KR100722980B1 - Method for production of houttuynia decoction fermentation product using Leuconostoc sp. - Google Patents

Abstract

The present invention is a genus Luuconostoc sp. Isolated from dried persimmon The present invention relates to a fermentation broth production of Echochocho extract using a strain, and more particularly, to a method for producing a fermentation broth of Echochocho extract containing yeast extract as a carbon source and a sugar as a carbon source by using G5 bacteria of Lukonostok.

Eoseongcho, Dried Persimmon, Lukonostok

Description

Method for producing fermented broth of Echochocho extract using Luconosstock strain {Method for production of houttuynia decoction fermentation product using Leuconostoc sp.}

1 is a genus Luconostok isolated ( Leuconostoc sp.) In the present invention 16s rDNA sequence of the strain.

Figure 2 is a genus Luconostok isolated ( Leuconostoc sp.) In the present invention The phylogeny of the strain.

The present invention relates to a method of preparing a fermented broth of Echo. More specifically, the present invention relates to a method of preparing fermented broth of Echochocho using Leuconostoc sp.

Echo vinegar has potent antibacterial activity derived from decanoyl acetaldehyde, diuretic action derived from curdrin, etc., antioxidant activity and immune enhancement action derived from phenolic substances, etc. It shows excellent pharmacological activity, and the interest is increasing as a functional food material. However, Eoseongcho has a strong taste and aroma, so it is not suitable for eating raw juice or extract as it is.

Thus, various inventions have been disclosed to improve the irritant fishy and unique taste peculiar to Eochocho. For example, Eochocho fermented beverage composition is disclosed in Korean Patent Application Publication No. 10-2005-0043325 (May 11, 2005). Is disclosed. The present invention provides a fermented beverage that is easy to drink by removing the odor peculiar to Eochocho, but only performs natural fermentation without specific reference to the fermentation bacteria, and the odor unique to Echochocho is substantially added by sugar or honey to be added. It is only offset.

In addition, Korea Patent Office Registration No. 10-0369638 (January 29, 2003) in order to eliminate fishy fishy fish smell by using a natural vegetable material by mixing aloe, corn starch, sugar, honey, etc. Although fermented beverages are disclosed, there is no specific reference to the fermentation bacteria, but only the fishy taste of fish vinegar is offset by the added sugar component.

As described above, in the case of Eochocho beverages prepared by adding sugar to the Echochocho juice for the purpose of improving functionality and sensuality, or by adding various additives including oligosaccharides to the dried Echochocho hot water extract, the products are uniform. Difficulty in obtaining alcohol, alcohol is excessively generated and classified as liquor under the State Tax Law, so that the law requires a liquor license to distribute legally.

Accordingly, an object of the present invention is to provide a method for preparing a fermentation broth of Echochocho by separating the strains suitable for fermentation of Echochocho extract and inoculating fermentation of Echochocho extract with appropriate composition.

In order to achieve the above object, the present invention is a method for producing a fermented broth extract by fermenting Echo extract, Leukonostoc sp. Provided is a method of preparing fermented broth of Echochocho, characterized by using KACC 91156P strain.

Hereinafter, the present invention will be described in more detail.

The present invention is Leuconostoc sp. Provide the KACC 91156P strain. The Leuconostoc G5 strain is isolated from dried persimmon and is generally a microorganism secured to be applicable to food fermentation. The strain is aerobic anaerobic, gram-positive, non-motile, streptococcus, showing the culture characteristics of the microorganisms of the genus Lukonostok, and identified by the phylogenetic analysis based on 16S rDNA sequence to confirm that the bacteria of the genus Lukonostok (Fig. 1 and 2, which was named Leuconostoc G5 and was deposited with the Institute of Agricultural Biotechnology on March 7, 2005 (accession number: KACC 91156P).

On the other hand, when the strain is inoculated into Eochocho extract to induce lactic acid fermentation, odor of peculiar to Echochocho due to lactic acid fermentation is removed, and the flavor of fermented Echochocho extract is made.

In the present invention, the eochochocho extract can be used as a raw vinegar extract raw water if it is extracted by the usual method by hot water extraction, preferably 1 hour by adding tap water at a rate of 4L per 100g of dried eochochocho. After boiling and filtering, the filtrate was evaporated under reduced pressure, and the extract of Echochocho was concentrated under reduced pressure so that the solid content was 3 Brix.

On the other hand, the present invention can be a Preferably seed culture prior to the lactic acid bacteria of the incubation, the seed culture is Lu Pocono stock (Leuconostoc) isolated from the invention G5 KACC 91156P strains MRS medium 40 ml This example 250 ml Erlenmeyer flask It is good to obtain by inoculating in and incubated for 2 days while shaking at 120 rpm in a 25 ℃ shaking incubator.

On the other hand, the lactic acid fermentation of Echochocho extract of the present invention is preferably carried out as follows. First, purified water is added to the extract of Eoseongcho so that the solid content is adjusted to 2 Brix, and 1% (w / v) of sugar is added to form a fermentation broth, followed by sterilization at 80 ° C. for 15 minutes. Lactic acid bacteria species culture inoculated 1% (v / v) and incubated at 25 ℃ for 3 days to proceed with lactic acid fermentation. In this case, when the sugar is added, the amount of lactic acid bacteria growth and lactic acid production is much increased as compared with the case where it is not, and preferably, the amount of sugar added is 3% (w / v).

On the other hand, when the nitrogen source is added, lactic acid fermentation of the Echo extract is enhanced. When yeast extract is added as a nitrogen source, the lactic acid bacteria growth and lactic acid production are the highest, and as the amount of added yeast extract increases, the growth and lactic acid production of lactic acid bacteria is increased. Is increased.

On the other hand, as a result of investigating the lactic acid fermentation pattern of Eochochocho extract according to fermentation temperature, the cell proliferation rate is the fastest at 35 ℃ as the temperature increases, but the final lactic acid production is the highest at 30 ℃.

Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited to the following examples.

Example 1-Lactic Acid Bacteria Separation

Lactobacillus Screening

The collected kimchi, vegetables, and fruits were mixed with sterile saline in a weight ratio of 1: 1 and homogenized with a mixer to use as a fungal sample. The fungal samples were diluted appropriately and plated on MRS agar medium, which is a medium for separating lactic acid bacteria with platinum, and the plate was incubated for 72 hours at 30 ° C incubator. Purely isolated lactic acid bacteria were inoculated with sterilized sugar 1% (w / v) containing Echochocho extract, and after incubation for 72 hours at 25 ° C. in a thermostat, the taste and aroma were examined sensually to select and isolate the excellent strains.

Finally, the isolated strain was isolated from dried persimmon and named G5 and selected as the fermented strain of Eochochocho extract of the present invention. Selected strains were stored subcultured in MRS agar medium.

Microbiological Characteristics of Isolated Strains

The isolated G5 strain was inoculated in MRS agar medium and incubated at 30 ° C. for 2 days to form a milky white opaque colony with a diameter of 1 to 1.5 mm, and was observed as a typical streptococcus microscopically. Response was observed to be gram positive.

16S rDNA sequences were analyzed to clarify which genus G5 strains were included (see FIG. 1). This sequence was subjected to phylogenetic analysis using the 16S rDNA base sequence of the related species suspected of being bacteriologically derived from the US GenBank database and the CRUSTAL W software. As a result, as shown in Figure 2 it can be seen that the strain belonging to the genus ( Leuconostoc genus). Therefore, the G5 strain was named as Leuconostoc G5 as a lactic acid bacterium belonging to Leuconostoc sp. And it was deposited on March 7, 2005 at the Institute of Agricultural Biotechnology (Accession Number: KCTC 91156P).

On the other hand, experiments that reveal the microbiological characteristics of Cappuccino and Sherman (Cappuccino, James G. and Sherman, Natalie, MICROBIOLOGY, A LABORATORY MANUAL, 4th ed.The Benjamin / Cumming Publishing Inc., Menlo Park, California, 1996 ) And Yoon et al . (Yoon et al ., Int. J. Sys. Bacteriol. 48, 833-837, 1998).

Example 2 Lactic Acid Fermentation

Preparation of Eoseongcho Extract

4 L of tap water was added to 100 g of dried fish vinegar, and the mixture was boiled for 1 hour and filtered. Thereafter, the filtrate was evaporated under reduced pressure, concentrated under reduced pressure so as to have a solid content of 3 Brix, to prepare a stock solution of the extract.

Lactobacillus Species Culture

Example 1 A base Pocono separated from the stock (Leuconostoc) G5 KACC 91156P strains MRS medium 40 ml is 2 days and inoculated into a 250 ml Erlenmeyer flask containing and shaken at 120 rpm at 25 ℃ shaking incubator culturing of Houttuynia cordata extract, lactic acid Species cultures were made for fermentation.

Lactic Acid Fermentation of Estuary Extracts by Sugar Addition

Purified water was added to the extract of Eoseongcho so that the solid content was adjusted to 2 Brix, and 1% (w / v) of sugar was added to form a fermentation broth and sterilized at 80 ° C. for 15 minutes. The lactic acid bacteria species culture was inoculated with 1% (v / v), and the lactic acid fermentation was performed in a stationary culture at 25 ° C. for 3 days.

Compared to the case where no sugar was added, it was found that lactic acid bacteria growth and lactic acid production were much increased, thereby promoting lactic acid fermentation (Table 1). The amount of lactic acid produced was calculated by counting the colonies formed by smearing the fermentation broth with 0.1N-NaOH and converting the amount of NaOH consumed into lactic acid and the number of viable cells diluted on a properly diluted sample on MRS agar medium.

Effect of Sugar Addition on Lactic Acid Fermentation pH Lactic acid (g / l) Viable cell count (cfu / ml)  No sugar added 4.75 29.3 1.03 x 10 ⁷  1% sugar added 4.03 47.3 5.20 x 10 ⁷

Lactic Acid Fermentation of Estuary Extracts by Addition of Nitrogen Sources

As a result of proceeding the lactic acid fermentation by adding several nitrogen sources to 0.1% (w / v) in the Echo extract extracted with the 1% (w / v) sugar was as shown in Table 2. As shown in Table 2, when yeast extract was added as a nitrogen source, lactic acid bacteria growth and lactic acid production were the highest.

Effect of Nitrogen Source on Lactic Acid Fermentation Nitrogen source pH Lactic acid (g / l) Viable cell count (cfu / ml)  No nitrogen source added 4.02 47.0 6.27 x 10 ⁷  Yeast extract added 3.96 53.1 1.05 x 10 ⁸  Add malt extract 4.01 48.4 4.05 x 10 ⁷  Peptone addition 4.01 49.1 3.65 x 10 ⁷  Soyton addition 4.01 47.9 3.43 x 10 ⁷

Lactic Acid Fermentation of Estuary Extracts with Different Sugar Concentrations

As a result of the lactic acid fermentation by adding the sugar concentration differently to the yeast extract added with 0.1% (w / v) yeast extract as a nitrogen source, it was as shown in Table 3. As shown in Table 3, the addition of 3% (w / v) of sugar showed the highest lactic acid bacteria growth and lactic acid production.

Effect of Sugar Concentration on Lactic Acid Fermentation Sugar Concentration (% w / v) pH Lactic acid (g / l) Viable cell count (cfu / ml)  One 4.01 49.5 3.15 x 10 ⁸  2 3.95 56.3 6.90 x 10 ⁸  3 3.89 65.3 1.27 x 10 ⁹  4 3.93 65.2 1.21 x 10 ⁹  5 3.92 64.5 1.18 x 10 ⁹

Lactic Acid Fermentation of Estuary Extracts with Different Yeast Extract Concentrations

As a result of proceeding the lactic acid fermentation by adding the yeast extract differently as a nitrogen source to the extracted Echoseongcho added sugar 3% (w / v) as shown in Table 4 below. As shown in Table 4, as the amount of yeast extract added increased, the growth of lactic acid bacteria and the amount of lactic acid increased.

Effect of Yeast Extract Concentration on Lactic Acid Fermentation Yeast Extract Concentration (% w / v) pH Lactic acid (g / l) Viable cell count (cfu / ml)  0 4.11 50.4 3.01 x 10 ⁸ 0.5 3.88 86.4 1.80 x 10 ⁹  One 3.91 102.6 2.11 x 10 ⁹  2 3.98 135.9 1.31 x 10 ¹⁰  3 4.05 155.7 5.01 x 10 ¹⁰  4 4.12 157.5 3.00 x 10 ¹⁰

Lactic Acid Fermentation of Echinacea Extracts with Different Fermentation Temperatures

The fermentation of lactic acid fermentation of Echo extract containing 3% (w / v) sugar and 0.5% (w / v) yeast extract was carried out at 25 ° C, 30 ° C, and 35 ° C by varying the temperature, respectively. It was as Table 6 and Table 7. As shown in the table, the cell proliferation rate was the fastest at 35 ℃ as the temperature increased, but the final lactic acid production was the highest at 30 ℃.

Lactic acid production and cell proliferation at 25 ℃ Fermentation time (hours) pH Lactic acid (g / l) Viable cell count (cfu / ml)  0 4.71 36.9 2.3 x 10 ⁷ 12 4.53 52.2 1.9 x 10 ⁸ 24 4.05  71.1 3.8 x 10 ⁸ 36 3.88 85.5 9.1 x 10 ⁸ 48 3.80 91.4 2.9 x 10 ⁹ 60 3.80 92.7 1.4 x 10 ⁹ 72 3.97 93.1 1.3 x 10 ⁹

Lactic Acid Production and Cell Proliferation According to Fermentation at 30 ℃ Fermentation time (hours) pH Lactic acid (g / l) Viable cell count (cfu / ml)  0 4.71 36.9 2.3 x 10 ⁷ 12 4.30 60.8 7.4 x 10 ⁸ 24 3.98  90.9 9.7 x 10 ⁸ 36 3.93 103.5 1.2 x 10 ⁹ 48 3.89 103.9 1.6 x 10 ⁹ 60 3.89 104.4 3.3 x 10 ⁸ 72 3.86 106.7 2.2 x 10 ⁸

Lactic Acid Production and Cell Proliferation at 35 ℃ by Fermentation Fermentation time (hours) pH Lactic acid (g / l) Viable cell count (cfu / ml)  0 4.71 36.9 2.3 x 10 ⁷ 12 4.36 58.1 7.1 x 10 ⁸ 24 3.97 76.5 6.9 x 10 ⁹ 36 3.86 89.1 7.0 x 10 ⁸ 48 4.03 90.1 5.0 x 10 ⁸ 60 3.99 90.1 4.8 x 10 ⁸ 72 4.01 90.1 1.1 x 10 ⁸

Lactic acid fermentation solution of the extract of Echo vinegar prepared according to the method of the present invention is characterized by excellent functionality due to the functionality imparted by the functionality and lactic acid fermentation compared to conventional echo extract.

Claims (4)

In the method of producing a fermented broth fermented by the Echo extract, Leuconostoc sp. Isolated from dried persimmons as spawn Method for producing a fermented broth of Echochocho characterized by using the KACC 91156P strain. The method of claim 1, wherein Method of producing a fermented broth of Echochocho characterized by adding 3% (w / v) sugar to the medium. The method of claim 1, wherein Yeast extract fermentation broth characterized in that the yeast extract is added to the medium as a nitrogen source. The method of claim 1, wherein Method for producing a fermented broth of Echochocho characterized in that the fermentation temperature is carried out at 30 ℃.

Images