0.3 U or more, and even more preferably 1 U or more, 3 U or more, 5 U or more, or 7 U or more.

[0025] The lower limit of the amount of cyclodextrin-forming enzyme to be added can be set to, for example, 0.02 U or more per gram of polysaccharide (preferably starch) contained in the plant-based raw material, and from the viewpoint of further enhancing the foaming properties of the plant-based food or beverage, the amount can be set to preferably 0.05 U or more, more preferably 0.5 U or more, even more preferably 2 U or more, and even more preferably 5 U or more, or 10 U or more.

[0026] The upper limit of the amount of cyclodextrin-forming enzyme to be added is not particularly limited as long as it does not impair the effects of the present technology, but 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 plant-derived raw material.

[0027] The upper limit of the amount of cyclodextrin-forming enzyme to be added can be set to, for example, 300 U or less, 200 U or less, 100 U or less, 50 U or less, 30 U or less, or 20 U or less per gram of polysaccharides (preferably starch) contained in the plant raw material.

[0028] In the present technology, the activity of the cyclodextrin-forming enzyme is a value measured by the measurement method described in the Examples below.

[0029] The various conditions for the cyclodextrin-forming enzyme reaction step can be freely set as long as they do not impair the effects of the present technology. For example, the pH, temperature, reaction time, etc. can be set depending on the physicochemical properties of the cyclodextrin-forming enzyme used, such as the optimal pH, stable pH range, optimal temperature, and temperature stability. The pH can be set, for example, to 5.0 to 8.0, preferably 5.5 to 7.5, and more preferably 5.5 to 7.0. The temperature can be set, for example, to 30°C to 90°C, preferably 40°C to 80°C, and more preferably 50°C to 70°C. The reaction time, in terms of total reaction time, can be set, for example, to 5 minutes to 12 hours, preferably 10 minutes to 6 hours, and more preferably 20 minutes to 120 minutes. The optimal reaction conditions can be determined through preliminary experiments.

[0030] (3) Protein deamidating step The protein deamidating step is a step of treating a plant-based raw material with a protein deamidating enzyme. In this technology, treating a plant-based raw material with a protein deamidating enzyme can further improve the foaming properties of the produced plant-based food or beverage.

[0031] The timing of the protein deamidating enzyme action step is not particularly limited, and the protein deamidating enzyme action step may be carried out before or after the cyclodextrin-forming enzyme action step. The protein deamidating enzyme action step may be carried out simultaneously with the cyclodextrin-forming enzyme action step. Alternatively, the protein deamidating enzyme action step may be carried out in multiple batches. It is preferable to proceed with the protein deamidating enzyme action step by adding a protein deamidating enzyme without inactivating the cyclodextrin-forming enzyme after the cyclodextrin-forming enzyme action step has been carried out.

[0032] Protein deamidases that can be used in the present technology are enzymes that exhibit the action of degrading amide group-containing side chains of proteins without cleaving peptide bonds or crosslinking proteins. Protein deamidases that can be used in the present technology may also have other actions as long as they have protein deamidase activity. It is preferable to use an enzyme whose main activity is protein deamidase activity.

[0033] Examples of protein deamidating enzymes include enzymes that deamidate glutamine residues in proteins to convert them to glutamic acid (e.g., protein glutaminase) and enzymes that deamidate asparagine residues in proteins to convert them to aspartic acid (e.g., protein asparaginase). These protein deamidating enzymes may be used alone or in combination of two or more. Among these protein deamidating enzymes, protein glutaminase is preferred from the viewpoint of further enhancing the foaming property-improving effect.

[0034] The origin of protein deamidase that can be used in the present technology is not particularly limited, and examples thereof include protein deamidases derived from the genus Chryseobacterium, Flavobacterium, Empedobacter, Sphingobacterium, Aureobacterium, Myroides, Luteimicrobium, Agromyces, Microbacterium, or Leifsonia. These protein deamidating enzymes are known, and reference can be made to, for example, JP2000-50887A, JP2001-218590A, WO2006 / 075772A1, WO2015 / 133590, etc.

[0035] Protein deamidase that can be used in the present technology can be prepared from a culture medium of the microorganism from which the above-mentioned protein deamidase is derived. Specific preparation methods include culturing a protein deamidase-producing bacterium and isolating the protein deamidase by known means, or by using genetic recombination technology, and the like.

[0036] In the present technology, a commercially available protein deamidase can also be used, and a preferred example of a commercially available protein deamidase is protein glutaminase (derived from the species Chryseobacterium proteolyticum) manufactured by Amano Enzyme Inc.

[0037] The amount of protein deamidase added in the protein deamidase reaction step can be freely set as long as it does not impair the effects of the present technology. A specific lower limit of the amount of protein deamidase added in the present technology can be set to, for example, 0.01 U or more, preferably 0.1 U or more, 0.5 U or more, and more preferably 1 U or more per 1 g of plant-derived raw material.

[0038] The lower limit of the amount of protein deamidase to be added can be set to, for example, 0.05 U or more, preferably 0.2 U or more, 0.5 U or more, or 1 U or more, more preferably 3 U or more, 5 U or more, or 7 U or more, relative to 1 g of protein contained in the plant raw material.

[0039] The upper limit of the amount of protein deamidase to be added is not particularly limited as long as it does not impair the effects of the present technology, and can be set to, for example, 1000 U or less, 200 U or less, 100 U or less, preferably 50 U or less, 20 U or less, more preferably 10 U or less, and even more preferably 5 U or less, 3 U or less, or 2 U or less per 1 g of plant raw material.

[0040] The upper limit of the content of protein deamidase can be set to, for example, 2000 U or less, 400 U or less, 200 U or less, preferably 100 U or less, more preferably 50 U or less, and even more preferably 30 U or less, 20 U or less, or 15 U or less, per 1 g of protein contained in the plant raw material.

[0041] In the present technology, the activity of protein deamidase is a value measured by the measurement method described in the Examples below.

[0042] Various conditions for the protein deamidase reaction step can be freely set as long as they do not impair the effects of the present technology. For example, the pH, temperature, reaction time, etc. can be set depending on the physicochemical properties of the protein deamidase used, such as the optimal pH, stable pH range, optimal temperature, and temperature stability. The pH can be set, for example, to 5.0 to 8.0, preferably 5.5 to 7.5, and more preferably 6.0 to 7.0. The temperature can be set, for example, to 30°C to 90°C, preferably 35°C to 80°C, and more preferably 40°C to 70°C. The reaction time can be set, for example, to 5 minutes to 12 hours, preferably 10 minutes to 6 hours, and more preferably 20 minutes to 180 minutes. Optimal reaction conditions can be determined through preliminary experiments.

[0043] (4) α-amylase treatment step The α-amylase treatment step is a step in which α-amylase is treated with a plant-based raw material. In this technology, by treating the plant-based raw material with α-amylase, the viscosity of the produced plant-based food or drink can be reduced.

[0044] The timing of the α-amylase action step is not particularly limited, and it may be performed before or after the cyclodextrin-forming enzyme action step. It may be performed simultaneously with the cyclodextrin-forming enzyme action step. It may also be performed before or after the protein deamidating enzyme action step. It may also be performed simultaneously with the protein deamidating enzyme action step. Furthermore, it may be performed in multiple separate steps. After the cyclodextrin-forming enzyme action step has been performed, it is preferable to proceed with the protein deamidating enzyme action step without inactivating the cyclodextrin-forming enzyme by adding α-amylase at the same time.

[0045] The α-amylase that can be used in this technology is an enzyme that acts on starch and mainly hydrolyzes α-1,4 glycosidic bonds. The α-amylase that can be used in this technology may also have other functions as long as it has α-amylase activity. It is preferable to use an enzyme whose main activity is α-amylase activity.

[0046] The origin 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.) and the genus Bacillus (e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.), preferably α-amylases derived from the genus Bacillus or Aspergillus, more preferably α-amylases derived from Bacillus amyloliquefaciens or Bacillus licheniformis. Examples of suitable α-amylases include those derived from Bacillus licheniformis.

[0047] The α-amylase that can be used in the present technology can be prepared from the culture medium of the microorganism from which the α-amylase is derived. Specific preparation methods include culturing an α-amylase-producing bacterium and isolating the α-amylase using known means, or by using genetic recombination techniques.

[0048] In the present technology, commercially available α-amylase can also be used, and a preferred example of a commercially available α-amylase is α-amylase (derived from Bacillus amyloliquefaciens) manufactured by Amano Enzyme Inc.

[0049] The amount of α-amylase added in the α-amylase reaction step can be freely set as long as it does not impair the effects of the present technology. A specific lower limit of the amount of α-amylase added in the present technology can be set to, for example, 0.01 U or more, preferably 0.1 U or more, 0.5 U or more, 1 U or more, more preferably 2 U or more, 3 U or more per 1 g of plant raw material.

[0050] The lower limit of the amount of α-amylase to be added can be set to, for example, 0.05 U or more, preferably 0.2 U or more, 0.5 U or more, or 1 U or more, more preferably 2 U or more, 3 U or more, or 4 U or more, per 1 g of polysaccharides (preferably starch) contained in the plant raw material.

[0051] The upper limit of the amount of α-amylase to be added is not particularly limited as long as it does not impair the effects of the present technology, and can be set to, for example, 1000 U or less, 200 U or less, 100 U or less, preferably 50 U or less, 20 U or less, more preferably 10 U or less, and even more preferably 5 U or less, 3 U or less, or 1 U or less per 1 g of plant raw material.

[0052] The upper limit of the α-amylase content can be set to, for example, 2000 U or less, 400 U or less, 200 U or less, preferably 100 U or less, more preferably 30 U or less, and even more preferably 10 U or less, 4 U or less, or 2 U or less per gram of polysaccharides (preferably starch) contained in the plant raw material.

[0053] In the present technology, the α-amylase activity is a value measured by the measurement method described in the Examples below.

[0054] Various conditions for the α-amylase reaction step can be freely set as long as they do not impair the effects of the present technology. For example, the pH, temperature, reaction time, etc. can be set depending on the physicochemical properties of the α-amylase used, such as the optimal pH, stable pH range, optimal temperature, and temperature stability. The pH can be set, for example, to 5.0 to 8.0, preferably 5.5 to 7.5, and more preferably 6.0 to 7.0. The temperature can be set, for example, to 30°C to 90°C, preferably 35°C to 80°C, and more preferably 40°C to 70°C. The reaction time can be set, for example, to 5 minutes to 12 hours, preferably 10 minutes to 6 hours, and more preferably 20 minutes to 180 minutes. The optimal reaction conditions can be determined through preliminary experiments.

[0055] (5) Oil / Fat Addition Step The oil / fat addition step is a step of adding oil / fat. The oil / fat addition step also includes using a raw material containing oil / fat as the plant-based raw material. The added oil / fat may be of the same origin as the plant-based raw material or may be of a different origin. The inclusion of oil / fat can further improve foaming properties. The origin, type, etc. of the oil / fat are not particularly limited and can be freely selected depending on the desired plant-based food or drink. Examples include soybean oil, canola oil, sunflower oil, corn oil, olive oil, rice oil, palm oil, safflower oil, cottonseed oil, etc.

[0056] The timing of the fat / oil addition step is not particularly limited, and the fat / oil addition step may be carried out before or after the cyclodextrin-forming enzyme action step. It may also be carried out simultaneously with the cyclodextrin-forming enzyme action step. Furthermore, the fat / oil addition step may be carried out before or after the protein deamidating enzyme action step. It may also be carried out simultaneously with the protein deamidating enzyme action step. Furthermore, when an α-amylase action step is carried out, the fat / oil addition step may be carried out before or after the α-amylase action step. It may also be carried out simultaneously with the α-amylase action step.

[0057] (6) Recovery Step The recovery step is a step of recovering the plant-based food or beverage with improved foaming properties produced through the cyclodextrin-forming enzyme action step and the protein deamidating enzyme action step. Specific recovery methods can be selected from one or a combination of two or more general recovery methods used in the production of plant-based food or beverage, depending on the type of plant-based food or beverage to be produced.

[0058] Plant-based foods and beverages produced through a cyclodextrin-forming enzyme action step and a protein deamidating enzyme action step are characterized by improved foaming properties. Specifically, for example, the foaming properties of a plant-based food and beverage produced through a cyclodextrin-forming enzyme action step and a protein deamidating enzyme action step are improved by 10% or more, preferably 20% or more, and more preferably 30% or more, compared to a plant-based food and beverage produced by a similar method except that the cyclodextrin-forming enzyme action step and the protein deamidating enzyme action step are not performed. That is, the present technology can perform a recovery step to recover a plant-based food and beverage with improved foaming properties of 10% or more, preferably 20% or more, and more preferably 30% or more.

[0059] One embodiment of the method for improving the foaming properties of a plant-based food or drink according to the present technology includes the following steps (1) to (3). Note that a step (4) of treating with α-amylase may be carried out before, after, or simultaneously with step (2) or (3). Furthermore, a step (5) of inactivating the enzyme may be added after steps (2) to (4). A step (6) of adding oils or fats may be added before or after steps (2) to (5). A step (7) of recovering the plant-based food or drink with improved foaming properties may be added after steps (2) to (6). (1) A step of preparing a raw material containing plant carbohydrates; (2) A step of treating the prepared raw material with a cyclodextrin-forming enzyme; and (3) A step of treating the prepared raw material with a protein deamidating enzyme.

[0060] One embodiment of the method for improving the foaming property of a plant-based food or beverage according to the present technology includes the following steps (1) to (3). After step (3), the following steps may be added: (4) a step of inactivating an enzyme; (5) a step of adding an oil or fat; and / or (6) a step of recovering a plant-based food or beverage with improved foaming property. The timing of the enzyme inactivation step (4) is not limited as long as it is performed after step (3). The timing of the oil or fat addition step (5) is not limited as long as it is performed after step (1). (1) A step of preparing a raw material containing plant-based carbohydrates and an oil or fat; (2) A step of treating the prepared raw material with a cyclodextrin-forming enzyme; and (3) A step of adding a protein deamidating enzyme and an α-amylase to the raw material being treated with the cyclodextrin-forming enzyme while continuing step (2), and treating the raw material with the protein deamidating enzyme and the α-amylase. In step (6), a plant-based food or beverage with improved foaming property of 10% or more can also be recovered.

[0061] 2. Foaming Enhancer for Plant-Based Foods and Beverages The foaming enhancer for plant-based foods and beverages according to the present technology contains a cyclodextrin-forming enzyme and a protein deamidating enzyme. For example, by adding a foaming enhancer containing a cyclodextrin-forming enzyme and a protein deamidating enzyme to the plant-based raw materials used in the production of plant-based foods and beverages, the cyclodextrin-forming enzyme action step and the protein deamidating enzyme action step can be promoted, thereby improving the foaming properties of the plant-based foods and beverages produced, as described above. The composition of the foaming enhancer is described in detail below.

[0062] (1) Cyclodextrin-forming enzyme The foaming improver for plant-based foods and beverages according to the present technology contains a cyclodextrin-forming enzyme. Note that the details of the cyclodextrin-forming enzyme are the same as those described in the method for improving the foaming property of plant-based foods and beverages, and therefore will not be described here.

[0063] The amount of cyclodextrin-forming enzyme in the foaming enhancer for plant-based foods and beverages according to the present technology can be freely set as long as it does not impair the effects of the present technology. The amount of cyclodextrin-forming enzyme added is the same as the amount of cyclodextrin-forming enzyme added in the above-mentioned cyclodextrin-forming enzyme action step, so further explanation is omitted here.

[0064] (2) Protein deamidating enzyme The foaming enhancer for plant-based foods and beverages according to the present technology contains a protein deamidating enzyme. Note that the details of the protein deamidating enzyme are the same as those described in the method for improving the foaming property of plant-based foods and beverages, and therefore will not be described here.

[0065] The content of protein deamidating enzyme in the foaming enhancer for plant-based foods and beverages according to the present technology can be freely set as long as it does not impair the effects of the present technology. The content of protein deamidating enzyme is the same as the amount of protein deamidating enzyme added in the above-mentioned protein deamidating enzyme action step, and therefore, a description thereof will be omitted here.

[0066] (3) α-amylase The foaming property improver for plant-based foods and beverages according to the present technology can also contain α-amylase. The use of α-amylase can reduce the viscosity of the plant-based foods and beverages. Details of the α-amylase are the same as those described in the method for improving the foaming property of plant-based foods and beverages, and therefore will not be described here.

[0067] The amount of α-amylase contained in the foaming enhancer for the plant-based food or beverage according to the present technology can be freely set as long as it does not impair the effects of the present technology. The amount of α-amylase contained is the same as the amount of α-amylase added in the above-mentioned α-amylase action step, so a detailed explanation is omitted here.

[0068] The foaming improver for plant-based foods and beverages according to the present technology preferably comprises a first agent containing a cyclodextrin-forming enzyme, a second agent containing a protein deamidase, and a third agent containing α-amylase. By providing the cyclodextrin-forming enzyme, protein deamidase, and α-amylase as separate agents, it becomes easier to adjust the order in which they act on a composition containing a plant-based raw material, etc., according to the purpose.

[0069] In this case, the first agent may contain only the cyclodextrin-forming enzyme alone. That is, the first agent may be the cyclodextrin-forming enzyme itself. The second agent may contain only the protein deamidase alone. That is, the second agent may be the protein deamidase itself. Furthermore, the second agent may contain a cyclodextrin-forming enzyme in addition to the protein deamidase. The third agent may contain only α-amylase alone. That is, the third agent may be the α-amylase itself. Furthermore, the third agent may contain a cyclodextrin-forming enzyme and / or a protein deamidase in addition to the α-amylase. In addition, the first agent, the second agent, and the third agent may contain one or more selected enzymes that can be used in the production of plant-based foods and beverages, as long as the addition does not affect the improvement in foamability.

[0070] (4) Others The foaming enhancer for plant-based foods and beverages according to the present technology can contain one or more other ingredients selected freely, as long as the effects of the present technology are not impaired. Examples of other ingredients that can be used include excipients, pH adjusters, colorants, flavoring agents, disintegrants, lubricants, stabilizers, and other ingredients commonly used in formulations. Furthermore, ingredients with known or future functions can also be used in combination, as appropriate, depending on the purpose.

[0071] In addition, the foaming enhancer for plant-based foods and beverages according to the present technology may further contain, in addition to the aforementioned first, second, and third agents, other enzymes that can be used in the production of plant-based foods and beverages, or fourth, fifth, etc. agents that contain one or more selected components with known or future functions, as long as the foaming enhancer does not affect the foaming effect.

[0072] 3. Plant-based Food and Drink The plant-based food and drink according to the present technology is a plant-based food and drink produced using the foaming enhancer described above. A specific example of the plant-based drink is an oat drink, more specifically, oat milk (also known as "oat milk").

[0073] The plant-based food and drink according to the present technology has improved foaming properties. The foaming properties of the plant-based food and drink according to the present technology are not particularly limited. The foaming properties of the plant-based food and drink according to the present technology are improved by 10% or more, preferably 20% or more, and more preferably 30% or more, compared to a plant-based food and drink produced by the same method except that no foaming enhancer is used.

[0074] The present invention will be described in more detail below with reference to examples. Note that the examples described below are representative examples of the present invention and should not be construed as narrowing the scope of the present invention.

[0075] 1. Enzymes used Cyclodextrin-forming enzyme (CGT): Cyclodextrin glucanotransferase derived from Anoxybacillus caldiproteolyticus (formerly known as Geobacillus stearothermophilus), manufactured by Amano Enzyme Inc. Protein deamidase (PG): Protein glutaminase derived from Chryseo- bacterium proteolyticum, manufactured by Amano Enzyme Inc. α-Amylase (AMY): α-Amylase derived from Bacillus amyloliquefaciens, manufactured by Amano Enzyme Inc.

[0076] 2. Enzyme Activity Measurement Method [Cyclodextrin-forming Enzyme Activity Measurement Method] Cyclodextrin-forming enzyme activity can be measured using a method based on the cyclodextrin-forming enzyme activity test method in the 9th edition of the Japanese Standards for Food Additives. 20 mL of water was added to 1.0 g of potato starch, and 5 mL of sodium hydroxide TS (1 mol / L) was gradually added with stirring to form a paste. After heating in a boiling water bath for 3 minutes with stirring, 25 mL of water was added, cooled under running water, and the pH was adjusted to 5.5 with acetic acid TS (1 mol / L). Further water was added to make a 100 mL solution, which was used as the substrate solution. 10 mL of the substrate solution was measured and heated at 40°C for 10 minutes, followed by the addition and mixing of 1 mL of enzyme solution. After incubation at 40°C for 10 minutes, the reaction was stopped by adding 10 mL of hydrochloric acid TS (0.1 mol / L). To 1 mL of this reaction solution, 10 mL of iodine-potassium iodide test solution (0.4 mmol / L) was added and mixed to prepare a test solution. The iodine-potassium iodide test solution was prepared by dissolving 10.0 g of potassium iodide and 1.0 g of iodine in water to make 100 mL, and then diluting 200 times with water. A control solution was prepared using water instead of the reaction solution, and the absorbance at 660 nm of the test solution and the control solution was measured. The amount of enzyme that reduces the blue iodine coloration of starch by 1% in 1 minute was defined as 1 unit (U).

[0077] [Method for measuring protein deamidase activity] 0.1 mL of a sample solution containing protein deamidase was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and the mixture was allowed to stand at 37°C for 10 minutes, after which 1 mL of 0.4 M TCA solution was added to terminate the reaction. As a blank, 1 mL of 0.4 M TCA solution was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and 0.1 mL of the sample solution containing protein deamidase was further added, followed by standing at 37°C for 10 minutes.

[0078] The amount of ammonia generated in the reaction solution was measured using an Ammonia Test Wako (Fujifilm Wako Pure Chemical Industries, Ltd.). The ammonia concentration in the reaction solution was determined from a calibration curve showing the relationship between ammonia concentration and absorbance (630 nm) prepared using an ammonia standard solution (ammonium chloride).

[0079] The activity of protein deamidase was calculated using the following formula, where 1 unit (1 U) is the amount of enzyme that produces 1 μmol of ammonia per minute. In the formula, the volume of the reaction solution is 2.1, the volume of the enzyme solution is 0.1, Df is the dilution factor of the enzyme solution, and 17.03 is the molecular weight of ammonia.

[0080]

[0081] [Method for measuring α-amylase activity] 10 mL of a 1% potato starch substrate solution (0.1 mol / L acetic acid (pH 5.0)) was heated at 37°C for 10 minutes, after which 1 mL of a sample solution containing α-amylase was added and immediately shaken. This solution was left to stand at 37°C for 10 minutes, after which 1 mL of this solution was added to 10 mL of 0.1 mol / L hydrochloric acid test solution and immediately shaken. Next, 0.5 mL of this solution was measured, and 10 mL of 0.0002 mol / L iodine test solution (Japanese Pharmacopoeia) was added. After shaking, the absorbance (AT) at a wavelength of 660 nm was measured using water as a control. Separately, 1 mL of water was added instead of the sample solution, and the procedure was repeated to measure the absorbance (AB). The 0.0002 mol / L iodine test solution (Japanese Pharmacopoeia) was prepared by adding 10 mL of water to 12.7 g of iodine and 25 g of potassium iodide, mixing thoroughly, adding water to make 100 mL, and then diluting 2500 times with water. α-Amylase activity was calculated using the following formula. The amount of enzyme that reduces the color of potato starch caused by iodine by 10% in 1 minute was defined as 1 unit (1 U).

[0082] α-Amylase activity (U / g, U / mL) = {(AB - AT) / AB} x 1 / W AT: absorbance of reaction solution AB: absorbance of blank solution W: amount of sample in 1 mL of sample solution (g or mL)

[0083] 3. Experimental Example 1 (1) Experimental Method 18.1 g of oat flour was mixed with 150 g of water and suspended in a 300 mL Erlenmeyer flask. This suspension (oat flour content 10.8 wt%, protein content 1.4 wt%, dietary fiber content 1.1 wt%, starch content 7.3 wt%) was completely gelatinized. The enzyme amount shown in Table 1 was added, and the mixture was reacted at 60°C for 0.5 hours. Upon completion of the reaction, the mixture was boiled for 5 minutes. After filtering through a 100-mesh filter, canola oil was added to the resulting oat base drink to a final concentration of 2%, and the mixture was homogenized at 200 bar. The mixture was then stirred in a milk foamer at 65°C for 3 minutes to foam the oat drink. Foamability and foam stability were calculated using the following formula: Foamability (%) = (VF0 / 60) x 100 Foam stability (%) = (VF30 / VF0) x ​​100 60: Volume of liquid VF0: Volume of foam + liquid immediately after foaming VF30: Volume of foam + liquid after leaving to stand for 30 minutes after foaming

[0084] (2) Results The results are shown in Table 1 below.

[0085] (3) Discussion As shown in Reference Examples 2 to 5 in Table 1, it was confirmed that the foaming ability was improved by the cyclodextrin-forming enzyme treatment. It was also confirmed that the foam stability was equivalent.

[0086] 4. Experimental Example 2 (1) Experimental Method: 18.1 g of oat flour was mixed with 150 g of water and suspended in a 300 mL Erlenmeyer flask. This suspension (oat flour content: 10.8 wt%, protein content: 1.4 wt%, dietary fiber content: 1.1 wt%, starch content: 7.3 wt%) was completely gelatinized. The enzyme amounts shown in Table 2 were added in the first-stage reaction (60°C, 0.5 hours) and the second-stage reaction (50°C, 2.0 hours). After completion of the reaction, the mixture was boiled for 5 minutes. After filtering through a 100-mesh filter, canola oil was added to the resulting oat base drink to a final concentration of 2%, and the mixture was homogenized at 200 bar. The mixture was then stirred in a milk foamer at 65°C for 3 minutes to foam the oat drink. Foamability and foam stability were calculated using the same method as in Experimental Example 1.

[0087] (2) Results The results are shown in Table 2 below.

[0088] (3) Discussion As shown in Reference Examples 7 to 10 in Table 2, it was confirmed that foaming properties were improved even when α-amylase treatment was performed after cyclodextrin-forming enzyme treatment. It was also confirmed that α-amylase treatment reduced viscosity.

[0089] 5. Experimental Example 3 (1) Experimental Method: 18.1 g of oat flour was mixed with 150 g of water and suspended in a 300 mL Erlenmeyer flask. This suspension (oat flour content: 10.8 wt%, protein content: 1.4 wt%, dietary fiber content: 1.1 wt%, starch content: 7.3 wt%) was completely gelatinized. The enzyme amounts shown in Table 3 were added and reacted in the first-stage reaction (60°C, 0.5 hours) and the second-stage reaction (50°C, 2.0 hours). At the end of the reaction, the mixture was boiled for 5 minutes. After filtering through a 100-mesh filter, canola oil was added to the resulting oat base drink to a final concentration of 2%, and the mixture was homogenized at 200 bar. The mixture was then stirred in a milk foamer at 65°C for 3 minutes to foam the oat drink. Foamability and foam stability were calculated using the same method as in Experimental Example 1.

[0090] (2) Results The results are shown in Table 3 below.

[0091] (3) Discussion As shown in Table 3, it was found that Comparative Examples 1 and 2, which were treated with a cyclodextrin-forming enzyme or a protein deamidating enzyme, had improved foaming properties compared to Reference Example 11, which was treated with α-amylase. Furthermore, it was confirmed that the combined use of a cyclodextrin-forming enzyme and a protein deamidating enzyme synergistically improved foaming properties. Furthermore, it was confirmed that the foam stability was equivalent and there was no effect on stability.

Claims

1. A method for improving the foaming properties of a plant-based food or beverage, comprising: a cyclodextrin-forming enzyme action step of acting on a composition containing a plant-based raw material with a cyclodextrin-forming enzyme; and a protein deamidating enzyme action step of acting on a composition containing a plant-based raw material with a protein deamidating enzyme.

2. A method for improving the foaming properties of a plant-based food or beverage as described in claim 1, comprising an α-amylase action step of acting on a composition containing a plant-based raw material with α-amylase.

3. A method for improving the foaming properties of the plant-based food or beverage described in claim 1, comprising an oil / fat addition step of adding oil / fat.

4. A foaming enhancer for plant-based foods and beverages, which contains a cyclodextrin-forming enzyme and a protein deamidating enzyme.

5. A plant-based food or drink in which the foaming enhancer according to claim 4 is used.

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