Enzyme agent for improving shape retention

Abstract

Provided is a technique for improving the shape retention of vegetable ice cream. The present technology provides: an enzyme agent for improving the shape retention of a vegetable ice cream, said enzyme agent containing one or more enzymes selected from the group consisting of a maltotriose-producing amylase, a lipase, and a beta-amylase; and a vegetable ice cream in which the enzyme agent for improving the shape retention is used. Furthermore, according to the present technology, a method for producing vegetable ice cream and a method for improving the shape retention of vegetable ice cream are provided, the method for producing the vegetable ice cream and the method for improving the shape retention of the vegetable ice cream comprise a maltotriose-producing amylase action step of causing a maltotriose-producing amylase to act on a material containing a vegetable sugar, a lipase action step of causing a lipase to act on a material containing a vegetable fat and oil, and a maltotriose-producing amylase action step of causing a maltotriose-producing amylase to act on a material containing a vegetable fat and oil. And one or more steps among a beta-amylase action step for causing a beta-amylase to act.

Description

Technical Field

[0001] The present invention relates to an enzyme for improving the shape retention of plant-based ice cream. More specifically, the present invention relates to an enzyme for improving the shape retention for improving the quality of plant-based ice cream, plant-based ice cream, a method for producing plant-based ice cream, and a method for improving the shape retention of plant-based ice cream. Background Art

[0002] In recent years, the dairy alternative market and the meat alternative market are expanding against the backdrop of sustainability and health awareness in the food market. Plant-based ice cream occupies an important category in the dairy alternative market and is an area of ​​continued attention. In its production, it is standard to mix a protein source and a lipid source in plant-based milk as needed and emulsify them.

[0003] Under such circumstances, technology for improving the quality of plant-based ice cream is also being developed. For example, Patent Document 1 discloses a production technology for reducing the amount of plant protein used in order to improve the flavor of plant-based ice cream while minimizing animal-derived ingredients such as milk ingredients.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: U.S. Patent Publication No. 2018 / 0184684 Summary of the invention

[0007] As mentioned above, plant-based foods and beverages have attracted attention in recent years due to the increase in health awareness, but since the composition of plant-based ice cream is fundamentally different from that of dairy ice cream, there is still room for improvement in obtaining satisfactory properties. For example, in relation to product stability, shape retention is an important property for preventing plant-based ice cream from melting or deforming due to heat shock during storage, and it is expected to be further improved.

[0008] Therefore, in the present technology, a main object is to provide a technology for improving the shape retention of plant-based ice cream.

[0009] The inventors of the present application conducted intensive research on a technology for improving the shape retention of plant-based ice cream and found that the shape retention of plant-based ice cream can be improved by allowing maltotriose-generating amylase, lipase or β-amylase to act on plant materials, thereby completing the present technology.

[0010] That is, in the present technology, first, an enzyme agent for improving the shape retention of plant-based ice cream is provided, the enzyme agent for improving the shape retention of plant-based ice cream containing one or more enzymes selected from the group consisting of maltotriose-producing amylase, lipase, and β-amylase.

[0011] Next, in the present technology, there is provided a vegetable-based ice cream using the enzyme agent for improving the shape retention according to the present technology.

[0012] The present technology also provides a method for producing plant-based ice cream and a method for improving the shape retention of plant-based ice cream, which include one or more steps selected from a maltotriose-producing amylase action step of allowing maltotriose-producing amylase to act on a material containing plant saccharides, a lipase action step of allowing lipase to act on a material containing plant oils and fats, and a β-amylase action step of allowing β-amylase to act on a material containing plant saccharides.

[0013] According to the present technology, it is possible to provide a plant-based ice cream that can improve the shape retention of the plant-based ice cream, has high stability during storage, and is not easily melted. DETAILED DESCRIPTION

[0014] Hereinafter, a preferred mode for carrying out the present invention will be described. In addition, the embodiment described below is an example of a representative embodiment of the present invention, and the scope of the present invention is not to be interpreted narrowly thereby.

[0015] 1. Enzymes for improving shape retention

[0016] The enzyme agent for improving shape retention involved in the present technology contains one or more enzymes selected from the group consisting of maltotriose-generating amylase, lipase and β-amylase as an active ingredient. In addition, it may also contain α-amylase, monoglyceride, diglyceride, other components, etc. as needed. Below, each component is described in detail.

[0017] (1) Maltotriose produces amylase

[0018] The maltotriose-producing amylase that can be used in the present technology is an enzyme that has an activity of acting on starch to mainly produce maltotriose. The maltotriose-producing amylase that can be used in the present technology may be an enzyme that has other actions as long as it has an activity of producing maltotriose.

[0019] In the present technology, by allowing maltotriose-producing amylase to act on a plant material, the shape retention of the produced plant-based ice cream can be improved.

[0020] The source of the maltotriose-generating amylase that can be used in the present technology is not particularly limited, and examples thereof include maltotriose-generating amylases from Streptomyces (Streptomyces griseus, etc.), Bacillus (Bacillus subtilis, etc.), Microbacterium sp. disclosed in Japanese Patent Laid-Open No. 03-251173, and Cellulosimicrobium disclosed in WO2020 / 090734. These maltotriose-generating amylases can be used alone or in combination. Among these maltotriose-generating amylases, from the viewpoint of further improving the shape retention of plant-based ice cream, it is preferred to select a maltotriose-generating amylase from a Microbacterium organism, and it is more preferred to select a maltotriose-generating amylase from a Microbacterium organism.

[0021] Here, the "maltotriose-producing amylase derived from the genus Microbacterium" refers to maltotriose-producing amylase produced by a microorganism classified as the genus Microbacterium (which may be a wild strain or a mutant strain), or maltotriose-producing amylase obtained by genetic engineering using a maltotriose-producing amylase gene. Therefore, a recombinant produced by a host microorganism into which a maltotriose-producing amylase gene obtained from a Microbacterium (or a gene in which the gene is modified) is introduced also corresponds to the "maltotriose-producing amylase derived from the genus Microbacterium".

[0022] The maltotriose generating amylase used in the present technology can be prepared from the culture solution of the microorganism that is the source of the above-mentioned maltotriose generating amylase. As a specific preparation method, a method for recovering maltotriose generating amylase from the culture solution or bacterial cell of the above-mentioned microorganism can be cited. For example, in the case of using a maltotriose generating amylase secretory microorganism, the bacterial cell can be recovered from the culture solution by filtering, centrifugation, etc. in advance as needed, and then the enzyme can be separated and / or purified. In addition, in the case of using a maltotriose generating amylase non-secreting microorganism, the bacterial cell can be recovered from the culture solution in advance as needed, and then the enzyme can be exposed by crushing the bacterial cell by pressure treatment, ultrasonic treatment, etc., and then the enzyme can be separated and / or purified. As an enzyme separation and / or purification method, a disclosed protein separation and / or purification method can be used without particular limitation, for example, a centrifugal separation method, a UF concentration method, a salting-out method, various chromatography methods using ion exchange resins, etc. can be cited. The enzyme after separation and / or purification can be powdered by drying methods such as freeze drying and reduced pressure drying, and can also be powdered by using suitable excipients and / or drying aids in the drying method. In addition, the enzyme after separation and / or purification can also be liquefied by adding suitable additives, filtering and sterilizing.

[0023] In the present technique, a commercially available product may be used as the maltotriose-producing amylase, and a preferred example of the commercially available product is maltotriose-producing amylase manufactured by Amano Enzyme Products Co., Ltd.

[0024] As long as the effect of the present technology is not impaired, the content of maltotriose generating amylase in the enzyme agent for improving shape retention involved in the present technology can be freely set. The content of maltotriose generating amylase can be set to 0.05U or more relative to 1g of the material containing plant saccharides that allows maltotriose generating amylase to function, for example, and can be preferably set to 0.3U or more from the viewpoint of further improving shape retention, more preferably set to 0.6U or more, further preferably set to 3U or more, and further preferably set to 6U or more.

[0025] The content of maltotriose generating amylase can be set to, for example, 0.08 U or more relative to 1 g of the polysaccharide (preferably starch) on which maltotriose generating amylase acts. From the viewpoint of further improving the shape retention, it can be preferably set to 0.5 U or more, more preferably set to 1 U or more, further preferably set to 5 U or more, and further preferably set to 10 U or more.

[0026] As long as the effect of the present technology is not impaired, the upper limit of the content of maltotriose-generating amylase is not particularly limited, and can be set, for example, to 250 U or less, 50 U or less, 25 U or less, 20 U or less, 15 U or less, or 10 U or less relative to 1 g of the material containing plant sugars that enables maltotriose-generating amylase to function.

[0027] The upper limit of the content of the maltotriose generating amylase can be set to, for example, 400 U or less, 80 U or less, 40 U or less, 30 U or less, 20 U or less, or 15 U or less relative to 1 g of the polysaccharide (preferably starch) on which the maltotriose generating amylase acts.

[0028] In the present technology, the activity of maltotriose-producing amylase is a value defined by the following method.

[0029] [Definition of maltotriose-generating amylase activity]

[0030] An appropriate amount of enzyme is added to 0.5 ml of 2% soluble starch dissolved in 0.1 M phosphate buffer (pH 7.0), and a total of 1.0 ml is reacted at 40°C. The generated maltotriose and other reducing sugars are quantified using the Somogyi-Nelson method. Under these conditions, the amount of enzyme that generates reducing sugars equivalent to 1 micromole of glucose in 1 minute is considered 1 unit (1U).

[0031] (2) Lipase

[0032] The lipase that can be used in the present technology is an enzyme that has the activity of hydrolyzing triglycerides to generate diglycerides, monoglycerides, and fatty acids. As the lipase that can be used in the present technology, as long as it has the activity of hydrolyzing triglycerides to generate diglycerides, monoglycerides, and fatty acids, it may also be an enzyme that has other functions.

[0033] In the present technology, by allowing lipase to act on the plant material, the shape retention of the produced plant-based ice cream can be improved, the stability during storage can be improved, and the ice cream can be made less likely to melt.

[0034] The source of the lipase that can be used in the present technology is not particularly limited, and examples thereof include lipases from Aspergillus, Candida, Rhizopus, Mucor, and Penicillium. These lipases may be used alone or in combination. Among these lipases, from the viewpoint of further improving the shape retention of the vegetable-based ice cream, it is preferred to select a lipase from a Penicillium organism, and more preferably a lipase from a Penicillium roqueforti organism.

[0035] The "lipase from Penicillium roquefort" herein refers to a lipase produced by a microorganism classified as Penicillium roquefort (which may be a wild strain or a mutant strain), or a lipase obtained by genetic engineering using a lipase gene. Therefore, a recombinant produced by a host microorganism into which a lipase gene obtained from Penicillium roquefort (or a gene that has modified the gene) is also equivalent to a "lipase from Penicillium roquefort".

[0036] The lipase used in the present technology can be prepared from the culture solution of the microorganism as the above-mentioned lipase source. As a specific preparation method, a method for recovering lipase from the culture solution or bacterial cell of the above-mentioned microorganism can be cited. For example, when using a lipase secreting microorganism, the bacterial cell can be recovered from the culture solution by filtering, centrifugation, etc. in advance as needed, and then the enzyme can be separated and / or purified. In addition, when using a lipase non-secreting microorganism, the bacterial cell can be recovered from the culture solution in advance as needed, and then the enzyme can be exposed by crushing the bacterial cell by pressure treatment, ultrasonic treatment, etc., and then the enzyme can be separated and / or purified. As an enzyme separation and / or purification method, a disclosed protein separation and / or purification method can be used without particular limitation, for example, centrifugation, UF concentration method, salting-out method, various chromatography methods using ion exchange resins, etc. can be cited. The separated and / or purified enzyme can be powdered by drying methods such as freeze drying and reduced pressure drying, and 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 can be liquefied by adding appropriate additives and sterilizing by filtration.

[0037] In the present technique, commercially available products may be used as lipases, and examples of preferred commercial products include lipases derived from organisms of the genus Penicillium manufactured by Amano Enzyme Products Co., Ltd.

[0038] As long as the effect of the present technology is not impaired, the content of lipase in the enzyme agent for improving shape retention involved in the present technology can be freely set. The content of lipase can be set to 0.01U or more relative to 1g of the material containing vegetable oils and fats for causing the lipase to act, for example, and from the viewpoint of further improving shape retention, it can be preferably set to 0.1U or more, more preferably set to 0.4U or more, further preferably set to 1U or more, and further preferably set to 2U or more.

[0039] In addition, the content of lipase can be set to, for example, 0.09U or more relative to 1g of fat or oil that causes the lipase to function. From the viewpoint of further improving the shape retention, it can be preferably set to 0.4U or more, more preferably set to 0.9U or more, further preferably set to 1U or more, and further preferably set to 2U or more.

[0040] As long as the effect of the present technology is not impaired, the upper limit of the lipase content is not particularly limited, and can be set to, for example, less than 100U, less than 20U, less than 15U, less than 10U, less than 5U or less than 3U relative to 1g of material containing vegetable oil that enables the lipase to function.

[0041] The upper limit of the lipase content can be set to, for example, 100 U or less, 20 U or less, 15 U or less, 10 U or less, 5 U or less, or 3 U or less, based on 1 g of fat or oil for causing the lipase to act.

[0042] In the present technology, the lipase activity is a value defined by the following method.

[0043] [Definition of lipase activity]

[0044] 45g of olive oil was mixed with 18.5g / L emulsion (polyvinyl alcohol (completely saponified, saponification degree 98.8±0.2), 150mL polyvinyl alcohol (partially saponified, saponification degree 88.8±1.0) 1.5g / L), and emulsified using a homogenizer to prepare a matrix solution. 1mL of lipase solution was added to 5mL of matrix solution and 4mL of 0.1mol / L phosphate buffer (pH7.0), and after reacting at 37°C for 30 minutes, 10mL of ethanol-acetone mixture was added to stop the enzyme reaction. Next, 10mL of 0.05mol / L sodium hydroxide solution and 10mL of ethanol-acetone mixture were added, and the pH was titrated to 10 with 0.05mol / L hydrochloric acid. In addition, the amount of enzyme that brings about an increase of 1μmol of fatty acids in 1 minute is taken as 1 unit (1U).

[0045] (3) β-amylase

[0046] The β-amylase that can be used in the present technology is an exo-type enzyme that successively decomposes α-1,4-glucosidic bonds from the non-reducing ends of starch in maltose units. The β-amylase that can be used in the present technology may be an enzyme that has other functions as long as it has the above-mentioned β-amylase activity.

[0047] In the present technology, by allowing β-amylase to act on a plant-based material, the shape retention of the produced plant-based ice cream can be improved.

[0048] The source of the β-amylase that can be used in the present technology is not particularly limited, and examples thereof include β-amylases from Bacillus (Bacillus flexus, Bacillus megaterium, Bacillus polymyxa, Bacillus circulans, etc.), Pseudomonas, and Streptomyces. These β-amylases may be used alone or in combination. Among these β-amylases, from the viewpoint of further improving the shape retention of vegetable ice cream, it is preferred to select β-amylases from Bacillus, and more preferably to select β-amylases from Bacillus flexus.

[0049] Here, "β-amylase derived from Bacillus flexus organisms" refers to β-amylase produced by a microorganism classified as Bacillus flexus (which may be a wild strain or a mutant strain), or β-amylase obtained by genetic engineering using a β-amylase gene. Therefore, a recombinant produced by a host microorganism into which a β-amylase gene obtained from Bacillus flexus organisms (or a gene in which the gene is modified) is also equivalent to "β-amylase derived from Bacillus flexus organisms".

[0050] The β-amylase used in the present technology can be prepared from the culture solution of the microorganism that is the source of the above-mentioned β-amylase. As a specific preparation method, a method for recovering β-amylase from the culture solution or bacterial cells of the above-mentioned microorganisms can be cited. For example, when using a β-amylase secretory microorganism, the bacterial cells can be recovered from the culture solution by filtering, centrifugation, etc. in advance as needed, and then the enzyme can be separated and / or purified. In addition, when using a β-amylase non-secreting microorganism, the bacterial cells can be recovered from the culture solution in advance as needed, and then the enzyme can be exposed by crushing the bacterial cells by pressure treatment, ultrasonic treatment, etc., and then the enzyme can be separated and / or purified. As an enzyme separation and / or purification method, a disclosed protein separation and / or purification method can be used without particular limitation, for example, centrifugation, UF concentration method, salting-out method, various chromatography methods using ion exchange resins, etc. can be cited. The separated and / or purified enzyme can be powdered by a drying method such as freeze drying and reduced pressure drying, and in addition, it can also be powdered using an appropriate excipient and / or drying aid in the drying method. Alternatively, the separated and / or purified enzyme can be liquefied by adding appropriate additives and sterilizing by filtration.

[0051] In addition, in the present technology, β-amylases derived from plants, such as soybeans, wheat, and barley, can be used without being limited to the above-mentioned β-amylases derived from microorganisms.

[0052] In the present technique, commercially available products may be used as β-amylase, and preferred examples of commercially available products include β-amylase derived from Bacillus flexus manufactured by Amano Enzyme Products Co., Ltd.

[0053] As long as the effect of the present technology is not impaired, the content of β-amylase in the enzyme agent for improving shape retention involved in the present technology can be freely set. The content of β-amylase can be set to 0.01U or more relative to 1g of the material containing plant saccharides on which β-amylase acts, for example, and can be preferably set to 0.10U or more from the viewpoint of further improving shape retention, more preferably set to 0.50U or more, further preferably set to 1.0U or more, and further preferably set to 1.5U or more.

[0054] The content of β-amylase can be set to, for example, 0.016 U or more relative to 1 g of the polysaccharide (preferably starch) that enables the β-amylase to function. From the viewpoint of further improving the shape retention, it can be preferably set to 0.16 U or more, more preferably set to 0.80 U or more, further preferably set to 1.6 U or more, and further preferably set to 2.4 U or more.

[0055] As long as the effect of the present technology is not impaired, the upper limit of the β-amylase content is not particularly limited. It can be set to, for example, less than 100 U, less than 50 U, or less than 10 U, preferably less than 7.5 U, more preferably less than 4.0 U, and further preferably less than 2.5 U, relative to 1 g of material containing plant sugars that enables β-amylase to function.

[0056] The upper limit of the β-amylase content can be set to, for example, 160 U or less, 80 U or less, or 16 U or less, preferably 12 U or less, more preferably 6.4 U or less, and further preferably 4.0 U or less, relative to 1 g of the polysaccharide (preferably starch) that enables the β-amylase to function.

[0057] In the present technology, the activity of β-amylase is a value defined by the following method.

[0058] [Definition of β-amylase activity]

[0059] Using potato starch as a substrate, the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose within 1 minute is defined as 1 unit (1U).

[0060] (4) α-amylase

[0061] In the present technology, α-amylase can be used. 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.

[0062] The source of the α-amylase that can be used in the present technology is not particularly limited. For example, 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.) can be mentioned. Preferably, α-amylases derived from organisms of the genus Bacillus are mentioned, and more preferably, α-amylases derived from organisms of the genus Bacillus are mentioned.

[0063] The α-amylase that can be used in the present technology can be prepared from 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, using a method of isolating α-amylase using a public means, and using a method of genetic recombination technology.

[0064] In the present technique, commercially available α-amylase may be used, and preferred examples of commercially available α-amylase (derived from Bacillus amyloliquefaciens) manufactured by Amano Enzyme Products Co., Ltd. may be mentioned.

[0065] As long as the effect of the present technology is not impaired, the content of α-amylase in the enzyme agent for improving shape retention involved in the present technology can be freely set. The content of α-amylase can be set to 0.5U or more relative to 1g of the material containing plant saccharides on which the α-amylase acts, for example, and from the viewpoint of further improving shape retention, it can be preferably set to 2U or more, more preferably set to 4U or more, further preferably set to 7U or more, and further preferably set to 10U or more.

[0066] In addition, the content of α-amylase can be set to, for example, 0.8 U or more relative to 1 g of the polysaccharide (preferably starch) that allows the α-amylase to function. From the viewpoint of further improving the shape retention, it can be preferably set to 3 U or more, more preferably set to 6 U or more, further preferably set to 10 U or more, and further preferably set to 15 U or more.

[0067] As long as the effect of the present technology is not impaired, the upper limit of the α-amylase content is not particularly limited. It can be set to, for example, less than 500U, less than 100U, or less than 50U, preferably less than 30U, more preferably less than 20U, and further preferably less than 15U, relative to 1g of material containing plant sugars that allows α-amylase to function.

[0068] In addition, the upper limit of the content of α-amylase relative to 1 g of polysaccharide (preferably starch) on which α-amylase acts can be set to, for example, 800 U or less, 160 U or less, or 80 U or less, preferably 40 U or less, more preferably 30 U or less, and further preferably 20 U or less.

[0069] In the present technology, the activity of α-amylase is a value defined by the following method.

[0070] [Definition of α-amylase activity]

[0071] The activity of amylase is measured by iodine reaction using potato starch as the substrate. In other words, the activity of amylase is measured by the amount of enzyme that reduces the color by 10% in 1 minute by iodine reaction using potato starch as the substrate by conventional methods.

[0072] In the present technology, when maltotriose generating amylase and α-amylase are used together, the content ratio thereof is determined based on the aforementioned content of each enzyme, but from the viewpoint of further improving the effect, the content of α-amylase per 1U of maltotriose generating amylase can be preferably set to 0.001U or more, more preferably to 0.01U or more, further preferably to 0.05U or more, further preferably to 0.08U or more, further preferably to 0.1U or more, further further preferably to 0.2U or more, and further further preferably to 0.5U or more.

[0073] The upper limit of the range of the content of α-amylase per 1U of maltotriose generating amylase can be, for example, 100U or less. From the viewpoint of further improving the effect of improving shape retention, it is preferably 20U or less, more preferably 10U or less, further preferably 5U or less, further preferably 4U or less, still further preferably 3U or less, further further preferably 2U or less, and further further preferably 1U or less.

[0074] In the present technology, when β-amylase and α-amylase are used in combination, the content ratio is determined based on the aforementioned content of each enzyme, but from the viewpoint of further improving the effect, the content of α-amylase per 1U of β-amylase can be preferably set to 0.001U or more, more preferably set to 0.01U or more, further preferably set to 0.05U or more, further preferably set to 0.08U or more, further preferably set to 0.1U or more, and further more preferably set to 0.12U or more.

[0075] The upper limit of the range of the content of β-amylase per 1U of α-amylase can be, for example, set to 100U or less. From the viewpoint of further improving the effect of improving shape retention, it can be preferably set to 20U or less, more preferably set to 10U or less, further preferably set to 5.0U or less, further preferably set to 2.0U or less, still further preferably set to 1U or less, further further preferably set to 0.5U or less, and further further preferably set to 0.2U or less.

[0076] (5) Monoglyceride and diglyceride

[0077] The enzyme agent for improving shape retention according to the present technology can use monoglyceride and / or diglyceride. By using monoglyceride and / or diglyceride, the effect of improving the shape retention of the produced vegetable-based ice cream can be further enhanced.

[0078] As long as the effect of the present technology is not impaired, one or more monoglycerides and / or diglycerides that can be used in the food field can be freely selected and used as the monoglyceride and / or diglyceride that can be used in the present technology. Examples of monoglyceride include glyceryl caprylate, glyceryl caprate, glyceryl laurate, glyceryl stearate, glyceryl oleate, glyceryl behenate, etc. Examples of diglyceride include diglyceride formed by combining two identical or different fatty acids of these monoglycerides.

[0079] The content of monoglyceride and / or diglyceride in the enzyme agent for improving shape retention according to the present technology can be freely set as long as the effect of the present technology is not impaired. The content of monoglyceride and / or diglyceride can be set to 0.5 parts by weight or more, for example, based on 100 parts by weight of the vegetable fat used in the vegetable ice cream produced, and can be preferably set to 1 part by weight or more, and more preferably set to 2 parts by weight or more, from the viewpoint of further improving shape retention.

[0080] As long as the effect of the present technology is not impaired, the upper limit of the content of monoglyceride and / or diglyceride in the enzyme agent for improving shape retention involved in the present technology is not particularly limited, and can be set to, for example, 20 parts by weight or less, preferably 10 parts by weight or less, and more preferably 8 parts by weight or less, relative to 100 parts by weight of vegetable oil used in the vegetable ice cream produced.

[0081] (6) Other ingredients

[0082] The enzyme agent for improving shape retention involved in the present technology can be composed of only one or more enzymes selected from the group consisting of the aforementioned maltotriose generating amylase, lipase and β-amylase, as long as it contains one or more enzymes selected from the group consisting of the aforementioned maltotriose generating amylase, lipase and β-amylase, or can be used in combination with the aforementioned α-amylase, monoglyceride, diglyceride, etc. As long as the effect of the present technology is not impaired, it is also possible to freely choose to contain one or more other ingredients. As other ingredients, for example, components such as excipients, pH adjusters, colorants, flavoring agents, disintegrants, lubricants, stabilizers, etc. used in conventional formulations can be used. Furthermore, it is also possible to appropriately use functional components disclosed or discovered in the future according to the purpose.

[0083] 2. Plant-based ice cream

[0084] The plant-based ice cream involved in the present technology is a plant-based ice cream produced using the aforementioned enzyme for improving shape retention involved in the present technology. The plant-based ice cream involved in the present technology is characterized in that in its production process, by treating the plant material with one or more enzymes selected from the group consisting of maltotriose-generating amylase, lipase and β-amylase, the content of one or more selected from maltotriose, monoglyceride and diglyceride is increased, thereby compared with the plant-based ice cream produced without using the enzyme for improving shape retention involved in the present technology, the shape retention is high, the stability during storage is high, and it is not easy to melt. That is, the plant-based ice cream involved in the present technology is characterized in that it contains one or more materials selected from the material containing plant sugars after maltotriose-generating amylase treatment, the material containing plant oils and fats after lipase treatment, and the material containing plant sugars after β-amylase treatment.

[0085] The content of maltotriose contained in the plant-based ice cream involved in the present technology is, for example, 10 parts by weight or more, preferably 15 parts by weight or more, more preferably 20 parts by weight or more, and further preferably 30 parts by weight or more per 100 parts by weight of the sugar derived from the material containing the plant-based sugar.

[0086] The upper limit of the content of maltotriose contained in the plant-based ice cream involved in the present technology is, for example, 60 parts by weight or less, preferably 50 parts by weight or less, more preferably 45 parts by weight or less, further preferably 40 parts by weight or less, and further preferably 35 parts by weight or less per 100 parts by weight of the sugar derived from the material containing the plant-based sugar.

[0087] The content of maltotriose in the plant-based ice cream of the present technology may be, for example, 0.1 parts by weight or more, preferably 0.3 parts by weight or more, more preferably 1 part by weight or more, and further preferably 1.5 parts by weight or more per 100 parts by weight of the plant-based ice cream.

[0088] The upper limit of the content of maltotriose contained in the plant-based ice cream involved in the present technology is, for example, 10 parts by weight or less, preferably 5 parts by weight or less, more preferably 4 parts by weight or less, further preferably 3 parts by weight or less, and further preferably 2.5 parts by weight or less per 100 parts by weight of the plant-based ice cream.

[0089] The content of the monoglyceride and / or diglyceride in the vegetable ice cream of the present invention is, for example, 1 part by weight or more, preferably 2 parts by weight or more, more preferably 5 parts by weight or more, and further preferably 6 parts by weight or more per 100 parts by weight of fats and oils.

[0090] From the viewpoint of preventing a decrease in flavor due to an increase in the free amount of saturated fatty acids, the upper limit of the content of monoglyceride and / or diglyceride contained in the vegetable ice cream involved in the present technology may be, for example, 70 parts by weight or less, 50 parts by weight or less, 30 parts by weight or less, 20 parts by weight or less, 15 parts by weight or less, or 10 parts by weight or less per 100 parts by weight of fats and oils.

[0091] The content of the monoglyceride and / or diglyceride in the vegetable ice cream of the present invention is, for example, 0.1 parts by weight or more, preferably 0.2 parts by weight or more, more preferably 0.3 parts by weight or more, and further preferably 0.35 parts by weight or more per 100 parts by weight of the vegetable ice cream.

[0092] From the viewpoint of preventing flavor degradation due to an increase in the free amount of saturated fatty acids, the upper limit of the content of monoglyceride and / or diglyceride contained in the plant-based ice cream involved in the present technology may be, for example, 5 parts by weight or less, 3 parts by weight or less, 2 parts by weight or less, 1 part by weight or less, 0.8 parts by weight or less, or 0.5 parts by weight or less per 100 parts by weight of the plant-based ice cream.

[0093] 3. Method for producing plant-based ice cream and method for improving its shape retention

[0094] The manufacturing method and shape retention improvement method of the plant-based food and beverage involved in the present technology are methods of performing at least one of the following steps: a maltotriose-generating amylase action step in which maltotriose-generating amylase is acted on a material containing plant saccharides, a lipase action step in which lipase is acted on a material containing plant oils and fats, and a β-amylase action step in which β-amylase is acted on a material containing plant saccharides. In addition, an α-amylase action step in which α-amylase is acted on a material containing plant saccharides, a monoglyceride and / or diglyceride addition step in which monoglyceride and / or diglyceride are added, a recovery step, etc. can also be further performed. In addition, depending on the type of plant-based ice cream, before and after each step or at the same time as each step, a general plant-based ice cream manufacturing process can also be performed within the scope of not impairing the effect of the present technology. Below, each step, etc. are described in detail.

[0095] (1) Materials

[0096] As the plant material used in the method for producing a plant-based food and drink and the method for improving shape retention according to the present technique, a material containing plant-based sugars and a material containing plant-based oils and fats are used.

[0097] (1-1) Materials containing plant sugars

[0098] The material containing plant saccharides that can be used in the present technology is not particularly limited as long as it contains saccharides from plants, and a typical example is a material obtained by mechanically crushing cereals (oats, etc.) with a mill, etc. As saccharides, polysaccharides (starch, dextrin, etc.) are preferably contained.

[0099] The material containing vegetable saccharides and the material containing vegetable oils and fats described later may be different materials or the same material.

[0100] The shape of the material containing plant sugar is not limited unless the effects of the present technology are impaired, and examples thereof include powder, flake, and granular shapes.

[0101] As long as the effects of the present technology are not impaired, the content of the sugar (preferably starch) contained in the material containing the plant sugar (based on the weight of the material containing the plant sugar in a dried state) is not particularly limited, and for example, it can be 20% by weight or more. From the viewpoint of further improving the shape retention, it can be preferably 50% by weight or more, 55% by weight or more, more preferably 60% by weight or more, and even more preferably 65% ​​by weight or more.

[0102] As long as the effect of the present technology is not impaired, the upper limit of the content of sugars (preferably starch) contained in the material containing plant sugars is not particularly limited, and for example, it can be 95% by weight or less, preferably 90% by weight or less, more preferably 85% by weight or less, and further preferably 80% by weight or less.

[0103] As long as the effect of the present technology is not impaired, there is no particular limitation on the source of plant sugars, for example, oats, barley (glutinous wheat, glutinous wheat), rice, wheat, rye, buckwheat, barnyard grass, millet, teff, quinoa, corn and other cereals; adzuki beans, broad beans, peas, chickpeas, mung beans, red beans, kidney beans and other cereals; almonds, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts, coconuts, pili nuts, chestnuts, sesame seeds, industrial hemp seeds, canary seeds, flax seeds, pine nuts and other seeds; algae, etc., and one of these plant sugars can be used alone, or multiple combinations can be used. Among them, in the present technology, it is particularly preferred to use materials containing sugars from cereals, and it is more preferred to use materials containing sugars from oats.

[0104] (1-2) Materials containing vegetable oils and fats

[0105] The material containing vegetable oils that can be used in the present technology is not particularly limited as long as it contains oils from plants. Typical examples include oils obtained from plant raw materials by physical pressing, extraction in a solvent, or extraction by a combination of these and then undergoing various refining steps.

[0106] The shape of the material containing vegetable oil and fat is not limited unless the effects of the present technique are impaired, and examples thereof include solid and liquid forms.

[0107] As long as the effect of the present technology is not impaired, the content of oil contained in the material containing vegetable oil (based on the weight of the material containing vegetable oil used) is not particularly limited, and for example, it can be 20% by weight or more. From the viewpoint of further improving the shape retention, it can be preferably 60% by weight or more, more preferably 80% by weight or more, and even more preferably 95% by weight or more.

[0108] Unless the effects of the present technique are impaired, the upper limit of the content of fats and oils contained in the vegetable fat-containing material is not particularly limited, and examples thereof include 99% by weight or less, 97% by weight or less, or 95% by weight or less.

[0109] As long as the effect of the present technology is not impaired, there is no particular limitation on the source of the vegetable oil, and examples thereof include coconut oil, soybean oil, rapeseed oil, rice oil, corn oil, sunflower oil, cottonseed oil, peanut oil, safflower oil, olive oil, palm oil, palm kernel oil, coconut oil, cocoa butter, etc., and one of these vegetable oils can be used alone or in combination. Among them, in the present technology, it is particularly preferred to use a material containing coconut oil.

[0110] (2) Maltotriose-producing amylase process

[0111] The maltotriose generating amylase action step is a step of allowing maltotriose generating amylase to act on a material containing plant saccharides. As long as the effect of the present technology is not impaired, the amount of maltotriose generating amylase added in the maltotriose generating amylase action step can be freely set. The specific amount of maltotriose generating amylase added in the present technology is the same as the content of maltotriose generating amylase in the aforementioned enzyme agent for improving shape retention, so the description is omitted here.

[0112] As long as the effect of the present technology is not impaired, various conditions of the maltotriose generating amylase action process can be freely set. For example, the 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 maltotriose generating amylase used. The pH can be set to pH5 to 9, preferably pH6 to 8, and more preferably pH6.5 to 7.5. The temperature can be set to 20 to 80°C, preferably 30 to 70°C, and more preferably 40 to 60°C. The action time can be set to 5 minutes to 24 hours, preferably 15 minutes to 12 hours, and more preferably 30 minutes to 2 hours. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0113] The material containing plant saccharides that has been subjected to the step of maltotriose generating amylase has an increased content of maltotriose under the action of maltotriose generating amylase. The content of maltotriose contained in the material containing plant saccharides that has been subjected to the step of maltotriose generating amylase (hereinafter also referred to as "the material containing plant saccharides treated with maltotriose generating amylase") can be, for example, 10 parts by weight or more per 100 parts by weight of saccharides. Preferably, it is 15 parts by weight or more, more preferably, 20 parts by weight or more, and even more preferably, 30 parts by weight or more.

[0114] In addition, as an upper limit of the content of maltotriose contained in the material containing plant saccharides that has undergone the step of maltotriose generating amylase, for example, 60 parts by weight or less per 100 parts by weight of saccharides can be cited, preferably 50 parts by weight or less, more preferably 45 parts by weight or less, further preferably 40 parts by weight or less, and further preferably 35 parts by weight or less.

[0115] (3) Lipase action process

[0116] The lipase action step is a step of allowing lipase to act on a material containing vegetable oil. The amount of lipase added in the lipase action step can be freely set as long as the effect of the present technology is not impaired. The specific amount of lipase added in the present technology is the same as the content of lipase in the aforementioned enzyme agent for improving shape retention, so the description is omitted here.

[0117] As long as the effect of the present technology is not impaired, various conditions of the lipase 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 lipase used. pH can be set to, for example, pH5-9, preferably pH6-8, and more preferably pH6.5-7.5. Temperature can be set to, for example, 20-80°C, preferably 30-70°C, and more preferably 40-60°C. Action time can be set to, for example, 5 minutes to 24 hours, preferably 10 minutes to 12 hours, and more preferably 20 minutes to 1 hour. In addition, the best reaction conditions can be determined by preliminary experiments.

[0118] The content of monoglyceride and / or diglyceride in the material containing vegetable oils and fats that has been subjected to the lipase action step is increased under the action of lipase. The content of monoglyceride and / or diglyceride contained in the material containing vegetable oils and fats that has been subjected to the lipase action step (hereinafter also referred to as "lipase-treated vegetable oils and fats-containing material") can be, for example, 0.6 parts by weight or more, preferably 0.7 parts by weight or more, more preferably 0.8 parts by weight or more, further preferably 0.9 parts by weight or more, and further preferably 1 part by weight or more per 100 parts by weight of oils and fats.

[0119] In addition, the upper limit of the content of monoglyceride and / or diglyceride contained in the material containing vegetable oil and fat that has undergone the lipase action process can be, for example, 10 parts by weight or less, 8 parts by weight or less, 6 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, or 2 parts by weight or less per 100 parts by weight of oil and fat.

[0120] The acid value of the material containing vegetable oils and fats after the lipase action step increases under the action of lipase. The acid value of the material containing vegetable oils and fats after the lipase action step may be, for example, 1.1 or more, preferably 1.2 or more, more preferably 1.3 or more, further preferably 1.5 or more, and further preferably 1.7 or more.

[0121] The upper limit of the acid value contained in the vegetable oil-containing material subjected to the lipase action step is, for example, 5 or less, preferably 4 or less, more preferably 3 or less, and further preferably 2 or less.

[0122] The maltotriose generating amylase action step and the lipase action step can be performed on either side or on both sides. In the case of performing both sides, the order of the maltotriose generating amylase action step and the lipase action step is not particularly limited. For example, by performing each step separately, the material containing vegetable saccharide after the maltotriose generating amylase action step and the material containing vegetable oil after the lipase action step are mixed, and vegetable ice cream can be manufactured. In addition, a method in which maltotriose generating amylase is acted on a material containing vegetable saccharide and vegetable oil and then lipase is acted on a material containing vegetable saccharide and vegetable oil and then maltotriose generating amylase is acted, a method in which maltotriose generating amylase and lipase are simultaneously acted on a material containing vegetable saccharide and vegetable oil and the like can be cited. In the present technology, it is particularly preferred that a method of manufacturing vegetable ice cream by performing each step separately, mixing a material containing vegetable saccharide after the maltotriose generating amylase action step and a material containing vegetable oil after the lipase action step is mixed.

[0123] (4) β-amylase action process

[0124] The β-amylase action step is a step of allowing β-amylase to act on a material containing plant saccharides. As long as the effect of the present technology is not impaired, the amount of β-amylase added in the β-amylase action step can be freely set. The specific amount of β-amylase added in the present technology is the same as the content of β-amylase in the aforementioned enzyme agent for improving shape retention, so the description is omitted here.

[0125] As long as the effect of the present technology is not impaired, the various conditions of the β-amylase action process can be freely set. For example, the 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 β-amylase used. The pH can be set to pH5 to 9, preferably pH6 to 8, and more preferably pH6.5 to 7.5. The temperature can be set to 20 to 80°C, preferably 30 to 70°C, and more preferably 40 to 60°C. The action time can be set to 5 minutes to 24 hours, preferably 15 minutes to 12 hours, and more preferably 30 minutes to 2 hours. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0126] (5) α-amylase action process

[0127] The α-amylase action step is a step of allowing α-amylase to act on a material containing vegetable saccharides. By allowing α-amylase to act on a material containing vegetable saccharides, starch in the material containing vegetable saccharides can be liquefied. In the present technology, the α-amylase action step is not an essential step. For example, by using a material containing vegetable saccharides that has been treated with α-amylase in advance, the same effect as that of performing the α-amylase action step can be obtained.

[0128] The amount of α-amylase added in the α-amylase action step can be freely set as long as the effect of the present technology is not impaired. The specific amount of α-amylase added in the present technology is the same as the content of α-amylase in the aforementioned enzyme agent for improving shape retention, so the description is omitted here.

[0129] As long as the effect of the present technology is not impaired, the various conditions of the α-amylase action process can be freely set. For example, the 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. The pH can be set to pH5.0-8.0, preferably pH5.5-7.5, and more preferably pH6.0-7.0. The temperature can be set to 30°C-80°C, preferably 40°C-75°C, and more preferably 50°C-70°C. The action time can be set to 10 minutes-12 hours, preferably 30 minutes-6 hours, and more preferably 1 hour-3 hours. In addition, the optimal reaction conditions can be determined by preliminary experiments.

[0130] The order of performing the α-amylase action step is not limited unless the effects of the present technology are impaired, but it is preferably performed before the maltotriose generating amylase action step and / or the β-amylase action step, or simultaneously with the maltotriose generating amylase action step and / or the β-amylase action step.

[0131] An enzyme inactivation step may be performed after each enzyme action step described above.

[0132] (6) Monoglyceride and / or diglyceride addition step

[0133] The monoglyceride and / or diglyceride adding step is a step of adding monoglyceride and / or diglyceride in the manufacturing step of vegetable ice cream. In the present technology, the monoglyceride and / or diglyceride adding step is not an essential step, and for example, by using vegetable materials containing monoglyceride and / or diglyceride, the same effect as performing the monoglyceride and / or diglyceride adding step can be obtained.

[0134] The amount of monoglyceride and / or diglyceride added in the monoglyceride and / or diglyceride addition step can be freely set as long as the effect of the present technology is not impaired. The specific amount of monoglyceride and / or diglyceride added in the present technology is the same as the content of monoglyceride and / or diglyceride in the aforementioned enzyme agent for improving shape retention, so the description is omitted here.

[0135] As long as the effect of the present technology is not impaired, the order of performing the monoglyceride and / or diglyceride addition process is not limited. For example, after adding monoglyceride and / or diglyceride to a material containing vegetable sugars and / or a material containing vegetable oils and fats, one can freely choose a method of performing a maltotriose-generating amylase action process and / or a lipase action process, a method of adding monoglyceride and / or diglyceride to a material containing vegetable sugars and / or a material containing vegetable oils and fats that has undergone a maltotriose-generating amylase action process and / or a lipase action process, etc. In the present technology, it is particularly preferred to adopt a method of adding monoglyceride and / or diglyceride to a material containing vegetable oils and fats and then performing a lipase action process.

[0136] (7) Recycling process

[0137] The recovery step is a step of recovering the vegetable ice cream produced through one or more steps selected from the group consisting of the maltotriose generating amylase action step, the lipase action step, and the β-amylase action step, and, if necessary, the α-amylase action step, the monoglyceride and / or diglyceride addition step, etc. The specific recovery method can be one or more recovery methods used in the production of vegetable ice cream in general, depending on the type or method of the vegetable ice cream produced.

[0138] The method for producing the plant-based ice cream and the method for improving the shape retention according to the present technique described above can adopt an embodiment including the following steps (1) to (8).

[0139] (1) Process of preparing a material containing vegetable sugars and / or a material containing vegetable oils and fats

[0140] (2) Step of allowing maltotriose-generating amylase to act on the prepared material containing plant sugars

[0141] (3) Step of allowing lipase to act on the prepared material containing vegetable oil

[0142] (4) Step of allowing β-amylase to act on the prepared material containing plant sugars

[0143] (5) Step of allowing α-amylase to act on the prepared material containing plant sugars

[0144] (6) Step of adding monoglyceride and / or diglyceride

[0145] (7) Enzyme inactivation process

[0146] (8) Process of recycling plant-based ice cream

[0147] In addition, any one of steps (2) to step (4) may be performed, two steps may be performed from them, or all steps may be performed. When performing two or more steps, the order is not particularly limited, and they may be performed separately or simultaneously. When they are performed separately, the enzyme in step (7) may be inactivated as needed between each step. In addition, steps (5) to step (8) are not required and may be performed as needed. When performing step (5), the order of steps (2) to step (4) is not particularly limited, and they may be performed separately or simultaneously. When they are performed separately, the enzyme in step (7) may be inactivated as needed between each step. In addition, when performing step (6), it may be performed between any one of steps (1) to step (7), or it may be performed simultaneously with any one of steps (2) to step (5).

[0148] Example

[0149] Hereinafter, the present invention will be described in more detail based on examples. In addition, the examples described below are examples of representative examples of the present invention, and the scope of the present invention is not to be construed narrowly thereby.

[0150] 1. Materials and enzymes used

[0151] The materials and enzymes used in the Examples are shown in Table 1 below.

[0152] [Table 1]

[0153]

[0154] 2. Enzyme activity assay

[0155] [Method for determining the activity of maltotriose-producing amylase]

[0156] An appropriate amount of enzyme is added to 0.5 ml of 2% soluble starch dissolved in 0.1 M phosphate buffer (pH 7.0), and a total of 1.0 ml is reacted at 40°C. The generated maltotriose and other reducing sugars are quantified using the Somogyi-Nelson method. Under these conditions, the amount of enzyme that generates reducing sugars equivalent to 1 micromole of glucose in 1 minute is considered 1 unit (1U).

[0157] [Lipase activity determination method]

[0158] First, 45g of olive oil was mixed with 18.5g / L emulsion (polyvinyl alcohol (completely saponified, saponification degree 98.8±0.2), 150mL polyvinyl alcohol (partially saponified, saponification degree 88.8±1.0) 1.5g / L), and emulsified using a homogenizer to prepare a matrix solution. 1mL of lipase enzyme solution was added to 5mL of the prepared matrix solution and 4mL of 0.1mol / L phosphate buffer (pH7.0), and the reaction was carried out at 37°C. After 30 minutes, 10mL of ethanol-acetone mixture was added to stop the enzyme reaction. Next, 10mL of 0.05mol / L sodium hydroxide solution and 10mL of ethanol-acetone mixture were added, and the pH was titrated to 10 with 0.05mol / L hydrochloric acid. In addition, the amount of enzyme that brings about an increase of 1μmol of fatty acids in 1 minute is taken as 1 unit (1U).

[0159] [β-amylase activity determination method]

[0160] The determination was carried out according to the method described in the official book of food additives (9th edition). Specifically, potato starch was used as the matrix, dried at 105°C for 2 hours in advance, 1.0g of the dried product was measured, 20mL of water was added, and 5mL of sodium hydroxide test solution (2mol / L) was slowly added while stirring to make a paste. Then, after heating in a water bath for 3 minutes while stirring, 25mL of water was added. After cooling, hydrochloric acid test solution (2mol / L) and hydrochloric acid test solution (0.1mol / L) were added for neutralization, 10mL of 1mol / L acetic acid-sodium acetate buffer (pH5.0) was added, and water was added to 100mL to make this as the matrix solution. 10 mL of the matrix solution was measured and heated at 37°C for 10 minutes, 1 mL of the sample solution was added and immediately shaken to mix, and after heating at the same temperature for 10 minutes or 30 minutes, 4 mL of Fehling's test solution was further added and gently shaken to mix, and after heating in a boiling water bath for 15 minutes, cooled to below 25°C, 2 mL of 30% potassium iodide solution and 2 mL of sulfuric acid (1→6) were added to prepare the test solution. In addition, the Fehling's test solution was prepared by measuring 34.66 g of fine crystals of copper (II) sulfate pentahydrate and dissolving them in water to prepare 500 mL of copper solution and measuring 173 g of (+)-sodium potassium tartrate tetrahydrate and 50 g of sodium hydroxide and dissolving them in water to prepare 500 mL of alkaline tartrate solution, and mixing them at a ratio of 1 volume of copper solution to 1 volume of alkaline tartrate solution. In addition, 10 mL of water was used instead of the matrix solution, and the same operation as the test solution was performed to prepare the comparative solution. For the test solution and the comparative solution, the free iodine is titrated with a 0.05 mol / L sodium thiosulfate solution. The end time is when the blue color produced by adding 1 to 2 drops of soluble starch test solution disappears near the end time of the titration. The amount of enzyme that increases the reducing power equivalent to 1 mg of glucose in 1 minute is taken as 1 unit (1U) and calculated by the following formula.

[0161] β-amylase activity (U / g, U / mL) = amount of glucose (mg) × 1 / 10 × 1 / M

[0162] Amount of glucose (mg) = (ba) × 1.6 × f

[0163] a: Titration value of enzyme reaction solution (mL)

[0164] b: titration value of blank solution (mL)

[0165] 1.6: 1 mL of 0.05 mol / L sodium thiosulfate solution is equivalent to 1.6 mg of glucose

[0166] 1 / 10: Unit conversion factor for reaction time (minutes)

[0167] M: The amount of sample in 1 mL of sample solution (g or mL)

[0168] f: Factor of 0.05 mol / L sodium thiosulfate solution (for quantitative use)

[0169] [Method for determining the activity of α-amylase]

[0170] After 10 mL of 1% dextrin potato is pretreated at 37°C for 15 minutes, 1 mL of amylase solution is added and mixed immediately. After 10 minutes at 37°C, 1 mL is taken out and mixed with 10 mL of 0.1M hydrochloric acid. 0.5 mL is taken out from the solution and mixed with 10 mL of 0.2 mM iodine solution, and the absorbance is measured (wavelength 660 nm). The amount of enzyme that reduces the color development caused by iodine reaction by 10% within 1 minute is defined as 1 unit (1U).

[0171] 3. Experimental Examples

[0172] <Experimental Example 1>

[0173] In Experimental Example 1, the effect of improving the shape retention of vegetable-based ice cream by maltotriose-producing amylase and β-amylase was investigated.

[0174] (1) Production of plant-based ice cream

[0175] 120 g of oat powder was added to 1 L of purified water, and mixed at 50° C. for 20 minutes at a speed of 2 using a heating mixer (“Thermomix TM6”, manufactured by Vorwerk). Then, the enzymes shown in Table 2 below were added, and the mixture was stirred for another hour. Then, the mixture was passed through a cheese filter cloth to separate the solid content, which was used as an oat base.

[0176] 4.48 g of mono- and diglycerides were added to 89.6 g of refined coconut oil, and mixed at 70° C. for 20 minutes at speed 2 using a heating mixer (“ThermomixTM6”, manufactured by Vorwerk). Next, 89.6 g of glucose and 78.4 g of sugar were added, and mixed for another 10 minutes to prepare a coconut oil base.

[0177] The prepared oat base was added to the prepared coconut oil base, heated to 85°C, and stirred for 15 minutes. Then, the mixture was transferred to a bowl, emulsified with a hand mixer for 5 minutes, and stored in a refrigerator overnight. The next day, 2 g of vanilla flavor and 2 g of salt were added, mixed for 30 to 45 seconds, and plant-based ice cream was manufactured by an ice cream maker (NOSTALGIA PRODUCTS, model: PICM20DBL).

[0178] (2) Sugar composition analysis

[0179] The above-obtained 0.5 mL oat base was diluted 5 times with purified water, centrifuged (13,000 rpm, 5 minutes), the supernatant was taken out, and the sugar composition of the membrane filter (0.2 μm) was analyzed by high performance liquid chromatography (detector: RID-20A, manufactured by Shimadzu Corporation). The column used was CK04S (manufactured by Mitsubishi Chemical Corporation), and the degree of polymerization of sugars was analyzed in particular. The ratio of each sugar component to the total peak area is shown in Table 2 below.

[0180] (3) Conformity evaluation

[0181] The prepared vegetable ice cream is filled into paper cups and left overnight at -15°C before being frozen and formed. The shape retention is evaluated by placing the ice cream with the paper cup removed horizontally on a mesh plate on a scale at room temperature. The scale value is read every 5 to 90 minutes, starting from the time when the first drop of ice cream falls on the scale when it starts to melt.

[0182] Then, the value calculated using the following mathematical formula (1) was used as the evaluation value of the shape retention.

[0183] Shape retention (%) = {((weight of ice cream at the beginning) - (value of the scale after 90 minutes)) ÷ (weight of ice cream at the beginning)} × 100… (1)

[0184] The shape retention rate of increase was calculated using the following mathematical formula (2) for the obtained evaluation value (shape retention (%)).

[0185] Shape retention increase rate (%) = (shape retention of each test area / shape retention of the control product) × 100… (2)

[0186] (4) Results

[0187] The results are shown in Table 2 below.

[0188] [Table 2]

[0189]

[0190] (5) Investigation

[0191] As shown in Table 2, the test area 1 in which the plant-based ice cream was produced using β-amylase and the test area 2 in which the plant-based ice cream was produced using maltotriose-producing amylase showed high shape retention compared to the control 1 in which the plant-based ice cream was produced without using maltotriose-producing amylase. In particular, the shape retention was significantly improved in the test area 2 in which the plant-based ice cream was produced using maltotriose-producing amylase.

[0192] <Experimental Example 2>

[0193] In Experimental Example 2, the dependency of maltotriose-producing amylase on the amount of addition was investigated.

[0194] (1) Production of plant-based ice cream

[0195] In the same manner as in Experimental Example 1, plant-based ice creams of Control 2 and Test Groups 3 and 4 shown in Table 3 were produced.

[0196] (2) Conformity evaluation

[0197] The shape retention rate was evaluated in the same manner as in Experimental Example 1.

[0198] (3) Results

[0199] The results are shown in Table 3 below.

[0200] [Table 3]

[0201]

[0202] (4) Investigation

[0203] As shown in Table 3, the test plots 3 and 4 in which the plant-based ice cream was produced using maltotriose-producing amylase showed high shape retention compared to the control product 2 in which the plant-based ice cream was produced without using maltotriose-producing amylase. In addition, the results of the test plots 3 and 4 show that the maltotriose-producing amylase exhibits a sufficient shape retention-enhancing effect even at about 1.2 U.

[0204] <Experimental Example 3>

[0205] In Experimental Example 3, the effect of improving the shape retention of vegetable-based ice cream by lipase was investigated.

[0206] (1) Production of plant-based ice cream

[0207] 120 g of oat powder was added to 1 L of purified water, and mixed at 50° C. for 20 minutes at a speed of 2 using a heating mixer (“Thermomix TM6”, manufactured by Vorwerk). Next, α-amylase in the amount shown in Table 4 below was added, and the mixture was stirred for another hour. The mixture was then passed through a cheese filter cloth to separate the solid content, which was used as an oat base.

[0208] 4.48 g of mono- and diglycerides were added to 89.6 g of refined coconut oil, and mixed at 70° C. for 20 minutes at speed 2 using a heating mixer (Thermomix TM6, manufactured by Vorwerk). Next, lipase in the amount shown in Table 4 below was added to test plot 5, and the mixture was stirred at 50° C. for 30 minutes. Then, 89.6 g of glucose and 78.4 g of granulated sugar were added, and mixed for another 10 minutes to prepare a coconut oil base.

[0209] Using the prepared oatmeal base and coconut oil base, plant-based ice cream was manufactured in the same manner as in Experimental Example 1.

[0210] (2) Conformity evaluation

[0211] The shape retention rate was evaluated in the same manner as in Experimental Example 1.

[0212] (3) Results

[0213] The results are shown in Table 4 below.

[0214] [Table 4]

[0215]

[0216] (4) Investigation

[0217] As shown in Table 4, Test Section 5 in which plant-based ice cream was produced using lipase showed higher shape retention than Control Section 3 in which plant-based ice cream was produced without using lipase.

[0218] <Experimental Example 4>

[0219] In Experimental Example 4, the dependency of the amount of lipase added was examined.

[0220] (1) Production of plant-based ice cream

[0221] In the same manner as in Experimental Example 3, plant-based ice creams of Control 4 and Test Sections 6 and 7 shown in Table 5 were produced.

[0222] (2) Conformity evaluation

[0223] The shape retention rate was evaluated in the same manner as in Experimental Example 1.

[0224] (3) Results

[0225] The results are shown in Table 5 below.

[0226] [Table 5]

[0227]

[0228] (5) Investigation

[0229] As shown in Table 5, it is known that the shape retention is improved by allowing lipase to act on the material containing vegetable oil. In addition, it is known that the shape retention is improved depending on the amount of lipase added.

Claims

1. An enzyme agent for improving the shape retention of plant-based ice cream, comprising one or more enzymes selected from the group consisting of maltotriose-producing amylase, lipase, and β-amylase. 2 . A vegetable-based ice cream comprising the enzyme agent for improving shape retention according to claim 1 .

3. A method for producing plant-based ice cream, comprising one or more steps selected from a maltotriose-generating amylase action step of allowing maltotriose-generating amylase to act on a material containing plant saccharides, a lipase action step of allowing lipase to act on a material containing plant oils and fats, and a β-amylase action step of allowing β-amylase to act on a material containing plant saccharides.

4. A method for improving the shape retention of plant-based ice cream, comprising one or more steps selected from a maltotriose-generating amylase action step of allowing maltotriose-generating amylase to act on a material containing plant saccharides, a lipase action step of allowing lipase to act on a material containing plant oils and fats, and a β-amylase action step of allowing β-amylase to act on a material containing plant saccharides.

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