KR101338526B1 - Reduction of biogenic amines in the fermented food by the plants extracts - Google Patents

Abstract

The present invention provides an Enterococcus spp. Extract comprising an extract of at least one substance selected from the group consisting of Schisandra chinensis, Plum, Ome, Rosemary and Licorice. It relates to a composition for inhibiting growth.

Description

Reduction of biogenic amines in the fermented food by the plants extracts}

The present invention provides Enterococcus to produce bio amines Growth inhibition composition of spp., fermented foods containing the composition and Enterococcus It relates to a method of inhibiting the growth of spp.

Cheonggukjang is one of the Korean Jangjang, and is a type of fermented soybeans, depending on the province. The Cheonggukjang is made from autumn to spring in each family eat, has a unique smell, is a high quality food rich in protein.

Cheonggukjang is boiled with beans and wrapped in rice straw, and fermented for one to three days in a warm lower neck of 40-42 ° C., the surface becomes grayish white and sticky threads between the beans. It is mixed with salt, garlic, and red pepper powder, put it in a mortar and put it in a jar and used it as a stew ingredient. Unlike our traditional soy sauce, soybean paste, and red pepper paste, it is the only traditional Korean sauce that does not add salt. It is fermented food.

Such Cheonggukjang contains various nutrients and functional ingredients such as isoflavonoids, and has useful functions such as antioxidant, anti-cancer, cholesterol reduction and hypertension prevention.

However, recent studies have shown that Cheonggukjang contains 0.7 to 483.1 mg / kg of tyramine and 1.3 to 54.3 mg / kg of histamine, and the tyramine content of Cheonggukjang is Nato's tyramine content. It was measured to be higher. The tyramine is a substance that causes mutation as a kind of biological amine that is produced by decomposing amino acids by decarboxylase during microbial metabolism during fermentation.

The tyramine is generally Enterococcus spp. Known to be produced by Enterococcus spp. Is found over 10 ⁶ in Cheonggukjang.

Research Report (Korea Food and Drug Administration, 'Reduction of Biogenic Amine in Jangjang', 2008)

In the present invention, it is possible to improve hygiene and safety by reducing the mutant tyramine, and to include a composition containing natural products (Schisandra chinensis, Plum, Ome, Rosemary and Licorice) extract, and fermented food containing the composition. Can provide.

The present invention comprises Enterococcus comprising an extract of one or more substances selected from the group consisting of Schisandra chinensis, Plum, Ome, Rosemary and Licorice as active ingredients spp. It relates to a composition for inhibiting growth.

Hereinafter, Enterococcus according to the present invention spp. The growth inhibiting composition will be described in more detail.

Enterococcus in the present invention spp. is known to produce bioamines. In the present invention, the Enterococcus spp., for example, Enterococcus faecium Can be used.

Enterococcus in the present invention Examples of the bioamines produced by spp. include, for example, tyramine, histamine, putrescine, cardaberine, spermine, spermidine, phenylethylamine, tryptamine, akmadine, or mixtures thereof. These may be mentioned, Preferably tyramine or histamine is mentioned, More preferably, tyramine is mentioned. It has been reported that poisoning with the above-described types of biological amines can cause toxicity in the human body.

Enterococcus in the present invention spp. Growth inhibiting composition comprises an extract of one or more substances selected from the group consisting of Schisandra chinensis, plum, ume, rosemary and licorice as an active ingredient. The extract is Enterococcus By inhibiting the growth of spp. can inhibit the production of biological amines, especially tyramine.

In the present invention, when the extract is prepared, dried or undried Schisandra chinensis, plum, ume, rosemary and licorice can be used as the extracting material, and these materials can be crushed finely for the efficiency of extraction. In addition, in order to obtain an effective extract, it may be subjected to a process of incubating for about 5 minutes at a temperature of about 100 ℃ before the material is ground to inactivate the enzyme in the living cell.

Extraction in the present invention can be extracted according to conventional extraction methods using 3 to 10 times the extraction solvent of the material.

In this case, as the extraction solvent, water, an organic solvent or a mixed solvent thereof that is commonly used in natural product extraction may be used. As said organic solvent, methanol or ethanol is preferable. In this case, methanol or ethanol can be used in admixture with water, but it is preferable to use it without mixing with water for easy elution of the active ingredient.

The extract first extracted by the extraction solvent may be further fractionated using an organic solvent having a different polarity from the solvent used in the first extraction, for example, nucleic acid, chloroform, ethyl acetate and the like. In addition, fractions further purified may be obtained using various chromatography such as silica gel column chromatography, thin layer chromatography or high performance liquid chromatography. For example, Schisandra chinensis, plum, ume, rosemary and licorice may be extracted with ethanol at room temperature, concentrated under reduced pressure, and further fractionated with various organic solvents such as nucleic acid, chloroform and ethyl acetate. It may be further purified by chromatography. In the present invention, the 'extract' includes all extracts, fractions and purified products obtained in each step of extraction, fractionation or purification as described above, their dilutions, concentrates or dried products.

The present invention also relates to fermented foods comprising extracts of one or more substances selected from the group consisting of Schisandra chinensis, Plum, Ome, Rosemary and Licorice.

Extract in the present invention is the same as the aforementioned extract.

In the present invention, the fermented foods include jangjang, salted fish, fermented liquor or fermented milk. Specifically, examples of jangjang include Cheonggukjang, Doenjang, Gochujang, Chunjang, Ssamjang, or Soy Sauce, and preferably Cheonggukjang.

Extract in the present invention may include 0.1 to 5.0 parts by weight, preferably 0.5 to 1.0 parts by weight based on 100 parts by weight of dried white beans. The composition has excellent Enterococcus at a content in the aforementioned range. spp. Has growth inhibition effect.

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In addition, the present invention comprises Enterococcus spp. Mixed with fermented food extract of at least one substance selected from the group consisting of Schisandra chinensis, plum, ume, rosemary and licorice. It relates to a method of inhibiting growth.

In the present invention, the extract may be used in the aforementioned content. In addition, as the fermented food can be used without limitation the above-described type, preferably may be used Cheonggukjang.

In the present invention, after mixing the extract with fermented food, it may further comprise the step of ripening.

At this time, the ripening can be made at a temperature of less than 10 ℃, salinity of 7% or more and less than pH 4.

Fermented foods are Enterococcus , which produces tyramine and the like after ripening at salinity of 40-45 ° C., less than 6% and pH 4 or higher. Since the growth of spp. is active, it is preferable to maintain the above-mentioned temperature, salinity and pH conditions in the above-described ripening, respectively.

Fermented food prepared by the present invention can improve the hygiene safety due to the reduction of tyramine, a mutagenic substance by the natural product extract, it is possible to improve the functionality. In addition, it is possible to isolate the antimicrobial substances by checking the antimicrobial components of Schizandra chinensis, plum, ume, rosemary and licorice.

1 is a graph showing the total number of bacteria, Enterococcus bacteria and Bacillus bacteria of the Cheonggukjang samples used in the embodiment of the present invention.
Figure 2 is a graph showing the tyramine content of the Cheonggukjang sample used in the embodiment of the present invention.
Figure 3 Enterococcus to produce tyramine It is a graph showing the growth characteristics against the pH of spp.
Figure 4 Enterococcus to produce tyramine It is a graph showing the growth characteristics against the temperature of spp.
Figure 5 Enterococcus to produce tyramine This graph shows the growth characteristics of salinity of spp.
Figure 6 Enterococcus producing tyramine It is a graph showing the growth characteristics against the pressure of spp.
7 is a graph showing the total bacterial counts of Cheonggukjang, Omija Cheonggukjang, Omae Cheonggukjang, Plum Plum Cheonggukjang, Rosemary Claim and Licorice Cheonggukjang.
8 is a graph showing the B. subtilis bacterial counts of Cheonggukjang, Omija Cheonggukjang, Omae Cheonggukjang, Plum Plum Cheonggukjang, Rosemary Claim and Licorice Cheonggukjang.
9 is a graph showing the E. faecium bacterial counts of Cheonggukjang, Omija Cheonggukjang, Omae Cheonggukjang, Plum Plum Cheonggukjang, Rosemary Claim and Licorice Cheonggukjang.
Figure 10 is a graph showing the tyramine content of Chungkookjang treated with herbal extracts.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. However, it is to be understood that the present invention is not limited to the disclosed embodiments, but may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

Example

1. Sample

Beans purchased Korean Baekdu in 2009, Schizandra chinensis, plums, plums and licorices were purchased at Gyeongdong Market. Rosemary was purchased at Cheonan Herb Farm.

Schisandra chinensis, plum, ume, licorice and rosemary were extracted with 70% ethanol and lyophilized to use as powder. Enterococcus spp. Has separated from the 13 kinds of Soybean (CA, CB, CC, CD , CE, CF, CG, CP, CQ, CR, JA, JB and JC) that can be easily purchased in Mart, Bacillus subtilus (KCTC 3014) was purchased from the Korean Collection for Type Cultures (KCTC).

2. Determination of Total Bacteria, Bacillus Bacteria, and Enterococcus Bacteria in Cheonggukjang

After collecting 1g of 13 kinds of Cheonggukjang (CA to JC) on the market, each plate was plated agar, Bacillus medium agar and Enterococcus agar to check the number of bacteria. Total bacteria, Bacillus bacteria and Enterococcus The number of bacteria is shown graphically in FIG.

As shown in Fig. 1, Cheonggukjang contains Enterococcus spp. Specifically, the total number of bacteria, Bacillus number and Enterococcus number in Cheonggukjang were 4.5 X 10 ⁸ to 2.54 X 10 ¹⁰ CFU / g, 2.9 X 10 ⁸ to 1.12 X 10 ¹⁰ CFU / g and ND (below 10 ¹ ) to 7.0 X, respectively. 10 ¹⁰ CFU / g.

3. Determination of Tyramine Content in Cheonggukjang

13 kinds of Cheonggukjang (CA to JC) on the market were collected 5g each, and the content of tyramine was confirmed by HPLC. The content of the tyramine is shown graphically in FIG.

* As shown in Figure 2, Cheonggukjang contains tyramine, specifically, the content of tyramine was 117.6 to 2467.8 mg / kg.

4. Identification and Identification of Tyranamine-producing Strains in Cheonggukjang

After spreading Cheonggukjang on Enterococcus agar and collecting various strains, it was dropped on Bover-cid's agar containing 0.5% tyrosine and qualitatively experimented with the change of color of the medium. Enterococcus The tyramine production ability of the spp. Thereafter, strains were identified using API 20 Strep.

Enterococcus spp. Tyramine (ppm) JB18 2777 JB20 2650 JB24 3106 CQ10 2685 CQ22 3010 CQ30 2723

Table 1 shows the identified Enterococcus spp. Strain and the tyramine generating ability of the strain. As shown in Table 1, it can be confirmed that the tyramine generating ability of Enterococcus spp. Is significantly high, it is possible to produce more than 2500mg / kg tyramine E nterococc us spp. Six strains can be identified. In addition, the identification of the strain was Enterococcus faecium .

5. High producing tyramine Enterococus Growth characteristics of faecium -pH, temperature, salinity and high pressure

pH was adjusted to pH 2.0, 2.5, 3.0, 3.5, 4.0, 7.0, 8.0, 10.0 and 12.0 of deMan Rogosa Sharp (MRS) liquid medium, respectively, and then inoculated with E. faecium for 24 hours at 37 ° C. Experiment.

The incubation temperature was 4, 15, 20, 25, 30, 37, 45, 50, 51, 52, 53, 54, 55, 60 and 65 after inoculation of E. faecium in deMan Rogosa Sharp (MRS) liquid medium. Experiment after incubation for 24 hours after adjusting to ℃.

Salinity was adjusted to 0, 5, 6, 7, 8, 9, 10 and 15% of deMan Rogosa Sharp (MRS) liquid medium, respectively, and then inoculated with E. faecium and incubated for 24 hours at 37 ° C. It was. The culture medium was placed in a spectrophotometer (Lambda 35, Perkin elmer, USA) and measured at 600nm.

In addition, the high pressure test was carried out in a deMan Rogosa Sharp (MRS) liquid medium incubated with E. faecium at 37 ° C for 24 hours to change the conditions of the high pressure machine (TOYO KOATSU Co., LTD, Japan) to 0, 15, 30, 45 and 60. The growth of E. faecium was confirmed in minutes.

Enterococcus at the pH, temperature, salinity and high pressure conditions The results of growth experiments of faecium are shown in FIGS. 3 to 6.

3 is a graph showing the growth characteristics for pH, Enterococcus faecium has excellent growth characteristics from pH 4 to pH 10.

4 is a graph showing growth characteristics with respect to temperature, Enterococcus faecium has excellent growth characteristics at 37-53 ℃.

5 is a graph showing the growth characteristics for salinity, Enterococcus Faecium has excellent growth characteristics at less than 6%, and growth is lowered at more than 7%.

In addition, Figure 6 is a graph showing the growth characteristics for high pressure, Enterococcus faecium has excellent growth characteristics for up to 60h at high pressure.

6. Antimicrobial Activity of Natural Extracts of E. faecium Producing Tyramine -Agar Diffusion Test (ADT), Minimum Inhibitory Concentration (MIC)

The natural extract was added dropwise to an 8 mm paper disk, dried, and placed on E. faecium smeared medium, followed by incubation at 37 ° C. for 24 hours. After incubation, the size of the transparent ring around the paper disk was measured, and the antimicrobial natural product was added dropwise to the deMan Rogosa Sharp (MRS) liquid medium to check the antimicrobial activity, and then cultured with E. faecium . Thereafter, the growth of E. faecium was confirmed and the minimum inhibitory concentration was measured.

Name of Medical Substance Extract Agar Diffusion Test (ADT) Minimum Inhibitory Concentration (MIC) 10 (mg / ml) 50 (mg / ml) 100 (mg / ml) MIC MIC 90 (mg / ml) (mg / ml) CON ND ND ND ND ND Schisandra ND 16 ± 0 19.5 ± 0.5 <5 <5 plum ND 17.5 ± 0.5 19.5 ± 0.5 <5 <5 Oh 9 ± 0 18.5 ± 0.5 20.5 ± 0.5 10 5 Rosemary 9 ± 0 11.5 ± 0.5 13.5 ± 0.5 <5 <5 licorice 13 ± 1 17 ± 0 17 ± 0 10 5

Table 2 shows the size and minimum inhibitory concentration of the paper disk ring by the concentration of natural products. As shown in Table 2, the ring size of the paper disk dropping 100 mg / ㎖ of Schisandra chinensis, plum, ume, rosemary and licorice extract was 19.5 ± 0.5 mm, 19.5 ± 0.5, 20.5 ± 0.5, 13.5 ± 0.5, 17.0 ± 0 and minimum inhibitory concentrations of Schisandra chinensis, plum and rosemary were below 5 mg / ml, and the minimum inhibitory concentrations of licorice and ume were 10 mg / ml.

7. Preparation of Cheonggukjang Added with Natural Extracts, Number of Bacteria and Tyramine

Steam and cool white beans, then extract natural (Schisandra chinensis, Plum, Ome, Rosemary and Licorice extract), E. faecium And B. subtilus (KCTC 3014) was incubated at 10 ° C. or lower after inoculation. And, when stored in the refrigerator 7% salt was added.

Confirmation of the number of bacteria and tyramine content of the finished Chunggukjang is the same as the measuring method described above.

7 to 9 show the total number of bacteria (Fig. 7), B. subtilis bacteria (Fig. 8) and E. faecium bacteria (Fig. 9) of Cheonggukjang, Omija Cheonggukjang, Omae Cheonggukjang, Maeyang Cheonggukjang, Rosemary Claims and Licorice Cheonggukjang, respectively. It is a graph.

As shown in FIG. 7 to FIG. 9, the Cheonggukjang containing the extracts of Schisandra chinensis, ume, plum and rosemary has a total bacterial count and B. subtilis. The number of bacteria was not different from that of general Cheonggukjang. However, the E. faecium bacteria were found to have a 10 ¹ to 10 ⁴ reduced bacteria compared to the general Cheonggukjang. In particular, Cheonggukjang containing Omae extract was found to have the lowest E. faecium bacteria.

In addition, Figure 10 is a graph showing the tyramine content of Cheonggukjang containing the extract of Schisandra chinensis, ume, plum and rosemary. As shown in FIG. 10, the tyramine content of the Chungkookjang containing the extract was reduced by 50 to 95%.

8. Sensory Evaluation of Cheonggukjang Containing Natural Extracts

Cheonggukjang containing natural extracts (Schisandra chinensis, ume, plum, licorice or rosemary) was subjected to sensory evaluation on 15 adults (male and female) and five points for fragrance, color, savory taste, sweetness, bitterness and total rating. Using the scale method, the rating was very good at 5 points, usually 3 points and very bad 1 point.

At this time, Cheonggukjang containing no natural extract was used as a control.

Sensory evaluation results are shown in Table 3 below.

Scent color A sweet taste sweetness bitter Overall Control group 3.53 ± 0.14 ^bc 4.13 ± 0.15 ^a 3.40 ± 0.19 ^b 2.47 ± 0.20 ^bd 2.30 ± 0.20 ^a 3.83 ± 0.18 ^b Schisandra 3.53 ± 0.19 ^bc 4.10 ± 0.15 ^a 3.13 ± 0.20 ^bc 2.20 ± 0.19 ^bc 3.03 ± 0.23 ^bc 3.33 ± 0.23 ^b Oh 3.87 ± 0.17 ^c 4.10 ± 0.15 ^a 3.33 ± 0.21 ^b 2.87 ± 0.21 ^d 2.43 ± 0.21 ^ac 3.90 ± 0.18 ^b plum 3.47 ± 0.18 ^bc 4.10 ± 0.16 ^a 3.03 ± 0.19 ^bc 2.50 ± 0.21 ^bd 2.70 ± 0.23 ^ac 3.57 ± 0.20 ^b licorice 3.20 ± 0.19 ^b 4.13 + 0.14 ^a 2.67 ± 0.19 ^c 3.77 ± 0.25 ^a 2.70 ± 0.24 ^ac 2.63 ± 0.26 ^a Rosemary 2.07 ± 0.20 ^a 3.87 ± 0.21 ^a 2.00 ± 0.17 ^a 1.83 ± 0.18 ^c 3.43 ± 0.23 ^b 2.30 ± 0.24 ^a

The data collected above were computed by using SPSS WIN 12.0 program, and one way ANOVA and Duncan multiple comparison verification method were used.

As shown in Table 3, natural extracts, especially Cheonggukjang containing Schisandra chinensis, Ome or plum, have similar values in aroma, color, savory taste, sweetness and bitterness as the general Cheonggukjang used as a control.

Claims (4)

Enterococcus spp. Comprising extracts of one or more substances selected from the group consisting of Schisandra chinensis, Plum, Schisandra chinensis and Rosemary as active ingredients . After mixing the growth inhibiting composition with fermented food,
Enterococcus spp. Method for producing fermented foods in which growth is suppressed.
The method of claim 1,
Fermented foods are enterococcus spp. Method for producing fermented foods in which growth is suppressed.
3. The method of claim 2,
The soy sauce is Enterococcus spp. Method for producing fermented foods in which growth is suppressed.
The method of claim 1,
Enterococcus spp. Comprising 0.1 to 5 parts by weight based on 100 parts by weight of Baekdu . Method for producing fermented foods in which growth is suppressed.

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