KR20200013367A - Fermented composition of protaetia brevitarsis seulensis larvae and the preparation method thereof - Google Patents

Abstract

Disclosed by the present invention is a method for producing a fermented Protaetia brevitarsis seulensis larva composition, which comprises: a step of preparing a Protaetia brevitarsis seulensis larva extract; a step of producing a first mixture by adding ginseng powder to the Protaetia brevitarsis seulensis larva extract; a step of producing a second mixture by inoculating the first mixture with lactic acid bacteria; and a step of fermenting the second mixture, wherein the lactic acid bacteria are selected from Lactobacillus plantarum, Lactobacillus brevis, and a mixture of the same. The fermented Protaetia brevitarsis seulensis larva composition produced by the production method: combines larvae of Protaetia brevitarsis seulensis and ginseng; is effective by being absorbed into a body more quickly and efficiently since various polymers contained in larvae of Protaetia brevitarsis seulensis and ginseng are bioconverted by lactic acid bacteria and are decomposed into a form having small molecular weight; and has more improved antioxidant effects and antibacterial effects against various bacteria.

Description

Flower fermentation composition and its manufacturing method {FERMENTED COMPOSITION OF PROTAETIA BREVITARSIS SEULENSIS LARVAE AND THE PREPARATION METHOD THEREOF}

The present invention relates to a flower fermentation composition and a method for preparing the same, and more specifically, to prepare a flower extract, add a mixture of the prepared extract and ginseng powder, fermentation of the flower comprising a step of inoculating and fermenting the lactic acid bacteria It relates to a method for producing the composition and to a composition prepared by the above method.

White spotted radish ( Protaetia brevitarsis seulensis ) is an insect belonging to the species Coleoptera ( Ceopterina ), the larvae of the larvae are used for food and medicinal purposes. Flower bud is called medicinal herb and has been used as a medicine since ancient times. It is effective in treating adult diseases such as liver disease, dysmenorrhea, decreased vision, cataracts, gold swelling, stomatitis, tetanus, hemorrhoids and stroke. This is known to be. Recent studies have shown that zinnia has various pharmacological effects such as antioxidant, anti-cancer, antibacterial, thrombolytic, blood lipid improvement, whitening, skin moisturizing, liver function improvement, anti-dementia and anti-obesity.

Panax ginseng is a perennial herb belonging to the genus Araliaceae Panax ginseng . It is a medicinal herb that has traditionally been used in oriental medicine because it has little toxicity and has various effects. Ginseng roots are known to have various effects such as antioxidant, anti-cancer, anti-fatigue and stress, liver function protection, anti-inflammatory, anti-aging, analgesic effect, such as saponins, phenolic compounds, alkaloids, polysaccharides, etc. It is known to be due to ingredients, of which saponins are particularly important.

Republic of Korea Patent No. 10-1616688 discloses a functional food using a slug, where the white spotted radish larvae or beetle larvae powder, marshmallow powder, and hawthorn powder to make a mixture, the mixture powder The present invention relates to a functional food which can be reprocessed as an additive of a pill, a beverage, a tea or a general food. However, there are not many prior arts on functional foods and pharmaceutical compositions including flower clusters, and in particular, there is no disclosure about a method of preparing a composition that combines flower clusters (ginseng) with ginseng and enhances their pharmacological effects.

The present invention has been made to solve the above problems, an embodiment of the present invention is to prepare a flower extract, the step of adding a ginseng powder to the flower extract to produce a first mixture, the first mixture Inoculating a lactic acid bacterium to produce a second mixture and fermenting the second mixture, wherein the lactic acid bacteria is selected from Lactobacillus plantarum, Lactobacillus brevis, or a mixture thereof. Provide a method.

In addition, one embodiment of the present invention provides a flower fermentation composition prepared by the above method.

The technical problem to be achieved by the present invention is not limited to the technical problem mentioned above, and other technical problems not mentioned above may be clearly understood by those skilled in the art from the following description. There will be.

As a technical means for achieving the above technical problem, in accordance with an aspect of the present invention, the method for producing a flower fermentation composition comprises the steps of preparing a flower extract; Adding a ginseng powder to the flower extract to produce a first mixture; Inoculating the first mixture with lactic acid bacteria to produce a second mixture; Fermenting the second mixture; wherein the lactic acid bacteria are selected from Lactobacillus plantarum , Lactobacillus brevis , or a mixture thereof.

Here, the flower extract may be characterized in that the powder is solvent-extracted, concentrated under reduced pressure and then lyophilized powder.

The first mixture production step may have an additional sterilization step immediately after.

The second mixture may be characterized in that 1 to 15% by weight of the flower extract, 0.5 to 5% by weight of ginseng powder, 2 to 10% by weight of lactic acid bacteria.

The fermentation step may be performed for 20 to 60 hours at 30 to 40 ℃.

The fermentation step may have an additional sterilization step immediately after.

Flower pot fermentation composition according to another aspect of the present invention is characterized in that it is prepared by the above production method.

The above-described problem solving means are merely exemplary, and should not be construed as limiting the present invention. In addition to the above-described exemplary embodiments, additional embodiments may exist in the drawings and detailed description of the invention.

In the method for preparing a flower fermentation composition according to an embodiment of the present invention, by combining ginseng and ginseng, bioconversion of various polymers contained in the flower and ginseng by bioconversion by lactic acid bacteria (bioconversion) It is possible to obtain a composition that can be absorbed more quickly and efficiently in the body to have an effect.

Flower fermentation composition according to an embodiment of the present invention is further improved the antioxidant effect of the flower and ginseng, and has antibacterial properties against various bacteria.

The effects of the present invention are not limited to the above-described effects, but should be understood to include all the effects deduced from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a result of 16s rRNA sequence similarity analysis of lactic acid bacteria according to Example 2 of the present invention.
Figure 2 is a method for producing a flower fermentation composition according to an embodiment of the present invention.
Figure 3 is a TLC analysis of the flower fermentation composition according to Example 3 of the present invention (Lane 1: L. plantarum ; Lane 2: L. brevis ; Lane 3: two kinds of mixing).
Figure 4 is the antimicrobial activity analysis of the flower fermentation composition according to Example 3 of the present invention (1) L. plantarum ; 2) two kinds of mixing; 3) L. brevis ).

Hereinafter, the present invention will be described in more detail. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

In addition, the terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting of the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. "Including" any component throughout the specification of the present invention means that it may further include other components, except to exclude other components unless specifically stated otherwise.

In accordance with an aspect of the present invention, a method for producing a flower fermentation composition comprises preparing a flower extract; Adding a ginseng powder to the flower extract to produce a first mixture; Inoculating the first mixture with lactic acid bacteria to produce a second mixture; And fermenting the second mixture; wherein the lactic acid bacteria is selected from Lactobacillus plantarum, Lactobacillus brevis, or a mixture thereof.

Here, the flower extract may be characterized in that the powder of the flower extract and concentrated under reduced pressure by solvent extraction and then lyophilized powder. The flower powder may be obtained by fasting the flower and then grinding and drying. Fasting the blooms can remove feces inside the blooms. The milling and drying of the flower cluster may be repeated until the desired particle size is obtained. Preferably, the particle size of the powder may be 100 μm or less, and the smaller the particle size of the flower powder is, the more efficient later extraction may be. More preferably, it may be less than 10 μm. The extraction solvent may be selected from water, ethanol, hexane, ethyl acetate and isopropyl alcohol, and water may be preferably used. The extraction temperature may be at least room temperature, preferably at room temperature. Concentration under reduced pressure may be carried out at room temperature or higher, preferably at 50 ° C or higher. The concentrated extract under reduced pressure may be dried by lyophilization, hot air drying, spray drying, vacuum drying, drum drying, foam drying, and preferably freeze drying.

The ginseng may be selected from the processed state is raw ginseng, white ginseng, red ginseng, taegeuk ginseng, black ginseng, the type may be selected from the ginseng, wild ginseng, camphor ginseng, goats. The ginseng may preferably be a goat ginseng in a raw ginseng state.

The first mixture may be in the form of a powder mixture of the above-described flower extract and ginseng powder, it may be in addition to the liquid commonly mixed with a solvent, such as water commonly used for fermentation of food.

The first mixture production step may have an additional sterilization step immediately after. The sterilization step may be to remove bacteria other than lactic acid bacteria to be added to the first mixture. When other bacteria are present, they interact with the lactic acid bacteria and the components in the first mixture, thereby inhibiting chemical reactions that produce unwanted by-products or producing a desired effect, or inhibiting the growth of the lactic acid bacteria. Because side effects can exist. The sterilization can be carried out by a conventional sterilization, sterilization or sterilization method without addition of chemicals used in the food manufacturing process, such as heating, electrosterilization, microwave sterilization, filtration and the like. The sterilization may preferably be carried out by filtration, wherein the first mixture may be liquid.

The second mixture may be characterized in that it comprises 1 to 15% by weight of the flower extract, 0.5 to 5% by weight of ginseng powder, 2 to 10% by weight of lactic acid bacteria, more preferably based on the ingredients before the fermentation proceeds. The extract may be characterized in that it comprises 3 to 10% by weight, ginseng powder 2 to 4% by weight, lactic acid bacteria 3 to 7% by weight. In this case, the balance may be water.

The lactic acid bacteria may be selected to be isolated and identified as beta glucosidase (β-glucosidase) positive lactic acid bacteria. The lactic acid bacteria may preferably be selected from Lactobacillus. Lactobacillus may preferably be selected from Lactobacillus plantarum, Lactobacillus brevis, or a mixture thereof, and most preferably may be selected from Lactobacillus plantarum SM50, Lactobacillus brevis SM61, or a mixture thereof, isolated and identified in one embodiment of the present invention.

It may be most preferable to use the two strains mixed for the fermentation, the mixing ratio of the two strains may be mixed in a ratio of 1:10 to 10: 1, more preferably 1: 2 to 2 It may be a ratio of 1: 1, and most preferably, it may be used by mixing 1: 1.

The fermentation step may be performed for 20 to 60 hours at 30 to 40 ℃. If the temperature is lower than 30 ℃ or higher than 40 ℃ during fermentation, the growth rate of lactic acid bacteria may not be sufficient or not suitable for growth, and the activity of enzymes that act on bioconversion during fermentation may be inhibited. If the fermentation period is shorter than 20 hours, the desired fermentation reaction may not be sufficient. If the fermentation period is longer than 60 hours, the lactic acid bacteria may become excessively dense, causing side effects such as a change in the combination of enzymes expressed by quorum sensing. Can be.

The fermentation step may have an additional sterilization step immediately after. This can stop further fermentation. The sterilization may be carried out by the conventional sterilization, sterilization or sterilization method of the above-described food manufacturing process, preferably may be carried out by a filtration method.

Flower pot fermentation composition according to another aspect of the present invention is characterized in that it is prepared by the above production method.

It is obvious that the composition may be added variously to the composition, in addition to the contents mentioned in the above method, other additives that may further enhance the absorption or effect of the pharmaceutically acceptable carrier and / or the active ingredient. The detailed description thereof will be omitted.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

Example 1 Preparation of Flower Extract.

The flower buds were bred for 6 days, dried and ground with a homogenizer. 1000 mL of water per 100 g of flower powder was added using water as the extraction solvent, and extracted by stirring and leaching at room temperature for 2 hours. This was filtered (Whatman No. 2), and the extract filtrate was concentrated under reduced pressure on a 60 ° C. water bath using a rotary vacuum evaporator, and the concentrate was lyophilized to obtain a flower extract.

Example 2. Lactobacillus Isolation and Identification.

The culture of lactic acid bacteria was anaerobicly performed at 37 ° C. for 24 hours in Lactobacilli MRS broth or MRS agar prepared with 0.05% L-cysteine. To isolate the lactic acid bacteria, 25 g of fermented food samples were mixed in 225 mL of 0.1% peptone water, homogenized by using a stomacher, and then diluted in decimal, and incubated at 37 ° C for 48 to 72 hours after smearing in MRS agar. And morphologically different among the cultured colonies were selected and delineated in culture in MRS agar to obtain a single colony. After inoculating each of the isolated strains into esculin agar to confirm β activity, bacteria that transformed the medium to dark brown by hydrolyzing esculin, ie, β-producing strains, were isolated.

16s rRNA sequencing was performed on the selected strains. The lactic acid bacteria isolated for sequencing were cultured in MRS broth, 1 mL of the culture solution was centrifuged, and washed with 0.8% sterile saline. DNA was extracted using genomic DNA kit and used as template DNA for PCR, and 785F (GGATTAGATACCCTGGTA) and 907R (CCGTCAATTCMTTTRAGTTT), universal primers for bacterial 16s rRNA, were used. After PCR, the amplification product was confirmed by electrophoresis and purified using a QIAquick PCR purification kit. The nucleotide sequence was determined by Dye Terminator Cycle Sequencing Ready Reaction Kit and ABI 3700 sequencer. At this time, the same primers were used. Similarity of 16S rRNA sequences was analyzed by comparing the DNA database of the European Nucleotide Archive of the European Molecular Biology Laboratory (https://www.embl.org/) (FIG. 1). As a result of confirming the similarity of the nucleotide sequence to 16s rRNA of the isolated two-week lactic acid strain, SM50 was identified as Lactobacillus plantarum and SM61 as Lactobacillus brevis . Since the fermentation step in the manufacture of the flower fermentation composition was used as a starter L. plantarum SM50 and L. brevis SM61 isolated as described above.

Example 3. Preparation of Flower Fermentation Composition.

Using the flower extract, goat ginseng and lactic acid bacteria isolated according to Example 2, L. plantarum SM50 and L. brevis SM61 obtained according to Example 1, the flower fermentation composition was prepared as follows. 5% of flower extract and 1% goat ginseng powder were mixed to produce a first mixture, which was then sterilized using a 0.2 μm syringe filter. Next, the pre-cultivated lactic acid bacteria were mixed with a single strain or two weeks to inoculate a mixed mixture at a weight ratio of 5% to produce a second mixture, and fermentation was performed at 37 ° C. for 24 hours or 48 hours. Thereafter, the fermentation product was centrifuged at 8000 g for 10 minutes, and the supernatant was sterilized using a syringe filter to obtain a flowery fermentation composition (FIG. 2).

Experimental Example 1. Sugar Composition Analysis of Flower Fermentation Composition

Thin layer chromatography (TLC) was performed on the flower fermentation composition prepared according to the method for preparing the flower fermentation composition of Example 3. At this time, glucose polymer (G1-G7) was used as a standard material. As a result, sugars of various sizes were present in the composition, and in particular, it could be confirmed that a large amount of glucose, maltose, maltotriose, etc. were mainly included (FIG. 3). This is believed to result from the decomposition of various polymers present in the first mixture including the flower extract and goat ginseng powder through fermentation.

Experimental Example 2. Antioxidant Activity of Fermented Fermented Composition

DHHP free radical scavenging ability was evaluated during the fermentation period by using the Blois method to confirm the antioxidant capacity of the flower fermentation composition prepared according to the method for producing a flower fermentation composition of Example 3. At this time, the fermentation step was carried out using L, plantarum SM50, L. brevis SM61, respectively, and ascorbic acid was used as a standard material. As a result, it could be confirmed that the antioxidant capacity, which was high even before the fermentation, was further improved as the fermentation proceeded.

DPPH radical scavenging activity (%) 0h 24h 48h L. plantarum SM50 86.85 ± 0.13 98.19 ± 0.03 98.20 ± 0.17 L. brevis SM61 89.75 ± 0.08 99.49 ± 0.04 96.82 ± 0.22

Experimental Example 3. Antimicrobial Activity Assays of Flower Fermentation Compositions

In order to confirm the antibacterial activity of the flower fermentation composition prepared according to the method for preparing a flower fermentation composition of Example 3, a well diffusion method was performed. At this time, the fermentation step was carried out using L. plantarum SM50, L. brevis SM61 and the mixed strain of the two strains, respectively, each composition is clear zone appearing in each microorganism of E. coli JM109, L. monocytogenes KCTC3569 and S. aureus KCTC3881 It was confirmed whether the antimicrobial activity. As a result, it showed the greatest antimicrobial activity against Gram negative bacteria E. coli , L. monocytogenes , S. aureus in order (Fig. 4). There was no significant difference in antimicrobial activity between fermented lactic acid bacteria strains.

The above description of the present invention is for illustrative purposes, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. There will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present invention is represented by the following claims, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present invention.

Claims (7)

Preparing a flower extract;
Adding a ginseng powder to the flower extract to produce a first mixture;
Inoculating the first mixture with lactic acid bacteria to produce a second mixture;
Fermenting the second mixture;
Including,
The lactic acid bacteria is Lactobacillus plantarum , Lactobacillus brevis or characterized in that selected from the mixture, a method for producing a flower fermentation composition. The method of claim 1,
The flower extract is,
Solvent powder, solvent extraction, and concentrated under reduced pressure, characterized in that the lyophilized powder, the method for producing a flower fermentation composition. The method of claim 1,
Characterized in that it has an additional sterilization step immediately after the step of producing the first mixture, a method for producing a flower fermentation composition. The method of claim 1,
The second mixture is 1 to 15% by weight of the extract of the flower, 0.5 to 5% by weight of ginseng powder, 2 to 10% by weight of lactic acid bacteria, a method for producing a flower fermentation composition. The method of claim 1,
The fermentation step is characterized in that it is carried out for 20 to 60 hours at 30 to 40 ℃, method of producing a flower fermentation composition. The method of claim 1,
Characterized in that it has an additional bactericidal step immediately after the fermentation step. Flower fermentation composition prepared according to the method of claim 1.

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