JP2011148724A - Plant material saccharified product, brewed product and method for producing these products - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for producing a plant material saccharified product which can increase and reinforce an oligosaccharide and a polyphenol of health functional components present in the plant material saccharified product and a brewed product such as table vinegar in the form derived from the raw material. <P>SOLUTION: The plant material saccharified product is obtained by subjecting a plant material to mixed enzyme treatment and can be made in a state where the concentration of at least either of the raw material-derived oligosaccharide and the raw material-derived polyphenol is increased by the enzyme treatment even without undergoing concentration treatment. As the plant material, squeezed lees of a fruit such as an apple can be used. Further, as the mixed enzyme, cellulase and pectinase are used. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a saccharified plant material, a brewed product, and a method for producing the same, and in particular, a saccharified plant material, a brewed product, and a saccharified plant material capable of increasing oligosaccharides and polyphenols as health functional ingredients. It relates to a manufacturing method.

Conventionally, various technical proposals have been made on plant agricultural and marine product processed foods (for example, vinegar) and their production methods.
JP-A-2001-61434 (Patent Document 1 below) efficiently decomposes plant tissues by adding water and plant tissue-degrading enzymes to plant agricultural and marine products and grinding them, and as a result, discards them. There has been disclosed a technique for obtaining a processed food product of plant-based agricultural and marine products excellent in flavor and texture and containing a large amount of useful components without any ingredients. JP-A-6-105661 (Patent Document 2 below) discloses that plant tissue degrading enzymes such as pectinase, hemicellulase, and cellulase are allowed to act on finely ground raw or dried plants to separate and decompose plant tissues. Thus, a technique for obtaining a plant unicellular food is disclosed.

  In JP-A-5-23162 (Patent Document 3), vinegar production obtained by acetic acid fermentation of a mixture of branched oligosaccharide-containing saccharides such as isomaltose, panose, and isomaltotriose and alcohol for vinegar production. Technology is disclosed. Japanese Patent Laid-Open No. 5-23163 (Patent Document 4) discloses a vinegar production technique obtained by subjecting a mixture of a galactooligosaccharide-containing saccharide or fructooligosaccharide-containing saccharide and alcohol for vinegar production to acetic acid fermentation. .

Japanese Patent Laid-Open No. 5-111374 (Patent Document 5) discloses a powdered vinegar technology containing 10 g or more of galactooligosaccharide or fructooligosaccharide per 100 g, and Japanese Patent Laid-Open No. 5-111375 (Patent Document 6). Powdered vinegar technologies containing 10 g or more of branched oligosaccharides such as maltose, panose and isomaltotriose per 100 g are disclosed.

In addition, in JP-A-6-14762 (Patent Document 7), potassium salt, magnesium salt, citric acid and the like are added to provide an excellent mineral balance and acid-alkali balance, and have a salt reducing effect and taste. Also disclosed are vinegar and ume vinegar composition techniques that are also excellent. The disclosed techniques in Patent Documents 3 to 7 relate to vinegar that has been subjected to acetic acid fermentation using a branched oligosaccharide obtained by enzymatic treatment of starch for saccharification, such as corn starch, potato starch, sweet potato starch, and tapioca starch.

  JP 2003-24006 (Patent Document 8) describes fermenting fruit juice and vegetable juice using lactic acid and lactic acid bacteria capable of producing acetic acid, and presents an unpleasant odor such as diacetyl odor. In addition, a method for producing a vinegar or sour food paste that is mellow and mild and has a good flavor is disclosed. Japanese Patent Application Laid-Open No. 2005-27535 (Patent Document 9) discloses a healthy vinegar utilizing fructooligosaccharides, polyphenols, and the like, which are useful components contained in yacon tuberous root, and a method for producing the same.

The disclosed technique in Patent Document 8 relates to vinegar made functional by adding minerals, and the disclosed technique in Patent Document 9 uses the fructooligosaccharide and polyphenol contained in the yacon itself without any enzyme treatment until the end of fermentation. It relates to the remaining vinegar.

Japanese Patent Laid-Open No. 2001-61434 “Process for producing vegetable processed agricultural and fishery products and processed vegetable agricultural and fishery products” Japanese Patent Application Laid-Open No. 6-105661 “Method for producing plant unicellular food” Japanese Patent Laid-Open No. 5-23162 “New vinegar” Japanese Patent Laid-Open No. 5-23163 “New Food Vinegar” Japanese Patent Laid-Open No. 5-111374 "New powdered vinegar" Japanese Patent Laid-Open No. 5-111375 “New powdered vinegar” JP-A-6-14762 “Functional vinegar and ume vinegar composition” JP 2003-24006 “New vinegar, sour food paste, and production method thereof” Japanese Patent Application Laid-Open No. 2005-27535 “Mellow Healthy Vinegar and Method for Producing the Same”

  By the way, according to the disclosure techniques of Patent Documents 1 and 2 described above, it is possible to produce a pureed food by disrupting the agricultural and marine product tissue by enzyme treatment. However, functional components such as oligosaccharides are increased or decreased depending on production conditions such as enzyme treatment, but no investigation has been made. Also, including all other patent documents, functionalities derived from raw materials, such as oligosaccharides produced from cell wall polysaccharides in raw materials and polyphenols derived from skins, etc., by enzymatic treatment of fruit and vegetable raw materials There are no examples of vinegar with enhanced ingredients.

  As described above, the conventional technologies are limited to technologies for pureing agricultural and marine products by enzyme treatment and fermenting functional components such as oligosaccharide liquids and minerals. Enzymatic saccharification to increase functional components Until the verification of the process and the study of fermented products in which oligosaccharides or polyphenols derived from the raw materials were reinforced by fermenting them, nothing was done.

  Therefore, the problem to be solved by the present invention is based on the problems of the prior art, in the brewed products such as saccharified plant material and vinegar, in the form derived from the raw materials such as oligosaccharides and polyphenols as health functional ingredients. It is to provide a plant material saccharified product, a brewed product, and a method for producing them.

  As a result of studying the above problems, the present inventors have increased the functional components such as oligosaccharides and polyphenols derived from the raw materials by repeating the enzyme treatment, and the functional components remain in fermented products using the same. Through clarifying the conditions, it was found that functional components derived from raw materials can be strengthened, and the present invention has been completed. That is, the invention claimed in the present application, or at least the disclosed invention, as means for solving the above-described problems is as follows.

[1] A plant material obtained by an enzyme treatment of a plant material as a raw material and having a concentration of at least one of a raw material-derived oligosaccharide or a raw material-derived polyphenol increased by the enzyme treatment without going through a concentration treatment Saccharified product.
[2] The plant material saccharified product according to [1], wherein the plant material is a juice-treated plant material or other plant tissue separation-treated plant material.
[3] The plant material saccharified product according to [1] or [2], wherein a mixed enzyme in which cellulase and pectinase are mixed is used for the enzyme treatment.
[4] The saccharified plant material according to any one of [1] to [3], wherein the plant material is a juice residue of apples or other fruits.
[5] A brewed product obtained by an acetic acid fermentation treatment of the saccharified plant material according to any one of [1] to [4].
[6] A carbohydrate material with a high concentration of oligosaccharides obtained by alcohol fermentation of the saccharified plant material according to any one of [1] to [4].
[7] After subjecting the plant material, which is a raw material, to hydrotreatment, pulverization treatment, and heat treatment, at least one of the raw material-derived oligosaccharide or the raw material-derived polyphenol is subjected to an enzyme treatment once or twice or more without undergoing a concentration treatment. A method for producing a saccharified plant material, wherein a saccharified plant material having an increased concentration is obtained.

[8] The method for producing a saccharified plant material according to [7], wherein the plant material is a juice-treated plant material or other plant tissue separation-treated plant material.
[9] The method for producing a saccharified plant material according to [7] or [8], wherein a mixed enzyme in which cellulase and pectinase are mixed is used for the enzyme treatment.
[10] The enzyme treatment is performed twice or more, the primary saccharified solution obtained by the preceding enzyme treatment is added to the untreated raw material, and after further pulverization treatment and heat treatment, the subsequent enzyme treatment is performed. The method for producing a saccharified plant material according to [9], characterized in that it is made.
[11] The method for producing a saccharified plant material according to any one of [7] to [10], wherein the plant material is a juice residue of apples or other fruits.
[12] Brewing, wherein the saccharified plant material obtained by the production method according to any one of [7] to [11] is subjected to an alcoholic fermentation treatment or an alcohol addition treatment, followed by an acetic acid fermentation treatment to obtain a brewed product Manufacturing method.

  Since the plant material saccharified product, the brewed product and the production method thereof according to the present invention are configured as described above, according to this, in the brewed product of the plant material saccharified product or vinegar, the health functional ingredient is included in these. Oligosaccharides and polyphenols can be increased and strengthened in a form derived from raw materials.

That is, according to the present invention, the following specific effects can be obtained.
1) A method for producing a saccharified product in which components of oligosaccharide and polyphenol are increased, and a brewed product thereof.
As a method for producing a saccharified product having an increased amount of oligosaccharide and polyphenol, the present inventors have found that the enzyme treatment step is repeated twice and then the step shown in FIG. 1 is possible. Therefore, according to the production method of the present invention, a saccharified product having increased functional components can be produced by an enzyme treatment step without using a concentrator or a concentration step. Moreover, the oligosaccharide and polyphenol derived from the raw material obtained by decomposing the plant tissue as the raw material can be sufficiently left even after the brewing process.

2) The technology can be used for agricultural products and plants in general.
Cellulose and pectin are common components constituting plant tissues. Therefore, various agricultural products or plants in general including unused and low-utilized plants can be widely used as the raw material of the present invention. In other words, the present invention is an invention that has a wide range of applications and has extremely high applicability and suitability for commercialization.

3) A method for producing a characteristic saccharified product and a brewed product thereof, in which the raw material-derived components are strongly left.
In the second and subsequent enzyme treatment steps, by using the primary saccharified solution that accompanies the first enzyme treatment, there is no dilution due to hydration, and as a result, the resulting product that strongly retains ingredients and flavors derived from the raw materials. It is possible to obtain.

It is a flowchart which shows the structure of the typical manufacturing method of the saccharified plant material of this invention. It is a flowchart which shows the structure of the specific example of a manufacturing method of the plant material saccharified material (secondary saccharified liquid) and brewed material (vinegar) using the enzyme treatment of this invention. It is a flowchart which shows the structure of the example of the specific manufacturing method of the brewed product (vinegar) when not using an enzyme process. It is explanatory drawing which put together the usability of the product by each manufacturing method of this invention when using squeezed residue as plant material compared with the prior art without an enzyme treatment. The following figures are all related to the examples, and this figure is a graph showing changes in the amount of sugar caused by the washing operation with apple juice residue. It is a flowchart which shows the water washing of a squeezed residue, and an enzyme treatment process. It is a graph which shows the constituent sugar analysis result of the final supernatant after a Sumiteam AC (trademark) process. It is a graph which shows the constituent sugar analysis result of the final supernatant after a Sumi team PTE (trademark) process. It is a graph which shows the structural saccharide | sugar analysis result of the final supernatant after a mixed enzyme process. It is a graph which shows the molecular weight distribution by gel filtration chromatography about each sample. It is a flowchart which shows the process of the vinegar production test by an apple juice residue. It is a graph which shows the molecular weight distribution by gel filtration chromatography about each vinegar sample. It is a graph which shows the structural sugar analysis result performed about each vinegar sample. It is a graph which shows the antioxidant test result by DPPH radical scavenging activity about each sample. It is a graph which shows the cancer cell proliferation suppression test result about each sample. It is a graph which shows the relationship between the number of enzyme treatments and the amount of oligosaccharide produced. It is a graph which shows the relationship between the number of enzyme treatments, and the amount of recoverable supernatants. It is a graph which shows the relationship between the number of enzyme treatments, and the concentration of the produced oligosaccharide. It is a graph which shows the molecular weight distribution (absorbance 490 nm) by the gel filtration chromatography of a primary saccharified liquid and a secondary saccharified liquid. It is a graph which shows the molecular weight distribution (absorbance 490nm) by the gel filtration chromatography of the vinegar 3 and the vinegar 5. It is a graph which shows the molecular weight distribution (absorbance 490nm) by the gel filtration chromatography of the vinegar 4 and the vinegar 6. It is the graph which showed the amount of oligosaccharides which can be collect | recovered from each vinegar of vinegar 1 to 6 according to the frequency | count of enzyme treatment when the residue usage-amount was assumed to be 1 kg. It is the graph which showed the total polyphenol amount which can be collect | recovered from each vinegar of vinegar 1 to 6 according to the frequency | count of enzyme treatment at the time of assuming squeeze residue usage-amount to 1 kg.

The present invention will be described in further detail.
The plant material saccharified product of the present invention is obtained by enzyme treatment of plant material as a raw material, and does not undergo concentration treatment as in the prior art, by enzyme treatment, and does not depend on later addition or the like. It is a saccharified plant material in which the concentration of the oligosaccharide or the polyphenol derived from the raw material, or both is increased.

  As the plant material, the plant material that has been subjected to fruit fruit juice processing, which will be described later in the examples, can be used, which means effective treatment, effective use, and added value creation of industrially produced waste. It is realized. However, the plant material of the present invention is not limited to this, so long as the functional component derived from the raw material can be increased by enzyme treatment, it can be widely used, including those that have been conventionally used and unused. It can be used as a raw material of the present invention.

Cellulase or pectinase can be used for the enzyme treatment, and more preferably, a mixed enzyme obtained by mixing these can be used. This is because plant tissues are generally composed of a large amount of cellulose and pectin, so that the use of these enzymes, particularly the combined use, is effective. Other enzymes are not excluded.

  As the plant material, juice residue of apples and other fruits can be used. Of course, not only fruits of apples, oysters, peaches, pears, and other fruit trees but also fruits of plants other than fruit trees such as tomatoes and watermelons are within the fruit range. In addition, a plant material other than this is not excluded.

  In addition, a brewed product obtained by subjecting the saccharified plant material to an acetic acid fermentation treatment is also within the scope of the present invention. Therefore, the vinegar obtained from the saccharified plant material is within the scope of the present invention.

  In addition, a carbohydrate material having a high oligosaccharide concentration obtained by subjecting the saccharified plant material to an alcoholic fermentation treatment is also within the scope of the present invention. Sugar materials can be used in a wide variety of ways as carbohydrate materials in various food processing, pharmaceutical manufacturing, and others.

  The plant material saccharified product production method of the present invention is mainly composed of subjecting a plant material as a raw material to a hydration treatment, a pulverization treatment, and a heat treatment, and then performing an enzyme treatment once or twice or more. By such a method, a saccharified plant material having an increased concentration of at least one of the raw material-derived oligosaccharide or the raw material-derived polyphenol can be obtained without going through a concentration treatment.

  In this production method, plant material that has been squeezed or other plant tissue separation-treated plant material can be used as the plant material. Here, “other plant tissue-separated plant material” refers to a treatment other than the squeezing treatment, which is a treatment capable of separating plant tissues mainly composed of components such as cellulose and pectin, Any separation method can be used as long as it is a separation method that is conventionally performed or can be carried out in the food-related industrial field, whether it is a simple method or a chemical method. It should be noted that plant materials other than squeezed processed plant materials or other plant tissue separated processed plant materials are not excluded.

  In the production method of the present invention, a mixed enzyme in which cellulase and pectinase are mixed can be used for the enzyme treatment. This is because the mixed enzyme having such a structure can provide a saccharified product or brewed product in which a functional component derived from a plant tissue as a raw material is reinforced, and can sufficiently achieve the object of the invention. Other enzymes are not excluded.

  Particularly in the production method of the present invention, the enzyme treatment is performed twice or more, and after the primary saccharified solution obtained by the preceding enzyme treatment is added to the untreated raw material, and further subjected to pulverization treatment and heat treatment, A configuration in which subsequent enzyme treatment is performed can be employed. As described in detail in the examples, a more useful result can be obtained through such a step. In addition, it is not excluded from the present invention that the number of times of enzyme treatment and the method of proceeding with the enzyme treatment step are other than this method.

As plant material, you may use the squeezed residue of an apple or other fruits as detailed in an Example. The squeezed residue of apples and other fruits is industrial waste, but it can be effectively processed and effectively used by the present invention.
In addition, the brewed product can be obtained by subjecting the saccharified plant material obtained by the above-described production method to an alcoholic fermentation treatment or an alcohol addition treatment and then an acetic acid fermentation treatment.

FIG. 1 is a flow diagram showing the configuration of a typical method for producing a saccharified plant material of the present invention. Also,
FIG. 1-2 is a flowchart showing the configuration of a specific example of a method for producing a saccharified plant material (secondary saccharified solution) and a brewed product (vinegar) using the enzyme treatment of the present invention,
FIG. 1-X is a flow diagram showing the configuration of an example of a specific method for producing a brew (vinegar) when no enzyme treatment is used. In this invention, in order to decompose | disassemble plant tissue with an enzyme, the mixed enzyme which mixed the cellulase and the pectinase is used, and by the manufacturing method as shown in FIG. 1, FIG. It is possible to increase and strengthen oligosaccharides and polyphenols in brewed products (vinegar). In relation to the examples to be described later, “primary saccharified solution” corresponds to “supernatant” in FIG. 1, and “liquid addition residue” corresponds to “liquid addition agricultural product” in FIG. To do.

By subjecting the secondary saccharified product to alcohol fermentation, monosaccharides are consumed and oligosaccharides remain, so that a brewed product (secondary alcohol fermented product) having a high ratio of oligosaccharides, which are functional components, can be obtained. it can. Moreover, a brewed product (secondary acetic acid fermented product) can be obtained by subjecting a secondary alcohol fermented product, a secondary saccharified product or a secondary saccharified product to which a brewing alcohol is added to acetic acid fermentation.

1-3 is explanatory drawing which put together the usability of the product by each manufacturing method of this invention in the case of using squeezed residue as plant material compared with the prior art without an enzyme treatment. As shown in the drawing, according to each production method of the present invention using plant materials such as squeezed residue as raw materials, vinegar and sugar materials reinforced with functional ingredients derived from the raw materials can be obtained.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited to this Example. In addition, an Example describes the experimental progress and experimental result which reach this invention.

1) Examination of enzyme treatment conditions (reason for using mixed enzyme in the present invention)
In order to confirm the oligosaccharide produced | generated by an enzyme treatment, the apple sugar residue discharged | emitted from an apple juice manufacturing process was used as a raw material, the free sugar contained in it was removed by water washing, and the enzyme treatment test was done using this. That is, after doubling the amount of apple juice residue, it is pulverized with a desktop universal mixer (Aikosha, Kenmix KM-600), and separated into a supernatant and a precipitate residue by centrifugation (9000 rpm, 15 minutes). The precipitation residue was repeatedly washed with water until no sugar was detected during the cleaning. By repeating the washing with water 5 times, it was confirmed that the residual sugar in the supernatant was almost zero. Therefore, the precipitation residue after the 5 times washing with water was defined as the washing residue. Further, the same amount of water was added to the water washing residue, which was pulverized and heated (121 ° C., 20 minutes) to obtain a water washing water residue.

FIG. 2 is a graph showing changes in the amount of sugar by the apple juice residue washing operation. As shown in the figure, it was confirmed that the acid polysaccharide increased in the water-washed water residue. The total sugar amount was calculated as glucose equivalent by the phenol-sulfuric acid method, and the acidic sugar amount was calculated as galacturonic acid equivalent by the carbazole-sulfuric acid method.

Using this as a raw material, three types of enzyme treatment tests were conducted as follows.
・ Sumiteam AC (registered trademark) (0.5% by weight of apple juice residue),
・ Sumi Team PTE (registered trademark) (0.5%),
・ Sumiteam AC (registered trademark) (0.25%) + Sumiteam PTE (registered trademark) (0.25%))
The saccharified product after each enzyme treatment was heated (121 ° C., 5 minutes) for enzyme inactivation, and separated into a final precipitate residue and a final supernatant by centrifugation (9000 rpm, 15 minutes). Hereinafter, Sumiteam AC (registered trademark) + Sumiteam PTE (registered trademark) will be referred to as a mixed enzyme. In addition,
FIG. 3 is a flow diagram showing the water washing and enzyme treatment process of the apple juice residue described above. In addition, the “water-washed water residue” in the figure is also simply referred to as a “water residue”. The “final supernatant” corresponds to the “primary saccharified solution” in the present description.

Each final supernatant after the enzyme treatment (equivalent amount of sugar 10 mg) was subjected to gel filtration chromatography of Bio-Gel P-2, and absorbance 490 nm by phenol sulfate method and absorbance 530 nm by carbazole sulfate method were measured.
FIG. 4 is a graph showing the constituent sugar analysis results of the final supernatant after treatment with Sumiteam AC (registered trademark);
FIG. 5 is a graph showing the constituent sugar analysis results of the final supernatant after the Sumiteam PTE® treatment, and
FIG. 6 is a graph showing the constituent sugar analysis results of the final supernatant after the mixed enzyme treatment. In any measurement, glucose (G1), maltose (G2), maltotriose (G3), maltopentaose (G5), maltoheptaose (G7), and blue dextran 2000 (V0) were used as markers.

  Furthermore, the total sugar (by phenol sulfuric acid method), acidic sugar (by carbazole sulfuric acid method), and oligosaccharide amount (calculated from gel filtration distribution) contained in each sample were analyzed. Table 1 shows the results of analyzing the amount of sugar in each sample. From these results, in Sumiteam AC (registered trademark), oligosaccharides and monosaccharides increased, in Sumiteam PTE (registered trademark), polysaccharides increased, and in Sumiteam AC (registered trademark) + Sumiteam PTE (registered trademark), Sumiteam AC ( It was clarified that the polysaccharide increased slightly as well. In the table, Sumiteam PTE (registered trademark) and Sumiteam AC (registered trademark). The same applies when displayed in the following drawings.

In the present invention, it is assumed that the amount of sugar is increased by repeating the enzyme treatment, but it was assumed that when the Sumiteam AC (registered trademark) treatment is repeated twice, monosaccharideization proceeds and oligosaccharide decreases. And since only the Sumiteam PTE (registered trademark) treatment has a small increase in oligosaccharide, a method of treating with a mixed enzyme of Sumiteam AC (registered trademark) and Sumiteam PTE (registered trademark) was selected.

2) Apple squeeze residue oligo saccharification test The squeeze residue of apple squeezes as it is because the amount of water is insufficient and the enzymatic reaction does not proceed. In the present invention, the same weight of water was added to the juice residue, and after pulverization, autoclaved (121 ° C., 20 minutes) was used as the water residue. Next, in a state where the water residue is heated to 40 ° C., Sumiteam AC (registered trademark) and Sumiteam PTE (registered trademark) are respectively added in a residue wet weight ratio of 0.25% (w / w). Using a mentor (Tokyo Science Instrument, Jarfa Mentor MBF250-M), an enzyme treatment was performed by stirring for 24 hours at 40 ° C. to obtain a primary saccharified product. Subsequently, the primary saccharified product is centrifuged (9000 rpm, 30 minutes), and the supernatant is used as a primary saccharified solution, and the same weight is added to a fresh apple juice residue, followed by pulverization and autoclaving (121 ° C., 20 The second enzyme treatment was carried out in the same manner as the above-mentioned hydrated residue, using the lysate as the added residue. The solubilized product obtained by this operation was used as a secondary saccharified product.

Among the oligosaccharide saccharification tests, the total sugar, acidic sugar, oligosaccharide amount, and polyphenol amount (by the foreign thiocult method) for the centrifugal supernatant of each of the squeezed residue, the hydrolyzed residue, the primary saccharified product, and the secondary saccharified product It was measured. Table 2 shows the analysis results of each sample in the oligosaccharification test.

From this result, it was confirmed that the amount of oligosaccharide increased about 4.4 times in the secondary saccharified product compared to the water residue. Polyphenols also increased with the enzyme treatment, and it was considered that they were eluted from the skin. Further, each final supernatant (equivalent amount of sugar 10 mg) was subjected to Bio-Gel P-2 gel filtration chromatography, and the absorbance at 490 nm by phenol sulfuric acid method and the absorbance at 530 nm by carbazole sulfuric acid method were measured (FIG. 7). In the oligosaccharide region (fractions No. 33 to No. 64), an oligosaccharide was detected with the enzyme treatment.
FIG. 7 is a graph showing the molecular weight distribution of each sample by gel filtration chromatography. In the figure, black circles: total sugar (490 nm), white circles: acidic sugar (530 nm).

3) Apple juice residue saccharification test / alcohol fermentation In addition to the primary saccharified product and secondary saccharified product, the supernatant obtained by centrifuging the hydrolyzed residue (9000 rpm, 30 minutes) (the hydrolyzed residue supernatant) was added to a 500 ml glass. 200 g each was dispensed into the container, and a breathable silicosene was attached to the mouth of the glass container, which was further covered with an aluminum foil to make a semi-sealed state. This was sterilized in an autoclave (121 ° C., 20 minutes), allowed to cool to room temperature, inoculated with 500 μl (0.25% (v / w)) of the restored yeast, and allowed to stand at 25 ° C. Alcohol fermentation was started. Since CO ₂ is generated along with alcohol fermentation, and the total weight of the CO ₂ is reduced when it comes out of the container, the weight of the entire container is measured once a day, and the time when the weight reduction stops is judged as the end of fermentation. did.

-Acetic acid fermentation Next, a portion of each saccharified product of primary saccharified product, secondary saccharified product and hydrolyzed residue was sampled at the end of alcoholic fermentation, and the alcohol concentration was measured for each supernatant (10000 rpm, 10 minutes) (RIKEN) Instrument, Alcomate AL-2 type). Based on the results, as a non-alcohol fermentation test, brewing alcohol was added to the primary saccharified product, secondary saccharified product, and water residue supernatant in the clean bench so that each alcohol fermentation test group had the same concentration. did.

Next, 2% (v / w) equivalent of each of the acetic acid bacteria expanded and cultured in a total of 6 test sections of alcohol fermentation test and non-alcohol fermentation test was inoculated. Thereafter, the aluminum foil covering the silicosene was removed, and it was allowed to ventilate and left still at 30 ° C., sampled every 3 to 4 days to measure the acidity, and when the increase in acidity stopped It was judged.
FIG. 8 is a flowchart showing the steps of the vinegar production test using the apple juice residue. Table 3 shows an outline of the manufacturing process for each vinegar.

In order to compare the test-produced vinegar with commercially available vinegar, two items of over-the-counter apple vinegar were obtained. As the commercial vinegar, vinegar produced by alcohol fermentation (commercial vinegar 1) and vinegar produced by alcohol addition (commercial vinegar 2) were selected from the notation of raw materials without adding oligosaccharides. Both are marked with JAS (Japanese Agricultural Standards) mark.

Among these, about each vinegar 1, 2, 5, 6 and commercial vinegars 1, 2, each centrifugation supernatant (sugar amount equivalent to 10 mg) is used for the gel filtration chromatography of Bio-Gel P-2, and a phenol sulfuric acid method is used. Absorbance of 490 nm and absorbance by the carbazole sulfate method were measured.
FIG. 9 is a graph showing the molecular weight distribution by gel filtration chromatography for each vinegar sample. In the figure, black circles: total sugar (490 nm), white circles: acidic sugar (530 nm).

Furthermore, when the constituent sugar analysis was performed on the gel filtration fractions of the squeezed residue supernatant, the secondary saccharified solution, vinegars 1, 2, 5, 6 and commercial vinegars 1 and 2, it was derived from plant polysaccharides increased by enzyme treatment. It was confirmed that a plurality of types of oligosaccharides remained after brewing without being utilized by microorganisms.
FIG. 10 is a graph showing the results of the constituent sugar analysis performed on each vinegar sample.

4) Component analysis and functionality evaluation test Two commercial vinegars were added to six vinegars produced as described above, and the total sugar amount, acidic sugar amount, oligosaccharide amount and polyphenol amount were measured. Table 4 shows the component analysis results for each vinegar.

From this result, it was confirmed that the amounts of oligosaccharides and polyphenols increased in the vinegar (vinegars 5 and 6) subjected to the enzyme treatment twice. In vinegars 3 and 5, the ratio of oligosaccharides to total sugars increased to 80% due to the consumption of monosaccharides accompanying alcohol fermentation, and commercialization was expected as a low monosaccharide and high oligosaccharide vinegar. Since the total amount of sugar in the commercial vinegar 1 exceeded the amount of sugar contained in the apple raw material, it was suggested that it could be produced using apple juice concentrate.

Next, Oki et al. (Tomoyuki Oki, Mami Masuda, Osamu Furuta, Yoichi Nishiba, Ikuo Suda, Radical scavenging activity of chips made from purple sweet potato, food industry, 48, pp.926-932 (2001) ) To measure DPPH radical scavenging activity.
FIG. 11 is a graph showing an antioxidant test result by DPPH radical scavenging activity for each sample. From this result, it was confirmed that the antioxidant effect became stronger by repeating the enzyme treatment. Although it does not reach the commercial vinegar 1 in which the use of concentrated fruit juice is presumed, the antioxidant activity has increased. Therefore, the vinegar produced in the present invention can be expected to have sufficient antioxidant functionality.

Furthermore, human colon cancer cells DLD-1 were used, and Kato et al.'S method (: Kato, Y., Uchida, J., Ito, S. and Mitsuishi, Y., Structural analysis of the oligosaccharide units of xyloglucan and their effects). on growth of COLO201.
Human tumor cells. International Congress Series, 1223, pp. 161-164 (2001)) was used as a reference for cell growth inhibition tests. Each sample was used after serial dilution after lyophilization. After cell culture, the number of viable cells was calculated by absorbance measurement using Tetra Color ONE reagent (manufactured by Biochemical Biobusiness), and suppression of cell proliferation was recognized in a concentration-dependent manner. The sample concentration at which this was suppressed by 50% was defined as IC50.

FIG. 12 is a graph showing the results of cancer cell proliferation inhibition test for each sample. From this result, as the number of enzyme treatments increased, and by carrying out acetic acid fermentation, the growth inhibitory effect of cancer cells was observed respectively.

About 8 points of these vinegars, sensory evaluation by blinds with the product name, production method, etc. faced down was performed by 38 panels (15 males, 23 females, 21-60 years old, average 37.3 years old). About each item of color, fragrance, and flavor and overall evaluation, scoring was made in five stages (excellent 5, good 4, generally good 3, inferior 2, bad 1), and the average value (mean) and standard deviation (SD) were calculated. Asked. In Table 5, the sensory test result of vinegar is shown.

As a result, although it was a 5-point evaluation, a remarkable difference was not seen, but comparing alcohol-fermented vinegar (vinegar 1, 3, 5) and vinegar with alcohol addition (vinegar 2, 4, 6), The latter was more highly evaluated in terms of color, fragrance, flavor, and overall. This was thought to be due to the lack of sweetness due to the decrease in monosaccharides due to yeast by alcohol fermentation, and the change in aroma associated with fermentation. In addition, the evaluation tends to be higher when the enzyme treatment is repeated. That is, the vinegar produced using the test juice brewed apple juice as a raw material and subjected to the enzyme treatment twice has obtained the same evaluation as commercially available vinegar, and not only in terms of functionality but also in terms of flavor. It was able to be evaluated that it was the method which can manufacture a more preferable vinegar.

5) Others Now, agricultural products are composed of plant polysaccharides, and there are several ways to decompose them. For example, Japanese Patent Application Laid-Open No. 2009-195189 “Method for Producing Monosaccharide or Oligosaccharide from Polysaccharide”, Japanese Patent Application Laid-Open No. 2009-11317 “Method for Producing Monosaccharide or Oligosaccharide from Polysaccharide Using Acidic Sugar”, No. 2005-110675 “Method for producing low-sugar and / or oligosaccharide and functional low-sugar and / or oligosaccharide”, and the like.

In the present invention, a mixed enzyme, specifically, cellulase and pectinase are allowed to act to decompose by a method of increasing monosaccharides and oligosaccharides derived from plant tissues. However, in the method of simply increasing the number of enzyme treatments, it was considered that the oligosaccharide itself would have a low molecular weight. Therefore, an oligoglycation test and a vinegar production test using a saccharified product were conducted to verify the oligosaccharide low molecular weight.

Here, based on the yield obtained in the above test, assuming that the amount of residue to be used is 1 kg, the amount of oligosaccharide that can be recovered was calculated. However, the oligosaccharides that could be recovered were small (under the conditions of this test, from about 1 g of the residue to about 7 g of oligosaccharides (sucrose)), but the oligosaccharides that could be recovered by the enzyme treatment increased about 4 times. However, there was no large difference in the total amount of oligosaccharides that could be recovered between the enzyme treatment once and twice, and the amount that could be recovered was considered to be constant depending on the amount of residue to be treated.
FIG. 13 is a graph showing the relationship between the number of enzyme treatments and the amount of oligosaccharide produced. As shown in the figure, 27 to 28 g of oligosaccharide was recovered from 1 kg of the residue under the test conditions.

FIG. 14 is a graph showing the relationship between the number of enzyme treatments and the amount of recoverable supernatant. Also,
FIG. 15 is a graph showing the relationship between the number of enzyme treatments and the concentration of oligosaccharide produced. As shown in these figures, when focusing on the amount of supernatant (liquid amount) that can be recovered, it increases as liquefaction progresses in the first enzyme treatment, but less in the second enzyme treatment than in the untreated enzyme. Therefore, the amount recovered was reduced. That is, it was confirmed that the oligosaccharide concentration increased with the number of enzyme treatments by the production method of the present invention (FIG. 15).

Thus, for example, if the initial amount of water can be produced with 1/3 water instead of the same weight, it was considered that only one treatment is required. However, if at least an equal amount of water is not added, the pulverization process is difficult, and if the pulverization is not performed, the enzyme treatment does not proceed. Thus, a concentrated saccharified solution is produced by repeating the enzyme treatment as in the production method of the present invention. It is considered desirable to do so. Industrially, it is not necessary to carry out the treatment twice in this way, and a primary saccharified solution produced by equal amount of water is prepared in advance, and an enzyme treatment with an equal amount of addition using this is performed once. Should be done.

FIG. 16 is a graph showing the molecular weight distribution (absorbance 490 nm) of the primary saccharified solution and the secondary saccharified solution by gel filtration chromatography. Also,
FIG. 17 is a graph showing molecular weight distribution (absorbance 490 nm) of vinegar 3 and vinegar 5 by gel filtration chromatography.
FIG. 18 is a graph showing the molecular weight distribution (absorbance 490 nm) of vinegar 4 and vinegar 6 by gel filtration chromatography. As can be seen from these figures, the oligosaccharide polymer region No. 2 can be obtained by performing enzyme treatment twice. It can be seen that the ratio from 33 to 45 is decreasing.

Furthermore, when the vinegar subjected to acetic acid fermentation using the saccharified liquid is divided into vinegar obtained by alcohol fermentation (FIG. 17) and vinegar obtained by addition of alcohol (FIG. 18), the ratio reduction of the oligosaccharide polymer region is more remarkable. became. In this case, vinegars 3 and 4 were subjected to two heat treatments for enzyme treatment and sterilization, but vinegars 5 and 6 were subjected to a total of three heat treatments, so that acid sugar increased 2 It was considered that decomposition of the polymer oligosaccharide progressed during the heating of the secondary saccharified product.

FIG. 19 is a graph showing the amount of oligosaccharide that can be recovered from each vinegar of vinegars 1 to 6 according to the number of enzyme treatments, assuming that the amount of juice residue used is 1 kg. As a result, the vinegar produced without enzyme treatment originally had a small amount of oligosaccharides because there were few oligosaccharides, and the vinegar using the saccharified product after one enzyme treatment was used between alcohol fermented vinegar and alcohol-added vinegar. There was almost no difference (both the oligosaccharides were increased by the enzyme treatment, and it was assumed that 27 to 28 g of oligosaccharides could be produced from 1 kg of the residue).

In the vinegar using the saccharified product after the enzyme treatment two times, the alcohol fermented vinegar slightly increased from the enzyme treatment once, but decreased in the alcohol-added vinegar. The decrease in alcohol-added vinegar was thought to be due to the degradation of oligosaccharides as described above, but for alcohol-fermented vinegar, an increase in oligosaccharides around 2-3 sugars, which is thought to be due to the pectinase of yeast itself, was observed. From this, it was clarified that the total oligosaccharide amount obtained from vinegar using a secondary saccharified product varies depending on the fermentation method.

FIG. 20 is a graph showing the total amount of polyphenols that can be recovered from each vinegar of vinegars 1 to 6 according to the number of enzyme treatments, assuming that the amount of juice residue used is 1 kg. From the above, repeating the enzyme treatment multiple times is a factor that promotes a reduction in the molecular weight of the produced oligosaccharides, and in particular, vinegar produced by the alcohol addition method has a high monosaccharide ratio, and the functional component oligosaccharides are Although it was assumed that it decreased, about the amount of total polyphenols as shown in FIG. 20, it was assumed that the quantity which can collect | recover 1 kg of residues as a raw material will increase with the number of enzyme treatments.

From this, it became clear that the target functional component can be strengthened by appropriately determining the number of enzyme treatments. On the other hand, when considering the enhancement of the oligosaccharide amount above a certain level, it was considered optimal to produce vinegar using a saccharified product concentrated twice by enzyme treatment.

According to the plant material saccharified product, the brewed product, and the production method thereof, in the brewed product of the plant material saccharified product or vinegar, the health function component oligosaccharide or polyphenol is derived from the raw materials. It is possible to obtain a brewed product such as saccharified plant material or vinegar that has been increased or strengthened in step (b). The plant material according to the present invention is not particularly limited as described above as long as it relates to a plant tissue from which a functional component such as an oligosaccharide can be obtained. Obtaining a high value-added product such as that of the present invention using waste is very meaningful in terms of effective waste disposal, effective use of low-use / unused resources, and high added-value. Therefore, the present invention is highly industrially applicable.

Claims (12)

A saccharified plant material obtained by an enzyme treatment of a plant material as a raw material and having a concentration of at least one of a raw material-derived oligosaccharide or a raw material-derived polyphenol increased by the enzyme treatment without passing through a concentration treatment. The saccharified plant material according to claim 1, wherein the plant material is a squeezed plant material or another plant tissue separation-treated plant material. The saccharified plant material according to claim 1 or 2, wherein a mixed enzyme obtained by mixing cellulase and pectinase is used for the enzyme treatment. The saccharified plant material according to any one of claims 1 to 3, wherein the plant material is a juice residue of apples or other fruits. A brewed product obtained by an acetic acid fermentation treatment of the saccharified plant material according to any one of claims 1 to 4. A carbohydrate material with a high concentration of oligosaccharides obtained by alcohol fermentation of the saccharified plant material according to any one of claims 1 to 4. After subjecting the plant material, which is a raw material, to hydrotreatment, pulverization treatment, and heat treatment, by performing enzyme treatment once or twice or more, at least one of raw material-derived oligosaccharides or raw material-derived polyphenols without undergoing concentration treatment A method for producing a saccharified plant material, wherein a saccharified plant material having an increased concentration can be obtained. The method for producing a saccharified plant material according to claim 7, wherein the plant material is a juice-treated plant material or other plant tissue separation-treated plant material. The method for producing a saccharified plant material according to claim 7 or 8, wherein a mixed enzyme obtained by mixing cellulase and pectinase is used for the enzyme treatment. The enzyme treatment is performed twice or more, and the primary saccharified solution obtained by the preceding enzyme treatment is added to the untreated raw material, followed by pulverization treatment and heat treatment, followed by subsequent enzyme treatment. The method for producing a saccharified plant material according to claim 9, wherein: The method for producing a saccharified plant material according to any one of claims 7 to 10, wherein the plant material is a juice residue of apples or other fruits. A method for producing a brewed product, wherein the saccharified plant material obtained by the production method according to any one of claims 7 to 11 is subjected to an alcoholic fermentation treatment or an alcohol addition treatment, followed by an acetic acid fermentation treatment to obtain a brewed product.

Images