KR102370532B1 - Compositions containing enzyme hydrosis of soybean sprout by-product - Google Patents

Abstract

The present invention relates to a composition comprising an enzyme decomposition product of bean sprouts by-product as an active ingredient. By containing an enzyme decomposition product of bean sprouts by-product hydrolyzed with a proteolytic enzyme as an active ingredient, it has excellent antioxidant and antibacterial effects, and thus a food composition, a natural preservative Or it can be used as a feed additive.

Description

Compositions containing enzyme hydrosis of soybean sprout by-product as an active ingredient

The present invention relates to a composition comprising, as an active ingredient, an enzyme-decomposed product of bean sprouts having excellent antioxidant and antibacterial effects.

Preservatives approved for human use are added to many products such as food, pharmaceuticals, daily necessities, and cosmetics widely used in our daily life to prevent changes in internal properties and preserve them for a certain period of time.

Most foods and cosmetics use chemical preservatives such as parabens to prevent product spoilage. As a result, there is a growing sense of rejection by consumers.

In a broad sense, preservatives refer to additives used to maintain nutritional value and freshness by preventing deterioration, spoilage, or chemical changes in food, pharmaceuticals, cosmetics, etc. As an additive for preserving food and pharmaceuticals by suppressing it, it is used to prevent or sterilize the growth of decaying microorganisms.

The ideal condition for such a preservative is that it should not harm the human body, its addition should not impair quality, and it should be effective even in a small amount.

Due to the above problems, consumers' reluctance to use chemical preservatives or additives has increased recently, and accordingly, interest in natural preservatives capable of inhibiting the growth of microorganisms and the like is rapidly increasing.

However, most of the natural active substances used as natural preservatives have not been commercialized due to color odor, reduced stability, narrow antibacterial spectrum, and reshaping problems.

Meanwhile, bean sprouts consumed in Korea amount to about 60,000 tons per year, and during the cultivation and processing of bean sprouts, 1-1.2 times of the bean sprouts produced are separated during washing and packaging and produced as by-products. The by-products of bean sprouts mainly consist of the head and tail, and it is known that they have the same nutritional content as the commercialized bean sprouts.

Bean sprouts by-products tend to decay easily due to their high moisture content, so some of them are used as feed after natural drying processes through compression and dehydration. In order to solve this problem, there is a need for a method for producing high value-added substances using the bean sprout by-products to utilize the by-products of the bean sprouts that are thrown away.

Republic of Korea Patent Registration No. 10-1952344 Republic of Korea Patent Registration No. 10-1950919

It is an object of the present invention to provide a food composition comprising, as an active ingredient, an enzyme decomposition product of bean sprouts with excellent antioxidant and antibacterial effects.

Another object of the present invention is to provide a natural preservative comprising, as an active ingredient, an enzyme decomposition product of bean sprouts with excellent antioxidant and antibacterial effects.

In addition, another object of the present invention is to provide a novel strain isolated from fermented food.

The food composition having antibacterial and antioxidant activity of the present invention for achieving the above object may include, as an active ingredient, an enzyme hydrolyzed product of bean sprouts by-products obtained by hydrolyzing them with proteolytic enzymes.

The proteolytic enzyme may be obtained from a strain isolated from fermented food.

The fermented food may be cheonggukjang, soybean paste or soy sauce.

The strain may be a coenzyme solution of Bacillus tequilensis SM18 [Accession No.: BP1429751].

The bean sprouts by-product and proteolytic enzyme may be mixed in a solid-liquid ratio of 1: 2-20.

The bean sprouts by-product may be at least one selected from the group consisting of roots, heads, and skins of bean sprouts germinated under light-blocked conditions.

The bean sprouts by-product may be dried with hot air at 40 to 80 °C.

In addition, the natural preservative of the present invention for achieving the above other object may include as an active ingredient an enzyme hydrolyzed product of bean sprouts by-products hydrolyzed by proteolytic enzymes.

In addition, the novel strain of the present invention for achieving the above another object may be Bacillus tequilensis SM18 [Accession No.: BP1429751] isolated from fermented food.

The composition comprising the enzyme decomposition product of bean sprouts of the present invention as an active ingredient can sufficiently function as a new antibacterial agent and antioxidant due to its excellent antibacterial and antioxidant activity, and is a functional material with excellent stability.

The composition of the present invention can be used not only as a food composition, but also can be applied as a preservative (antibacterial preservative) and feed additive in the food field, as well as cosmetics, pharmaceuticals and health functional foods with natural antibacterial properties.

1 is a comparison of protein molecular weights by using SDS-PAGE to confirm changes in protein molecular weight according to enzymatic hydrolysis when using different enzymes according to Examples and Comparative Examples, and FIG. 2 is an enlarged view of FIG. did it
3 is an antibacterial test performed using the disk diffusion method for the bean sprouts by-product enzyme lysate prepared according to Example 1, Comparative Example 1 and Comparative Example 3, and the growth inhibition rate of each cell was measured using the enzyme lysate 7.1 unit/disc. did it
4 is a graph measuring the growth inhibition rate of each cell according to the treatment volume (0-1.

The present invention relates to a composition comprising, as an active ingredient, an enzyme decomposition product of bean sprouts with excellent antioxidant and antibacterial effects.

Hereinafter, the present invention will be described in detail.

The food composition of the present invention contains, as an active ingredient, an enzyme hydrolyzed product of bean sprouts by-products hydrolyzed by proteolytic enzymes.

Bean sprouts are vegetables from soybeans, which are inexpensive, grow fast, and are easy to cultivate, so they can be grown in large quantities. Nutrient content of soybean is 20% fat, 40% protein, 35% carbohydrate and 5% other components such as vitamins and minerals, which are recognized as the main food source. The bean sprouts produce phytochemicals, which are chemical defense substances, in order to protect them from external mold or bacteria in the germination process of soybeans. In addition, as soybeans germinate, vitamins that were absent or low in soybeans are synthesized, and antioxidants such as polyphenols and flavonoids are synthesized in large amounts.

The polyphenols are precursors of lignin that form the cell wall of bean sprouts and are substances having two or more hydroxyl groups (-OH) in the benzene ring.

The bean sprouts by-product used in the present invention is at least one selected from the group consisting of roots, heads and skins of bean sprouts germinated under light-blocked conditions, and preferably used alone or in a ratio of 1: 0.5-1 It is mixed in a dry weight ratio.

If the content of the root based on the bean sprouts head in the bean sprouts by-product is less than the lower limit, the antibacterial effect of the enzyme decomposition may be lowered, and if it exceeds the upper limit, the antioxidant activity may be increased according to the increase of the enzyme decomposition product.

The bean sprouts by-product may be dried with hot air at 40 to 80 ℃, preferably 55 to 65 ℃ using a hot air dryer. If the drying temperature is less than the lower limit, hydrolysis may not be smoothly performed.

In addition, when the bean sprouts by-product is dried by another drying method such as cold air drying or natural drying in which the hot air drying method is different, not only may the bean sprouts deteriorate easily, but also the antibacterial effect may be reduced.

The proteolytic enzyme hydrolyzing the bean sprouts by-product of the present invention is a supernatant obtained by centrifugation after culturing a strain isolated from a fermented food, and is called a crude enzyme solution. Preferably, cheonggukjang is mentioned. The fermented food may be a food whose stability has been verified.

The strains isolated from the fermented food belong to Bacillus tequilensis as a result of identifying each strain, and it is named Bacillus tequilensis SM18 , and as of March 04, 2019, the Center for Biological Resources ( It was deposited with KCTC: Korean Collection for Type Cultures) and was given an accession number as BP1429751.

General proteolytic enzymes show maximum activity at 30 to 50 °C, and performing protein hydrolysis in the above temperature range can prevent denaturation of proteins due to temperature.

The bean sprouts by-product and proteolytic enzyme are mixed in a solid-liquid ratio of 1: 2-20, preferably 1: 4-10. Based on the content of bean sprouts by-products, if the content of the proteolytic enzyme is less than the lower limit, hydrolysis of the protein may not be performed.

As used herein, the term 'contained as an active ingredient' means including an amount sufficient to achieve the efficacy or activity of the bean sprout by-product enzyme decomposition product. For example, the enzyme decomposition product of the bean sprouts by-product is used at a concentration of 20 to 400 unit/ml, preferably 100 to 200 unit/ml. Since the peptide, which is an enzymatic decomposition product of bean sprouts by-products, naturally decomposes over time in vivo, there is no side effect even when used in excess. there is.

The present invention provides a food composition having antibacterial and antioxidant activity containing the enzyme decomposition product of bean sprouts as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, sweets, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice cream, vitamin complexes, health supplements etc.

The food composition of the present invention may include not only bean sprout by-product enzyme decomposition product as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. do. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink or beverage, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included in addition to the enzyme decomposition product of the bean sprouts by-product of the present invention. there is.

In addition, the enzyme decomposition product of bean sprouts by-products having antibacterial and antioxidant activity of the present invention can be used as a natural preservative in food. In particular, Ps. aeruginosa, E. coli, and S. aureus , the causative agent of intestinal food poisoning, have excellent selective growth inhibitory effects and at the same time have excellent antioxidant functions, so a composition containing them can prevent or alleviate oxidation and bacterial infections. It can be developed as an active ingredient such as a food composition, health functional food, food additive, livestock feed additive, cosmetic composition, etc.

In the present invention, the term 'livestock' refers to all animals raised for food, and in the present invention, in particular, refers to pigs for meat, cattle for meat, dairy cows, meat for meat, or broilers for laying eggs.

The feed composition for livestock is, for example, 100 parts by weight of the feed additive for livestock of the present invention, 30 to 70 parts by weight of cashew nut meal, 1 to 10 parts by weight of peanut meal (compression type), 1 to 10 parts by weight of pineapple meal, mango seeds 1 to 10 parts by weight, 0.5 to 5 parts by weight of calcium lime, 0.1 to 1 parts by weight of salt, and 0.5 to 5 parts by weight of tannin may be included.

Hereinafter, preferred examples are presented to help the understanding of the present invention, but the following examples are merely illustrative of the present invention, and it will be apparent to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It goes without saying that such variations and modifications fall within the scope of the appended claims.

Preparation Example 1. Proteolytic enzyme

Isolation of fermented food-derived proteolytic enzyme strains

In order to isolate the protease-producing strain, the protease strain was separated from cheonggukjang.

A sample of Cheonggukjang was diluted with sterile physiological saline, spread on MRS flat plate solid medium to isolate bacteria, lactic acid bacteria, yeast and mold, and cultured at 30°C for 48 hours to obtain 12 single colonies. The obtained strain was inoculated in MRS liquid medium and cultured for 48 hours, and then centrifuged (10,000Xg, 20 minutes, 4 °C) to use as a crude enzyme solution for protein degradation. The proteolytic activity of the crude enzyme solution was measured by the modified Kunitz method, and the strain with the highest proteolytic activity was selected.

Identification of protease-producing strains

In order to identify the strain isolated above, 16S rDNA sequencing was performed. In order to amplify the 16S rRNA nucleotide sequence of the selected strain, genomic DNA was extracted (DNA purification kit, Promega), and Cosmogene Tech. As requested by the company, general bacterial primers (Universal PCR primer) 27F (5'-AGAGTTTTGATCCTGGCT CAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3') were used. The nucleotide sequence of the strain obtained through nucleotide sequence analysis was compared with the nucleotide sequence of the NCBI GenBank database using the Blast Network Service to compare homology. In the present invention, it was attempted to isolate microorganisms with high proteolytic efficacy from cheonggukjang. As a result of phylogenetic analysis of the 16S rRNA nucleotide sequence of two strains isolated from cheonggukjang as a strain that decomposes bean sprout protein, it was found to show 99% homology with 16S rRNA of B. tequilensis strain (SM18) and B. subtilis strain (SM8). appear. Therefore, the proteolytic strains SM18 and SM8 were deposited in KCTC (Korean Collection for Type Cultures) under the name Bacillus sp. 16S rRNA analysis was performed to identify a protease-producing strain isolated from No. 18 cheonggukjang selected by confirming the hydrolysis characteristics of the isolated strain. After diluting No. 18 cheonggukjang with sterilized distilled water, the supernatant was inoculated on MRS agar medium and the cheonggukjang strain colony was used as a sample. Table 1 shows the species that showed high similarity to the nucleotide sequence obtained through 16S rRNA sequencing with the microorganisms registered in NCBI Genbank through BLAST search. The finally selected strain was named B. tequilensis SM18 (Accession No. BP1429751, SEQ ID NO: 1).

division Closest relative GenBank
access no. Identity (%) Cheonggukjang Bacillus tequilensis (No. 18) MF767889.1 99 Bacillus
subtilis (numbers 8 and 17) JX293291.1
MH580158.1 99 Bacillus licheniformis CP021507.1 99

Example 1. Use of proteolytic enzyme derived from cheonggukjang

bean sprouts by-product

The bean sprouts heads and roots are mixed in a weight ratio of 1:0.5 and dried with a hot air dryer (OF-22GW, Jeiotech, Daejeon, Korea) at 60 ° C. 0505S, Korea), and then filtered through a sieve (100 mesh), and then the filtered particles were refrigerated and stored at 2 °C for use.

Enzyme activity measurement and proteolytic hydrolysis

Hydrolysis was performed by adding 1.2% MnCl to the MRS medium and culturing at 35° C. for 24 hours, followed by culturing the bean sprouts by-product and the strain selected in Preparation Example 1 ( B. tequilensis SM18 ). A supernatant containing a protease (Crude enzyme solution) was stirred at 150 rpm using a shaking incubator under conditions of a soybean sprout by-product and solid-liquid ratio of 1:20 (w/v), a reaction temperature of 35 °C, and a reaction time of 72 hours. After 24 hours of incubation of B. tequilensis SM18 , the protease of the produced crude enzyme solution was measured, and the activity was compared with that of an existing commercial enzyme and is shown in Table 2. After the enzymatic hydrolysis of the bean sprouts by-product protein was completed, the supernatant was recovered by standing still and centrifuged to recover the residual supernatant (enzyme decomposition product) to measure the enzyme activity.

The following [Table 2] compares the proteolytic activity of the existing commercial protease and the crude enzyme solution produced from Bacillus tequilensis SM18 .

enzyme Enzyme activity (unit/) Alcalase ¹ 353.0±15.2 Protamax ¹ 206.4±10.8 Neutrase ¹ 189.2±5.9 Flavorzyme ¹ 81.1±3.9 B. tequilensis (coenzyme solution) ² 222.3±6.9

¹⁴ Four kinds of commercial enzymes were diluted 200-fold in a buffer solution and proteolytic activity was measured.

² The proteolytic activity of the crude enzyme solution derived from B. tequilensis was measured using the stock solution without dilution.

Comparative Example 1. Use of Buffer

It was carried out in the same manner as in Example 1, but the hydrolysis was carried out by adding 20 ml of 0.2 M sodium phosphate buffer to 1 g of the dried bean sprouts by-product and enzymatic decomposition at pH 7 for 72 hours. For enzymatic decomposition, hydrolysis was performed at a reaction temperature of 50 °C in a shaking incubator to produce peptides or amino acids. After the hydrolysis was completed, the enzyme reaction was stopped by storing at -20 °C for 10 minutes, and then centrifuged at 10,000 rpm and 4 °C for 10 minutes to recover the obtained supernatant (enzyme lysate).

comparative example 2. Alcalase ( Alcalase ) using protease_24 hours

In order to compare the enzyme decomposition ability of the crude enzyme solution with a commercial enzyme, the same procedure as in Example 1 was performed, except that, instead of the B. tequilensis SM18 -derived crude enzyme solution as a proteolytic enzyme, alkalinease was diluted 200 times (v/v) compared to the buffer solution. It was added to the buffer solution and enzymatic digestion was performed for 24 hours to obtain an enzymatically digested product.

Comparative Example 3. Using Alcalase protease_72 hours

The same procedure as in Example 1 was performed, except that, instead of the coenzyme solution derived from Bacillus tequilensis SM18 as a proteolytic enzyme, alkalinease was diluted 200 times (v/v) compared to the buffer solution and added to the buffer solution to perform the enzymatic degradation reaction for 72 hours. to obtain an enzymatic lysate.

<Test Example>

Test Example 1. Electrophoretic measurement of bean sprouts by-product enzyme decomposition product

1 is a comparison of protein molecular weights by using SDS-PAGE to confirm changes in protein molecular weight according to enzymatic hydrolysis when using different enzymes according to Examples and Comparative Examples, and FIG. 2 is an enlarged view of FIG. did it

For electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used according to Laemmli's method, and 12%, 16.5% separating gel and 5% stacking gel were used for the gel used in the experiment. Prestained protein ladder (Thermo Scientific, IL, USA) and Ultra Low Range Molecular Weight Marker (Sigma, St Louis, MO, USA) were used as proteins. The electrophoresis sample was mixed with 5x sample buffer of 60 mM Tris-HCl (pH 6.8), 5% glycerol, 0.02% bromophenol blue, 1% 2-mercapto ethanol, and 0.4% SDS, and then treated at 100 °C for 10 minutes. Electrophoresis was performed under conditions of mA, 63 min. After the electrophoresis was completed, the stacking gel was separated, stained with coomasie blue for 2 hours, and destaining buffer was added, followed by reaction overnight.

As shown in FIGS. 1 and 2 , it was confirmed that two bands were present in the enzymatic lysate prepared according to Example 1 at 25 kDa and 14.2 kDa or less, and Comparative Examples 2 and 3 treated with Alcalase As compared to , the band at 6.5 kDa or less was not seen, so it is expected that all proteins were decomposed into single amino acids (FIG. 2).

In the enzyme lysate of Comparative Example 1, 10 or more bands were confirmed in the range of 6.5 to 250 kDa, and in the case of LB, no band was confirmed, so it was confirmed that there was no effect on the protein content result (FIG. 1). In addition, three bands were confirmed in the enzyme lysate of Comparative Example 2, and the band was not identified in the enzyme lysate of Comparative Example 3, indicating that the enzyme lysate of Comparative Example 3 exhibits higher proteolytic efficacy.

B. tequilensis SM18 -derived coenzyme solution used in Example 1 of the present invention can be used as a commercial enzyme because it shows similar or superior proteolytic ability to the commercially available enzyme Alcalase.

Test Example 2. Measurement of total polyphenol content and DPPH scavenging ability

2-1. Total polyphenol content (mg GAE/g DW): Measured by modifying the Folin-Denis method, folin-ciocalteu's phenol solution (Folin &Ciocalteu'sphenol; Sigma-Aldrich, USA) was added to the sample and reduced by a polyphenol compound The molybdenum blue color reaction generated by the process was based on the principle. After adding 0.2 N FC solution to 0.14 ml of bean sprouts by-product enzyme decomposition, and leaving it for 10 minutes, 0.56 ml of 7.5% Na ₂ CO ₃ was added and reacted for 1 hour, and the absorbance value was measured at 756 nm. Gallic acid (Sigma-Aldrich, USA) was used as a standard material, and the unit was expressed as mg gallic acid equivalent (GAE)/g dry weight (DW) compared with the prepared gallic acid calibration curve.

2-2. DPPH scavenging ability (%): The electron donating ability was measured by modifying the method of Blois, and DPPH (2,2-Diphenyl-1-picrylhydrazyl, Sigma -Aldrich) The reducing power of the sample was measured through free radical scavenging activity. After adding 1.25 ml of DPPH solution to 0.25 ml of bean sprout by-product enzyme decomposition, and reacting in the dark for 20 minutes, absorbance was measured at 517 nm. Activity was expressed as a percentage.

[Equation 1]

DPPH radical scavenging activity (%)={1-(Abs(test)-Abs(color))/Abs(control)}X100

division Total polyphenol content DPPH radical scavenging ability medium 18.62 53.45 Example 1 180.61 86.30 Comparative Example 1 48.70 0 Comparative Example 2 148.99 55.01 Comparative Example 3 148.78 57.68

As shown in Table 3 above, it was confirmed that the soybean sprout by-product enzyme lysate prepared according to Example 1 of the present invention showed significantly higher total polyphenol content and DPPH radical scavenging ability compared to the enzyme lysate of Comparative Examples 1 to 3. In the case of cheonggukjang culture, a small amount of total polyphenol content at 18.62 mg GAE/g DW and low DPPH radical scavenging ability were confirmed at 53.45%.

In addition, it was confirmed that Comparative Examples 2 and 3 using Alcalase had no significant difference in total polyphenol content and DPPH radical scavenging ability even when the decomposition time was increased.

In total polyphenol content and DPPH radical scavenging ability, it was confirmed that the enzyme decomposition product of Example 1 increased by 1.3 times or more, respectively, compared to Comparative Examples 2 and 3 using alkalinease. During the fermentation process of cheonggukjang, glycosides decreased and daidzein and genistein increased due to the change in total isoflavone component during the fermentation process. By comparing total polyphenol content and DPPH radical scavenging ability, Bacillus sp. Bacillus sp . It was confirmed that the proteolytic enzyme (coenzyme solution) of the can be used instead of the conventional commercial enzyme Alcalase.

Test Example 3. Measurement of antibacterial activity by disc diffusion method (Paper Disc Test)

Each test strain was pre-cultured on an LB agar plate for 24 hours, the OD of the culture solution was adjusted to 0.1 (600 nm), and 50 uL of 0.5% soft agar before coagulation (below 40 ℃) was added to the plate and mixed evenly. Pour to form a double medium. While the soft agar hardens, inoculate 20 uL of sample on a paper disc (6 mm), dry for 5 minutes, attach the paper disc to the medium inoculated with bacteria, and incubate at 35 ° C for 24 hours. Clear zone formed around the disc Check whether the antibacterial activity is through

Distilled water (DW) was used as a negative control, and the food harmful strains used in the experiment were purchased from the Korea Research Institute of Bioscience and Biotechnology and the Korea Microbial Conservation Center. Staphylococcus aureus subsp. aureus KCCM 11593 was used.

division Inhibition zone(mm) Distilled water Example 1
( B. tequilensis ) Comparative Example 1 Comparative Example 3
(alkarate) Pseudomonas aeruginosa KCCM 11328 - + - - Escherichia coli KCTC 2772 - + - - Staphylococcus aureus subsp . aureus KCCM 11593 - - - - -: No inhibition zone +: inhibition zone

As shown in Table 4 above, the soybean sprout by-product enzyme lysate prepared according to Example 1 of the present invention exhibited antibacterial activity against Gram-negative bacteria at a concentration of 7.1 unit/disc, but in distilled water, Comparative Example 1 and Comparative Example 3 It was confirmed that antibacterial activity against Gram-negative bacteria was not observed in the enzymatic digest.

In addition, in the gram-positive bacteria, it was confirmed that Example 1, distilled water, Comparative Example 1 and Comparative Example 3 did not show any antibacterial activity.

Test Example 4. Measurement of cell growth inhibition rate

3 is inoculated into 8 ml of LB medium with 0.09 ml of pre-cultured seed cells for measuring the cell growth inhibition rate of pathogenic bacteria according to Example 1, Comparative Example 1 and Comparative Example 3, and the volume of the soybean sprout protein digest, respectively. After the addition of 0 ml, 0.2 ml, 0.5 ml, and 1.0 ml, distilled water was added so that the total volume was 10 ml. The cell growth inhibition rate was measured as shown in [Equation 2] by measuring the absorbance at 600 nm every predetermined time by culturing in a culture tube at 35 °C conditions. 4 is a graph measuring the growth inhibition rate of each cell according to the concentration of the bean sprout by-product enzyme lysate prepared according to Example 1.

[Equation 2]

A: Absorbance of the strain-added group

B: Absorbance of strain-free group

division
Growth inhibition rate (%)
incubation time strain 0 ml 0.1 ml 0.25 ml 0.5 ml 17

Pseudomonas aeruginosa KCCM 11328 0 100 99.8 89.6 Escherichia coli KCTC 2772 0 99.4 100 98.5 Staphylococcus aureus subsp. aureus KCCM 11593 0 3.17 100 94.2

As shown in Figure 3, the bean sprouts by-product enzyme lysate prepared according to Example 1 ( B. tequilensis crude enzyme solution) of the present invention compared to the enzyme lysate of distilled water, Comparative Examples 1 and 3 (alkalease) for each strain Accordingly, it was confirmed that the growth inhibition rate was high.

In addition, as shown in FIG. 4 and Table 5 above, the enzyme decomposition product prepared according to Example 1 of the present invention was found to have an inhibition rate of 90 to 100% overall, so it is considered to have long-term antibacterial activity. On the other hand, when cultured for 17 hours, Staphylococcus aureus subsp. aureus KCCM 11593 showed a low growth inhibition rate of 3.17% at 0.1 ml, but increased to 100% at 0.25 ml.

Hereinafter, formulation examples of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit the present invention, but merely to describe it in detail.

Formulation Example 1. Preparation of granules

1,000 mg of the enzyme lysate powder obtained in Example 1

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

50 μg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules in a preferred embodiment, but the mixing ratio may be arbitrarily modified. It can be prepared and used in the preparation of a health functional food composition according to a conventional method.

Formulation Example 2. Preparation of functional beverage

1,000 mg of the enzyme lysate powder obtained in Example 1

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 ml by adding purified water

After mixing the above ingredients according to the usual health drink manufacturing method, after stirring and heating at 85 ° C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, then refrigerated. It is used to prepare the functional beverage composition of the present invention.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demanding country, and use.

Biological Resource Center (Korea) BP1429751 20190304

<110> Industry-Academic Cooperation Foundation, Sun Moon University <120> Compositions containing enzyme hydrosis of soybean sprout by-product <130> HPC-8924 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1455 <212> RNA <213> Unknown <220> <223> Bacillus tequilensis <400> 1 aaaatggggg gcgtgctaat acatgcaagt cgagcggaca gatgggagct tgctccctga 60 tgttagcggc ggacgggtga gtaacacgtg ggtaacctgc ctgtaagact gggataactc 120 cgggaaaccg gggctaatac cggatggttg tttgaaccgc atggttcaaa cataaaaggt 180 ggcttcggct accacttaca gatggacccg cggcgcatta gctagttggt gaggtaacgg 240 ctcaccaagg caacgatgcg tagccgacct gagagggtga tcggccacac tgggactgag 300 acacggccca gactcctacg ggaggcagca gtagggaatc ttccgcaatg gacgaaagtc 360 tgacggagca acgccgcgtg agtgatgaag gttttcggat cgtaaagctc tgttgttagg 420 gaagaacaag taccgttcga atagggcggt accttgacgg tacctaacca gaaagccacg 480 gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc cggaattatt 540 gggcgtaaag ggctcgcagg cggtttctta agtctgatgt gaaagccccc ggctcaaccg 600 gggagggtca ttggaaactg gggaacttga gtgcagaaga ggagagtgga attccacgtg 660 tagcggtgaa atgcgtagag atgtggagga acaccagtgg cgaaggcgac tctctggtct 720 gtaactgacg ctgaggagcg aaagcgtggg gagcgaacag gattagatac cctggtagtc 780 cacgccgtaa acgatgagtg ctaagtgtta gggggtttcc gccccttagt gctgcagcta 840 acgcattaag cactccgcct ggggagtacg gtcgcaagac tgaaactcaa aggaattgac 900 gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga agaaccttac 960 caggtcttga catcctctga caatcctaga gataggacgt ccccttcggg ggcagagtga 1020 caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1080 agcgcaaccc ttgatcttag ttgccagcat tcagttgggc actctaaggt gactgccggt 1140 gacaaaccgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200 cacacgtgct acaatggaca gaacaaaggg cagcgaaacc gcgaggttaa gccaatccca 1260 caaatctgtt ctcagttcgg atcgcagtct gcaactcgac tgcgtgaagc tggaatcgct 1320 agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1380 tcacaccacg agagtttgta acacccgaag tcggtgaggt aacctttagg agccagccgc 1440 cgaaaggggg accca 1455

Claims (9)

It contains as an active ingredient an enzyme-decomposed product of bean sprouts by-products hydrolyzed with proteolytic enzymes,
The proteolytic enzyme is a food composition having antibacterial and antioxidant activity, characterized in that it is a coenzyme solution of B. tequilensis SM18 [Accession No.: BP1429751] as a strain isolated from fermented food. delete The food composition according to claim 1, wherein the fermented food is cheonggukjang, soybean paste or soy sauce. delete The food composition according to claim 1, wherein the bean sprouts by-product and the proteolytic enzyme are mixed in a solid-liquid ratio (w/v) of 1: 2-20. The food composition according to claim 1, wherein the bean sprouts by-product is at least one selected from the group consisting of roots, heads, and skins of bean sprouts germinated under light-blocked conditions. The food composition according to claim 1, wherein the bean sprouts by-product is dried with hot air at 40 to 80°C. It contains as an active ingredient an enzyme-decomposed product of bean sprouts by-products hydrolyzed with proteolytic enzymes,
The protease is a natural preservative, characterized in that it is a coenzyme solution of B. tequilensis SM18 [Accession No.: BP1429751] as a strain isolated from fermented food.

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