WO2022085847A1 - Antibacterial and antiviral composite natural extract composition containing wasabia japonica matsum fermented extract and preparation method therefor - Google Patents

Abstract

A method for preparing an antibacterial and antiviral composite natural extract composition containing a Wasabia japonica Matsum fermented extract comprises the steps of: preparing Wasabia japonica Matsum; producing an extract from the prepared Wasabia japonica Matsum through subcritical water extraction; fermentating the Wasabia japonica Matsum extract to generate a fermented extract; adding a natural extract to the fermented extract to form a natural fermented composite; and stabilizing the natural fermented composite through a microemulsion process.

Description

Complex natural extract composition for antibacterial and antiviral containing fermented horseradish extract and manufacturing method thereof

The present invention relates to a complex natural extract composition for antibacterial and antiviral containing a fermented wasabi extract, and specifically, it contains a fermented component of an active ingredient extracted from wasabi by a subcritical water extraction (SWE) process. It relates to a complex natural extract composition for antibacterial and antiviral containing a fermented horseradish extract and a method for manufacturing the same.

Wasabia japonica Matsum ( Wasabia japonica Matsum ) is a cruciferous perennial hard-rooted plant that grows wild in cold valleys and can be cultivated in cold areas. It has been found that horseradish has a strong antibacterial activity against pathogens such as food poisoning bacteria, pathogenic yeast, and mold, and is also known to have effects such as antioxidant activity, inhibitory activity of platelet aggregation, antimutagenic activity, or anticancer activity. Wasabi has its own pungent taste, sweetness, and aroma, so it can be used as a spice for foods such as sashimi, sushi, and noodles. The main component of the unique flavor of wasabi is ally isothiocyanate (AIT), which is a volatile component contained in wasabi, and this component exists in the form of sinigrin in the tissue. Such glycosides themselves do not have a pungent taste, but when the tissue is destroyed, the glycosides may be hydrolyzed by myrosinase present in the tissue to exhibit a pungent taste. Alkyl isothiocyanate (AIT) is distributed only in mustard radish, horseradish, white mustard, black mustard, rapeseed, radish, cabbage, Chinese cabbage, or similar cruciferous plants, and promotes appetite and digestion, promotes the synthesis of vitamin B1, and vitamins It is known to inhibit the oxidation of C, inhibit abnormal fermentation in the body after a meal, as an anthelmintic, antibacterial, antifungal, anticancer, and insecticide. Hot water extraction, organic solvent extraction, supercritical extraction or similar methods may be applied to extract an active ingredient such as AIT from horseradish. For example, Patent Publication No. 10-2005-0055644 or Patent Publication No. 10-2016-0044084 discloses a method of extracting an active ingredient from horseradish by distillation or an organic solvent. However, in the case of the method presented in the prior art or the hot water extraction method or the neutral salt extraction method corresponding to the known technology, impurities may be mixed, and a solvent that can become a toxic substance remains, causing problems with the quality of the product. In addition, since it goes through a multi-step process, the process is complicated and active ingredients may be lost, there is a possibility of foreign substances being mixed, and the purity of the extract is low. The hot water extraction method and the neutral salt extraction method also have the same disadvantages as described above. Therefore, there is a need to develop an extraction method capable of solving such a problem.

An object of the present invention is a composition comprising an extract obtained by subcritical extraction of horseradish, and while containing an alkyl isothiocyanate (AIT) component, horseradish having an antibacterial effect, an antifungal effect, a bactericidal effect, an antiviral effect and an antioxidant effect It is to provide a complex natural extract composition for antibacterial and antiviral containing a ferment extract and a method for preparing the same.

According to a suitable embodiment of the present invention, the method for preparing a complex natural extract composition for antibacterial and antiviral containing a fermented wasabi extract comprises the steps of preparing wasabi; preparing an extract from the prepared horseradish by a subcritical extraction process; fermenting the wasabi extract to produce a fermented extract; adding a natural extract to the fermented extract to form a natural fermentation complex; and stabilizing the natural fermentation complex by a micro-emulsification process.

According to another suitable embodiment of the present invention, the subcritical extraction is extracted under the conditions of a temperature of 130 to 200 ℃ and 15 to 30 atmospheres.

According to another suitable embodiment of the present invention, the natural extract is at least one component selected from the group consisting of broccoli extract, garlic extract, green tea extract, sesame leaf extract, goji berry extract, jujube extract and golden extract.

According to another suitable embodiment of the present invention, the fermentation step is a Lactobacillus plantarum culture medium and Saccharomyces cerevisiae by culturing for 48 to 96 hours at 25 to 35 ° C. includes steps.

According to another suitable embodiment of the present invention, the antibacterial and antiviral complex natural extract composition containing the fermented wasabi extract is a wasabi extract; and at least one component selected from the group consisting of broccoli extract, garlic extract, green tea extract, sesame leaf extract, goji berry extract, jujube extract and golden extract, and has antibacterial effect, antifungal effect, bactericidal effect, antiviral effect and antioxidant effect have

According to another suitable embodiment of the present invention, the extract is a fermented product by Lactobacillus plantarum or Saccharomyces Cerevisiae, characterized in that it contains a fermented extract of horseradish. do.

According to another suitable embodiment of the present invention, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, MRSA (methicillin-resistant Staphylococcus aureus), Bacillus subtilis It has antibacterial or bactericidal activity against Bacillus subtilis, Vibrio parahaemolyticus, Salmonella typhimurium, Listeria monocytogenes and Shigella flexneri, and Candida albicans (Candida) albicans), has antifungal activity against Aspergillus niger, and has antiviral activity against influenza virus.

According to another suitable embodiment of the present invention, the composite composition is a cosmetic, a skin cleanser, a hair cleanser, a vaginal cleanser, a bathroom cleanser, a mouthwash, a kitchen cleanser, a cosmetic preservative, a food preservative, a pharmaceutical preservative, a food additive and a feed additive. at least one composition selected from the group consisting of

The method for producing a composition comprising a complex natural extract for antibacterial and antiviral use containing a fermented wasabi extract according to the present invention is effective from wasabi by a subcritical water extraction (SWE) process and a microfluidizer. By extracting the components, the extraction efficiency is improved and the inflow of impurities is prevented. The composition according to the present invention has antibacterial, antifungal, antiviral and antioxidant effects, and thus can be applied to various uses, including food, cosmetics, or household products. Due to these effects, the composition according to the present invention can be used as a cosmetic, skin cleaner, hair cleaner, vaginal cleaner, bathroom cleaner, mouthwash, kitchen cleaner, cosmetic preservative, food preservative, pharmaceutical preservative, food additive or feed additive. .

1 shows an example of a method for preparing a complex natural extract composition for antibacterial and antiviral containing a fermented wasabi extract according to the present invention.

Figure 2 shows the results of the bacterial and fungal activity test by the composition according to the present invention.

Figure 3 shows the results of the fast puncture test for the composition according to the present invention.

Figure 4 shows the results of the persistence test for the composition according to the present invention.

Hereinafter, the present invention will be described in detail with reference to the embodiments shown in the accompanying drawings, but the embodiments are for a clear understanding of the present invention, and the present invention is not limited thereto. In the following description, components having the same reference numerals in different drawings have similar functions, so unless necessary for the understanding of the invention, the description will not be repeated and well-known components will be briefly described or omitted, but the present invention It should not be construed as being excluded from the embodiment of

1 shows an example of a method for preparing a complex natural extract composition for antibacterial and antiviral containing a fermented wasabi extract according to the present invention.

Referring to Figure 1, the method for preparing a complex natural extract composition for antibacterial and antiviral containing horseradish ferment extract includes the steps of preparing wasabi (P11); preparing an extract from the prepared horseradish by a subcritical extraction process (P12); fermenting the wasabi extract to produce a fermented extract (P13); Forming a natural fermentation complex by adding a natural extract to the fermented extract (P14); and stabilizing the natural fermentation complex by a micro-emulsification process (P15).

Horseradish ( Wasabia japonica Matsum ) is a cruciferous perennial mature plant, and can be a plant containing a component that produces singrin or alkyl isothiocyanate (AIT). Wasabi can be grown hydroponically or can be cultivated in the field, and horseradish grown for 1 to 2 years can be prepared (P11). The rhizome and leaf of wasabi can be isolated, and optionally the separated leaf can be prepared. The prepared rhizomes may be washed and dried so that the moisture content is 5 to 15 wt%. The dried rhizomes of horseradish may be pulverized, for example, pulverized to have an average diameter of 0.5 to 10 mm. An active ingredient can be extracted from the wasabi prepared in this way by a subcritical water extraction (SWE) (P12). A fluid that exists above temperature and pressure becomes a subcritical fluid, and the process of extracting active ingredients from various raw materials using the subcritical fluid as a solvent is a subcritical extraction process. Subcritical water can be made at a temperature of 200° C. and a pressure of 15 to 30. Since subcritical water has a high hydrolysis ability and can dissolve organic matter, it can be advantageously applied to extract an active ingredient from wasabi. The subcritical solvent may be water, ethanol, or methanol, or a mixed solution of water and alcohol may be used as the subcritical fluid in order to increase the decomposition ability of organic matter while facilitating the control of temperature and pressure. A mixed solvent in which ethanol has a weight ratio of water: methanol or ethanol = 100: 10 to 30 may be used as the subcritical solvent, and water, water and Ethanol or a mixed solvent of water and methanol may be prepared, and horseradish may be immersed in the solvent to adjust the temperature and pressure at a temperature of 130 to 200 ° C and a pressure of 15 to 30 atm. For example, it can proceed for 30 to 1000 seconds, and the extracted active ingredient can be separated by centrifugation or a similar method.When the separated ingredient is filtered and the wasabi extract is prepared (P12), natural additives can be mixed (P13).

The natural additive may be at least one component selected from the group consisting of broccoli extract, garlic extract, green tea extract, sesame leaf extract, goji berry extract, jujube extract and golden extract. Such natural additives may be extracted by various methods and may be extracted through a subcritical extraction process, a hot water extraction process, a supercritical extraction process, or a similar extraction process. The natural additive may be added in various amounts depending on the application use of the composition, for example, horseradish extract: natural additive = 10: 10 to 500 in a weight ratio. Depending on the type of natural additive to be added, the amount of different natural additives may be appropriately adjusted. For example, the green tea extract is added in an amount of golden extract: green tea extract = 100: 50 to 150 in weight ratio with respect to the golden extract. can be In addition, garlic extract may be added in a ratio of golden extract: green tea extract = 100: 10 to 30 by weight with respect to the golden extract. The type or amount of the natural additive may be appropriately adjusted depending on the use and is not limited to the examples presented. When these natural additives are added (P13), the complex can be fermented (P14). Fermentation is, for example, Lactobacillus plantarum, Saccharomyces Cerevisiae, Leuconostoc spp., Pichia spp., Latobacillus paracasei, Streptococcus thermophilus, Latobellius spp. Or it can be fermented by Sacharomyces spp. Specifically, it may be cultured for 48 to 96 hours in a liquid medium at a temperature of 25 to 35 °C, and the cultured fermented product may be filtered to obtain a fermented extract. When the fermented extract is obtained in this way (P14), it can be stabilized by the micro-emulsification process (P15). The micro-emulsification process may be made, for example, by a microfluidzer, specifically, by passing the microfluidizer 1 to 5 times under a pressure condition of 100 to 500 MPa to obtain an average diameter of 50 to 500 nm. Eggplants can be run to create a nanoemulsion. In order to make a nano-emulsion form by a microfluidizer, the natural fermentation complex may be made into a micro-emulsion form by a high speed mixer. Thereafter, it may be passed through a microfluidizer 1 to 5 times to have an average diameter of 50 to 500 nm, preferably 50 to 200 nm, and a particle dispersion index of 0.45 or less, preferably 0.35 or less. As such, at least one selected from the group consisting of cosmetics, skin cleaners, hair cleaners, vaginal cleaners, bathroom cleaners, mouthwashes, kitchen cleaners, cosmetic preservatives, food preservatives, pharmaceutical preservatives, food additives and feed additives made in the form of nano-emulsions. It may be added to the composition to exhibit antibacterial or antiviral properties. The complex natural extract composition can be prepared by various methods, and is not limited to the examples presented. Examples of the composition thus made will be described below.

Example

go. The growing period was about 15 months, and the rhizomes of wasabi having a weight of 120 to 150 g were prepared by pulverizing to a size of 0.8 to 1.2 mm in average diameter.

me. A subcritical solvent in which water and ethanol are 100: 20 in a weight ratio was prepared, and crushed horseradish was subjected to subcritical extraction at a temperature of 150 to 180° C. and a pressure of 20 atm for 120 seconds.

all. The extract was centrifuged to separate the active ingredient, and it was fermented in a Lactobacillus plantarum liquid medium at a temperature of 30° C. for 60 hours. Green tea extract and garlic extract were mixed in the ferment extract so that the ferment extract: green tea and garlic extract = 20: 100 by weight relative to the ferment extract, respectively.

la. The mixed natural extract is put into a high-speed mixer to make a microemulsion, and then passed through a microfluidizer three times under a pressure condition of 120 to 150 MPa to have an average diameter of 150 to 180 nm and a particle dispersion index of 0.40. It was made as a nano-emulsion as much as possible.

Efficacy tests were performed on the composite natural extract composition made in this way.

efficacy test

1. Bacterial and fungal activity test

Efficacy tests for food additives of the prepared complex natural extract composition were made, and the test results are presented in Tables 1 and 2 below, and the (Microdilution method) conditions of the MIC are as follows.

- NB (Nutrient broth, bacteria), PDB (Potato Dextrose broth, mold)

- Bacteria: 5.0x10 ⁴ CFU/mL, Mold: 6.0x10 ⁴ CFU/mL

- Bacteria: 35℃/48h, mold: 25℃/48h culture

- Adjusted to pH 6.0

strain	extract composition	Sodium Benzonate	Grapefruit Seed Extract
S. aureus	0.156	0.078	0.156
E. coli	0.313	0.156	0.156
P. aeruginosa	0.626	0.156	0.313
C. albicans	0.156	0.078	0.313
A.niger	0.156	0.039	0.313

<Antibacterial and fungal activity test>2. As an antibacterial test, a 30-second fast-acting test was performed, and the test results are presented in FIG. 2 .

As can be seen from the test results, it can be seen that at a concentration of 1.0 wt% to 2.0 wt% of the extract composition, 99.99% or more of bacteria are killed.

3. A persistence test was performed, and the test results for S. aureus and E. coli were presented in Tables 2 and 3 and (A) and (B) of FIG. 4, respectively. For the persistence test, each sample was injected into a paper disc, and after 0 / 30 / 60 minutes of stationary storage, microbial smearing was performed. In the case of fermentation alcohol, microbial control activity was observed immediately after sample injection, whereas microbial control effect was not observed during microbial smearing after standing for 30 / 60 minutes. .

Sample	Inhibition Zone(㎜)
	0 min	30 min	1 hour
Fermented alcohol 65 %	30	-	-
Composite composition 0.5 wt%	21	21	20

<Persistence test of S. aureus>

Sample	Inhibition Zone(㎜)
	0 min	30 min	1 hour
Fermented alcohol 65 %	27	-	-
Composite composition 0.5 wt%	24	24	22

<Results of persistence test of E. coli> As can be seen from the above test results, the complex natural extract composition for antibacterial and antiviral containing the fermented horseradish extract according to the present invention is a cosmetic, skin cleanser, hair cleanser, and vaginal cleanser. , bath cleaners, mouthwashes, kitchen cleaners, cosmetic preservatives, food preservatives, pharmaceutical preservatives, food additives and feed additives may be added to at least one composition selected from the group consisting of. For example, it may be added in an amount of 0.5 to 4.0 wt% of the total preservative for long-term storage of kimchi. The composition according to the present invention is Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, MRSA (methicillin-resistant Staphylococcus aureus), Bacillus subtilis (Bacillus subtilis) , Vibrio parahaemolyticus, Salmonella typhimurium, Listeria monocytogenes and Shigella flexneri have antibacterial or bactericidal activity against, Candida albicans, Aspergillus It has been shown to have antifungal activity against Aspergillus niger, and antiviral activity against influenza virus. The manufacturing method according to the present invention has a high extraction efficiency by a subcritical extraction process, and for example, 0.40 to 0.80 mg/g of alkyl isothiocyanate (AIT) can be extracted while using a macrofluidizer. It was shown that it was made in the form of a nano-emulsion and had excellent stability.

Although the present invention has been described in detail with reference to the presented embodiment, those skilled in the art will be able to make various modifications and variations of the invention without departing from the technical spirit of the present invention with reference to the presented embodiment. . The present invention is not limited by such variations and modifications, but only by the claims appended hereto.

The composition according to the present invention can be used as an additive to various cosmetic articles including cosmetics.

Claims (8)

1. preparing horseradish;

   preparing an extract from the prepared horseradish by a subcritical extraction process;

   fermenting the wasabi extract to produce a fermented extract;

   adding a natural extract to the fermented extract to form a natural fermentation complex; and

   A method for producing a complex natural extract composition for antibacterial and antiviral containing a horseradish ferment extract, comprising the step of stabilizing the natural fermentation complex by a micro-emulsification process.
2. The method according to claim 1, wherein the subcritical extraction is a method for producing a complex natural extract composition for antibacterial and antiviral containing a fermented wasabi extract, characterized in that the extraction is performed at a temperature of 130 to 200 ℃ and a pressure of 15 to 30 atmospheres.
3. The method according to claim 1, wherein the natural extract is at least one component selected from the group consisting of broccoli extract, garlic extract, green tea extract, sesame leaf extract, goji berry extract, jujube extract, and golden extract. Method for producing a complex natural extract composition for antibacterial and antiviral.
4. The horseradish according to claim 1, wherein the fermentation step includes filtering the culture solution obtained by culturing a Lactobacillus plantarum culture solution and Saccharomyces cerevisiae in a liquid medium at 25 to 35° C. for 48 to 96 hours. A method for producing a complex natural extract composition for antibacterial and antiviral containing a ferment extract.
5. horseradish extract; and at least one component selected from the group consisting of broccoli extract, garlic extract, green tea extract, sesame leaf extract, goji berry extract, jujube extract and golden extract, and has antibacterial effect, antifungal effect, bactericidal effect, antiviral effect and antioxidant effect A complex natural extract composition for antibacterial and antiviral containing eggplant wasabi fermented extract.
6. The antibacterial and antiviral use of claim 5, wherein the extract is a fermented product by Lactobacillus plantarum or Saccharomyces Cerevisiae. Complex natural extract composition.
7. The method according to claim 5, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, MRSA (methicillin-resistant Staphylococcus aureus), Bacillus subtilis (Bacillus subtilis) It has antibacterial or bactericidal activity against Vibrio parahaemolyticus, Salmonella typhimurium, Listeria monocytogenes and Shigella flexneri, and Candida albicans, Aspergillus (Aspergillus niger) has antifungal activity, and antibacterial and antiviral complex natural extract composition containing wasabi fermented extract, characterized in that it has antiviral activity against influenza virus.
8. The method according to claim 5, wherein the composite composition is at least one selected from the group consisting of cosmetics, skin cleaners, hair cleaners, vaginal cleaners, bathroom cleaners, mouthwashes, kitchen cleaners, cosmetic preservatives, food preservatives, pharmaceutical preservatives, food additives and feed additives. Antibacterial and antiviral complex natural extract composition containing fermented horseradish extract, characterized in that it is added to the composition.

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