KR101465696B1 - Antibacterial and insect resistance composition - Google Patents

Abstract

The present invention relates to an antibacterial and pest-control composition comprising a fermented extract extracted from a ferment obtained by fermenting, by using lactobacillus, a mixture in which Sophora Flavescens Ait., Meliae Fructus, Meliae Cortex and Syzygii Flos are mixed in a ratio of 1:1:1:2. The present invention is characterized by including, as an active ingredient, a fermented extract obtained by extracting a ferment obtained by: inoculating a mixture including Sophora Flavescens Ait., Meliae Fructus, Meliae Cortex and Syzygii Flos with one or more fermenting microorganisms selected from the groups consisting of lactobacillus; and fermenting the resultant for 3 to 7 days at a temperature range from 20 to 37°C in a pH maintained in 4.0 to 5.0. The present invention exhibits an excellent antibacterial activity on bacteria, fungi, and yeasts, and has a pest-control function.

Description

Antibacterial and insect resistance composition < RTI ID = 0.0 >

The present invention relates to an antibacterial and insecticidal composition, and more particularly, to an antibacterial and insecticidal composition comprising a fermented extract obtained by fermenting a fermented product obtained by fermenting a mixture of ginseng, celadon, loess, and clove with lactic acid bacteria .

To date, many studies on natural antimicrobial substances have reported natural antimicrobial substances from foods such as spices, milk and fish, essential oils and herbal medicines. Alkaloids, flavonoids, phytoalexins, Peptides have also been reported.

Antimicrobial properties of known natural preservatives such as organic acids, fatty acids, lower fatty acids, lysozyme, lactofine, and bacteriocin have also been reported.

One of the natural preservatives that has been put to practical use is the DF-100, made from Grandfruit Seed Extract (GFSE) developed by Chemi-research, USA. However, most of them are not commercialized due to physical and economical problems such as marginal adult odor, color, stability, narrow antibacterial spectrum, problem of form, and use amount as natural substances, and therefore research on natural antibacterial agents is required .

In addition, studies for developing antimicrobial agents using natural materials have been carried out and are still underway.

Korean Patent Laid-Open Publication No. 20100003046 discloses an antimicrobial composition comprising a sun-dried salt and a coconut tree extract.

Korean Patent Laid-Open Publication No. 20130114055 discloses an antimicrobial composition comprising a light extract of a fig tree extract or a fraction thereof.

In addition, studies for developing insect repellents using natural materials have been carried out and are underway.

Korean Patent Laid-Open Publication No. 20120021475 discloses an insecticidal or insecticidal composition for preserving cultural properties containing an extract or fraction of oak wood vinegar as an active ingredient.

Korean Patent Laid-Open Publication No. 20120021507 discloses an insecticidal and insecticidal composition for preserving a cultural heritage containing an Aspicidae extract or a fraction as an active ingredient.

However, there is a demand for the development of a new concept composition that can perform both functions safely and effectively using antimicrobial extracts derived from natural materials, especially natural materials, and antibacterial and insecticidal action together.

One object of the present invention is to provide a multifunctional preservative which can be used as a preservative in order to develop a multifunctional preservative from a natural substance capable of supplementing irritation which is a disadvantage of a chemical preservative widely used as a preservative. The present invention provides a method for producing a natural antimicrobial insecticidal material exhibiting a very excellent antimicrobial activity through fermentation.

It is another object of the present invention to provide a fermented extract prepared by fermenting a fermented natural product with the fermented extract and an antibacterial insecticidal composition containing the fermented extract as an active ingredient, A natural fermented product having an excellent antibacterial and insecticidal effect through fermentation, and an antibacterial and insecticidal material containing the natural fermented product.

The antimicrobial and insecticidal composition according to the present invention is prepared by inoculating lactic acid bacteria into a mixture containing ginseng, cinnabar, sesame, and clove, fermenting the mixture for 3 to 7 days at a pH within a range of from 4.0 to 5.0 within a temperature range of from 20 to 37 캜 And the fermented extract obtained by extracting the obtained fermented product as an active ingredient.

The mixture containing the ginseng, celadon, loess, and clove contains 0.01 to 35% by weight of ginseng, 0.01 to 35% by weight of cinnabar, 0.01 to 35% by weight of spinach, and 0.01 to 70% 1: 1: 1: 2 ratio.

The lactic acid bacteria may be Leuconostoc mesenteroides (KCCM 35471).

The lactic acid bacterium may be in addition to the Leuconostoc mesenteroides (KCCM 35471), such as Enterococcus faecium , Enterococcus faecalis , Lactobacillus plantarum , Lactobacillus plantarum , Lactobacillus fermentum , Lactobacillus lactis , Lactobacillus rhamnosus , Lactobacillus sakei , Lactobacillus brevis , Lactobacillus graminis , Lactobacillus spp ., Lactobacillus spp ., Lactobacillus spp . Leuconostoc citreum , Weissella hellenica , Lactococcus garvieae , Pediococcus pentosaceus , Pediococcus cedilactisis ( Pseudomonas spp. ), Pediococcus acidilactici), Bifidobacterium No Marlies (Bifidobacterium animalis), bifidobacteria Te may be Solarium Infante tooth (Bifidobacterium infantis) and those of the lactic acid bacteria is selected from the group consisting of one or more mixture.

The fermented extract obtained by extracting the fermented product can be obtained by mixing the fermented product with an extractant selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane, (40 kHz).

The antimicrobial and antidandruff composition may have antibacterial activity against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi.

The antimicrobial and antidandruff composition may have insecticidal action against house dust mites and large leg dust mites.

The antimicrobial and antifungal composition may be formulated into nanocapsules, microcapsules, and nanosomes.

Also provided are paints, cosmetics or foods comprising said antimicrobial and antifungal composition.

In addition, the present invention provides a method for preparing a ginseng, a cinnabar, a spirulina, and an oriental fermentation extract, comprising the steps of: (1) inoculating the ginseng, cinnabar, And (2) fermenting the mixture inoculated with the lactic acid bacteria.

According to the present invention, when researching not only the antimicrobial activity but also the insecticidal activity against natural plants, the antimicrobial activity against fungi including bacteria, fungi and yeast when fermented with proper combination of Gosam, Tianjin, And it has an effect of providing a natural antimicrobial and insecticidal composition having a multifunctional antiseptic activity which is excellent in the insecticidal effect and has a side effect of the conventional chemical preservative, in particular, a very low possibility of human stimulation compared with a chemical preservative.

Hereinafter, the present invention will be described in detail with reference to specific examples.

The present invention relates to a process for producing a fermented product of fermented ginseng, celadon, spirulina, and oriental fermented product using a lactic acid bacterium, and using this fermented product as a multifunctional preservative in paints, food, cosmetics, To provide multifunctional natural materials.

The ginseng, cinnabar, spinach, and oriental fermented product according to the present invention are characterized by being obtained by fermenting the mixture with lactic acid bacteria.

The antimicrobial and insecticidal composition according to the present invention is prepared by inoculating lactic acid bacteria into a mixture containing ginseng, cinnabar, sesame, and clove, fermenting the mixture for 3 to 7 days at a pH within a range of from 4.0 to 5.0 within a temperature range of from 20 to 37 캜 And a fermentation extract obtained by extracting the obtained fermented product as an active ingredient.

The raw materials used for the production of the fermented extract are raw materials derived from natural sources, especially plants.

Sophora flavescens is a perennial herbaceous plant belonging to the leguminosae family . It has a strong bitter taste and is said to have the same effect as ginseng. The roots and stems of Gosam have long been used as traditional medicines in Korea, China and Japan. Their pharmacological properties are herbal medicines used for diseases such as fever, diuretic, gastroenteritis, enteritis, skin eczema and eczema. Most of the useful components of gosam are (+) - matrine ((+) - matrine), (+) - matrine N-oxide, -) - allomatrine, (+) - isomatrine, (+) - sophoranol, (+) - sophoranol N-oxide N-oxide, (-) - sophoramine, (-) - N-methylcytosine, (-) - anagyrine ), Alkaloid compounds such as (-) - baptifoline and daidzein, genistein, formononetin, etc. known as phytoestrogen, and the like Of isoflavones. ≪ / RTI >

The above-mentioned tianjin is mature fruit of late autumn matured of Melilla azedarach L. var. Japonica Makino ( Meliaceae ), and it is good that it is large and crumbly, the husk is golden color, and the flesh is yellowish white. Appearance is spherical to ovate with diameter of 10-15mm, outer surface is yellowish brown, glossy and wrinkled. The cut side is pale yellow, divided into 4 to 5 chambers, with one seed in it. The major components are the limonoids toosendanin and the alkaloid azaridine and the other components are resin, tannin and benzoic acid. The main pharmacological action is used for antifungal action and antimicrobial action.

Meliae Cortex is the bark of Meliaceae, its origin is China, and it is subtropical plant native to southern provinces in Korea. It is known that the major components of goryeonggi are alkaloid components such as Azadirachtin, Margosin and Nimbin. Alcoholic extract of goryeonggi is known to have antimicrobial action against Gram-negative bacteria and insecticidal action against intestinal parasites . The height of the tree is 15 to 20m and it grows in the northern part of Korea. The curved tubular or tubular flesh is a plant with a length of 15 to 20 cm, a width of 3 to 10 cm, and a thickness of 3 to 5 mm. Outer side is grayish brown, with a long torn pattern and horizontal shrubs and reddish brown when peeled off the cork layer. The folded surface is yellowish white and fibrous, and the vagina is multi-tiered but easy to break. In general, the flavor of this bloom is written and there is a peculiar smell, and it is also called a common tree. The bitter taste of goryeon bloom is mainly alkaloid material and it has been used for many years as a medicinal product for various purposes in oriental medicine because it contains resin such as margosine and resin.

The clove refers to a dried clustering of clusters of blue-green clusters belonging to cloves. Clove (Syringa velutina) is a deciduous shrub belonging to the clove (Myrtaceae). There are 16 to 19% essential oils in the peaks. Essential oils include 78-95% of eugenol, 3% of acetyleneguanol, alpha and beta-cariophenyl, and the like. Flower peaks are oriental tendon placebo, old-fashioned medicine. In consent therapy, when you vomit or diarrhea, you will use it when you feel stomachache or stomachache. In addition, cloves are used in the food processing sector, while essential oils and eugenol are used as dental preservatives and topical paralytics. As a study using conventional cloves, Korean Patent Laid-Open Publication No. 1999-0055304 discloses a 5α-reductase inhibiting composition containing clove extract, Korean Patent Registration No. 017394 discloses a perfume using clove extract, . However, the above-mentioned known methods have not been reported on insecticidal and antibacterial activity by using clove and clove extract as fragrances or as a 5α-reductase inhibiting active composition for preventing dandruff and for treating acne, Furthermore, there is no description about the development of the antimicrobial insecticidal compositions through the fermentation of the above-mentioned ginseng, cinnabar, and oriental clove extracts.

In the present invention, lactic acid bacteria are inoculated into a mixture containing the above-mentioned ginseng, celadon, loess, and clove, and the fermented product obtained by fermenting the lactic acid bacterium while maintaining the pH within 4.0 to 5.0 within a temperature range of 20 to 37 캜 for 3 to 7 days And extracting the extracted data.

The mixture containing the ginseng, celadon, loess, and clove contains 0.01 to 35% by weight of ginseng, 0.01 to 35% by weight of cinnabar, 0.01 to 35% by weight of spinach, and 0.01 to 70% 1: 1: 2 ratio. In particular, it exhibits excellent antimicrobial and insecticidal activity due to mutual cooperative action between extracts through fermentation, and thus can be effectively used as a multifunctional natural preservative.

Preferably, each plant (otherwise referred to as a " raw material ") constituting the mixture comprising the above-mentioned ginseng, cinnabar, clavulanum and clove is subjected to a treatment such as removal of surface contaminants, washing, freeze- It will be understood by those skilled in the art that the present invention is not limited thereto and may be suitably processed for fermentation by pulverization in raw material state and mixing with water without performing lyophilization .

Preferably, the mixture containing the ginseng, cinnabar, spinach, and clove may further include a step of performing fermentation and then extracting the mixture.

The term "fermented extract" as used herein means a fermented extract obtained by fermenting a mixture of ginseng, cinnabar, persimmon, and clove, preferably fermenting lactic acid bacteria.

The lactic acid bacterium for fermentation may be Leuconostoc mesenteroides (KCCM 35471), preferably Enterococcus faecium in addition to Leuconostoc mesenteroides (KCCM 35471) There is provided a method for producing a recombinant Lactobacillus species selected from the group consisting of Enterococcus faecalis , Lactobacillus plantarum , Lactobacillus fermentum , Lactobacillus lactis , Lactobacillus rhamnosus , Lactobacillus sakei , Lactobacillus brevis , Lactobacillus graminis , Leuconostoc citreum , Weissella helenica, Lactococcus garvieae , Lactobacillus spp. ), Pediococcus pentosaceus ( Pediococcus pentosaceus ), Pediococcus A bacterium selected from the group consisting of Pediococcus acidilactici , Bifidobacterium animalis , Bifidobacterium infantis , and mixtures of at least one of the foregoing.

The microorganisms to be used in the present invention are known microorganisms and are available from the BRC, Belgian Co-ordinated Collections of Microorganisms (BCCM) or the American Type Culture Collection (ATCC) of the Korea Research Institute of Bioscience and Biotechnology.

Fermentation refers to a process in which a microorganism uses its own enzyme to decompose or synthesize an organic material. Representative fermenting microorganisms include Bacillus subtilis, lactic acid bacteria, aspergillus, yeast, mold, and acetic acid bacteria.

The fermented extract of the fermented product is fermented and fermented without using any other solvent, and the sugar content of the natural material is changed to alcohol and the flavonoid, vitamin, alkaloid, catechin, etc. contained in the natural material are effective The extraction efficiency for the component can be increased. In addition, natural materials, which can sometimes be irritating or poisonous, can be used as a method of stabilizing natural materials through fermentation extraction, reducing toxicity or converting them into stable derivatives. In addition, the fermentation process of lactic acid bacteria and yeast can maximize the efficacy of natural materials and maximize the antibacterial and insecticidal effect in the process of conversion to a low molecular substance which is easily absorbed, in an unstable or inactive form, have.

The fermentation in the present invention can be carried out by inoculating one or more fermenting microorganisms and fermenting the fermented microorganism at a pH of 4.0 to 5.0 within a temperature range of 20 to 37 DEG C for 3 to 7 days, Fermentation conditions. Depending on the type of lactic acid bacteria used, fermentation may be possible even when the fermentation conditions are out of the above range of time, temperature and pH. Therefore, the present invention is not limited to the fermentation conditions described above. It can be understood.

The antimicrobial and insecticidal composition according to the present invention may contain other components which can give synergy to the main effect without impairing the intended main effect of the present invention, in addition to the above extract.

As the extraction method for extracting the fermented product, a conventional extraction method can be used. Examples of the extraction method include extraction with an extraction solvent, supercritical fluid extraction, ultrasonic extraction, and hot water extraction.

That is, the fermented extract obtained by extracting the fermented product can be obtained by extracting the fermented product from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures of two or more thereof It may be an ultrasonic wave (40 kHz) extracted as an extraction solvent.

Alternatively, according to the present invention, it may be a pre-fermentation and a post-extraction process consisting of extracting a fermented product by fermenting the raw material again to obtain a fermented extract. Alternatively, the raw material may be pre-extracted and post-fermented. That is, in order to improve the yield and physiological activity in the fermentation step, the mixture of the raw materials is first extracted by adding a solvent of 10 to 20 times the amount of the sample at a temperature of 40 to 100 ° C, , Followed by filtration under reduced pressure and concentration-freeze-drying to prepare an extract.

The fermented extract according to the present invention may be provided in the form of a solution or a powder. Alternatively, the fermented extract may be formulated into a formulation such as a nanocapsule, a microcapsule, a nanosome, etc., The amount of the antibacterial and insect repellent composition according to the present invention can be adjusted. As described above, a composition having an antibacterial and insect repellent effect can be provided in paints, cosmetics, foods and the like.

The antimicrobial and antifungal composition has antibacterial activity against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi.

The antimicrobial and antifungal compositions also have insecticidal action against house dust mites and large leg dust mites.

Cosmetic or food products comprising said antimicrobial and antifungal composition are also provided according to the present invention.

Although the ginseng, cinnabar, sesame, and clove fermentation extracts of the present invention are produced from natural products, they have high antibacterial activity and high antibacterial activity against resistant bacteria and fungi without causing resistance, It is a natural material with antioxidant activity as well. Therefore, it can be widely used as a multifunctional natural preservative in paints, foods, cosmetics and the like.

As described above, the antimicrobial and insecticidal composition according to the present invention contains an appropriate ratio of ginseng, cinnabar, spearmint, clove extract to exhibit a superior antimicrobial activity to that of a single extract and has a broad antibacterial spectrum. It is a biocompatible antimicrobial and insecticidal composition having excellent antiseptic ability and low possibility of inducing skin irritation without adding a chemical preservative.

A method for producing an antibacterial and insect repellent composition according to the present invention comprises the steps of: (1) inoculating a ginseng, a cinnabar, And (2) fermenting the mixture inoculated with the lactic acid bacteria.

The ratio of the blend weight to the blend ratio is within a range of from 0.01 to 35% by weight, from 0.01 to 35% by weight of cinnabar, from 0.01 to 35% by weight and from 0.01 to 70% by weight of cinnabar, 1: 2 ratio.

The inoculation and fermentation step is inoculated with the lactic acid bacteria (pre-culture solution) cultured in the fermentation tank containing the culture solution, and the lactic acid bacteria are inoculated to the mixture of the above-mentioned ginseng, cinnabar, sesame seeds and clove in 5 to 15 wt%.

The lactic acid bacteria inoculated in the inoculation step is Leuconostoc mesenteroides (KCCM35471), and preferably the lactic acid bacteria is Enterococcus faecium (KCCM35471) in addition to Leuconostoc mesenteroides (KCCM35471) ), Enterococcus faecalis , Lactobacillus plantarum , Lactobacillus fermentum , Lactobacillus lactis , Lactobacillus rhamnosus , Lactobacillus spp ., Lactobacillus spp ., Lactobacillus spp . Lactobacillus sakei , Lactobacillus brevis , Lactobacillus graminis , Leuconostoc citreum , Weissella helenica , Lactococcus garbiera ( Lactobacillus spp. Lactococcus garvieae , Pediococcus pentosaceu s ), Pediococcus acidilactici , Bifidobacterium animalis , Bifidobacterium infantis , and mixtures of at least one of the foregoing. have.

The fermentation process is characterized by being one or two or more selected from the group consisting of mixed strains thereof. The culture broth of lactic acid bacteria was adjusted to pH 4.0 to 5.0 and cultured at a temperature of 20 to 37 ° C for 3 to 7 days. The antimicrobial and insecticidal composition according to the present invention may contain other components which can give synergy to the main effect without impairing the intended main effect of the present invention, in addition to the above-mentioned extract. The mixed fermentation extract may be present in the composition in an amount of 0.001 to 80% by weight based on the freeze-dried weight of the whole composition.

The fermentation step is a step of fermenting the mixture inoculated with the lactic acid bacteria and a step of adding the fermented material to the fermented material in the range of 3 to 10 Deg.] C for 1 to 3 days.

The mixed fermented product according to the present invention can be processed in the form of a solution or a powder in the form of nanocapsules, microcapsules, nanosomes and the like using a known technique in the art, The amount of the solution or powder in the composition according to the present invention. As described above, antimicrobial and insect repellent effects are provided to paints, cosmetics, foods and the like which are processed into a composition.

Before the inoculation step, a pre-treatment step of rapidly cooling and lyophilizing the ginseng, cinnabar, loess, and cloves may be further performed. Such quenching and freeze-drying can be easily performed by those skilled in the art.

A culture medium for preparing a culture medium capable of culturing the lactic acid bacteria by pulverizing the dried ginseng, cinnabar, persimmon and cloves obtained in the pretreatment step between the pretreatment step and the inoculation step and mixing the resultant with water is further performed . At this time, the ginseng, cinnabar, spinach, clove can be mixed with 10 to 20 times water based on the weight of the dried powder to form a culture solution.

The fermentation step may be followed by an aging step of aging the fermentation broth obtained in the fermentation step. The aging step may be performed by allowing the fermentation broth to stand at room temperature in a sealed container in which air or oxygen is blocked. Preferably, the fermentation broth can be performed by storing the fermentation broth at 3 to 10 ° C for 1 to 3 days have.

A post-treatment step may be further performed in which the fermentation liquid obtained after the fermentation step or the aging step is concentrated under reduced pressure and lyophilized to a water content of 10% to obtain a lyophilized fermentation product. It will be appreciated that the post-treatment steps described above can also be easily performed by those skilled in the art.

The antimicrobial and insect repellent composition according to the present invention contains an amount of 0.001 to 80% by weight of a lyophilized fermented product obtained by lyophilization as an active ingredient based on the total weight.

The method for producing the antimicrobial and insect repellent composition according to the present invention preferably comprises the steps of: (1) washing a raw material of ginseng, cinnabar, sesame, clove, quenching and freeze-drying; (2) a culture liquid preparation step in which the pulverized material of the dried raw material is mixed with water to culture the lactic acid bacteria; (3) fermenting lactic acid bacteria by inoculation alone; And (4) aging the fermentation broth, or a concentrate obtained by partially or wholly decompressing and lyophilizing the fermentation broth obtained from the fermentation broth or the fermentation broth, or a chemical substance exhibiting the main effect contained in the fermentation broth or concentrate Including itself.

The mixture was separated from roots and stem, washed with water, stored at -80 ° C for rapid cryopreservation, lyophilized and pulverized to prepare a fine powder.

The mixture was lyophilized and pulverized, and the powdery sample was mixed with water at a stable temperature. The lactic acid bacteria were inoculated alone, and the fermentation broth was obtained through a fermentation process. After the fermentation, the fermentation broth was concentrated under reduced pressure and freeze- And may include a low-temperature aging process or a high-temperature aging process.

Hereinafter, preferred embodiments and comparative examples of the present invention will be described.

The following examples are intended to illustrate the invention and should not be construed as limiting the scope of the invention.

Example 1: Preparation of mixed fermented extract

The raw materials such as ginseng, cinnabar, sesame seeds and cloves were removed and the foreign materials attached to the outside were removed, and then the whole was washed. The roots and the stem were separately prepared and washed once more. Dried at -80 ° C, lyophilized, mixed at a blending ratio of 1: 1: 1: 2, and pulverized to prepare a powder.

10 times as much as the dry weight of the root and stem powder was added and mixed thoroughly. Leuconostoc mesenteroides (KCCM35471) (pre-culture solution; optimized for culture to inoculate the lactic acid bacterium into the mixed extract as a pre-cultured lactic acid bacterium Lactic acid bacteria was cultured for 3 days at a temperature of 25 캜 and a shaking rate of 120 RPM to prepare a lactic acid bacterium.) Was inoculated at 10% by weight. Thereafter, liquid fermentation was carried out for 5 days under optimum conditions (temperature 26 ° C). The fermentation was terminated by heating at 80 ° C for 10 minutes to induce loss of function of the lactic acid bacteria. After evaporating the water content of the culture obtained in the above section, the dried fermented extract was extracted with 70% ethanol. The extracted fermented mixed extract was placed in an ultrasonic extraction apparatus and sonicated (40 kHz) was extracted at an extraction temperature of 60 ° C for 60 minutes. The fermented material was aged at a low temperature of 4 캜 for 2 days for aging. The precipitate was removed using a centrifuge, the supernatant was collected, or the filtered supernatant was concentrated under reduced pressure to a water content of 10% or less at 60 ° C or lower and then freeze-dried.

Comparative Example 1: Preparation of mixed general extracts

Mixed common extracts used as control were obtained through the same ultrasonic extraction process without fermentation process. That is, 70% ethanol of 10 times the dry weight was added, mixed well, and sonicated (40 kHz) was extracted at an extraction temperature of 60 ° C for 60 minutes. Each extract was air-cooled and then filtered with a filter paper having a permeation size of 8 mu m. The filtered extract was concentrated under reduced pressure at 60 ° C or lower and completely dried using a vacuum dryer to obtain a mixed general extract.

Experimental Example 1: Antimicrobial activity test

The Kirby-Bauer method was used to investigate the antimicrobial activity spectrum of the extracts. Pseudomonas aeruginosa by positive bacteria are used in strain Gram (Pseudomonas aeruginosa: ATCC 9027), Staphylococcus aureus (Staphylococcus aureus: ATCC 6538P), E. coli gram-negative bacteria (Escherichia coli: ATCC 8739), yeast is Candida yeast (Candida albicans: ATCC 10231 ) And Aspergillus niger (ATCC 22343) as a fungus were purchased from the BRC at Korea Research Institute of Bioscience and Biotechnology.

The bacterium was cultured in a liquid culture medium (Letheen broth) and inoculated with the culture medium for 24 hours in a shaking incubator at 37 ° C. YM broth was used as a yeast strain, and the bacteria were inoculated on a culture medium and cultured for 2 days in a 25 ° C shaking incubator.

For the antibacterial test, bacteria are inoculated in a liquid of Letheen broth and incubated at 35 ° C for 24 hours. In case of yeast, yeast is inoculated into YM broth at 25 ° C , The bacteria are diluted in sterilized saline and the bacteria are plated on each agar plate medium at a concentration of about 1 × 10 ⁶ CFU and the yeast is plated on an agar plate medium at a bacterial concentration of about 1 × 10 ⁵ CFU. After saturated with 0, 1, 5, and 10% concentration of extract solution, 8 mm paper discs are placed on the solid medium at regular intervals. Then, the bacteria were cultured at 35 ° C. for 48 hours, and the yeast was cultured at 25 ° C. for 72 hours, and the size (mm) of the clear zone around the disk was measured and compared. The antimicrobial activity of the strains of Example 1 and Comparative Example 1 was measured and the results are shown in Table 1 below.

division Antimicrobial spectrum (mm) Staphylococcus aureus Escherichia coli P. aeruginosa Candida yeast Black mold Comparative Example 1 16 18 15 9 8 Example 1 28 29 18 13 13

Experimental Example 2: Antimicrobial Activity Test (MIC)

In order to evaluate the antimicrobial activity of the extracts, the minimum inhibitory concentration (MIC) was determined by measuring the inhibitory activity by concentration. Gram positive bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Gram negative bacteria, Escherichia coli, Candida yeast as yeast, and Black fungi as fungi were purchased from Korea Biotechnology Research Institute. The culture preparation for each strain is the same as described above.

Each strain was inoculated into a tryptic soy broth and cultured in a shaking incubator at 35 ° C. for 24 hours. The optical density (OD) was measured at 600 nm, and the final concentration was 1 × 10 ⁶ cells / Ml < / RTI > was inoculated into a medium containing the extracts in concentrations. After incubation at 35 ° C for 48 hours, the minimum inhibitory concentration (MIC) was determined by determining the concentration at which microbial growth was not visually observed. The fungi were cultured at 30 ° C. for 72 hours, and the minimum inhibitory concentration test (MIC test) was performed in the same manner as described above. The results of MIC measurement of mixed fermented extracts for each strain are shown in Table 2 below.

division Minimum inhibitory concentration (ppm) Staphylococcus aureus Escherichia coli P. aeruginosa Candida yeast Black mold Comparative Example 1 25 100 100 300 300 Example 1 25 25 100 150 300

Experimental Example 3: Insect test

3-1: Intrusion Inoculation Test (FC-TM-21)

The penetration inhibition method (FC-TM-21) was performed to measure the rejection rate of the insecticidal composition. The test method of the invasion inhibition method is as follows, and the dermatophagoides farina was used as the test bed .

① Place a test and control sample on the bottom of a plastic round dish (3.5 cm in diameter x 1 cm in height) cut into a circular plate size, and add 0.05 g of manure substance on it.

② Place a plate of 1) in the middle of a plastic circular plate (9 cm in diameter x 1.5 cm in height), and spread the voids between the circular plates of 1) and 2) uniformly.

③ The density of the insecticide shall be 30,000 grains / g or more.

④ Place a rectangular plastic sealed container in a thermostat (25 ± 2 ℃) and keep it in full darkness for 24 hours.

⑤ After 24 hours, measure the number of openings buried in a plastic round dish (diameter 3.5 cm x height 1 cm) with a microscope (magnification: 100).

[Equation 1]

x 100Avoidance rate (%) =

x 100

The results of the experiment of avoiding the large legged mites of the mixed fermented extract are shown in Table 3 below.

division Example 1 Comparative Example 1 N number #One #2 # 3 Number of invaders 1,970 2,230 2,135 4,458 Avoidance rate (%) 84.0 81.8 82.6 62.7

3-2: House dust mite salivation effect test

Direct spray experiments were carried out to test the salivary efficacy of house dust mites. Fifteen house dust mites were irradiated on a 20 x 20 cm black cloth, and Example 1 was diluted 50 times at a distance of about 30 cm, and the percentages (%) after 1, 3, and 5 injections were measured. Spraying conditions are shown in Table 4 below. In addition, the activity and the percentage of house dust mites were measured at 5, 15 and 30 minutes after injection according to the number of injections, and the results are shown in Table 5 below.

division Result Time 1 time throughput
(Ml / m 2) 3 times throughput
(Ml / m 2) 5 times throughput
(Ml / m 2) Total population
(Marie) Control group 5 minutes, 15 minutes
Every 30 minutes 0.6 1.8 3.0 45 Experimental group - - - - 15

&Quot; (2) "

x 100Percentage (%) =

x 100

The percentages were calculated using the Abbott formula (1925).

jet
observe One time processing
Percentage (%) 3 times processing
Percentage (%) 5 times processing
Percentage (%)
Comparative Example 1
5 minutes later 79.0 85.0 97.0 After 15 minutes 85.0 97.0 100.0 30 minutes later 97.0 100.0 100.0
Example 1
5 minutes later 89.0 98.0 100.0 After 15 minutes 97.0 100.0 100.0 30 minutes later 100.0 100.0 100.0

As a result of the above-mentioned examples, it was confirmed that the fermented extracts of ginseng, cinnabar, sesame and clove according to the present invention provide a higher antimicrobial and insecticidal effect than the non-fermented general extracts.

The present invention can be used in the field of fermentation industry utilizing natural products. While the invention has been shown and described with reference to certain embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (12)

( Leuconostoc mesenteroides : KCCM 35471) as a lactic acid bacterium is inoculated to a mixture containing ginseng, celery, persimmon, and clove, and the mixture is kept at a pH of 4.0 to 5.0 within a temperature range of 20 to 37 캜 for 3 to 7 days And the fermented extract obtained by extracting the fermentation product obtained by fermentation is contained as an active ingredient.
The method according to claim 1,
Wherein the mixture comprises 0.01 to 20% by weight of the ginseng, 0.01 to 20% by weight of the cinnabarin, 0.01 to 20% by weight of the sponge, and 0.01 to 40% by weight of the clove containing the ginseng, cinnabar, Wherein the antimicrobial and insecticidal composition is used.

delete The method according to claim 1,
Wherein the lactic acid bacteria are selected from the group consisting of Enterococcus faecium , Enterococcus faecalis , Lactobacillus plantarum , Lactobacillus fermentum, Lactobacillus spp. In addition to Leuconostoc mesenteroides (KCCM 35471) Lactobacillus sp., Lactobacillus fermentum , Lactobacillus lactis , Lactobacillus rhamnosus , Lactobacillus sakei , Lactobacillus brevis , Lactobacillus graminis , flow Pocono stock sheet reum (Leuconostoc citreum), Bar Sela helre NIKA (Weissella hellenica), Lactococcus teaching ratio in (Lactococcus garvieae), Phedi O Lactococcus pen soil three mouse (Pediococcus pentosaceus), the Phedi O Lactococcus CD rakti when (Pediococcus acidilactici), Bifidobacterium No Marlies (Bifidobacterium animalis), Bifidobacterium Titanium Infante tooth (Bifidobacterium infantis), and antibacterial and insecticide compositions characterized in that it further comprises a lactic acid bacteria is selected from the group consisting of at least one mixture thereof.
The method according to claim 1,
Wherein the fermented extract obtained by extracting the fermented product is a fermented product obtained by dissolving the fermented product in an extraction solvent selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane, (40 kHz). The antimicrobial and insecticidal composition according to claim 1,
The method according to claim 1,
Wherein said antimicrobial and antifungal composition has antibacterial activity against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi.
The method according to claim 1,
Wherein said antimicrobial and antifungal composition has insecticidal action against house dust mites and large legged mites.
3. The method according to claim 1 or 2,
Wherein said antimicrobial and insecticidal composition is formulated into a formulation of nanocapsules, microcapsules, and nanosomes.
7. A cosmetic comprising the antibacterial and antifungal composition according to any one of claims 1 to 6.
A food comprising the antimicrobial and antifungal composition according to claim 1 or 2.
A paint comprising the antimicrobial and antifungal composition according to any one of claims 1 and 2.
(1) inoculation step of inoculating the ginseng, cinnabar, sesame seeds and cloves with lactic acid bacteria; And
(2) a fermentation step of fermenting the mixture inoculated with the lactic acid bacteria.