JP2709289B2 - How to make plants completely unicellular - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for completely transforming plants such as vegetables, wild grasses, fruits, medicinal plants, tree leaves, and bamboo grass leaves into single cells using an intercellular substance-decomposing enzyme.

[0002]

2. Description of the Related Art In order to develop effective uses of agricultural products, processed residues thereof, herbs and the like, it is necessary to selectively recover only useful parts. In order to recover this useful part, there is a method of mechanically grinding using a mixer or the like, but according to this method, the cell wall of the agricultural product is destroyed, and the useful components contained in the cells are lost or changed. In particular, aroma components, pigment components, nutrient components and the like in the cells are lost or deteriorated due to the destruction of the cell wall, and the useful components cannot be efficiently recovered.

[0003] The present inventor has proposed a method for easily producing a single-cellular plant-containing food by using a method for selectively and efficiently recovering only useful components by eliminating the problems in the conventional mechanical grinding method. As a result, a plant cell single cell obtained by treating a plant with a plant tissue single cell enzyme at pH 4 to 5 to decompose the intercellular substance of the plant to convert the plant tissue into a single cell, and further removing undegraded substances A method for producing a single-cellular plant-containing food, in which the suspension is added to the food, was previously proposed, and this method was published as a patent application as Japanese Patent Publication No. 6-71413.

[0004]

SUMMARY OF THE INVENTION The above-mentioned Japanese Patent Publication No. 6-71.
As described above, the method of No. 413 eliminates the problems of the conventional mechanical grinding method and selectively and efficiently recovers only useful components. 7
When the method of No. 1413 is applied, plant materials that can be food include leaf vegetables, root vegetables, potatoes, legumes, cereals, wild grasses, medicinal herbs, tree leaves, and the like. Because of the myriad counts, unicellularization is impossible even if enzymatic action is performed by the same method, or only a small portion can be unicellularized, or unicellularization varies. Therefore, according to the method of Japanese Patent Publication No. Hei 6-71413, some vegetables and fruits can be made into single cells and can be used as food materials. In order to put plant single cell technology on an industrial scale, This method is still insufficient. The plant tissue single cell-forming enzyme (cell separating enzyme) used in the method of Japanese Patent Publication No. 6-71413 is an enzyme isolated from a filamentous fungus of the genus Rhizopus, and is endo-polygala.
cturonase and pectin-trans-el
iminase is the main component and does not contain cellulase.

[0005] In such a cell separating enzyme, as described above,
Only some plants can be converted into single cells, and it is impossible to decompose all of these plants, which are made up of numerous plant parenchyma, into single cells. To be specific, protopectin molecules that form a plant while maintaining a certain degree of hardness by agglutinating innumerable cells that make up soft tissue include:
There are various forms. That is, a relatively simple protopectin in which one pectin molecule is bound to another pectin molecule, a hemicellulose molecule or a cellulose molecule via Ca to which one pectin molecule is bound, and a free -COOH group is contained in one pectin molecule. There is a case in which protopectin is present in which 2 to 3 are linked and Ca is linked and linked to 2 or 3 other pectin molecules, hemicellulose molecules or cellulose molecules and the like, In the latter case, the agglutination between cells becomes stronger than in the former case in which they are linked via a single Ca, and is therefore less susceptible to enzymatic action. The enzyme used in the method of Japanese Patent Publication No. 6-71413 described above has an extremely weak decomposing ability on protopectin having such a strong bond.

Further, in the conventional single cell technology,
Exactly the same pretreatment and other treatments were performed even when the plant types were different. However, different types of plants, for example, hard tissue plants or soft tissue plants, plants whose surface is coated with a wax substance, plants containing a discoloration causing substance, plants containing astringency, astringency components, moisture, Conventionally, there are no proposals for a method of completely transforming a plant into a single cell by applying the above-described effective treatment or a method for transforming a single cell to a plant having a low content, a plant containing a component having a viscous property, or the like. Not done.

[0007] It is an object of the present invention to solve the above problems and to provide a method for decomposing all plant parenchyma to completely convert it into a single cell.

[0008]

Means for Solving the Problems The method for completely transforming a plant into single cells according to the present invention has been made in order to achieve the above-mentioned object. The method comprises the steps of providing a plant with an enzyme isolated from a filamentous fungus of the genus Rhizopus; The present invention is characterized in that an intercellular substance-degrading enzyme mainly composed of protopectinase, which dissociates natural protopectin, is acted on to decompose intercellular substances in plant parenchyma to completely convert a plant into a single cell.

[0009] The intercellular substance-decomposing enzyme used in the present invention is an enzyme capable of decomposing even extremely strong plant tissues such as tree leaves and bamboo grass leaves. It also has a decomposing effect on insoluble protopectin obtained by completely removing all pectin, and is also strong against Kumasa basal leaves, which the cell-separating enzyme used in the method of Japanese Patent Publication No. 6-71413 shows almost no degrading effect. Shows decomposition action.

In the present invention, when a plant parenchyma is dissociated into a single cell state by an enzyme, the protopectin and the protopectin, in which a myriad of cells constituting the plant tissue are stuck to each other to maintain a certain hardness. It is necessary to decompose agglutinates consisting of small amounts of hemicellulose and other compounds. Among these intercellular agglutinants, the fact that the main component is protopectin suggests that if there is an enzyme that mainly degrades protopectin, it is possible to break down the plant parenchyma and release single cells. Although, other hemicellulosic substances, as a result of examining what relationship the activity of cellulose or pectinolytic enzyme to the strength and rate of single cell dissociation, the present inventors have found that vegetables, wildflowers, fruits,
Rhiz, an intercellular substance-degrading enzyme that completely dissociates various plant parenchyma tissues such as herbs, tree leaves and kuma bamboo leaves into single cells.
An enzyme isolated from filamentous fungi of the genus opus, which is a proteopectinase that dissociates strong protopectin, has been found to be an intercellular substance-degrading enzyme of the present invention, and this enzyme is used in the method of Japanese Patent Publication No. 6-71413. It was confirmed that the action mechanism was different from that of the cell separating enzyme.

The fresh plant material is a multicellular body, not a single cell aggregate, but a tissue and an organ, each of which is a group of cells having a unique function. In such tissues and organs, countless cells surrounded by a double cell wall and cell membrane adhere to each other with intercellular adhesive substances (middle lamella, middle lobe) mainly composed of protopectin and containing a small amount of hemicellulose, pectin and lignin. It is configured. In this case, the cell wall is mainly composed of cellulose, is formed of hemicellulose, pectin protein, and the like, and is made strong to protect cell contents. In addition, protopectin, which is a main component of the intercellular adhesive substance that forms an organ or tissue by gluing innumerable cells, is composed of a large number of galacturonic acids bound linearly, and 12 to 12 of the COOH group of the galacturonic acid. 17% is methyl ester (CO
OCH ₃ ), but free COOH groups include Ca
Binds and insolubilizes, or binds to other pectin molecules, cellulose molecules, and hemicellulose molecules via Ca to form a hardened tissue or organ as an insoluble strong polymer complex. Lignin also deposits between these cells. Depending on the plant, there are a few cell-cell adhesives mainly composed of hemicellulose such as araban and xylan.

In addition, several free COOH groups of protopectin are present in linkage, and Ca binds to each COOH, and binds to other pectin molecules, cellulose molecules, and hemicellulose molecules via the Ca. It forms a stronger organization. It is presumed that the intercellular adhesion of hard tissues such as tree leaves and bamboo grass leaves is stuck with such complex protopectin.

[0013] Enzymes that dissociate plant parenchyma into single cells have been studied and published in patent publications and research reports. However, although these enzymes are indicated as the titers of the enzymes involved in them, it is not clear what percentage of plants actually dissociate into single cells. And, to date, no enzyme has been published yet that has the ability to completely dissociate single cells even in any robust plant tissue.

The inventor of the present invention differs from the single cell-forming action of plant parenchyma by enzymes disclosed so far, in addition to vegetables, wild grasses, fruits, herbs, etc., as well as hard plant parenchyma such as tree leaves and kuma bamboo leaves. In order to create a single-cell suspension or powder in a state that is completely industrialized and profitable, it completely dissociates and dissociates all of the composed plant fibers such as leaf veins, stems and the like and cells stuck to it. Without destroying the cells at all
We have developed an intercellular substance-degrading enzyme (hereinafter, referred to as the present intercellular substance-degrading enzyme) that has the action of completely dissociating even strong plant tissues into single cells.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The present intercellular substance degrading enzyme is Rhi.
An enzyme isolated from a filamentous fungus of the genus zoopus, which is an enzyme mainly composed of protopectinase that dissociates strong protopectin, and is produced by a mixed solid culture of a bran of a filamentous fungus belonging to the genus Rhizopus, a peel of orange, and the like. A clear liquid obtained by extracting the substance with water is precipitated by adding 25 to 75% of saturated alcohol to obtain a powder. Cellulose-based substances, pectin-based substances, hemicellulose-based substances, protopectin-based substances, and the degree of disintegration single cells (plant-tissue-degrading enzyme activities) of plant tissues, etc. were measured for the present intercellular substance-degrading enzyme, and the results are shown in the following table. 1, Table 2 and Table 3. For comparison, the cell-separating enzyme, cellulase (cellula) used in the method of JP-B-6-71413 was used.
se) and endopolygalacturonase (endo-p
Polygalacturonase) was measured in the same manner, and the results are shown in Tables 1, 2 and 3 below.

The present intercellular substance-degrading enzyme, which decomposes protopectin, which is an intercellular agglutinating substance, dissociates plant parenchyma and releases single cells, and the above-mentioned cell-separating enzymes are different from other cellulases and endopolygalacturonase. In contrast, the cellulase-based enzyme activity is almost non-existent. Among them, the present intercellular substance-decomposing enzyme has a lower activity than the above-mentioned cell-separating enzyme except for the β-cellobiose-degrading activity, and contains almost no cellulase-based enzyme activity. This means that the cell wall of the generated single cell is not degraded to reduce the cell number. Table 4 below shows the results of the decomposition of the cell wall by the four enzymes shown in Tables 1 to 3 for a single cell suspension of ginseng. Even if the results of Table 4 below in which the present intercellular substance-decomposing enzyme was added to the ginseng single cell suspension at a high concentration were not found, there was no difference in the number of cells.

As shown in Tables 1, 2 and 3 below,
Regarding the hemicellulase activity of the present intercellular substance-degrading enzyme, although xylanase activity is slightly observed, hemicellulose binds to or coexists with protopectin in intercellular agglutinants of plant parenchyma. Although slightly observed, xylan is often present, and araban is contained only in limited plants such as sugar beet. The fact that the present intercellular substance degrading enzyme contains only a small amount of xylanase activity seems to have a slightly advantageous effect on the decomposition of intercellular agglutinants.

Further, the pectic substance-degrading activity does not include any pectic acid liquefaction activity (endo-polygalacturonase), and the pectin esterase activity is slightly contained in the present intercellular substance degrading enzyme. Furthermore, it does not contain any pectin-trans-eliminase activity at all.
Unlike the cell-separating enzyme in the method described in Japanese Patent Publication No. 6-71413, no unicellular action by endo-polygalacturonase or pectin-trans-eliminase is performed at all.

Next, not only water-soluble pectic substances such as pectin and pectic acid but also protopectin which constitutes a strong plant tissue which is completely insoluble in hot water at 60 ° C.
The protopectin-degrading activity (protopectinase activity) that releases water-soluble pectin is the most potent of this intercellular substance-degrading enzyme, and the above-mentioned cell-separating enzyme is less active than the present intercellular substance-degrading enzyme, and cellulase is more active And endo-polygalacturonase has little activity. The results of dissociating a 1 cm square ginseng tissue into single cells by the four enzymes shown in Tables 1 to 3 below are shown in Tables 1 to 3 below.
The results are shown in Table 5 below together with Table 3. As shown in the index of protopectinase activity, the ginseng disintegration rate of this intercellular substance-decomposing enzyme is the highest, and the number of single cells in 1 ml of the obtained single cell suspension is as large as 5.9 × 10 ^6. . on the other hand,
Since the protopectinase activity of the above-mentioned cell-separating enzyme is considerably lower than that of the present intercellular substance-degrading enzyme, the rate of unicellularization of ginseng tissue is about 15% lower than that of the present intercellular substance-degrading enzyme. The number of single cells in 1 ml of the suspension is 3.2 × 10 ⁶ , which is about half that of the present intercellular substance-decomposing enzyme. In particular, in plants having strong tissues such as kuma bamboo leaves, the structure of protopectin is considered to be different from vegetables, wildflowers, and tree leaves.
%, And has little effect on cellulase and endo-polygalacturonase. As described above, the present intercellular substance-degrading enzyme has two or three galacturonic acids having a free COOH group constituting a pectin molecule of protopectin linked to each other, and further connected to the linked COOH group. Even if it has a strong structure due to the presence of other cellulose, pectin and hemicellulose molecules via Ca bound thereto, it has the ability to decompose this insoluble and bound strong pectin molecule Therefore, it was found that intercellular agglutinants in any plant parenchyma were randomly decomposed and dissociated into single cells. Therefore, this enzyme is used to completely dissociate the cells that make up any plant parenchyma, such as vegetables, wildflowers, fruits, medicinal plants, tree leaves, and bamboo grass leaves, into single cells and separate long fibers such as leaf veins and stems. The single-cell suspension can be fractionated at a high yield to produce food.

Next, a case where a tree leaf or the like whose leaf surface is covered with a wax substance is made into a single cell will be described.

When tea leaves are converted into single cells using the present intercellular substance-decomposing enzyme, the leaves may be untreated in the case of young leaves from May to August, but the leaves are protected by wax in the old leaves from September to April. As it is, there is no permeation of the enzyme solution as it is, and no single-cell dissociation action occurs. Then, the temperature 40 ~
Tea leaves are immersed in a NaOH solution of about 0.01 to 1% at 50 ° C. and stirred for about 30 seconds to about 3 minutes to dissolve and remove the wax material on the surface to lose luster, and wash with water to remove alkali. 90 ~
The tea leaves are immersed in hot water at a temperature of 95 ° C. for 1 to 2 minutes to perform a blanching process. The tea leaves are pulled out of the hot water, cooled with water, cut into 1 cm widths, put into a stirring tank, and placed in a stirring tank.
% Of water to reduce the intercellular substance-degrading enzyme to about 0.3 to
When 0.5% is added and stirred at 30-40 ° C for 90-120 minutes, countless cells stuck to veins and petiole are all dissociated into single cells, and long and short fibers such as veins and petiole and green Of a single cell suspension. This is removed with a sieve of about 20 mesh to obtain a green single-cell suspension. Table 6 below shows the yield of the tea cell single cell suspension, the number of single cells in 1 ml, and other measurement results. The yield of the single cell suspension with respect to all the raw materials was about 85 to 90%. The number of single cells in 1 ml of the solution is about 1.0 × 10 ⁷ .

Even if the temperature is below 5 ° C., the leaves of the bamboo grass turn black and dry and do not receive any enzymatic action. In addition, even when treated with alkali, no enzymatic action is received and no single cells are formed. Therefore, when stored in running water of 15 ° C. or less for long-term storage and swelling of the tissue, storage can be easily performed while maintaining freshness for one month or more. Besides, for moderate water absorption,
The bamboo grass leaves soften and become susceptible to the action of the present intercellular substance-decomposing enzyme, so that a single cell can be obtained with a recovery rate of about 50 to 60%. In addition, overwintering and dilapidated Kumasa bamboo leaves have a temperature of 40.degree.
It is immersed in 1% HCl solution for 30 minutes to swell the tissue, then immersed in 0.1-1% NaOH solution at 40 ° C. for 2-3 minutes, washed well with water and cut, and hydrated to about 100-200% of the raw material. And add 1 to 3% of the present intercellular substance degrading enzyme,
Work at 40 ° C. with stirring. Table 7 below shows the results obtained by converting Kuma-sasa leaves into single cells using the present intercellular substance-decomposing enzyme. Depending on the pretreatment method, the decomposition rate of Kuma-basa bamboo leaves differs, but the running water immersion method (water exposure) shows the best decomposition rate, and it can be decomposed by about 50%. Of course, the obtained single cells show a vivid color tone of dark green. The ginkgo leaves are immersed overnight in a 0.01% NaOH aqueous solution to remove the waxy substances on the leaves, or the raw material is immersed in running water (exposed to running water).
If the present intercellular substance-decomposing enzyme is allowed to act under stirring for 2 hours at a temperature of 30 ° C. with a% of water, a single cell suspension can be obtained with a recovery rate (yield) of about 60 to 65%.
When blanching is performed simultaneously with dewaxing at 90 ° C. for 1 minute with an aqueous solution of 0.05% NaOH or the enzyme is allowed to act without any treatment, the recovery (yield) of the single cell suspension is 70%.
As described above, the color tone of a single cell deteriorates, and the commercial value decreases or disappears. Table 8 below shows the results of the above-mentioned single cell formation of Ginkgo biloba.

In addition, cells such as green tea, sencha, and tencha are subjected to heating, rolling, and drying treatments such as steaming, roasting, and mild fermentation to kill cells, and are transformed and dried to form intercellular agglutinants. Table 9 below shows an example in which Tencha was decomposed, deformed and dissociated to separate dead cells and decomposed into single cell particles. In the case of Tencha, a single cell suspension can be obtained at a yield of about 75% with only one enzymatic treatment, but once the residue is enzymatically treated, a yield of 85% with respect to the first raw material is eventually achieved. To recover a single cell suspension, leaving about 10% of fiber residue.

Thus, like tree leaves, bear bamboo leaves, etc.
The leaves are hard-glued and shaped by an extremely strong intercellular agglutinant, and the leaves are hard-coated with a waxy material and protected, even if the leaves are protected by appropriate pretreatment without intracellular chlorophyll or other degradation. , The countless cells stuck to the stem and petiole can be dissociated into single cells, and all can be recovered.

Next, a method for inactivating oxidative enzymes in vegetables, wild plants and fruits and preventing discoloration due to polyphenols contained therein will be described.

The parenchyma of vegetables, fruits, and wild plants oxidizes chlorogenic acid, caffeic acid, catechol catechin, and many other polyphenols, as well as amino acids such as polyphenol oxidase, peroxidase, and tyrosine, which oxidize and discolor them. Catecholase to discolor,
Some include oxidases such as ascorbate oxidase that oxidize and discolor vitamin C. As for vegetables, spring chrysanthemum, Takana, radish leaf, spinach, burdock,
Butterflies, eggplants, potatoes, yams, lotus roots, and the like, and fruits include apples, peaches, and pears. Therefore, the discoloration due to oxidation occurs during the pretreatment operation or during the storage, as well as during the operation of forming a single cell such as stirring, and the quality deteriorates. Furthermore, it contains chlorophyllase A, which degrades chlorophyll A, which is the main component of chlorophyll. From this fact, if raw leaves or fruits are converted into single cells as they are under the condition of promoting oxidation as in the stirring operation with the present intercellular substance-decomposing enzyme, the oxidase or chlorophyllase A works during the operation, for example, polyphenols Is oxidized and changes its color from yellow to brown, and then from brown to black, and chlorophyll is decomposed to a brown pigment, deteriorating the quality and losing commercial value. Therefore, blanching is performed in advance in hot water at a temperature of, for example, 80 ° C. or more to such an extent that the cell wall is not destroyed for 1 to 5 minutes to inactivate these enzymes, and then the cells are decomposed into single cells by the present intercellular substance-decomposing enzyme.

When a light-colored product such as a fruit is cut off by peeling, cutting, or the like, the oxidase immediately oxidizes the polyphenol contained therein and changes its color to yellow or brown. In such fruits, antioxidants such as L-ascorbic acid, sodium L-ascorbate, erythorbic acid, and glutathione are required to stop the action of oxidase immediately after peeling and cutting, and to prevent autoxidation of polyphenol itself. Therefore, it is necessary to prevent the raw fruit from being oxidized and discolored before the treatment with the present intercellular substance-decomposing enzyme by immersion in a solution in which a pH-lowering agent such as citric acid is dissolved. Further, when the fruit and the like are converted into single cells under stirring with the present intercellular substance-decomposing enzyme, an antioxidant is similarly added in a slightly excessive amount in order to prevent the action of the oxidase and the autoxidation of the polyphenol itself. If the amount of the antioxidant is insufficient, the oxidation of the polyphenol proceeds during the action of the enzyme and browns. In addition, depending on the type of the raw material plant, it can be carried out only by any of the above-mentioned methods of blanching and adding an antioxidant. In the case of cucumber in Tables 10 and 11 below,
In the case of apple (Wang Lin) in Table 12 below and in the case of lotus root, sweet potato and potato in Table 13 below, these raw materials were dissociated by the above method using the present intercellular substance-decomposing enzyme. The result of single cell formation is shown. As shown in Table 12, when the chlorophyll content is low, such as apple, when blanching is performed by heating with hot water or steam to deactivate the oxidase, the chlorophyll is decomposed and discolored. Would. Of course, the antioxidant L
-Without ascorbic acid, it oxidizes strongly and turns brown.

Next, the astringency contained in vegetables and wild plants,
This paper describes the application of blanching method for removal of savory components and removal of polyphenol runoff.

Table 14 below shows the polyphenol content inside and outside the cells of vegetables. In parenchyma such as vegetables and wild grass, as shown in Table 14, almost the same amount of polyphenols was found inside and between cells. Contains polyphenols, causing oxidative discoloration. In Table 14, for intracellular polyphenols and intercellular polyphenols, for each vegetable, a precipitate obtained by centrifuging a single cell suspension obtained by converting the single cell with the present intercellular substance degrading enzyme at 3000 rpm for 5 minutes and separating the same ( The content of intracellular polyphenol and intercellular polyphenol were measured from the single cell particles) and the supernatant, respectively. Furthermore, the parenchyma of vegetables and wild plants contains astringent components such as chlorogenic acid and tannic acid, and harsh substances such as homogentisic acid, oxalic acid, alkaloids, and magnesium salts, both extracellularly and intracellularly. ing. In this way, it is necessary to reduce the outflow of polyphenols, astringency, and astringency components that cause oxidative discoloration from inside and outside the cell as much as possible, and to provide a single-cell suspension with little astringency and astringency without aroma. is there.

Further, various vegetables such as pumpkins are separated into single cell particles and intercellular fluid by the present intercellular substance-decomposing enzyme, and inorganic components (Ca, Mg, K, F) existing inside and between cells are separated.
The results of measuring the content (mg / 100 g) of e) are shown in Table 15 below. In the case of an inorganic component, there is a large difference in the amount of distribution within a cell and between cells depending on a measurement sample, a timing, and the like.
In order to elute and remove excess inorganic components such as Ca and Mg that cause such harshness and astringency, blanching is performed for at least 5 minutes in hot water at about 80 to 95 ° C, and the cells are exposed to water. In addition to eluting and removing these inorganic components from between and inside the cells, polyphenols that cause oxidative discoloration are also removed by leaching, reducing the content of inorganic components such as polyphenols, Ca, and Mg. Astringency can be reduced. Tables 16 and 17 below show blanching of a single cell suspension obtained by converting each of Takana (Table 16) and spinach (Table 17) into a single cell with the present intercellular substance degrading enzyme and a mechanically ground liquid. The polyphenol content (mg / 100 g) in the case of the treatment is shown. In Tables 16 and 17, the supernatant is defined as
For each of the single cell suspension and the mechanical attrition, 30
This refers to the supernatant obtained by centrifugation at 00 rpm for 10 minutes. Table 18 below shows the content of the inorganic components in the intracellular and intercellular fluids of the vegetables (spinach and carrot) after the blanching treatment.

As shown in Tables 16, 17 and 18, the amount of polyphenols eluted and removed by blanching reaches about 70-80% in the case of Takana but slightly lower in the case of spinach which is about 40-50%. It can be seen that the amount of elution differs depending on the structure and structure of the vegetables. On the other hand, the elution of the inorganic component is up to about 55 to 60% intracellularly and intercellularly in spinach, and up to about 50 to 70% intracellularly and intercellularly in the case of carrot. After all, 90-9 for hard vegetables
It is clear that if blanching is performed at 5 ° C and soft vegetables at 80 to 85 ° C for 1 to 5 or 6 minutes, substances causing discoloration such as polyphenols, Ca, and Mg, and astringent and astringent substances can be largely eluted and removed. is there. When distributing these as commercial products, as a result of examining the relationship between storage period and discoloration using spinach, Takana, kale radish leaves, etc. for discoloration during refrigeration and freezing, when raw leaves are directly used as single cell suspension It was found that there was no significant oxidative discoloration up to about 7 days in refrigerated storage, but the color changed markedly thereafter, losing commercial value. When a fresh leaf is blanched for 5 minutes or more to form a single cell suspension, there is no significant discoloration until about 27 days, and the commercial value can be maintained, but thereafter, oxidative discoloration proceeds. When frozen, a single-cell suspension prepared from fresh leaves shows little discoloration for one month. The single cell suspension prepared from the blanched fresh leaves has little discoloration for 2 to 3 months and does not lose its commercial value.

Next, vegetables having high viscosity or low water content,
The pretreatment method for converting wild grass and the like into single cells is described.

When vegetables, wildflowers, medicinal plants, fruits and the like are converted into single cells with the present intercellular substance-separating enzyme, in plants having a water content of 85% or more, single cells can be formed only by adding enzyme powder and stirring. On the other hand, extremely viscous materials such as Moroheiya (however, vegetable-type Moroheiya have a low viscosity) do not proceed with enzymatic action and turn black. In such a case, when blanching is performed, the water content reaches 90% and single cells can be formed. However, if the viscosity is reduced by adding water to perform better single cells, the enzyme action sufficiently proceeds. . Since viscous potatoes contain 83-85% water, they can be completely single-celled by adding enzyme powder after washing and cutting, but sweet potatoes have a low water content of 68%, equivalent to 90% water content. About 20% of water is required, and potato has a water content of 79%. In the case of squash, the water content is about 78%, and unless the water content is 10% or less, sufficient single cells cannot be formed. However, in a frozen pumpkin, the tissue is softened by thawing, and since water elutes on the surface of the cut tissue, single cells can be formed without adding water. Table 19 and Table 2 below show the results obtained by converting single cells of Moloheiya, red pumpkin (Kurisango), Ebisu pumpkin (dark green pumpkin), frozen Ebisu pumpkin, and frozen red pumpkin, respectively.
0, Table 21 and Table 22.

Further, in the case of tree leaves such as tea leaves and ginkgo leaves, and the leaves of kuma bamboo grass, the fresh leaves have a low water content of 60 to 70%, and the tissue is hard and contains hard and long fibers such as petiole. Enzyme action does not proceed as it is in short supply. Ginkgo leaves have a water content of about 50%, and those containing many petiole patterns such as tea leaves have a water content of about 100-200%. If it is possible to completely convert the cells into single cells, it is impossible to convert the cells into single cells without adding water or adding excessive water, because the stirring efficiency is deteriorated. In Table 23 below, tea leaves (water 67.5%, water content after blanching 78.5%),
Ginkgo biloba (water content 68.2%, moisture content after exposure to running water 75.2%)
And bear bamboo leaf (moisture 60.3%, moisture 60.6% after running water exposure)
%) Is used as a raw material leaf, and the results are shown for the case where these were made into single cells by the present intercellular substance-decomposing enzyme.

Next, in tea leaves having a water content of about 10% obtained by processing and drying Tencha, Sencha, etc., 500-70
Without 0% water, it is impossible to dissociate into single cells. Table 24 below shows an example in which Tencha leaves were hydrolyzed into single cells. In the case of dried defatted soybeans and dried tofu cake, it is impossible to make single cells unless 500-700% of the raw material is added with water. Raw soybeans are left as they are, and dried soybeans are cut off after sufficient water absorption, and 200% water is added to form single cells. In the case of an extremely hard material such as coffee beans, a single cell can be formed by an enzyme only after sufficient water absorption, cutting, and adding about 100% of water to the raw material. Sweet corn fresh matter has a lot of moisture, and it is possible to add enzyme powder to the raw material for cutting, dissociate and make single cells. Table 25 below shows an example of dried soybeans, Table 26 below shows coffee beans, and Table 27 below shows an example of sweet corn as single cells. In Table 27, "incomplete unicellularization" means that the unicellularization was discontinued halfway.

As described above, vegetables, wildflowers, and fruits are raw materials or blanched ones, and can be made into single cells even without water, and root vegetables can be made into single cells without water. Potatoes require a small amount of water (10-20%). As described above, in the method of the present invention, depending on the water content and physical properties of the raw material, when the cells are converted into single cells with the present intercellular substance-decomposing enzyme, they are added with water as needed.

Next, a mechanically milled liquid obtained by mechanically grinding a vegetable, wild grass, fruit, tree leaf, Kumabasa, etc., into a single cell suspension obtained by converting the cells into single cells with the present intercellular substance-decomposing enzyme. The differences from the above are described below.

A suspension obtained by converting plant parenchyma into a single cell with the present intercellular substance-degrading enzyme is clearly different from a trituration solution obtained by a general mechanical trituration method. In Tables 28 and 29 below, for carrots and strawberries, the particle size distributions of a suspension prepared as a single cell with the present intercellular substance-decomposing enzyme and a mechanically ground solution (pure) mechanically crushed using a homogenizer were measured. The results are shown. The particle size distribution was measured using a particle size distribution analyzer (LA-500 manufactured by Horiba, Ltd.).
V-1 rotation type). For carrots,
In the single cell suspension, cells having a particle size of 0.02 to 0.04 mm represent about 42%, and the remaining 50% consist of small cell particles having a particle size of 0.02 mm or less. 0.
It is as large as 018 mm. On the other hand, in the mechanical grinding solution, the crushed particles mainly consist of crushed particles of 0.03 mm or less, the average particle diameter is small as 0.009 mm, and no cells are found, and crushed intracellular granules and crushed cell walls are obtained. Consists of

Table 30 below shows carrots, spinach,
For each of Chinese cabbage, onion, apple and tangerine,
The result of measurement of the general components and the particle size distribution of the suspension made into a single cell by the present intercellular substance-decomposing enzyme and the mechanically ground mechanically ground solution (Purey) is shown. The particle size distribution varies depending on the type of vegetable, but unlike the mechanical attrition solution, where no cells are found, the single-cell suspension has more than 10 ⁶ oval or circular cells per ml. Occupied by. Table 31 below shows the comparison results between the single cell suspension for carrots and three commercially available mechanically milled liquids (freeze-milled, flattened liquid and micropaste). Not at all. In contrast, a single-cell suspension is a suspension of round or oval cells with no corners dispersed in the intercellular fluid, and therefore has irregular corners such as a mechanical trituration solution. The physical properties of the crushed particles are different from those of the grinding liquid.

The results of measuring the interfacial tension and the surface tension of the single-cell suspension and the cartilage solution for carrot are shown in Table 32 below. In Table 32, the single-cell suspension (A) and the single-cell suspension (B) were obtained using the present intercellular substance-decomposing enzyme and the cell-separating enzyme described in Japanese Patent Publication No. 6-71413, respectively. This is the obtained single cell suspension. The interfacial tension is a force that prevents liquid (water) and liquid (oil) from mixing with each other, and the surface tension is a force that prevents liquid (water) with air from mixing with each other. Therefore, a high interfacial tension means that it is not easily compatible with oil, and a low interfacial tension means that it is easily compatible with oil.
A high surface tension means that it is difficult to adapt to air, and a low surface tension means that it is easy to adapt to air. From the results in Table 32,
Single-cell suspension (A) has lower interfacial tension and surface tension than machine-milled liquid, and is easily mixed with oil and has good dispersibility in oil. Therefore, it is mixed with oil-dispersed dressing or cream. It is easy to make, easy to mix with air, easy to foam and fluffy.

As shown in Table 33 below, distilled water, soybean oil, and milk were added to the single-cell suspension (A), the single-cell suspension (B), and the mechanically ground suspension obtained from carrots. When the viscosity was measured (measured at 162 rpm / min at 26 ° C.), the viscosity of the single-cell suspension (A) was almost half lower than that of the mechanical grinding solution in each case, and the single-cell suspension was dispersed in water or oil. Due to the good properties, the flow is smooth and the viscosity is low, and the ease of the flow gives a good result to the smooth throat. On the other hand, the mechanical grinding liquid has a high viscosity, a large particle size, and poor dispersibility in water, so that it does not flow smoothly and becomes a rough throat.

Next, a method for confirming the fact that plant parenchyma such as vegetables, wildflowers, fruits, medicinal plants, tree leaves, and bamboo grass leaves have been converted into single cells with the present intercellular substance-degrading enzyme will be described below.

Even if it is reported that the plant parenchyma has been converted into a single cell,
However, the mere fact that cells were visible by microscopic observation does not prove that plant parenchyma has been converted into single cells. It must be ensured that the cell-cell adhesion of the plant parenchyma has been completely degraded and that countless individual cells have completely dissociated into single cells. As a method for confirming complete isolation, a single cell suspension is diluted in each stage of 2 to 10 times, sufficiently stirred and dispersed, and subjected to each dilution step with a microscope of 40 to 600 times magnification. Must be observed once or twice each to confirm that the cells are completely free without adhering to each other. Further, using a toma hemacytometer, the measurement is performed 5 times or more using a 2 to 10-fold dilution, and the number of cells in 1 g of the suspension is calculated. In this case, when the plant parenchyma used as the sample is converted into a single cell with the present intercellular substance degrading enzyme, when the water is added, the amount of water is converted to calculate the number of cells per 1 g of the raw parenchyma. Table 34 below shows an example in which the number of cells of a single cell suspension obtained by converting carrots into single cells with the present intercellular substance degrading enzyme was measured by the above method. In this way, the used raw material is completely dissociated into single cells without destroying the constituent cells by the present intercellular substance-decomposing enzyme, thereby forming a single cell suspension in which countless cells are suspended in the intercellular fluid. It can be said that for the first time plant parenchyma was converted into a single cell.

Next, the relationship between the viscosity of the single-cell suspension and the sedimentation of the single cells and measures for preventing sedimentation will be described below.

In plant parenchyma such as vegetables, wild grasses, fruits, medicinal plants, tree leaves, and bamboo grass, innumerable cells are generated from each other by pectic substances mainly composed of protopectin, a small amount of hemicellulose, trace amounts of lignin, etc. It is configured by being adhered to cellulose such as leaf veins. By continuously cutting countless protopectin molecules, etc., with this intercellular substance-decomposing enzyme, the adhesion between cells or cells and cellulose etc. is separated and dissociated into single cells to dissociate. The pectic substance and other water-soluble components are dissolved in the intercellular fluid interposed between the pectin molecules to form a viscous liquid. Countless single cells are suspended in this viscous liquid, and the viscosity is increasing. However, the viscosity of the single-cell suspension depends on the raw material, the variety, the soil of the production area and other cultivation areas, the climate, and countless other conditions. Taking carrots as an example, the present inventor made carrots produced in Japan, the United States, and China into single cells about 100 times over 4 years, and measured the viscosity of the obtained single cell suspension. The viscosity was 835 to 6350 cp. And there was a very large difference depending on the type of carrot. Specifically, it was found that the viscosity of each carrot was different. Table 35 below shows the change in viscosity depending on the degree of dilution of a single cell suspension of carrot and spinach. For carrots, adding 1.5 times the water of a single cell suspension and diluting 2.5 times reduces the viscosity to 180 cp and the initial viscosity of 1
/ 15. In spinach, similarly, when the water is diluted 1.5 times by adding 1.5 times the water of the single cell suspension, the viscosity decreases to 113 cp, and decreases to 1/15 of the initial viscosity.

Table 36 below shows, for the carrot and spinach plants, a decrease in viscosity due to dilution of these single cell suspensions and a sedimentation rate of cell particles upon standing (a rate of formation of a supernatant after standing at room temperature for 24 hours). Are shown). In the case of carrot, the same amount of water was added to a single-cell suspension of carrot, which initially exhibited a viscosity of 2700 cp, and the viscosity was reduced to 300 cp by diluting the suspension twice. There is no separation phenomenon in which the particles settle and produce a supernatant. In the case of spinach, 1
To a stock solution of a single cell suspension having a viscosity of 800 cp, 75% volume of water was added to dilute the solution to 1.75 times to obtain 2
Although the viscosity decreases to 90 cp, in this state, even if left at room temperature for 24 hours, no sedimentation of the cell particles occurs and no supernatant is generated.

Further, as shown in Table 37 below, a carrot single cell suspension having a viscosity of 2700 cp
A suspension having a viscosity of 145 cp, diluted 2.5 times with 0% water, causes sedimentation of cell particles after 24 hours at room temperature,
% Supernatant. Add agar to this liquid and add 0.1
% Or more, the viscosity shows 285 cp or more,
After standing for 4 hours, no sedimentation occurs and no supernatant is formed. Similarly, a 20 cp viscosity carrot suspension diluted 5 fold with 400% water yields a 36% supernatant, in which case 0.12% agar is added and the viscosity is 19%.
It only rises to 5 cp, producing 1% of supernatant in this state, and it is difficult to dissolve the agar any more, and eventually it is impossible to completely prevent sedimentation. A single-cell suspension of spinach is a stock solution having a viscosity of 1800 cp, and a suspension having a viscosity of 90 cp diluted 2.5-fold by adding 150% water to a stock solution having a viscosity of 1800 cp.
After 4 hours, sedimentation of the cell particles occurs, producing a 12% supernatant. When agar is added to this solution and 0.1% or more is added, the viscosity becomes 310 cp or more, and no sedimentation occurs even after standing for 24 hours, and no supernatant is generated. Likewise 400
A suspension of spinach with a viscosity of 15 cp, diluted 5 times with 1% water, produces a 38% supernatant, to which 0.12% of agar increases the viscosity only to 270 cp. But a slight settling occurs, producing 1.5% supernatant.

As shown in Table 38 below, a single-cell suspension of carrot at a 2.5-fold dilution had a viscosity of 145 cp and produced 8% of supernatant, which was 0.04% or more. When carrageenan is added, the viscosity increases to 270 cp or more, no sedimentation occurs even after standing for 24 hours, and no supernatant is generated. Similarly, a 5-fold dilution of a single-cell suspension of carrots exhibits a viscosity of 20 cp and yields 36% of the supernatant,
In this case, even if 0.12% of carrageenan is added, the viscosity becomes 3
It only rises to 80 cp and sedimentation of cell particles occurs, producing 12% supernatant. A single-cell suspension of spinach, diluted 2.5-fold, has a viscosity of 95 cp and has a viscosity of 1 cp.
3% of supernatant is generated, but when 0.12% of carrageenan is added thereto, the viscosity increases to 350 cp, no sedimentation occurs, and no supernatant is generated. Similarly, in the case of a 5-cell dilution of a single-cell suspension of spinach, a viscosity of 15 cp is exhibited and a 38% supernatant is generated. However, even if 0.12% of carrageenan is added thereto, the viscosity increases only to 160 cp. No, yielding 8% supernatant.

When the prepared single cell suspension is frozen and stored for a certain period of time after packaging and thawing, the suspension changes the physical state of the suspension due to freezing. Therefore, after thawing, it is necessary to mix well after stirring again to measure the cell dispersion and return the viscosity to the viscosity at the time of production.

Next, a method for forming a single cell by treating agricultural processing residues and wastes with the present intercellular substance degrading enzyme will be described below.

In the processing of agricultural products, the residue to be discharged may be juice residue made of crushed and rolled cells, such as non-uniform cutting pressure and non-uniform grinding pressure of fruits and vegetables, without heat treatment. In addition, there are also bean paste, tofu cake, etc., which are crushed and heated and extracted at an unbalanced pressure, and furthermore, there are jam residues sifted after adding a large amount of sugar to the raw materials and then heating and concentrating, and defatted soybeans which are solvent-extracted from the cut raw materials. If these are treated with the present intercellular substance-decomposing enzyme, the crushing pressure-biased and further heat-denatured tissues are separated into crushed and deformed cell units, which can be separated into fine particles of about 0.01 to 0.05 mm. Becomes possible, and can be used effectively. First, as shown in Table 40 below, when bean jam or adzuki bean jam was treated with the present intercellular substance degrading enzyme,
%, The bean jam can be effectively used. Table 4 below
As shown in FIG. 1, when dried tofu cake is treated with the intercellular substance-decomposing enzyme by adding 500% or more of water, fine particles separated into fine cell units with a recovery (yield) of approximately 80% or more are obtained. Can be recovered. As shown in Table 42 below, when about 10 to 20% of water is added to the citrus juice and treated with the present intercellular substance-decomposing enzyme, a recovery rate (yield) of 70 to 80% is obtained.
A crushed and deformed single cell suspension can be obtained. This liquid has a bitter taste belonging to limonoids when untreated juice lees is used, so twice the amount of water is added to the juice lees in advance, and the mixture is heated for a short time to extract the limonoids. After draining,
Enzyme treatment results in a single-cell suspension with low bitterness with a recovery of about 80%.

[0052] Carrot juice lees consist of 5 parts of raw carrot.
It reaches 0%, and its effective use is an important issue. As shown in Table 43 below, the same weight of water was added to carrot juice lees, and 0.25% of the present intercellular substance-degrading enzyme was added.
After stirring for 5 hours, the pressure-rupture-disrupted juice cake is completely dissociated in cell units and completely converted into single cells. Of course, 1
Since the fine particles have a diameter of not more than mm, even if the single-cell suspension is left alone, only slight water separation (separation of sap) occurs at the upper portion, and the fact that the suspension is fine particles per cell is apparent.

The residue on the pulper finisher during the production of strawberry jam is a residue having a particle size of 1 mm or more. As shown in Table 44 below, the residue was hydrolyzed by 25% to give the intercellular substance. When the degrading enzyme is added to form a single cell, strawberry fine particles in which the crushed strawberry tissue is dissociated into single cells can be obtained at a recovery rate (yield) of about 50 to 55%.
Further, in the case of defatted soybeans, as shown in Table 45 below,
When treated with the present intercellular substance-decomposing enzyme after about three times the amount of water, fine particles in a unit of single cell are recovered at a recovery rate (yield) of about 50%.

Next, the functionality of suspensions and particles obtained by converting cells, vegetables, wildflowers, fruits, herbs, and the like into single cells with the present intercellular substance-decomposing enzyme will be described below in comparison with mechanically ground products. .

The results of a general analysis and a free amino acid analysis of a suspension, a mechanically ground solution and a squeezed solution obtained by converting apples and peeled Satsuma mandarins into single cells using the present intercellular substance-decomposing enzyme are shown in the following table, respectively. 46 and Table 47. Regarding the content and characteristics of general components, the single cell suspension is the highest in both apple and Satsuma mandarin, and the value of the direct reducing sugar is particularly high in apples compared to the mechanically milled and squeezed liquids. Is about 3% larger. In addition, the single cell suspension is about 1.5 times as much in amino nitrogen and about 2.5 times or more in free amino acids in both apple and Satsuma mandarin compared to the other two. This is due to the fact that α-amylase (α-am
ylase), glucoamylase (glucoamyl)
ase) and the action of acidic protease (protease). As for the citrus peel, as shown in Table 48 below, the single-cell suspension has a higher vitamin B group content and, of course, a much higher free amino acid content than the mechanically ground solution.

As described above, the suspension treated with the present intercellular substance-decomposing enzyme for 60 to 120 minutes, such as a single-cell suspension, is readily digestible with direct reducing sugars, free amino acids, etc., including the vitamin B group. Nutrition component is 20-250% than machine grinding liquid
And the nutritional function of the food is clearly higher. Furthermore, chymotrypsin was added to each of the vegetable single-cell suspension and the machine-milled liquid, and the direct reducing sugar (glucose) and amino acid (glycine) released by decomposition were measured. , 132.5 mg for a single cell suspension and 6
More than twice as much as 0.12mg, and 1g of vegetables
The amount of amino acid (glycine) per unit was 86.23 μg in the case of a single cell suspension, and 79.23 in the case of a mechanically ground solution.
About 10% more than 66 μg. That is, since the single-cell suspension is a single-cell unit that easily undergoes digestion, it can be said that the single-cell suspension has a high nutritional function. However, as shown in Table 49 below, in the case of a strawberry-like substance that is softly separated into single cells within 30 minutes and is almost degraded by enzymes in fruits, the nutrition of the single-cell suspension can be maintained even if the single-cell processing is performed. The ingredients are the same as the nutrients of the machine grinding liquid (homogenizing liquid). However, single cell suspensions are all circular or elliptical, ie, round single cells, compared to the mechanical grinding solution, and the cell particles are almost uniform in size compared to the mechanical grinding solution. In, the over throat is good, and the single-cell suspension is smooth even when touched by hand,
Since the sizes of the cell particles are almost uniform, no sedimentation phenomenon occurs, and it is easy to drink as a beverage. In addition, since the single-cell suspension has an increased sugar content, amino acid, and acidity, the taste is rich, and the secondary functions as foods such as taste and taste are increased.

Next, with respect to the single cell suspension obtained by the action of the present intercellular substance-decomposing enzyme, the difficulty of inactivating the intracellular enzyme during heat sterilization will be described below.

In the case of general transparent or cloudy vegetable juice, fruit juice, etc., there is no obstacle such as a cell wall that blocks heat conduction at the time of heat sterilization. Therefore, heat conduction is carried out smoothly although there is a slight difference depending on the viscosity. It is easy to sterilize and deactivate suspended microorganisms and dissolved enzymes. On the other hand, like a single cell suspension, cellulose and hemicellulose, protopectin protein and other strong double cell walls surrounded by a triple obstacle between the cell wall and the cell membrane within 10
^{In the case where about 6} / ml cells are suspended, 10
Heat conduction into ⁶ / ml cells is poor, and it is impossible to completely inactivate hydrolases such as proteases, lipases and amylase in a normal sterilization time for clear liquids and turbid liquids. It is possible. Heat sterilization by adding a single-cell suspension of carrots to milk and skim milk,
Table 50 below shows the results of examining the generation of bitterness in milk due to the production of bitter peptides by intracellular proteases and the production of bitter fatty acids by intracellular lipase.
From these results, it can be seen that, unless sterilization is performed at 100 ° C. for 20 minutes or more, carrot protease acts to decompose and coagulate milk casein protein to produce a bitter peptide. The results of measuring the inactivation of the intercellular and intracellular proteases and lipase by heating are shown in FIGS. 1, 2 and 3. 1 and 2 show the acidic protease (pH 4.5) in the carrot single cell intercellular fluid and the carrot single cell suspension.
3 shows the residual activity after heat treatment. FIG. 3 shows the heat treatment of carrot single cells and intercellular fluid (the heat treatment time was 30 minutes at each temperature).
Lipase activity). The ml value of NaOH on the vertical axis of FIG.
This is a titration value obtained by titration with a 0N-NaOH solution. 1 to 3
As is clear from the above, the acidic protease and lipase in the cells are completely inactivated by heating at 100 ° C. for 30 minutes. However, in the case of acidic protease, at 90 ° C., 4
It was found that it was completely deactivated only after heating for a long time.

Next, a technique for completely transforming vegetables, wild plants, fruits, medicinal plants, tree leaves, bear bamboo leaves, etc. into single cells to produce food will be described.

According to the method of the present invention, soft tissues such as vegetables, wild grasses, fruits, medicinal plants, tree leaves, and bamboo grass leaves are completely dissociated into single cell units, separated from leaf veins, petiole and other stems, and all cells and cells are separated. It is possible to establish a technology for recovering the adhesive substance into food.

As a representative of wild grass, the upper leaves of wild large wormwood extending to about 0.3 to 1 m were mainly collected and made into single cells. For ordinary vegetables, the water content is 85-95%
Since it is contained, the water in the intercellular substance elutes during the single celling operation, and the present intercellular substance-degrading enzyme can be added to this as a raw material without adding water, followed by stirring to form a single cell. However, the large wormwood of wild grass has less moisture than the small wormwood, and the surface of the leaves is fluffy.
Unless 00% of water is added, single cells cannot be formed by enzymatic action. Table 51 below shows the results obtained by converting large wormwood that has grown to a plant height of 0.3 to 1 m into single cells. When the raw material is converted into a single cell, the cell wall is broken during the stirring of the single cell operation because the cell wall is hard, and the number of cells in the single cell suspension is about 1/10 of that when blanching, and the amount of centrifugal sedimentation is naturally small. . However, when the raw material is blanched at 90 to 95 ° C. for 1 to 2 minutes, the hard and bulky tissue of the raw material is softened, so that the cell wall is not broken by the impact of stirring, and the tissue is sequentially converted into single cells. . Thus, when the cells are completely unicellularized, only fibrous residues such as petiole and veins are 15 to 25%.
To some extent, the others are unicellularized and eventually become about 67-76.
A single cell suspension is recovered at a% yield.

Next, as a representative of root vegetables, radish, turnip, lotus root, burdock, bamboo shoot, and carrot were completely unicellularized with the present intercellular substance-decomposing enzyme, and the results are shown in Tables 52, 53, 54 and 54 below. 55 and Table 56. In white vegetables such as radishes and turnips, phenol or polyphenol substances contained in trace amounts during single cell formation are oxidized by oxidizing enzymes, and the resulting single cell suspension becomes grayish white, reducing commercial value. If ascorbic acid, sodium ascorbate, erythorbic acid and the like, which are inhibitors, are added in an amount of 0.3% or more, a white single-cell suspension similar to the raw vegetable can be obtained in high yield. Both lotus root and burdock thoroughly remove soil and sand, and after sufficient washing, blanch at 95 ° C or higher for 3 to 5 minutes to deactivate oxidases such as phenol oxidase and polyphenol oxidase, and after peeling, cut and cut into single cells. I do. In the case of lotus root, in order to prevent discoloration due to autoxidation during the operation of forming single cells with the present intercellular substance-decomposing enzyme, adding 0.4% or more of antioxidants such as ascorbic acid and sodium ascorbate, A white single-cell suspension similar to the raw vegetable is obtained. In the case of burdock, only blanching is sufficient and there is no need to add an antioxidant. In order to completely dissociate the lotus root into single cells, the residue after the first dissociation for about 90 minutes is further dissociated for another 3 hours, and the residue finally becomes 3.5%, with a final yield of 88%. A single cell suspension can be collected. In burdock, if only one dissociation lasts for 5 hours, 10% residue and 81%
The single cell suspension can be recovered at a yield of.

In the case of bamboo shoots, the first time was 2 hours,
After a third round of dissociation, the residue will ultimately be 17% and a single cell suspension can be recovered with a final yield of 76%. In the case of carrots, although not as much as mechanically milled liquids,
Since it has a strong viscosity of 000 to 5000 cp, the progress of the enzyme action is poor, and the formation of a single cell is strongly affected by the stirring operation and time. Therefore, in order to completely transform into a single cell, 60 cells are required.
When the single cell suspension can be recovered at a yield of 60% or more with an action time of about 180 minutes, once sieving about 20 mesh to collect the undecomposed residue, and then again When the enzyme powder is added to form a single cell, the cell can be almost completely converted into a single cell until the residue is about 5% of the whole raw material.

Table 57, Table 58, Table 59 and Table 60 below show the results obtained by converting pumpkins (Ebisu pumpkin and red-skinned pumpkin), cucumber and melon as representatives of fruit vegetables into single cells with the present intercellular substance-decomposing enzyme. . In the case of blue-skinned pumpkin (Western pumpkin), if the peeling is insufficient, the skin pigment, chlorophyll, is mixed with flavonoids and carotene-based yellow pigment of the flesh and turns grayish brown, losing commercial value. It is necessary to peel the skin so that the feel does not remain. However, the color tone is irrelevant even if the cotton and the seeds in the central part of the pulp occupying 30% of the squash are removed or not removed. Single cell suspensions have a slight reduction in sugar content, and the cell number is almost halved. Usually, in the processing of squash, heat treatment such as blanching is first performed with the whole fruit, but unlike a single fruit cell, the sugar content and the number of cells are slightly reduced. In the case of red-skinned squash (Kurisango), it is not necessary to remove the pericarp, and the whole fruit may be cut and made into a single cell after branching. The red-skinned pumpkin does not need to remove the pericarp like the blue-skinned pumpkin, so that a single-cell suspension can be obtained with a yield of about 90% of the raw material used. If the pumpkin stored frozen is naturally thawed and converted into single cells for storage of the raw materials, the single cells can be almost completely converted into single cells. Therefore, a single cell suspension is obtained with a yield of 90% or more of the raw materials, and the number of cells is approximately 10 ^6. / G.
In cucumber, part of the epidermis remains as a residue. As for melon, both raw materials and those stored frozen can be converted into single cells by enzymatic action for only 30 minutes. In the case of raw melon, seeds and fibrous material of "cotton" are about 10% as residue.
The single-cell suspension remains before and after and can be recovered with a yield of about 90%. Frozen melon has only 1-2% residue, 96%
A single cell suspension can be recovered at a yield of.

Tables 61 and 62 below show the results of converting apples and strawberries as typical fruits into single cells with the present intercellular substance-decomposing enzyme. In green apples, when subjected to pretreatment such as peeling and cutting, oxidized browning occurs due to polyphenols and their oxidases present in the pulp.
Since the commercial value is lost, the peeled and cut adjusted fruits are immediately treated with an antioxidant solution (for example, L-ascorbic acid 1%,
It is necessary to immerse the mixture in citric acid (0.5% or more) to inactivate oxidizing enzymes such as polyphenol oxidase and to prevent oxidation with L-ascorbic acid. after that,
When 0.3% of the present intercellular substance-decomposing enzyme and 0.5% or more of antioxidant L-ascorbic acid and the like are added to form single cells under stirring, the yield of green apple is about 90%. A single cell suspension of the same color as the pale green of the pulp can be collected. In the case of a red apple, a single-cell suspension of an amber color that is the same color as the color tone of the pulp of the red apple is obtained. In strawberries, the operation of removing waste is complicated, so that after washing thoroughly with water and removing the soil, microorganisms, and then draining, immediately convert to single cells,
There is no significant difference in the obtained single cell suspension between the case where the spine is removed and the single cell is formed. In the end, either method is acceptable due to labor and cost issues. 9% in case of looseness
In the case of waste removal, only about 3.5% of the residue is recovered as a single-cell suspension, and the time required for single-cell formation is about 30 minutes.

The results obtained by converting spinach, komatsuna, onion, garlic and light-colored vegetables (cucumber, cabbage, Chinese cabbage) as representatives of leaf stem vegetables into single cells with the present intercellular substance degrading enzyme are shown in Tables 63 and 64 below. Table 65, Table 66 and Table 67
Shown in When spinach is transformed into a single cell with the present intercellular substance-degrading enzyme as it is in a fresh leaf (untreated), the cell wall is broken due to the stirring operation, and the number of cells in the suspension is about 10 ⁵ / ml. / 10 or 1/100 or less, and the cells precipitated by centrifugation are 0.8 ml / 1
It is extremely small at 0 ml. With branching, the number of cells in the suspension, which can be obtained in 20 seconds or 5 minutes, is 1
0 ⁶ / ml increases above but, because of polyphenols contained, it is greenish yellow to green-brown to oxidation during refrigerated storage, since commercial value decreases, the water exposed for about 30 minutes after blanching, the polyphenol After elution and removal from inside and outside the cells, it is necessary to perform a single cell treatment.

Among the green and yellow vegetables, Komatsuna is a vegetable having less astringency, astringency components and oxidatively discoloring substances such as polyphenols. This is due to the fact that the difference in the polyphenol content between untreated raw vegetables and blanched Komatsuna is small. Is obvious from. Also, for both, there is no significant difference in the yield of the single cell suspension, there is no difference in the number of cells,
There is no significant difference in the color tone of the single cell suspension. However, in the case of no processing, the color tone tends to be slightly dark to some extent. In the case of onions, there are those with a high fiber content and those with a low fiber content.
A single-cell suspension can be obtained with the above recovery rate, while a variety with a large amount of fiber has a residue of about 10%, and a single-cell suspension can be obtained with a recovery rate of about 80%. In varieties with a high fiber content, the cell number of the single cell suspension is also lower than in varieties with a low fiber content.

When raw garlic is cut or ground, alliin, which is a sulfur-containing compound, is decomposed by alliinase to produce allicin, which gives an intense odor. is there. Therefore, first, garlic is blanched at 90 ° C. for about 2 minutes to inactivate the allinase, and then the present intercellular substance-degrading enzyme is actuated to form single cells. In this case, even if the garlic is cut,
Even if it is a circle, it has nothing to do with the single cell action. However, since the obtained single-cell suspension is viscous and the action does not progress at about 50 to 60% of dissociation, it is necessary to first act for 90 to 120 minutes and to obtain the single-cell suspension in order to completely convert to a single cell. Remove the turbid solution, re-single the residue into single cells.
A single cell suspension is obtained with a total recovery of 88% or more.
Light-colored vegetables, such as cucumber, cabbage, and Chinese cabbage, with little discoloration have very few polyphenol components and little discoloration occurs. Since cabbage has a core or stem, about 20% of undecomposed residue is generated, but only about 6% of fibrous residue remains in Chinese cabbage, and a single cell suspension can be obtained at a yield of about 90%. Can be

Table 68 below shows the results of single cells of the sweet potato and the potato as representatives of the potatoes by using the present intercellular substance-decomposing enzyme. Since these potatoes contain polyphenols and their oxidizing enzymes, immediately after cutting or peeling, they are immersed in a saline solution, acid solution or antioxidant solution to stop the action of the oxidizing enzymes, and then lifted to remove L-ascorbic acid. When a single cell is formed in addition to a slight excess, a single cell suspension is obtained with a yield of 86 to 92%.

Tables 69 and 27 show the results of soybeans and sweet corn, which are representative of beans, converted into single cells using the present intercellular substance-decomposing enzyme. After absorbing water sufficiently with soybeans,
Although cut into quarters and treated with the present intercellular substance-decomposing enzyme, only 45% can be dissociated in one dissociation. Even if the residue is collected and reprocessed, only 57% dissociates in total. And a final residue of 34%. In the case of sweet corn, when raw and quick-frozen products are thawed and completely converted into single cells, both raw materials have a large amount of skin and therefore have a residue of about 27%, and a single cell suspension with a yield of 53 to 54%. Is obtained.

The medicinal carrots (Korean carrots), aloe, kale leaves, safflower cotyledons and wheat young leaves as representatives of herbs are subjected to a single cell treatment with the present intercellular substance-decomposing enzyme,
The results are shown in Tables 70, 71, 72, 73, 74 and 75 below. In medicinal carrots, depending on the raw material, it is difficult to form a single cell without adding 30% water,
A single cell suspension can be recovered with 13-22% residue remaining and 73-87% yield. For medicinal carrots of 1st to 2nd grade and 4th grade and above, the roots are many and the soil smell is strong, so hydrogen peroxide decomposition treatment was performed to remove them, but since the earth smell could not be completely removed, it was completely oxidized with potassium permanganate. When the method of reducing and removing with oxalic acid is adopted, the earth odor can be completely removed, and about 20 to 30% of the residue and 60 to 7%
A single cell suspension can be recovered with a yield of about 0%. With aloe leaves, a single cell suspension is obtained with a 76% yield. Kale leaves have a high polyphenol content, and if not blanched, discoloration occurs due to oxidation of the polyphenol. However, when the single-cell suspension is sterilized by heat and stored for a long period of time, if it remains slightly acidic at a pH of about 5.5, it turns yellow due to the decomposition of chlorophyll, so the pH is neutralized to about 7.0. If heat sterilized, the decomposition of chlorophyll does not occur and the green single-cell suspension can be stored for a long time. Safflower cotyledons do not need to be blanched, and fresh leaves can be converted into single cells as they are, and the resulting single cell suspension can be neutralized to around pH 7.0, packaged, and heat-sterilized to preserve green for a long period of time. In wheat young leaves, the yield of the single-cell suspension is higher by 10% or more when the amount of water is 100% than when no water is added. As a matter of course, when the amount of water is 100%, the components of the single-cell suspension must be diluted by the amount of water. Also in this case, neutralize to pH 7.0, and
When sterilized for a minute, a dark green color can be maintained.

[0072]

【Example】

Example 1 After harvesting a suitably grown carrot, the leaves are cut off and washed sufficiently with water to remove the earth and sand and used immediately as a raw material. When left at room temperature, use within 2-3 days. When storing, it is normal to put it in a plastic bag or a sealable box, but if the humidity of the refrigerator can be adjusted to 85%, it may be unpackaged. Use within one month even if refrigerated below 5 ° C. During long-term storage, a reaction occurs that forms a melanin pigment in a part of the carrot epidermis.Using such a carrot results in a carrot with a slightly darker color tone, and its commercial value is greatly reduced. When producing a liquid, it is necessary to cultivate so that carrots suitable for production can be successively harvested by sequentially shifting the cultivation period of carrots. The raw material carrot thus obtained is put into a steam peeler in half volume, closed tightly, steam is immediately blown, and the carrot is strongly rotated 4 to 5 times to peel off the outer skin by rubbing between the carrots. Alternatively, it is put into hot water of 95 ° C. or more, blanched for 1 to 2 minutes, and the skin is rubbed off by polishing and washing to peel off. Carrot moisture is reduced by about 1% by steaming or blanching. The peeled carrot is cut into 0.5 cm squares by using a dicer as it is. In the case of steam pillar peeling, blanching is performed at 95 ° C. for about 30 seconds, immediately cooled to about 45 ° C., and cut with a dicer into 0.5 cm squares. If the cut is made smaller than this, the cells at the cut site will be broken, and the number of cells in 1 g of the single cell suspension of the product will be reduced by 1/3 or more of the raw material.

Since the temperature of the product immediately after being cut is kept at 30 ° C. or higher, the cut product is put into a stirring tank,
Add about 0.2-0.3% of the present intercellular substance separating enzyme to prevent oxidation and to lower the pH even slightly.
0.2% of ascorbic acid was added, and 9
70-80% when agitated for 0-120 minutes to form single cells
A single-cell suspension passing through a 20-mesh sieve is obtained at a recovery rate of 20 to 25% of the residue on the sieve, which is not fully unicellularized carrot granules. Then, a single cell is formed together with the following raw material for cutting. The resulting single cell suspension is obtained with a recovery of 70-80% of the charged raw material. The single-cell suspension is immediately sent to a pasteurizer, sterilized by heating at 95 ° C for 25 to 30 seconds, sent to a heat tank, and sealed in a packaging container while maintaining the product temperature of 80 to 75 ° C. Even if some microorganisms remain in the packaging container because it is heat-filled, a sterilized single-cell suspension at 75 ° C or higher is added, so even if there are microorganisms in the container or microorganisms that have fallen from the air at the time of filling, , Since the high temperature of 75 ° C or more is maintained for a long time even after sealing, the coliform group is negative, the general viable cell count is 300 / g or less, no mold, yeast, etc. are detected. A suspension was obtained, which was
When stored frozen at 5 ° C. or lower, a single-cell suspension without any abnormality can be maintained for 1 to 2 years. The final recovery is thus about 85% of the raw material used. The composition and properties of the single cell suspensions obtained when Chinese, American and Japanese carrots were used as raw materials were measured, and the minimum and maximum values are shown in Table 76 below. As shown in Table 76, the agricultural products are the same varieties, at the same time, and the ingredients cultivated in the same area all have different component compositions and characteristics. Strictly, the component compositions are different from each other, so each production batch It turned out that a component analysis was necessary.

Example 2 Under the conditions shown in Table 61 below, the raw green apples (Wang Lin) and the US green apples (Granny Smit) were used as raw materials.
h), domestic red apples (red balls) and Chinese green apples were each converted into single cells. Since the red or green apples have many yeasts on the surface, they are thoroughly washed with a brush washing machine, and if necessary, peeled and cored with a peeling machine. Apple contains phenolic compounds such as chlorogenic acid and catechol, and also contains oxidizing enzymes such as polyphenol oxidase, catecholase, and peroxidase, which oxidize this and turn brown. I do. Therefore, in order to prevent oxidative discoloration, L-ascorbic acid, 1.
Transport, transfer, etc. are performed in a state of being immersed in a mixed solution of 0 to 1.1% and citric acid 0.67%, further cut into 1 cm squares with a dicer, and similarly stored and transported in an antioxidant. .

Next, the apple cut into a dice is pulled up from the antioxidant solution, the surface is dried by blowing air, and then put into a stirring tank in a predetermined amount. To this, 0.3% of the present intercellular substance-decomposing enzyme and 0.5% of L-ascorbic acid as an antioxidant are added, and the mixture is stirred at 30 ° C for 60 to 60%.
The cells are converted into single cells for 120 minutes. This is sieved with a pulper finisher having a hole diameter of 0.7 to 0.6 mm to separate into a residue and a single cell suspension. Since green apples contain chlorophyll, they are 90-90
When a heat sterilization treatment at 95 ° C. is performed, a small amount of chlorophyll is decomposed to flofafen and browned. Therefore, if you really want to reduce the number of yeasts,
When the single cell suspension is sterilized at 65 ° C. for about 30 minutes instead of heat sterilization at 95 ° C., only the young yeast cells in the logarithmic growth phase of yeast cells are inactivated and the total number of yeast cells is reduced without decomposition of chlorophyll. It becomes possible. If the single-cell suspension is immediately frozen and stored at -25 ° C. after packaging and sealing, long-term storage is possible without discoloration. However, 10 ³ / g
It possesses the above yeast number, some mold and about 10 ⁴ / g general viable bacteria. The frozen single-cell suspension remains green and does not discolor even when exposed to air for 3 to 4 hours after spontaneous thawing, but discoloration starts and discoloration occurs when the suspension is left longer than that. The composition and properties of the obtained single cell suspension are shown in Table 61 below.

Example 3 A type of tree leaf that is most frequently used by Japanese is the evergreen tree, which has an annual leaf but is used in the number of 6 to
Until around September, the first to third teas are mainly used, and the others are slightly used as bancha. Since antioxidant components in tea leaves and polyphenol components having catechins, caffeine and other physiological functions are expected to be contained in a lot of old tea leaves until about October to May, effective use of unused tea leaves at this time is effective. For use, this intercellular substance-degrading enzyme dissociates cells from leaf veins, petiole, etc. into a single cell suspension, spray-dried and powdered, and is a natural antioxidant, chlorophyll green natural pigment It is considered that the range of use as a tea-like flavor imparting substance is wide.

From the above, the tea leaves from October to December are
When immersed in a 0.1-1.0% NaOH solution heated to 40 ° C. for 30-180 seconds and stirred to dissolve and remove waxy substances on the tea leaf surface, the natural gloss of the tea leaf surface is completely lost, It becomes clear that waxy matter has been removed. Alkali is sufficiently removed by washing with water, and 60 to 1 in hot water of 93 to 95 ° C.
Branch for 20 seconds. Immediately water-cool, pull up,
After cutting into 1 cm squares, add 1.75 times the weight of water,
0.5% of the present intercellular substance-degrading enzyme powder is added, and single cells are formed at 30 ° C. for 2 hours with stirring. In order to separate the petiole and leaf veins from the single-cell suspension, they are squeezed and separated on a pulper finisher or a sieve of about 20 mesh to separate the leaf veins and petiole residues and the single-cell suspension. The results obtained are shown in Table 6 below together with the conditions for forming a single cell. Although the residue amount accounts for 20 to 22% of the whole, it indicates about 55 to 60% of the blanching and draining raw material used as the sample.

The amount of the single-cell suspension accounts for 85 to 88% of the whole, and the number of cells in 1 ml is determined by the NaO used for dewaxing.
When the concentration of H is in the range of 0.1 to 0.5%, 10 ⁷ /
In the case of a 1% NaOH solution, the number of cells in 1 ml is as small as about 10 ⁵ to 10 ⁶ . This solution was adjusted to pH 7.0 and then spray-dried with a spray drier to obtain a single cell powder. The yield of single-cell powdered tea with respect to the first raw tea leaves is as large as 13 to 14% when the number of cells in the single-cell suspension is 10 ⁷ / ml,
In the case of 10 ⁶ / ml, it becomes 11%, and in the case of 10 ⁵ / ml, it becomes 10% or less. The color tone of the obtained single-cell powder is a green powder with a strong yellowish color when the number of cells in the single-cell suspension is 10 ⁶ / ml, and the color tone of the powder is green-brown when the number of cells in the suspension is 10 ⁵ / ml. This means that if there are many disrupted cells, chlorophyll will be exposed in the suspension and if this is sprayed onto a drum above 200 ° C during spray drying,
It is considered that the chlorophyll molecules were directly decomposed by heat and as a result, the chlorophyll content was reduced, and the yellow color of the flavonoids that had originally been intensified.

[0079]

[Table 1]

[0080]

[Table 2]

[0081]

[Table 3]

[0082]

[Table 4]

[0083]

[Table 5]

[0084]

[Table 6]

[0085]

[Table 7]

[0086]

[Table 8]

[0087]

[Table 9]

[0088]

[Table 10]

[0089]

[Table 11]

[0090]

[Table 12]

[0091]

[Table 13]

[0092]

[Table 14]

[0093]

[Table 15]

[0094]

[Table 16]

[0095]

[Table 17]

[0096]

[Table 18]

[0097]

[Table 19]

[0098]

[Table 20]

[0099]

[Table 21]

[0100]

[Table 22]

[0101]

[Table 23]

[0102]

[Table 24]

[0103]

[Table 25]

[0104]

[Table 26]

[0105]

[Table 27]

[0106]

[Table 28]

[0107]

[Table 29]

[0108]

[Table 30]

[0109]

[Table 31]

[0110]

[Table 32]

[0111]

[Table 33]

[0112]

[Table 34]

[0113]

[Table 35]

[0114]

[Table 36]

[0115]

[Table 37]

[0116]

[Table 38]

[0117]

[Table 39]

[0118]

[Table 40]

[0119]

[Table 41]

[0120]

[Table 42]

[0121]

[Table 43]

[0122]

[Table 44]

[0123]

[Table 45]

[0124]

[Table 46]

[0125]

[Table 47]

[0126]

[Table 48]

[0127]

[Table 49]

[0128]

[Table 50]

[0129]

[Table 51]

[0130]

[Table 52]

[0131]

[Table 53]

[0132]

[Table 54]

[0133]

[Table 55]

[0134]

[Table 56]

[0135]

[Table 57]

[0136]

[Table 58]

[0137]

[Table 59]

[0138]

[Table 60]

[0139]

[Table 61]

[0140]

[Table 62]

[0141]

[Table 63]

[0142]

[Table 64]

[0143]

[Table 65]

[0144]

[Table 66]

[0145]

[Table 67]

[0146]

[Table 68]

[0147]

[Table 69]

[0148]

[Table 70]

[0149]

[Table 71]

[0150]

[Table 72]

[0151]

[Table 73]

[0152]

[Table 74]

[0153]

[Table 75]

[0154]

[Table 76]

[0155]

According to the cell separating enzyme used in the method disclosed in Japanese Patent Publication No. 6-71413, hard tissue such as tree leaf tissue or kuma bamboo leaf made of protopectin which is a strong intercellular adhesive substance is used. Although it was almost impossible to separate cells into single cells, the use of the intercellular substance-decomposing enzyme used in the present invention easily decomposes the intercellular adhesive substance adhered by hard and strong protopectin to produce vegetables,
Wild grasses, fruits, medicinal plants, tree leaves, and the like can completely degrade the cells stuck to the petiole, stem, and vein and the adhesion between cells, and can completely convert all cells into single cells. Therefore, the method of the present invention is an industrially profitable and effective method.

The intercellular substance-degrading enzyme used in the present invention is the endo-polygalacturon of the cell separating enzyme disclosed in Japanese Patent Publication No. 6-71413.
case and pectin-trans-eliminas
e is mainly different from the enzyme which does not contain cellulase, and only protopectin which is double and triple linked via Ca to form a strong adhesive substance bonded to cellulose, hemicellulose and pectin molecules alone A great advantage is that it is a special enzyme that decomposes and dissociates into single cells. Furthermore, according to the method disclosed in Japanese Patent Publication No. 6-71413, only some of the vegetables and fruits can be made into single cells, but actually, the vegetables, wildflowers and fruits obtained by cutting the above-mentioned cell-separating enzyme powder are obtained. It is only a very small number of vegetables that can be made into single cells by agitation in addition to herbs, etc., and many of those that cannot be put into practical use by the method applied to only such a few vegetables .

In the method of the present invention, for vegetables and wildflowers having a water content of less than 85%, water is added up to a water content of 85 to 90%. Then, by adding 10 to 20% of water, it is possible to convert these vegetables and the like, which are difficult to convert into single cells, into single cells. In addition, vegetables containing a lot of polyphenols, Ca, Mg and other astringent tastes, vegetables containing astringent components, wild grasses, moderate blanching, deactivating the polyphenol oxidase by operations such as water exposure, polyphenols and The method of the present invention, which elutes and removes the astringent and astringent components such as Ca and Mg and then promotes the single cell action, does not cause discoloration such as browning and black discoloration, and is a single cell suspension of raw vegetables and wild grass as it is. A suspension can be obtained.

Furthermore, white vegetables such as radish and lotus root,
Vegetables and fruits, such as sweet potatoes, potatoes, apples, and cucumber, that contain a polyphenolic substance and its oxidase, polyphenol oxidase. In the case where the polyphenol itself is oxidized and discolored by the operation to make it impossible to commercialize at all, in the present invention,
From the moment of peeling and cutting, immersion in an antioxidant such as L-ascorbic acid, glutathione, sodium L-ascorbate, or a mixture of these antioxidants and an organic acid such as citric acid or acetic acid to form an oxidase. Inhibits oxidation by deactivating and promotes single celling. That is, in order to prevent the strong oxidization occurring during the stirring operation due to the action of the next intercellular substance-decomposing enzyme after the inactivation of the above oxidase, a slight excess amount of L-ascorbic acid or the like is added to the single cell. To Thus, in the present invention, single cells can be completely formed without any loss of the color tone of the raw material. At this time, if the added amount of the antioxidant is small, the antioxidant is completely oxidized during the action of the enzyme and turns into a colored substance, and the single-cell suspension turns into a strong brown substance, but in the present invention, By adopting an appropriate method for preventing discoloration according to such vegetables and fruits, a product having no discoloration and having commercial value can be obtained.

Further, according to the present invention, tree leaves can be completely converted into single cells by a pretreatment method for converting tree leaves into single cells or a watering method for converting single cells. Further, in the case of kuma bamboo leaves, ginkgo leaves, etc., discoloration or drying occurs due to refrigerated storage, but in the present invention, long-term storage for one month or more becomes possible by adopting a running water immersion method, and swelling The softened and hard tissue can be made into a single cell.

As described above, according to the present invention, any plant parenchyma such as vegetables, wild grasses, fruits, medicinal plants, tree leaves, bear bamboo leaves, etc. can be converted into single cells.

The single cell suspension obtained by the method of the present invention or the dry powder thereof can be used as a food material not only in terms of nutritional aspects as the first function, but also as delicious as the second function, that is, tongue texture. It has good physical properties such as over throat and flavor, and has a lower surface tension and interfacial tension than those obtained by mechanical grinding, and is very easy to mix with water and oil. It is of high value. In addition, the dry powder of the single cell suspension obtained by the method of the present invention has a hygroscopicity that is 1/2 that of the dry powder obtained by mechanical grinding.
The discoloration (discoloration) due to visible light irradiation is small.

[Brief description of the drawings]

FIG. 1 shows the acidic protease (p
H4.5) A curve diagram showing the residual activity after the heat treatment.

FIG. 2 is a curve diagram showing the residual activity after heat treatment of an acidic protease (pH 4.5) in a carrot single cell suspension.

FIG. 3 is a curve diagram showing residual lipase activity after heat treatment of a carrot single cell and an intercellular fluid.

Claims (13)

(57) [Claims] An enzyme isolated from a filamentous fungus of the genus Rhizopus, which acts on a plant with an intercellular substance-degrading enzyme mainly composed of protopectinase, which dissociates strong protopectin, to convert intercellular substances of plant parenchyma. A method for completely transforming a plant into single cells, comprising decomposing the plant to completely transform into a single cell. 2. immersing the plant in a warm alkaline solution in advance;
2. The method for completely transforming a plant into a single cell according to claim 1, wherein the waxy substance on the plant surface is dissolved off, further washed with water, and then blanched, and then the intercellular substance-degrading enzyme is acted on. 3. A plant which has been previously blanched to deactivate an oxidizing enzyme having a discoloring effect, an enzyme which degrades and discolors chlorophyll or an enzyme which causes an unpleasant odor contained in the plant. 2. The method for completely transforming a plant into a single cell according to claim 1, wherein an intercellular substance degrading enzyme is acted on. 4. The plant according to claim 1, wherein an antioxidant is added to the plant in advance to prevent oxidative discoloration caused by a polyphenol compound contained in the plant, and then the intercellular substance-degrading enzyme is acted on. Method of complete single cell. 5. The method according to claim 1, wherein the plant is previously blanched and exposed to water to remove the astringent and harsh components contained in the plant together with the polyphenol compound, and then act on the intercellular substance-degrading enzyme. A method for completely transforming a plant into a single cell as described above. 6. The water content of the plant is adjusted to 8 by adding water in advance.
2. The method according to claim 1, wherein the intercellular substance-degrading enzyme is allowed to act after adding 5% or more of water. 7. The plant is pre-watered with 20 to 50% water,
2. The method according to claim 1, wherein the intercellular substance-degrading enzyme is acted on after reducing the viscosity of the plant containing the component exhibiting viscous properties. 8. The method according to claim 1, wherein the single cell suspension obtained by completely converting the plant into single cells is powdered by spray drying. 9. The complete single cell of a plant according to claim 1, wherein the single cell suspension obtained by converting the plant into a single cell is maintained at a viscosity of 300 cp or more to prevent sedimentation of the single cell particles of the single cell suspension. Method. 10. A viscous liquid of carrageenan or agar is added to the single cell suspension to reduce the viscosity of the single cell suspension to 300 c.
The method for completely transforming a plant into a single cell according to claim 9, which is maintained at p or more. 11. The method according to claim 1, wherein a residue or waste discharged during processing of the plant is used as the plant. 12. The plant according to claim 1, wherein the single-cell suspension obtained by completely transforming the plant into a single cell is heated at 95 to 100 ° C. for 30 minutes or more to completely inactivate the intracellular hydrolase. Complete single cell method. 13. The method according to claim 1, wherein the intercellular substance degrading enzyme is allowed to act on the plant at least twice to completely convert the plant into a single cell.
A method for completely transforming a plant into a single cell as described above.