KR102318883B1 - Method for manufacturing companion animal feed containing burdock extract - Google Patents

Abstract

The present invention relates to a method for manufacturing a companion animal feed for alleviating depression including a burdock extract obtained through an enzyme treatment and lactic acid bacteria fermentation process. A method for manufacturing a companion animal feed for alleviating depression comprising a burdock extract according to the present invention includes the steps of: mixing rice bran powder and indigestible dextrin, and mixing the burdock extract thereto.

Description

Method for manufacturing companion animal feed containing burdock extract for improving depression {Method for manufacturing companion animal feed containing burdock extract}

The present invention relates to a method for manufacturing a companion animal feed for improving depression comprising a burdock extract, and more particularly, to a method for improving depression comprising a burdock extract comprising a burdock extract obtained through enzyme treatment and lactic acid bacteria fermentation process as an active ingredient It relates to a method for manufacturing pet food.

Recently, domestic and foreign companion animal breeding is in the midst of a rapid increase as income levels rise, aging, and nuclear families such as single-person households are becoming common. In the United States, in 2015, 53.5% of households with a dog or cat, 12.7% of households with a dog and a cat, and 10.5% of households with other companion animals, 76.7% of all households own a companion animal. .

In Korea, as the number of elderly and single-person households increases, the number of households raising companion animals is also increasing significantly. In 2015, 21.8% of all households in Korea were surveyed to have a companion animal, and this percentage continues to increase.

In particular, with the recent increase in single-person households in Korea, the proportion of companion animals raised by single-person households is also increasing. The problem of depression occurring in pets raised in the district is increasing.

As the companion animal market grows, many snacks and nutritional supplements for companion animals are being imported or developed, but nutritional supplements and feeds for the mental health of companion animals are still underdeveloped.

Domestic Registered Patent No. 10-1428263 (2014.08.01) Domestic Registered Patent No. 10-2172706 (2020.10.27)

An object of the present invention is to provide a method for preparing a companion animal feed for improving depression, including the burdock extract, so that a composition effective in alleviating depression in companion animals can be provided by using the burdock extract extracted from burdock as an active ingredient.

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The method for manufacturing a companion animal feed for improving depression containing a burdock extract of the present invention includes mixing rice bran powder and indigestible dextrin, and mixing the burdock extract thereto.

The burdock extract pulverizes burdock, an enzyme treatment process of enzymatic treatment through pectin-degrading enzyme and fibrin-degrading enzyme, a lactic acid bacteria fermentation process of fermenting the burdock enzyme-treated product formed through the enzyme treatment process with lactic acid bacteria, and the lactic acid bacteria fermentation It is preferable to extract through a freeze-drying process of freeze-drying the fermented burdock broth produced through the process.

The enzyme treatment process includes a grinding step of pulverizing the burdock after adding purified water, an enzyme addition step of adding a pectin-degrading enzyme and a fibrin-degrading enzyme to the pulverized burdock obtained through the grinding step, and the enzyme-added additive It comprises an enzyme reaction step of reacting, and an enzyme inactivation step of inactivating the enzyme after the completion of the enzyme reaction, wherein the lactic acid bacteria fermentation step is a first mixture by mixing purified water with the burdock enzyme treatment obtained through the enzyme treatment step. A purified water mixing step to form, an additive adding step of adding an additive to the first mixture in which the purified water is mixed to form a second mixture, and a lactic acid bacteria adding step of adding lactic acid bacteria to the second mixture to form a third mixture; , It is preferable to include a lactic acid bacteria culturing step of forming a fermentation broth while culturing the lactic acid bacteria in the third mixture, and a sterilization treatment step of heating and sterilizing the fermentation broth.

In addition, the companion animal feed for improving depression containing the burdock extract of the present invention as an active ingredient is selected from the group containing lactic acid bacteria, milk thistle, vitamin D, jujube, omega 3, minerals, noni, folic acid, vitamin B5, and vitamin B7. It is preferable to further include an additive consisting of one or a combination of two or more.

The present invention has the advantage of improving the depression of companion animals by burdock extract formed by extracting an active ingredient having an effect of improving depression from burdock.

1 is a flowchart showing the manufacturing process of a burdock extract contained in a companion animal nutritional supplement for improving depression comprising a burdock extract as an active ingredient according to the present invention;
Figure 2 is a flowchart showing the detailed process of the enzyme treatment process of Figure 1;
3 is a flowchart showing the detailed process of the lactic acid bacteria fermentation process of FIG. 1;
4 is a graph showing the cell viability of PC12 cells treated with burdock extract;
5 is a graph showing the cell viability of PC12 cells by corticosterone;
Figure 6 is a graph showing the inhibitory effect of burdock extract on the decrease in cell viability by corticosterone in PC12 cells;
7 is a graph showing the inhibitory effect of burdock extract on the increase in LDH production by corticorterone in PC12 cells;
8 is a graph showing the inhibitory effect of burdock extract on the reduction of the mitochondrial membrane potential by corticorterone in PC12 cells.

Hereinafter, a method for manufacturing a companion animal feed for improving depression including a burdock extract according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings. Since the present invention can have various changes and can have various forms, specific embodiments are illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to the specific disclosed form, it should be understood to include all modifications, equivalents and substitutes included in the spirit and scope of the present invention. In describing each figure, like reference numerals have been used for like elements. In the accompanying drawings, the dimensions of the structures are enlarged than the actual size for clarity of the present invention.

Terms such as first, second, etc. may be used to describe various elements, but the elements should not be limited by the terms. The above terms are used only for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, a first component may be referred to as a second component, and similarly, a second component may also be referred to as a first component.

The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly dictates otherwise. In the present application, terms such as “comprise” or “have” are intended to designate that a feature, number, step, operation, component, part, or combination thereof described in the specification exists, but one or more other features It is to be understood that it does not preclude the possibility of the presence or addition of numbers, steps, operations, components, parts, or combinations thereof.

Unless defined otherwise, all terms used herein, including technical and scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present application. does not

The companion animal feed for improving depression containing the burdock extract according to the present invention is formed by mixing rice bran powder, indigestible dextrin, burdock extract and additives.

Rice bran refers to the rice germ and rice bran that are peeled off during the milling process of brown rice, and milled to obtain rice bran powder.

The indigestible dextrin has been reported to have a lowering effect on visceral fat, blood sugar, triglyceride and cholesterol, and thus has an improvement effect on metabolic syndrome. In addition, it is known to facilitate bowel movements and to absorb minerals such as calcium, magnesium, iron and zinc, and is used in many foods.

When the rice bran powder is used as 100 parts by weight, 20 to 30 parts by weight of the indigestible dextrin and 1 to 3 parts by weight of the burdock extract are mixed to form a mixture.

The burdock extract is extracted by pulverizing the burdock root and fermenting it with enzyme treatment and lactic acid bacteria.

As shown in FIG. 1, the extraction method of the burdock extract of the present invention includes an enzyme treatment process of burdock, a lactic acid fermentation process of fermenting the enzyme-treated burdock with lactic acid bacteria, and a freeze-drying process of freeze-drying and powdering the fermented liquid material. includes

The enzymatic treatment process of burdock is a process of enzymatic treatment through pectin-degrading enzyme and fibrin-degrading enzyme to obtain optimal extraction efficiency from burdock.

Referring to FIG. 2, the enzyme treatment process includes a grinding step of adding purified water to burdock and pulverizing it, an enzyme addition step of adding pectin-degrading enzyme and fibrin-degrading enzyme to the ground burdock, and an enzyme-added additive under set environmental conditions. It includes an enzyme reaction step of reacting, and an enzyme inactivation step of inactivating the enzyme after completion of the enzyme reaction.

Each step is described in more detail. First, in the pulverizing step, purified water is added to the burdock and pulverized in a pulverizer to form a pulverized burdock. The purified water is preferably added in an amount of 50 to 100 parts by weight based on 100 parts by weight of burdock root.

When the amount of purified water is less than 50 parts by weight, the grinding of the burdock is not performed uniformly and the enzymatic reaction is not performed smoothly. And when the amount of purified water exceeds 100 parts by weight, the efficiency of the enzymatic reaction decreases because too much time is required for the enzymatic reaction.

The enzyme addition step is a step of adding an enzyme to the pulverized burdock root.

0.1 to 0.5 g of pectin-degrading enzyme and 0.2-0.5 g of fibrin-degrading enzyme per 1 kg of ground burdock.

In the enzymatic reaction step, the enzyme-added burdock pulverized material is stirred at a rate of 50 ppm or more while maintaining a temperature of 37 to 45° C. and an acidity of 4.5 to 6.5 to react the enzyme with the pulverized burdock. The enzyme reaction step is preferably maintained for 15 to 30 hours.

When the temperature at which the enzymatic reaction step is performed is less than 37° C., the enzyme does not function, and when the temperature exceeds 45° C., the enzyme is inactivated. And when the acidity is lower than pH 4.5 or higher than pH 6.5, the enzyme does not work.

If the enzymatic reaction time is also less than 15 hours, a sufficient enzyme reaction is not achieved, and if it exceeds 30 hours, the enzyme reaction is excessively performed, so that an appropriate extract cannot be obtained.

After the enzymatic reaction is completed, an enzyme inactivation step is performed to inactivate the enzyme to prevent further enzymatic reaction from proceeding. For enzyme inactivation, the enzyme reaction product on which the enzyme reaction is completed is heated to a temperature of 70 to 80° C., and the heating time is maintained for 1 to 2 hours.

The burdock enzyme-treated product obtained through the enzyme treatment process is fermented with lactic acid bacteria through the lactic acid bacteria fermentation process.

Referring to FIG. 3, the lactic acid bacteria fermentation process includes a purified water mixing step of mixing purified water with the burdock enzyme-treated product obtained through the enzyme treatment process to form a first mixture, and adding an additive to the first mixture mixed with purified water. An additive addition step of forming a second mixture, a lactic acid bacteria addition step of forming a third mixture by adding lactic acid bacteria to the second mixture, a lactic acid bacteria culturing step of forming a fermentation broth while culturing the lactic acid bacteria in the third mixture, and the fermentation broth It includes a sterilization treatment step of sterilizing by heating.

In the purified water mixing step, 30 to 50 parts by weight of purified water is added and mixed with respect to 100 parts by weight of the burdock enzyme-treated product obtained through the enzyme treatment process.

When the amount of purified water is added to less than 30 parts by weight, lactic acid bacteria cannot work evenly on the enzyme-treated product, so effective fermentation may not proceed, and if it exceeds 50 parts by weight, the fermentation time becomes long, which is inefficient.

In the additive addition step, an additive is added to the first mixture obtained through the purified water mixing step to form a second mixture. In the additive addition step, 1 to 3 parts by weight of lactose, 1 to 2 parts by weight of sucrose, and 1 to 2 parts by weight of the enzyme extract are added based on 100 parts by weight of the first mixture.

The lactose, sucrose, and enzyme extracts added in the additive addition step provide sugars necessary for culturing lactic acid bacteria to be described later, and if the amount of the additives is too small, it is difficult to properly culture the lactic acid bacteria, If it is exceeded, not only lactic acid bacteria but also general bacteria can grow, so there is a high probability of contamination of the fermented product.

The lactic acid bacteria addition step is a step of adding lactic acid bacteria to the second mixture obtained through the additive addition step. The lactic acid bacteria added in this step may be any one or a combination of two or more of Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus acidophilus.

The lactic acid bacteria is preferably added so that the volume ratio of the total volume of the second mixture is 0.2 to 0.5%.

The lactic acid bacteria culturing step is a step of culturing the lactic acid bacteria by allowing the third mixture obtained in the lactic acid bacteria addition step to stand for 48 to 60 hours at a temperature condition of 35 to 40 ℃.

The sterilization step is a step of sterilizing by heating the fermentation broth obtained in the lactic acid bacteria culture step to a temperature of 95 to 99 ℃, the heating time is preferably 2 to 3 hours.

Through this lactic acid fermentation process, a fermented broth that has been sterilized can be obtained.

The freeze-drying process is a process of drying the fermentation broth through a freeze dryer. Drying the fermentation broth through freeze-drying minimizes the loss of nutrients in the process of removing moisture.

In this way, powdered burdock extract can be obtained by subjecting the burdock to the enzyme treatment process, lactic acid bacteria fermentation process, and freeze-drying process.

A cell experiment was performed to confirm the depression improvement effect of the burdock extract extracted through the above extraction method.

First, cells for cell experiments are cultured as follows.

To culture PC 12 cells for the experiment, RPMI1640 medium containing 10% FBS and 1X antibiotic-antimycotic is used for PC 12 cells. All cells are cultured at 37°C and 5% CO2 conditions, and when they grow to about 70-80% in a culture flask, subculture using Trypsin-EDTA and use for the experiment.

[Experimental Example 1]

In order to examine the depression improvement effect of the burdock extract of the present invention, the change in cell viability by the burdock extract of the present invention and corticosterone was compared.

First, PC12 cells were cultured in 96 wells, and burdock extracts were treated at concentrations of 0, 10, 50, 100, 200, 300, 400, 500, 700, and 1000 μg/ml for 24 hours.

And after culturing PC12 cells in 96 well, corticosterone was treated at 30, 50, 100, 150, 200, 300, 500 μM concentration for 24 hours.

Cell viability was measured using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazoliu mbromide (MTT) reduction method. After dispensing the cells into 96-well plates and incubating for 12 hours, each sample is treated with the cells and incubated for 24 hours. Thereafter, the MTT solution (final concentration: 0.5 mg/mL) is added and the culture medium is further incubated at 37° C. for 4 hours to reduce the MTT, and the medium is removed so that the formazan does not fall off the medium. After dissolving 100 μL of DMSO to dissolve the formazan for 10 minutes, the absorbance is measured at 540 nm to calculate the cell viability.

4 is a graph of experimental results for confirming the effect of burdock extract on the viability of PC12 cells, and FIG. 5 is a graph of experimental results for confirming the effect of corticosterone on the viability of PC12 cells.

As can be seen from the graph, in the case of corticosterone, the cell viability rapidly decreases as the concentration increases, whereas in the burdock extract, there is no significant change in the cell viability.

[Experimental Example 2]

In addition, an experiment was further conducted to confirm the inhibitory effect of the burdock extract on apoptosis induced by corticosterone.

To check the PC12 cell protective effect of burdock extract from apoptosis induced by corticosterone, 200, 300, 400, μg/ml, and 200 μM of corticosterone were treated for 24 hours, and then MTT assay and LDH assay were used. to confirm.

As can be seen in the graph of FIG. 6 , as a result of confirming the change in cell viability through MTT assay, the cell viability of the corticosterone-treated group was reduced to 54%, but the burdock extract was treated at 200, 300, 400, μg/ml concentrations. In one group, the cell viability increased to 53, 73, and 87%.

[Experimental Example 3]

LDH (lactate dehydroenase) measurement

LDH, an enzyme present in the cytoplasm, increases the release of LDH present in the cell when the cell membrane is damaged. Therefore, the degree of damage to the cell membrane can be confirmed through LDH measurement.

For the LDH measurement experiment, cells were added to a 96-well plate at a concentration of 1×105 per well, incubated for 12 hours, then the sample was processed and cultured for 24 hours. Thereafter, LDH released into the medium was analyzed using an LDH assay kit.

As a result of confirming the change in LDH production through LDH assay, as can be seen in the graph of FIG. 7 , the corticosterone-treated group increased LDH production to 184%, but burdock extract was added at 200, 300, 400, μg/ml concentrations. In the treated group, LDH production was reduced to 162, 147, and 121%.

[Experimental Example 4]

Mitochondrial membrane potential measurement

When signaling related to apoptosis is induced through the mitochondrial pathway, calcium ions and apoptosis-related regulatory proteins present in the cytoplasm move to the mitochondria, causing mitochondrial fragmentation. Due to this, when depolarization of the mitochondrial membrane occurs, the permeability transition pore of the mitochondria is opened, and proteins acting on apoptosis are leaked out, resulting in nuclear aggregation and DNA fragmentation, and apoptosis occurs.

Mitochondrial membrane potential (MMP) is measured using JC-1 staining. After treatment, the cells are collected and washed once with PBS. To measure MMP, 600 μl of PBS diluted with JC-1 to a concentration of 100 nM is mixed and incubated in a dark place at 37°C for 30 minutes, and then absorbance is measured.

An experiment was conducted to confirm the inhibitory effect of the burdock extract on the reduction of MMP caused by corticosterone, and the inhibitory effect of the burdock extract on the MMP lowered by corticosterone was measured using JC-1.

As can be seen in the graph of Figure 8, the MMP of the group treated with corticosterone alone was reduced to 48%, but in the group treated with the burdock extract at 100, 200, 300, 400, μg/ml concentrations, it was 59, 61, and 74%. MMP was increased.

As a result of measuring LDH and mitochondrial membrane potential to confirm cell viability and inhibition of apoptosis on corticosterone by the burdock extract of the present invention, it can be confirmed that the burdock extract has an inhibitory effect on corticosterone, and brain related to depression The improvement effect of burdock extract can be expected for cells as well.

In addition, the companion animal feed for improving depression containing the burdock extract of the present invention can be formed by further mixing additives for improving the nutrition of companion animals in addition to the rice bran powder, indigestible dextrin and burdock extract.

The additive may consist of one or a combination of two or more selected from the group consisting of lactic acid bacteria, milk thistle, vitamin D, jujube, omega 3, minerals, noni, folic acid, vitamin B5, and vitamin B7.

In this embodiment, when lactic acid bacteria, milk thistle, vitamin D, jujube, omega 3, minerals, noni, folic acid, vitamin B5, and vitamin B7 are added as the additives, the amount of lactic acid bacteria is 1.8 to 2.2 with respect to 100 parts by weight of the rice bran powder. parts by weight, milk thistle 1-3 parts by weight, vitamin D 0.01 to 0.05 parts by weight, jujube 1-3 parts by weight, omega 3 0.01 to 0.05 parts by weight, minerals 0.01 to 0.05 parts by weight, noni 1.8 to 2.2 parts by weight, folic acid 0.01 to 0.05 parts by weight, and 0.01 to 0.05 parts by weight of vitamin B5 and vitamin B7 are mixed.

The rice bran powder, indigestible dextrin, burdock extract and additives are mixed according to the mixing ratio and then molded into a tablet shape of a predetermined size, or mixed with other feeds in a powder state and provided to companion animals.

The companion animal feed for improving depression containing the burdock extract according to the present invention described above contains the burdock extract extracted from burdock, which can be used as the natural antidepressant, and can be of great help in improving depression in companion animals.

The description of the presented embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the scope of the invention. Thus, the present invention is not intended to be limited to the embodiments presented herein, but is to be construed in the widest scope consistent with the principles and novel features presented herein.

Claims (5)

delete delete Based on 100 parts by weight of rice bran powder,
20 to 30 parts by weight of indigestible dextrin;
Including the step of mixing 1 to 3 parts by weight of the burdock extract,
The burdock extract is
an enzyme treatment process of pulverizing the burdock and enzymatically treating it with a pectin-degrading enzyme and a fibrin-degrading enzyme;
a lactic acid bacteria fermentation process of fermenting the burdock enzyme-treated product formed through the enzyme treatment process with lactic acid bacteria;
It characterized in that it is extracted through a freeze-drying process of freeze-drying the fermented burdock broth produced through the lactic acid bacteria fermentation process.
A method for manufacturing a pet food for improving depression containing burdock extract.
4. The method of claim 3,
The enzymatic treatment process includes a grinding step of adding purified water to the burdock and then grinding;
An enzyme addition step of adding a pectin-degrading enzyme and a fibrin-degrading enzyme to the ground burdock obtained through the grinding step;
An enzyme reaction step of reacting the additive to which the enzyme is added;
and an enzyme inactivation step of inactivating the enzyme after completion of the enzymatic reaction,
The lactic acid bacteria fermentation process includes a purified water mixing step of mixing purified water with the burdock enzyme-treated product obtained through the enzyme treatment process to form a first mixture;
An additive adding step of adding an additive to the first mixture in which the purified water is mixed to form a second mixture;
A lactic acid bacteria addition step of adding lactic acid bacteria to the second mixture to form a third mixture;
A lactic acid bacteria culture step of forming a fermentation broth while culturing the lactic acid bacteria in the third mixture, and
It characterized in that it comprises a sterilization treatment step of heating the fermentation broth to sterilize.
A method for manufacturing a pet food for improving depression containing burdock extract.
4. The method of claim 3,
Lactobacillus, milk thistle, vitamin D, jujube, omega 3, minerals, noni, folic acid, vitamin B5, vitamin B7, characterized by further mixing an additive consisting of one or a combination of two or more selected from the group consisting of
A method for manufacturing a pet food for improving depression containing burdock extract.

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