KR101981181B1 - Method for fermented Gastrodia elata BL using lactic acid bacteria - Google Patents

Abstract

The present invention relates to a fermented chicken meat using lactic acid bacteria, and more particularly, to a method for fermenting a horse chestnut using lactic acid bacteria to remove odor and unpleasantness of the horse meat. According to the present invention, the content of p-cresol which causes odor peculiar to the gum is reduced during the fermentation process, and the preference degree is improved. In addition, the fermented horse chestnut having the odor removed showed excellent antioxidative effect, and the useful components, the total flavonoid contents, and the total polyphenol contents in the fermented chestnut were significantly higher than those of the fermented non-fermented horse, Fermented horse chestnut can be usefully used as a raw material for high value-added food or health functional food.

Description

Method for manufacturing fermented cheonma using lactic acid bacteria {Method for fermented Gastrodia elata BL using lactic acid bacteria}

The present invention relates to a fermented cheonma using lactic acid bacteria, and specifically relates to a method of removing off-flavor of cheonma by fermenting cheonma using lactic acid bacteria.

Chunma is also called by other names such as Shincho (神草) and Jeongpungcho (定風草). In the Shinnongbonchogyeong, Cheonma is classified as a heavy product, and its medicinal properties are known as a medicinal material that is perfectly poisonous.

The clinical efficacy of Chunma is widely recorded in various herbal literatures including Bonchogangmok and Donguibogam. It is known to be effective against adult diseases such as high blood pressure, headache, paralysis, neurological diseases, diabetes, and symptoms such as stress and fatigue. . Most of the ingredients contained in Cheonma are phenolic compounds, and gastrodin, phenolic glycosides, sulfur-containing phenolic compounds, organic acids, sugars, and beta-sitosterol have been reported separately. Components such as (sterol), cholesterol, p-hydroxylbenvyl alcohol and vanillin are also known.

It is known that the water content of Saengcheonma is 81.2%, and the general components of the freeze-dried sample contain 6.21% crude protein, 1.50% crude fat, 2.55% crude ash, and 89.74% carbohydrates. Studies on Cheonma include investigation of general ingredients and functionality of Cheonma (Jeong Hyeon-seo, Ji Geun-eok, Korean Journal of Food Science and Technology, 28(1), 53-57, 1996), analysis of food ingredients of Cheonma and changes in characteristics according to drying method (Booyong Lee , Hyunseon Choi, Jinbong Hwang, Journal of the Korean Society of Food Science and Technology, 34(1), 37-42, 2002), A study on the composition analysis and in vitro biological activity of Chunma extract (Taesu Kang, Youngjun Kong, Hyejeong Kwon, Byunggon Choi, and Jeongki Hong , Park Yong-Gil, Journal of the Korean Mycology Society, 30(2), 136-141, 2002). Chunma began to be cultivated artificially in domestic farms from 78 years ago, and recently showed an oversupply phenomenon that exceeds the demand for medicinal purposes, so there is an urgent need to improve the use of Chunma. Due to restrictions on raw materials, the use of processed foods has been restricted. However, as the regulations were lifted from September 1, 2000 and can be used as food raw materials, basic data necessary for the development of processed foods are urgently needed.

In addition, Chunma has a peculiar off-flavor (pig feces, swine barn-like) that is unsuitable for use as a food or health functional material, but its main causative component is not specifically known.

Accordingly, the present inventors have completed the present invention by identifying the main causative components for off-flavor of Cheonma and developing a fermentation method for reducing the content thereof.

It is an object of the present invention to provide a method for producing fermented cheonma using lactic acid bacteria.

Another object of the present invention is to provide a fermented cheonma from which off-flavor is removed according to the method of the present invention.

In order to solve the above problems, the present invention is a method for producing fermented cheonma using lactic acid bacteria,

Increasing the cheonma powder;

Saccharifying the increased cheonma powder;

Sterilizing the saccharified cheonma powder; And

It provides a method for producing a fermented cheonma using lactic acid bacteria comprising; inoculating and fermenting lactic acid bacteria in the sterilized cheonma powder.

The lactic acid bacteria may be Lactobacillus plantarum VL-1, Pediococcus inopinatus BK-3 (accession number KACC91859P), Lactobacillus sakei C-11 (accession number KACC91863P).

The lactic acid bacteria may be Lactobacillus sakei C-11 (accession number KACC91863P).

In the fermenting step, the fermentation period may be 1 to 8 days.

The present invention provides a fermented cheonma from which off-flavor is removed by the method according to the present invention.

According to the present invention, the content of p-cresol, which causes off-flavor peculiar to Cheonma during fermentation, is reduced, thereby improving preference. In addition, since it was confirmed that the fermented cheonma from which off-flavor was removed exhibits excellent antioxidant effect, the useful components, total flavonoid content, and total polyphenol content in the fermented cheonma were significantly higher than that of unfermented cheonma. Fermented cheonma can be usefully used as a raw material for high value-added foods or health functional foods.

Figure 1 shows the GC obtained by analyzing the volatile fraction obtained by steam distillation of Cheonma (top) and the fraction (middle) expressing a strong off-flavor among the fractions obtained by silica gel chromatography and the standard p-cresol (bottom). It is a degree.
2 is a diagram comparing mass spectrum (MS) for GC analysis of Chunma and standard p-cresol; A: MS of p-cresol-like components detected in Chunma, B: MS of standard p-cresol.
3 is a diagram showing the result of preparing a calibration curve for quantifying p-cresol.
4 is a diagram showing the results of HPLC analysis of useful components in a control (cheonma powder) sample; 1: gastrodin, 2: 4-hydroxybenzyl alcohol.
5 is a diagram showing the results of HPLC analysis of useful components in a control (cheonma powder) and a fermented cheonma sample fermented for 8 days; 1: gastrodin, 2: 4-hydroxybenzyl alcohol.
6 is a partial nucleotide sequence of the 16s ribosomal RNA gene of Lactobacillus sakei C-11 strain (SEQ ID NO: 1).
7 is a partial nucleotide sequence of the 16s ribosomal RNA gene of Pediococcus inopinatus BK-3 strain (SEQ ID NO: 2).
8 is a partial nucleotide sequence of the 16s ribosomal RNA gene of Lactobacillus plantarum VL-1 strain (SEQ ID NO: 3).

The present invention is a method for producing fermented cheonma using lactic acid bacteria, comprising: increasing cheonma powder; Saccharifying the increased cheonma powder; Sterilizing the saccharified cheonma powder; And inoculating and fermenting the sterilized cheonma powder with lactic acid bacteria.

The lactic acid bacteria may be Lactobacillus plantarum VL-1, Pediococcus inopinatus BK-3 (accession number KACC91859P), Lactobacillus sakei C-11 (accession number KACC91863P), preferably Lactobacillus sakei C-11 (accession number KACC91863P). I can.

The Lactobacillus plantarum VL-1 was registered in NCBI under the name of Lactobacillus plantarum JBARES VL-1 under the registration number KX622695, and the Pediococcus inopinatus BK-3 (accession number KACC91859P) was registered as Pediococcus inopinatus BK-1 in the gene bank.

In one embodiment of the present invention, as a result of analyzing the p-cresol content in fermented cheonma, the p-cresol content was lower in the fermented cheonma sample compared to the control (cheonma powder), and in particular, in the fermented cheonma sample using C-11 The p-cresol content was the lowest.

The term "p-cresol" is a compound also named 4-methylphenol, and produces a characteristic and strong odor even at very low concentrations. In one embodiment of the present invention, a large peak coinciding with the standard p-cresol was detected in the fraction containing a strong off-flavor among the volatile fraction obtained by steam distillation of Cheonma and the silica gel chromatography fraction. In addition, as a result of comparing the odor characteristics of the p-cresol standard product and the fraction containing a strong off-odor by sensory evaluation, it was confirmed that the characteristic off-odor expressed in the fraction containing a strong off-odor had a odor similar to p-cresol. . Accordingly, the present invention provides a method of manufacturing a fermented cheonma, which can be easily eaten by a user because such off-flavor is removed.

The cheonma powder may include edible grains or powders of crops in addition to cheonma powder, and may preferably include any one or more powdered materials selected from the group consisting of germinated brown rice, black rice, adlay and purple sweet potatoes. Not limited.

With respect to the total weight of the powdered material, the cheonma powder may be 30 to 100% by weight, but is not limited thereto.

In the fermenting step, the fermentation period may be 1 to 8 days, preferably 1 to 4 days, and more preferably 1 to 2 days.

In one embodiment of the present invention, it was confirmed that the p-cresol content was significantly lower in the fermented cheonma powder sample fermented for 0 to 8 days compared to the non-fermented control (cheonma powder).

The present invention provides a fermented cheonma from which off-flavor is removed by the method according to the present invention.

In one embodiment of the present invention, the fermented cheonma from which off-flavor is removed by the method according to the present invention is gastrodin, 4-hydroxybenzyl alcohol, which are useful components in cheonma, total polyphenols and total The flavonoid content increased during the fermentation period, and the antioxidant effect was found to be significantly higher than that of unfermented Chunma.

Accordingly, the fermented cheonma from which off-flavor is removed according to the present invention can be usefully used as a raw material for high value-added foods or health functional foods.

In addition, the off-flavor removal method according to the present invention has a remarkable effect of increasing the useful components and thus the antioxidant effect while removing off-flavor in the cheonma.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely describe the present invention to those of ordinary skill in the art.

<Example 1> Isolation and identification of off-flavor components in Cheonma

1-1. Separation of volatile components

The volatile components of lyophilized cheonma are SDE (Likens & Nickerson type simultaneous steam distillation and extraction) method according to Schultz et al. (Isolation of volatile components from a model system, J. Agric. Food Chem. 25:446, 1997). Separated by. Specifically, 100 g of a sample and 2 L of distilled water were placed in a round-bottom flask, connected to an SDE device, and distilled for 2 hours. At this time, the solvent for recovering volatile components was n-pentane: diethyl ether. (1:1) 50 mL of the mixed solution was used.

1-2. Off-flavor fraction

In order to fractionate the volatile components separated by the SDE method in 1-1, silica gel column chromatography fractionation was performed. Specifically, after loading a volatile solvent into a column (3.0 cm x 20 cm) filled with 70 g of silica gel (230-400 mesh), n-hexane: diethyl Ether (50 ~ 95: 5 ~ 50, Table 1) 50 mL of a mixed solution or 50 mL of ethyl acetate was used to gradually increase the polarity, eluting and fractionation. Thereafter, the presence or absence of off-flavor was investigated through sensory evaluation, and the fraction in which off-flavor peculiar to Cheonma was strongly expressed was analyzed by GC (gas chromatography) method.

1-3. Sensory evaluation

Sensory evaluation personnel consist of 3 male and female college students over 20 years of age, and a small amount of each silica gel column chromatography fraction obtained in 1-2 was wetted in paper (smelling paper), and then the solvent was volatilized and presented to the sensory evaluation personnel. Fragrance characteristics were described by (profile test).

As a result, in the fraction eluted with a mixture of n-hexane:diethyl ether (80:20), a strong off-flavor peculiar to Chunma was expressed (Table 1).

Solvent ratio Fraction
number Odor profile Hex:ethyl ether (95:5) 1-7 Weak odor, not contained characteristic odor Hex:ethyl ether (90:10) 8-14 Slightly odor, waxy, green Hex:ethyl ether (80:20)
15-17 Off-odor, phenolic, medical 18-20 Strong off-odor, strong swine barn-like Hex:ethyl ether (60:40) 22-26 Weak odor Hex:ethyl ether (50:50) 27-32 Acidic, lactone-like Etyl acetate (100) 33-40 Odorless

1-4. Gas chromatography (GC) analysis

The volatile component fractions obtained by steam distillation by the SDE method and each fraction obtained by silica gel column chromatography obtained in 1-2 and 1-3, respectively, were analyzed by the GC method. Specifically, a fused silica capillary column (Supelcowax 10 fused silica capillary column, 25m x 0.32mm) was used, and the column temperature was heated from 50°C to 230°C at a rate of 2°C per minute, and then maintained at 230°C for 20 minutes. The injector and detector were set to 250°C, and the carrier gas was analyzed in a split ratio (= 20:1) mode using nitrogen gas (1.0 mL/min).

The volatile fraction obtained by steam distillation of Cheonma and the n-hexane:diethylether (80:20) fraction showing a strong off-odor peculiar to Cheonma among the results of silica gel column chromatography, p-cresol, 4-methylphenol ) (Wako, Japan) and a large peak was detected (Fig. 1). In addition, as a result of comparing the odor characteristics of each fraction with the standard p-cresol through sensory evaluation, the characteristic odor (swine barn-like) expressed in the n-hexane:diethyl ether (80:20) fraction was p-cresol. It had a smell similar to The odor threshold value of p-cresol is 1 ppb, which is known to generate a characteristic and strong odor even at very low concentrations.

<Example 2> Fermentation cheonma manufacturing method

Lactobacillus plantarum VL-1 (Figure 8, SEQ ID NO: 3) (NCBI registration, registration number KX622695), Pediococcus inopinatus BK-3 ( Pediococcus inopinatus BK-1 (gene bank), accession number KACC91859P, Figure 7, SEQ ID NO: 2), Lactobacillus sakei C-11 (Accession No. KACC91863P, Fig. 6, SEQ ID No. 1) was cultured in MRS medium at 30° C. for 2 days to obtain a culture solution.

Water was added in an amount of 0.5 times the total amount of cheonma powder (e.g., when the total amount of powdered material is 500g, the amount of water is 250ml). After mixing and kneading so that the ingredients and water were well mixed, it was steamed in a steamer for 40 minutes and increased. The steamed dough was put in an appropriate container, and malt extract was added (added equal to the total amount of the ingredients) to make it like porridge. After saccharification in a 65° C. constant temperature water bath with a water bath for 3 to 5 hours, sterilization was performed at 121° C. for 15 minutes. After cooling down to 30℃ or less, inoculate lactic acid bacteria (VL-1 (Lactobacillus plantarum ), BK-3 ( Pediococcus inopinatus ) and C-11 ( Lactobacillus sakei ) in a clean bench (clean room), respectively (inoculation amount is 1 of the total amount of the material). %, Example: When the total amount of the material is 500g, 5 ml of lactic acid bacteria culture solution was inoculated), fermented at 30° C. The fermented cheonma was dried at room temperature and powdered, each of the lactic acid bacteria of VL-1, BK-3 or C-11. The fermented cheonma powder sample fermented using was named as VL-1 sample, BK-3 sample, or C-11 sample below, and the non-fermented cheonma powder sample was used as a control.

<Example 3> Analysis of p-cresol in fermented cheonma

3-1. Preparation of calibration curve for p-cresol

50 mg of the standard p-cresol was taken and dissolved in 25 mL of ethyl acetate, and 105.6 mg of the internal standard o-cresol (Wako, Japan) was taken and dissolved in 100 mL of ethyl acetate to prepare a calibration curve (Table 2 and Fig. 3).

Solution for preparing calibration curve One 2 3 4 5 6 p-Cresol mL 0.02 0.05 0.1 0.2 0.3 0.4 o-Cresol(ISTD) mL 0.5 0.5 0.5 0.5 0.5 0.5 Ethyl acetate mL 0.48 0.45 0.4 0.3 0.2 0.1 Total volume mL 1.0 1.0 1.0 1.0 1.0 1.0 p -Cresol μg (A) 40 100 200 400 600 800 o -Cresol μg (B) 528 528 528 528 528 528 A/B 0.0758 0.189 0.379 0.758 1.136 1.515

3-2. Sample extraction

2.00 g of a powdery sample of Example 2 was taken, placed in a 50 mL conical tube, 10 mL of 1N-HCl solution, 0.5 mL of o-cresol solution used as an internal standard (containing 528 μg of o-cresol), and an extraction solvent After adding 29.5 mL of ethyl acetate, extraction was performed by shaking for 1 day and night (12 hours) at 120 rpm at room temperature. The extracted sample was centrifuged (4,500 rpm, 20 minutes), 20 mL of an ethyl acetate layer was taken into a 50 mL conical tube, 6 g of anhydrous sodium sulfate was added, shaken for 3 hours, and then filtered.

3-3. GC analysis

The filtrate was concentrated under reduced pressure, dissolved in 1.0 mL of acetone, and analyzed by GC method. Specifically, a fused silica capillary column (Supelcowax 10 fused silica capillary column, 25m x 0.32mm) was used, and the column temperature was maintained at 130°C for 3 minutes, and then heated to 230°C at a rate of 3°C per minute, and then 15 at 230°C. Held for a minute. The inlet and detector were set at 250°C, and the carrier gas was analyzed in a split ratio (= 20:1) mode using nitrogen gas (1.0 mL/min).

As a result of analyzing the p-cresol content in fermented cheonma, the p-cresol content in the fermented cheonma powder sample was lower than that of the non-fermented control (cheonma powder). In particular, the p-cresol content in the C-11 sample was the highest. Was low (Table 3).

Effective period (days) p -Cresol content (μg/g, dry base) VL-1 BK-3 C-11 Control 168.7 - - 2 128.6 107.9 76.3 4 118.1 89.9 92.3 6 130.3 127.7 90.5 8 124.4 103.5 90.2

<Example 4> Analysis of the content of useful ingredients in fermented cheonma

After adding 20 ml of 80% methanol to 2.00 g of a powdery sample of Example 2, extraction was performed by heating at 70 for 30 minutes, centrifugation (4,500 rpm, 20 minutes), and 20 ml of 80% methanol was added to the residue. Extracted by heating at 70° C. for 30 minutes and centrifuged (4,500 rpm, 20 minutes). The extracts were combined, concentrated under reduced pressure, dissolved in 5.0 ml of 70% methanol, filtered, and analyzed by HPLC (high performance liquid chromatography). Specifically, Eclipse XDB C18RS (4.5 x 25 cm, id 5 μm) was used as the column and PDA detector (220 nm) was used as the detector. As the solvent, 0.1% phosphoric acid aqueous solution (solvent A) and 100% methanol (solvent B) were used, and the gradient conditions were 85% (A) and 15% (B) (0-2 minutes), 45% (A) and 55% (B) (2-35 minutes), 85% (A) and 15% (B) (35-40 minutes), 85% (A) and 15% (B) (45- 50 minutes), the flow rate was 0.8 mL/min, and the injection volume was 10 μL. Each component was quantified after preparing a standard curve using a standard product.

As a result of analyzing the content of useful components in the standard and Chunma samples analyzed by the HPLC method, as shown in FIGS. 4 and 5, gastrodin (4-(hydroxymethyl)phenyl-β-D-glucopyranoside) and 4-hydroxybenzyl alcohol in Chunma are contained. It was confirmed that These ingredients have been found to be closely related to the physiological activity of Chunma.

In addition, the results of analyzing the content of useful components in the samples VL-1, BK-3, and C-11 were shown in Tables 4, 5, and 6. In the case of the VL-1 sample, gastrodin tended to increase as fermentation proceeded. The BK-3 sample also showed a similar tendency to that of the VL-1 sample, but the C-11 sample did not show a certain tendency in terms of the composition change due to fermentation.

Ingredient content in VL-1 sample (μg/g) Control Effective period (days) 2 4 6 8 Gastrodin 1280.2 2890.5 3007.2 3300.6 3306.9 4-Hydroxybenzyl alcohol 4069.1 3330.5 3303.3 3365.7 3356.4

Ingredient content in BK-3 sample (μg/g) Control Effective period (days) 2 4 6 8 Gastrodin 1280.2 1578.8 2157.6 2761.3 2320.7 4-Hydroxybenzyl alcohol 4069.1 3549.7 4309.0 4068.6 3944.7

C-11 Component content in sample (μg/g) Control Effective period (days) 2 4 6 8 Gastrodin 1280.2 2462.1 2185.0 2591.7 2276.1 4-Hydroxybenzyl alcohol 4069.1 4814.3 3697.1 3739.7 4773.9

<Example 5> Antioxidant effect of fermented cheonma, total flavonoid and total polyphenol content assay

5-1. DPPH (2,2-diphenyl-1-picrylhydrazyl) analysis

DPPH analysis was performed on the fermented cheonma powder sample and the control (cheonma powder) sample prepared in Example 2. DPPH radical scavenging activity was measured by modifying the method of Blois et al. (1958). Specifically, 0.4 ml of 0.1M Tri was added to 0.1 ml of a sample, mixed, 0.5 ml of a DPPH solution prepared in 500 μM was added in a 96-well plate, mixed well, reacted at room temperature for 20 minutes, and then 517 The absorbance was measured at nm. The negative control was measured by putting distilled water instead of the sample. DPPH radical scavenging activity was calculated by the following equation and expressed as a percentage (%), and is shown in Table 7. BHT (dibutyl hydroxy toluene) and BHA (butylated hydroxy anisole) were used as positive controls.

DPPH radical scavenging activity (%) =

[1-(Experimental area absorbance/Negative control area absorbance)] × 100

As a result, VL-1 sample showed 83.61% scavenging activity on the 6th day of fermentation, BK-3 showed 76.72% of scavenging activity on the 4th day of fermentation, and C-11 showed 82.88% scavenging activity on the 6th day of fermentation. I got it. From the second day of fermentation, it was confirmed that all fermented Cheonma powder samples showed superior DPPH scavenging activity compared to Cheonma powder or positive controls BHT and BHA.

Effective period (days) VL-1 BK-3 C-11 Control BHT BHA 2 73.27 71.23 70.06 51.42 68.28 68.10 4 75.71 76.72 66.67 6 83.61 72.17 82.88 8 69.99 63.00 70.02

5-2. FRAP(Ferric reducing antioxidant power) analysis

FRAP analysis was performed on the fermented cheonma powder sample and the control (cheonma powder) sample prepared in Example 2. Specifically, as a reaction solution, 300 mM acetate buffer (pH 3.6), 10 mM 2,4,6-tripyridyl-s-triazine dissolved in 40 mM hydrochloric acid (TPTZ, T1253, C ₁₈ H ₁₂ N ₆ , MW312.33 ) And 20mM FeCl ₃ (F7134, MW 162.20) were prepared, and the acetate buffer, TPTZ solution and FeCl ₃ solution were mixed at 10:1:1 (v/v/v) to pre-react at 37°C for 15 minutes. I put it. 0.15 ml of the sample and 2.85 ml of the pre-reacted FRAP reagent were mixed, reacted for 15 minutes, and absorbance was measured at 593 nm using a microplate reader (Biorad 3055, Sweden). The content (ppm) was obtained from the standard curve prepared using _{FeSO 4} ·7H _{2 O, and is shown in Table 8.}

As a result, on the 6th day of fermentation, the VL-1 sample was 532.7 ppm, the BK-3 sample was 375.0 ppm, and the C-11 sample was 584.6 ppm. I got it.

Effective period (days) VL-1 BK-3 C-11 Control 2 344.3 329.7 529.2 254.9 4 412.7 371.1 348.9 6 532.7 375.0 584.6 8 334.4 298.1 581.6

5-3. Total flavonoid content analysis

The total flavonoid content was analyzed for the fermented cheonma powder sample and the control (cheonma powder) sample prepared in Example 2. The total flavonoid content was mixed by adding 0.15 mL of diethylene glycol to 0.1 mL of a sample according to the Davis variant (2010), followed by adding 20 μl of a 1N-NaOH solution, followed by mixing and reacting at 37° C. for 1 hour, followed by 420 nm. The absorbance was measured at. The total flavonoid content was calculated from the calibration curve prepared using rutin and expressed in ppm, and is shown in Table 9.

As a result, the VL-1 sample was found to be 108.92 ppm on the 6th day of fermentation, the BK-3 sample was 78.06 ppm on the 4th day of fermentation, and the C-11 sample was 99.40 ppm on the 6th day of fermentation. It showed superior total flavonoid content compared to the powder sample.

Effective period (days) VL-1 BK-3 C-11 Control 2 68.68 64.40 70.15 40.97 4 77.25 78.06 65.61 6 108.92 77.00 99.40 8 68.30 63.53 67.85

5-4. Analysis of total polyphenol content

The total polyphenol content was analyzed for the fermented cheonma powder sample and the control (cheonma powder) sample prepared in Example 2. The total polyphenol content was determined by the Folins-Denis method. Specifically, 0.8 mL of distilled water was added to 0.2 mL of the sample, 0.1 mL of 2N Folin-ciocalteu reagent was added, mixed, allowed to stand for 3 minutes, and 0.7 mL of distilled water was added to 0.2 mL of a _{2% Na 2} CO _{3 solution.} . After reacting the mixture for 1 hour, the absorbance was measured in Abs750 using a spectrophotometer (UV/VIS spectrometer, Japan), and the total polyphenol content (ppm) was calculated from the standard curve prepared using the standard tannic acid. It is shown in 10.

As a result, the VL-1 sample was 193.5 ppm on the 6th day of fermentation, the BK-3 sample was 159.5 ppm on the 4th day of fermentation, and the C-11 sample was 204.2 ppm on the 4th day of fermentation. It showed superior total polyphenol content compared to the powder sample.

Effective period (days) VL-1 BK-3 C-11 Control 2 132.7 110.7 134.6 97.12 4 158.0 159.5 204.2 6 193.5 151.7 200.3 8 130.1 116.6 115.4

National Academy of Agricultural Sciences KACC91863P 20131108 National Academy of Agricultural Sciences KACC91859P 20131108

<110> Chonbuk-do Agricultural Research and Extension Services <120> Method for fermented Gastrodia elata BL using lactic acid bacteria <130> DHP16-576 <160> 3 <170> KoPatentIn 3.0 <210> 1 <211> 1474 <212> RNA <213> Unknown <220> <223> 16S ribosomal RNA gene of Lactobacillus sakei C-11 <400> 1 atgcgggtgc tataatgcag tcgaacgcac tctcgtttag attgaaggag cttgctcctg 60 attgataaac atttgagtga gtggcggacg ggtgagtaac acgtgggtaa cctgccctaa 120 agtgggggat aacatttgga aacagatgct aataccgcat aaaacctaac accgcatggt 180 gtagggttga aagatggttt cggctatcac tttaggatgg acccgcggtg cattagttag 240 ttggtgaggt aaaggctcac caagaccgtg atgcatagcc gacctgagag ggtaatcggc 300 cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtagg gaatcttcca 360 caatggacga aagtctgatg gagcaacgcc gcgtgagtga agaaggtttt cggatcgtaa 420 aactctgttg ttggagaaga atgtatctga tagtaactga tcaggtagtg acggtatcca 480 accagaaagc cacggctaac tacgtgccag cagccgcggt aatacgtagg tggcaagcgt 540 tgtccggatt tattgggcgt aaagcgagcg caggcggttt cttaagtctg atgtgaaagc 600 ctttcggctc aaccgaagaa gtgcatcgga aactgggaaa cttgagtgca gaagaggaca 660 gtggaactcc atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag 720 gcggctgtct ggtctgtaac tgacgctgag gctcgaaagc atgggtagca aacaggatta 780 gataccctgg tagtccatgc cgtaaacgat gagtgctagg tgttggaggg tttccgccct 840 tcagtgccgc agctaacgca ttaagcactc cgcctgggga gtacgaccgc aaggttgaaa 900 ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa 960 cgcgaagaac cttaccaggt cttgacatcc tttgaccact ctagagatag agctttccct 1020 tcggggacaa agtgacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg 1080 ttaagtcccg caacgagcgc aacccttatt actagttgcc agcatttagt tgggcactct 1140 agtgagactg ccggtgacaa accggaggaa ggtggggacg acgtcaaatc atcatgcccc 1200 ttatgacctg ggctacacac gtgctacaat ggatggtaca acgagttgcg agaccgcgag 1260 gtttagctaa tctcttaaaa ccattctcag ttcggattgt aggctgcaac tcgcctacat 1320 gaagccggaa tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct 1380 tgtacacacc gcccgtcaca ccatgagagt ttgtaacacc caaagccggt gaggtaaccc 1440 ttcggggagc cagccgtcta atgtgacaga tgtt 1474 <210> 2 <211> 1228 <212> RNA <213> Unknown <220> <223> 16S ribosomal RNA gene of Pediococcus inopinatus BK-3 <400> 2 gctagttggt gagataaagg cccaccaagg ctgtgatacg tagccgacct gagagggtaa 60 tcggccacat tgggactgag acacggccca gactcctacg ggaggcagca gtagggaatc 120 ttccacaatg gacgaaagtc tgatggagca acgccgcgtg agtgatgaag gctttagggt 180 cgtaaaactc tgttgntgga gaagaacgtg tgtgagagta actgctcatg cagtgacggt 240 atccaaccag aaagccacgg ctaactacgt gccagcagcc gcggtaatac gtaggtggca 300 agcgttatcc ggatttattg ggcgtaaagc gagcgcaggc ggtttcttaa gtctaatgtg 360 aaagccttcg gcttaaccga agaagtgcat tggaaactgg gaaacttgag tgcagaagag 420 gacagtggaa ctccatgtgt agcggtgaaa tgcgtagata tatggaagaa caccagtggc 480 gaaggcggct gtctggtctg taactgacgc tgaggctcga aagcatgggt agcgaacagg 540 attagatacc ctggtagtcc atgccgtaaa cgatgaatgc taagtgttgg agggtttccg 600 cccttcagtg ctgcagctaa cgcattaagc attccgcctg gggagtacga ccgcaaggtt 660 gaaactcaaa agaattgacg ggggcccgca caagcggtgg agcatgtggt ttaattcgaa 720 gctacgcgaa gaaccttacc aggtcttgac atcttctgct aacctaagag attaggcgtt 780 cccttcgggg acagaatgac aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt 840 tgggttaagt cccgcaacga gcgcaacccc tattattagt tgccagcatt aagttgggca 900 ctctagtgag actgccggtg ataaaccgga ggaaggtggg gacgacgtca aatcatcatg 960 ccccttatga cctgggctac acacgtgcta caatggacgg tacaacgagt tgcgagaccg 1020 cgaggttaag ctaatctctt aaaaccgttc tcagttcgga ctgcaggctg caactcgcct 1080 gcacgaagtt ggaatcgcta gtaatcgcgg atcagcatgc cgcggtgaat acgttcccgg 1140 gccttgtaca caccgcccgt cacaccatga gagtttgtaa cacccaaagc cggtggagta 1200 accttcggga gctagccgtc taagtgac 1228 <210> 3 <211> 1431 <212> RNA <213> Unknown <220> <223> 16S ribosomal RNA gene of Lactobacillus plantarum VL-1 <400> 3 tcgacgaact ctggtattga ttggtgcttg catcatgatt tacatttgag tgagtggcga 60 actggtgagt aacacgtggg aaacctgccc agaagcgggg gataacacct ggaaacagat 120 gctaataccg cataacaact tggaccgcat ggtccgagct tgaaagatgg cttcggctat 180 cacttttgga tggtcccgcg gcgtattagc tagatggtga ggtaacggct caccatggca 240 atgatacgta gccgacctga gagggtaatc ggccacattg ggactgagac acggcccaaa 300 ctcatacggg aggcagcagt agggaatctt ccacaatgga cgaaagtctg atggagcaac 360 gccgcgtgag tgaagaaggg tttcggctcg taaaactctg ttgttaaaga agaacatatc 420 tgagagtaac tgttcaggta ttgacggtat ttaaccagaa agccacggct aactacgtgc 480 cagcagccgc ggtaatacgt aggtggcaag cgttgtccgg atttattggg cgtaaagcga 540 gcgcaggcgg ttttttaagt ctgatgtgaa agccttcggc tcaaccgaag aagtgcatcg 600 gaaactggga aacttgagtg cagaagagga cagtggaact ccatgtgtag cggtgaaatg 660 cgtagatata tggaagaaca ccagtggcga aggcggctgt ctggtctgta actgacgctg 720 aggctcgaaa gtatgggtag caaacaggat tagataccct ggtagtccat accgtaaacg 780 atgaatgcta agtgttggag ggtttccgcc cttcagtgct gcagctaacg cattaagcat 840 tccgcctggg gagtacggcc gcaaggctga aactcaaagg aattgacggg ggcccgcaca 900 agcggtggag catgtggttt aattcgaagc tacgcgaaga accttaccag gtcttgacat 960 actatgcaaa tctaagagat tagacgttcc cttcggggac atggatacag gtggtgcatg 1020 gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaaccctta 1080 ttatcagttg ccagcattaa gttgggcact ctggtgagac tgccggtgac aaaccggagg 1140 aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac acgtgctaca 1200 atggatggta caacgagttg cgaactcgcg agagtaagct aatctcttaa agccattctc 1260 agttcggatt gtaggctgca actcgcctac atgaagtcgg aatcgctagt aatcgcggat 1320 cagcatgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga 1380 gtttgtaaca cccaaagtcg gtggggtaac cttttaggaa ccagccgcct a 1431

Claims (1)

As a method for producing a fermented horse meat using lactic acid bacteria,
Mixing the powder of rhizome with water at a weight ratio of 2: 1 and then adding the mixture;
Saccharifying the donated glucose-containing powder;
Sterilizing the saccharified chewing gum powder; And
And a step of inoculating 1% by weight of Lactobacillus sakei C-11 (accession number KACC91863P) to the sterilized chitosan powder and fermenting the mixture for 2 days to 4 days, and p-cresol A method for producing a fermented chicken meat having reduced content.

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