KR101451706B1 - Novel Acetobacter sp. SEA623-2, Citrus Mandarin vinegar and preparation method thereof - Google Patents

Abstract

The present invention relates to a novel Acetobacter sp. Strain SEA623-2, a mandarin vinegar, and a method for producing the same. The present invention relates to a mandarin vinegar rich in flavonoid components prepared using the above strain.
The tangerine vinegar of the present invention can provide a vinegar rich in flavonoid components such as naringenin and hesperetin, as well as a variety of health foods using the same, as compared with the case of using the strain used for the fermentation of acetic acid in the production of the conventional vinegar .

Description

SEA623-2 strain, a citrus vinegar, and a method for producing the same, which comprises a novel Acetobacter sp. SEA623-2, Citrus Mandarin vinegar and preparation method thereof}

More particularly, the present invention relates to a flavonoid-rich tangerine vinegar prepared using a novel strain and a method for producing the same.

In general, vinegar is used as a seasoning that takes up an important part in our daily life for a long time because it squeezes flavor of food with sour taste. These vinegar are divided into brewed vinegar (fermented vinegar) made by fermenting cereals, alcoholic beverages or fruit juices, and synthetic vinegar made from glacial acetic acid or acetic acid. Among them, the vinegar is divided into fruit vinegar and grape vinegar, and is divided into American cider vinegar fermented with apple juice, wine vinegar fermented with grape juice, UK fermented malt juice, It is deeply related to alcoholic beverages and cultivated fruits that are widely produced in many countries, such as malt vinegar in Germany and sake vinegar in Japan made from sake from sake. On the other hand, synthetic vinegar is obtained by diluting glacial acetic acid or acetic acid with water and adding amino acids or saccharides thereto, and it is mainly used in restaurants and the like. In recent years, the synthetic vinegar has been made using glacial acetic acid as a main ingredient, and thus it has been found that harmful substances are harmful to the human body. In recent years, the production of vinegar has become active.

On the other hand, tangerine contains a large amount of flavonoids, and such flavonoids can be used for various diseases such as liver disease, diabetes, rheumatoid arthritis, various cancers, analgesic, viral diseases, allergic diseases, vascular diseases such as arteriosclerosis, Lt; RTI ID = 0.0 & However, no effort has been made to develop a method for easily ingesting flavonoids rich in tangerine.

Accordingly, the inventors of the present invention have made intensive efforts to overcome the problems of the prior arts. As a result, they found new microorganisms and found that when tangerine vinegar was prepared, the content of flavonoid increased, especially the content of naringenin and hesperetin in flavonoid And thus the present invention has been completed.

The present invention for solving the above problems is to provide a strain of Acetobacter sp. SEA623-2, accession number KACC 91527P (Agricultural Genetic Resources Information Center), and a method for using this strain in the production of mandarin vinegar.

In addition, the present invention provides naringenin and / or tangerine vinegar having a high content of hesperetin using the strain SEA623-2.

The strains according to the present invention can rapidly convert (or digest) the glycoside flavonoid to the non-peptidic flavonoid, and thereby, health foods and medicines having excellent economic efficiency can be produced. In addition, the mandarin vinegar according to the present invention is useful for improving various diseases by containing flavonoid components contained in a large amount in mandarin orange in the vinegar during the production of vinegar vinegar, and can provide vinegar with enhanced flavor. Thus, it contributes to the consumption of mandarin orange, and it is not only harmless to human body but also can provide vinegar with excellent sugar-sugar efficiency as compared with synthetic vinegar produced by a chemical method.

FIG. 1 is a photograph of alcoholic fermentation of mandarin juice obtained in Example 2
2 is a schematic view of a process for producing a mandarin orange vinegar according to the present invention.
Fig. 3A shows a comparison of the results of the Acetobacter aceti) a picture and then culturing the strain KACC11978, B of FIG. 3 is a photograph of culture after SEA623-2 strain used in Example 3.
FIG. 4A shows the results of the measurement of the alcohol content change of the orange tangerine vinegar prepared in Experimental Example 2, and FIG. 5B shows the results of the measurement of the acid content change of the orange tangerine vinegar.
Figs. 5 and 6 are the results of HPLC (High Performance Liquid Chromatography) test for analyzing the functional components of the acetic acid fermentation broth of Experimental Example 3. Fig.
Figs. 7 and 8 show the results of the experiment conducted by the user of the citrus vinegar in Experimental Example 4.
9 is a nucleotide sequence 1 in which 18S rRNA of the strain SEA623-2 of the present invention was analyzed.

The term 'acetic acid fermentation broth' used in the present invention is prepared by fermenting acetic acid with a mixture of citrus juice with SEA623-2 strain (National Institute of Agricultural Science and Technology, Accession No .: KACC 91527P). Acetic acid fermentation broth itself may mean tangerine vinegar However, in order to clarify the present invention, before the liquid fermented with the strain SEA623-2 is diluted with carbonated beverage or water, it is defined as acetic acid fermentation broth.

Hereinafter, the present invention will be described in detail.

The present invention relates to a strain of Acetobacter sp. SEA623-2, accession number KACC 91527P (Agricultural Genetic Resource Information Center), which is characterized by comprising the nucleotide sequence shown in SEQ ID NO: 1. The strain is isolated from mandarin, and is characterized in rapidly converting glycosyl flavonoid into non-peptidic flavonoid. Thus, the strain can be used for the production of health foods and medicines by using the strain.

The present invention relates to a mandarina vinegar and is characterized in that it contains a high content of at least one flavonoid selected from naringenin and hesperetin.

The naringenin is a type of flavonoid, which has two benzene rings, which helps repair damaged DNA in cancer cells and is useful for the body known as an antioxidant component. And, the hesperetin is also a type of flavonoid, and is known as an antioxidant component.

The tangerine vinegar of the present invention is characterized in that the naringenin is contained in an amount of 10 ppm or more, preferably 1,500 to 3,000 ppm, more preferably 2,000 to 2,500 ppm, and the content of naringenin contained in the tangerine vinegar The acetic acid fermentation using SEA623-2 strain of the present invention is preferably about 10 days to 18 days, although there may be differences depending on acetic acid fermentation period and conditions in vinegar production.

The citrus vinegar of the present invention is characterized in that the above-mentioned hesperetin is 200 ppm or more, preferably 200 to 1,000 ppm, more preferably 300 to 1,700 ppm, and the content of the hesperetin contained in the citrus vinegar is There may be differences according to the fermentation period and conditions of acetic acid in the production of mandarin orange juice.

In addition, the naringenin and hesperetin are non-parasitic.

The tangerine vinegar of the present invention can be produced using the strain SEA623-2 in Acetobacter (National Institute of Agricultural Science and Technology, Accession No .: KACC 91527P).

Hereinafter, the manufacturing method will be described in detail.

The method of manufacturing a mandarin vinegar according to the present invention comprises: And a citrus extract containing a flavonoid, and acetic acid fermented with a mixture of citrus juice mixed with SEA 623-2 (National Institute of Agricultural Science and Technology, Accession No .: KACC 91527P) in Acetobacter.

The citrus juice fermentation solution is prepared by fermenting mandarin juice solution with alcohol to prepare a citrus fermentation broth, and mixing the water with water to prepare an appropriate alcohol concentration and acid content.

The citrus fruits used to obtain the mandarin orange juice are not particularly limited, but citrons cultivated in southern Europe and the like, lemons cultivated in the United States of America and the like, paragraphs cultivated in tropical regions and the like, Grape fruit, which is a variant of the paragraph cultivated in the United States of America, Florida, USA, sour orange cultivated in Europe and the like, citrus, kumquat, citron, hara bong, Citron, citrus, mandarin, sadou sensation, main texture, and tangerine. The citrus fruit juice preferably has a sugar content of 5 brix or more and an acidity of 0.2 to 0.8%, and preferably has a sugar content of 10 brix to 20 brix and an acidity of 0.2 to 0.6%.

The citrus fermentation broth is obtained by fermenting citrus juice with citrus-malomelo yeast (CMY-28 strain), preferably CMY-28 strain, with an alcohol concentration of 14-18% and an acid content of 0.5-1% 15 to 17% and an acid content of 0.6 to 0.8%. Here, the alcohol fermentation conditions are not particularly limited as fermentation conditions widely used in the art.

The fermented aqueous solution of citrus juice has an alcohol concentration of 5 to 10% and an acid content of 0.2 to 0.6%, preferably an alcohol concentration of 5 to 7% and an acid content of 0.3 to 0.5% Water may be mixed with the citrus fermentation broth fermented with alcohol to dilute it to the alcohol concentration and the acid content. When the fermentation aqueous solution of citrus juice is out of the above-mentioned alcohol concentration and acid content, acetic acid fermentation may not occur well, so that it is preferable to have an alcohol concentration and an acid content within the above range.

Preferably, the citrus mixture contains citrus juice fermentation aqueous solution and flavonoid-containing citrus extract at a volume ratio of 80 to 95: 5 to 20, more preferably 85 to 95: 5 to 15. The citrus vinegar of the present invention can be prepared by inoculating and culturing the above-mentioned citrus mixed solution with the strain SEA623-2 in Acetobacter and fermenting it with acetic acid. The acetic acid fermentation condition is not particularly limited as a fermentation condition widely used in the art, but it is preferable to cultivate the strain and to conduct acetic acid fermentation while supplying oxygen at 10 to 20 ° C.

And, the orange extract containing the flavonoid is extracted from Kilo-prep (eodydfid citrus flavonoid extraction method) in a citrus peel having a lot of flavonoid components.

The present invention is characterized in that the above-mentioned Acetobacter sp. Gel_SEA623-2 strain can be used for the production of mandarin vinegar containing a large amount of specific flavonoids, and said SEA623-2 strain is deposited in the microorganism bank of the Agricultural Genetic Resources Information Center Deposited on February 25, 2010 (Accession No .: KACC 91527P), can be obtained from the orange extract.

The tangerine vinegar of the present invention contains a large amount of naringenin and / or hesperetin, which are antioxidative components. The tangerine vinegar of the present invention can be used as a health food and a functional mainstream such as a salad dressing, a carbonated drink, a juice drink, .

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[ Example ]

Example 1 Acetic Acid Fermentation Strain Isolation

An initial citrus juice solution having a sugar content of 11.2 brix and an acidity of 0.4% was prepared. The citrus fruit juice was allowed to stand for 15 days at 15 ° C for 10 days or more through natural fermentation, and then cultured in a medium containing 1% ethanol-added gluconobacter oxydans (10% glucose, 1% yeast extract, 1.5% agar (pH 6.8) The isolate was named SEA623-2, and it was identified as an agronomic gene. The isolate was named as SEA623-2, (Accession No .: KACC 91527P) on February 25, 2010, and the gene sequence of this strain contains SEQ ID NO: 1 as shown in Fig.

Example 2: Preparation of citrus fermentation broth and alcohol fermentation

A citrus fruit juice having a sugar content of 11.2 brix and an acidity of 0.4% was prepared. The ratio of citrus fruit juice to water was 1: 2, and CMY (Citrus-malomelo yeast) -28 strain was inoculated. At this time, the amount of strains used was 1 × 10 ⁶ cells / ml.

The cells were incubated at 15 ° C for alcohol fermentation. The citrus fruit juice inoculated with CMY28 had an alcohol content of 16% and an acidity of 0.7% after 7 days. Fig. 1 is a photograph of alcoholic fermentation of mandarin orange juice.

Example 3: Preparation of acetic acid fermentation broth (mandarin vinegar)

The citrus fermentation broth prepared in Example 2 was diluted with water at a ratio of 1: 6 (v / v) to adjust the alcohol concentration to 6%, the acid content to 0.4%, and the flavor extract containing the citrus extract to 10% (v / v) to prepare a citrus mixture. The strain SEA623-2 is inoculated and cultured. Acetic acid fermentation broth was prepared by fermenting acetic acid while supplying oxygen to the bubble generator at 15 ℃. FIG. 2 shows a schematic diagram of a process for producing an acetic acid fermentation broth according to the present invention.

Comparative Example

Acetobacter aceti) was a pre-sale KACC11978 strains in the gene bank. In the case of the acetic acid fermentation according to Example 3, instead of the strain SEA623-2, the above Acetobacter aceti), and to perform the alcoholic fermentation and acetic acid fermentation according to Example 2 and Example 3 except that the strain was prepared KACC11978 acid fermentation broth.

Fig. 3 is a graph showing the results obtained after the cultivation of the strain SEA623-2 used in Example 3 and after the culture of Acetobacter aceti) a photograph showing the strains after the culture of KACC11978.

Experimental Example 1: Measurement of Alcohol and Acid Content of Acetic Acid Fermentation Solution

Alcohol and acid contents of the fermented acetic acid fermentation broths prepared in Example 3 and Comparative Example were measured. The alcohol content was measured by GC / MS at intervals of 2 days. The measurement was carried out by conversion to citric acid using a 0.1N titration method and the measurement results are shown in FIGS. 4A and 4B, respectively. As shown in FIG. 4, the alcohol content after the acetic acid fermentation according to Example 3 was not much different from that according to the comparative example, and the alcohol content was lowered to 1% or lower after 12 days, .

In addition, in the case of the acetic acid fermentation after inoculation of the strain SEA623-2 according to Example 3, the acid production rate was slowed by about 3 days, but the final acid production was not significantly different from the comparative example.

Experimental Example 2: Analysis of functional components of acetic acid fermentation broth

The HPLC (High Performance Liquid Chromatography) experiment was conducted to analyze the functional ingredient content of the acetic acid fermentation broth prepared in Example 3 and Comparative Example, and the results are shown in FIG. 5 and FIG. 6, respectively.

The HPLC analysis was performed using SCL-10A regulator, SPD-M10A UV spectrophotometer, LC-10AT liquid chromatography, FCV-10AL and DGU-14A (Shimadzu Scientific Co., Japan). The analytical column was eluted with a YMC C18 column (250 x 3.0 mm), mobile phase with 0.1% acetic acid / water and 100% MeOH under mobile phase conditions as shown in Table 1 below. The flow rate was maintained at 1 ml / min and detected at UV 280 nm.

minute 0.1% Acetic acid in Di-Water (%) Methanol (%) 12 60 40 19 60 40 30 0 100 33 0 100 35 80 20

5 and 6, in Example 3 and Comparative Example, the contents of glycerin and hesperidin in the form of glycosides were decreased, and the contents of naringenin and hesperetin in the form of non-parmeter were increased. In particular, when the naringenin (Fig. 6) was examined (Fig. 6), the conversion rate of acetic acid fermentation broth (Example 3) prepared by inoculating strain SEA623-2 was lower than that of the acetic acid fermentation broth of the comparative example and the rate of conversion of geranin and hesperidin to hesperetin 4 times faster, and sugar efficiency is better. Therefore, it is expected that it will be possible to produce foods that are very digestive.

Experimental Example 4: Consumer preference survey

The acetic acid fermentation broth of Example 3 and Comparative Example was put into a carbonated beverage to prepare a 5% by volume acetic acid component. That is, the citrus vinegar was added to the carbonated beverage so that the acetic acid component contained 5 vol%, respectively.

Each of the prepared citrus vinegar was used to examine the preference of the consumers of 40 university students in their twenties. The results are shown in FIG. 7 and FIG. 7 and 8, it was found that 47% of the citrus vinegar prepared using the acetic acid fermentation broth prepared according to Example 3 was good for taste and aroma, whereas the acetic acid fermentation broth prepared according to Comparative Example The tangerine vinegar produced by using only 27% and 15% of the taste and aroma was good.

In addition, when the citrus vinegar was prepared using the acetic acid fermentation broth prepared according to the comparative example, it was found that the number of people who answered that the taste and aroma was poor was 2.5 times or more higher than that of Example 3.

Experimental Example 5: Analysis of physiological characteristics and genetic tendency of strain SEA623-2

Acetobacter sp. In order to compare the biochemical properties of the strain SEA623-2, Acetobacter trophicus KACC 12236 (= IFO 16470) was purchased from the KACC, and Acetobacter xylinum ) KCCM 40407 (= IFO 3288) were purchased from KCCM, and the biochemical properties of these reference strains and the isolates were compared.

KACC
91527P ^{1 )} KACC
12236 ²⁾ KCCM
40407 ³⁾ ONPG
(β-galactosidase) - - - Arginine dehydrolaze - - - Li Xin
Decarboxylase - - - Ornithine
Decarboxylase - - - Using citric acid - - - Production of H ₂ S - - - Uraase - - - Tryptophan
Deaminase + - - Indole production - - - VP test + + + Gelatin liquefaction - - - Production of acid from glucose + + + Acid production from mannitol + - - Production of acid from inositol - - - Acid production from sorbitol + - - Production of acid from laminose + - + Acid production from Saccharose + - + From Delhi Bios
Generation of acid + + + Production of acid from amigalline - - - From arabinose
Generation of acid + + + NO ₂ production - + - N ₂ gas reduction - - - -, voice; + Positive
1) Acetobacter sp. SEA623-2
2) Acetobacter trophicus
3) Acetobacter xylinum

The strain SEA623-2 in Acetobacter showed a positive reaction with tryptophan deaminase in the VP test and showed a positive reaction in the fermentation with glucose, mannitol, sorbitol, rhamnose, saccharose, delhi bios and arabinose, No strain showing the same biochemical properties among the comparative strains of the genus Bacteria was identified.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

Agricultural Genetic Resources Information Center KACC91527P 20100225

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Claims (17)

delete delete delete delete delete delete delete delete delete Citrate juice fermentation solution and citrus extract containing 80: 95: 5 ~ 20 volume ratio of flavonoid were mixed with Acetobacter sp. SEA623-2 strain (National Institute of Agricultural Science and Technology, Accession No .: KACC 91527P) For 10 to 14 days at 10 ° C to 20 ° C to prepare an acetic acid fermentation broth,
Wherein the fermented aqueous solution of citrus juice comprises an alcohol concentration of 5 to 7% and an acid content of 0.3 to 0.5%, wherein the citrus juice fermentation solution comprises alcohol,
The citrus fermentation broth contains an alcohol concentration of 15 to 17% and an acid content of 0.6 to 0.8%
Wherein the nitric acid content of the acetic acid fermentation broth is 1,500 ppm to 2,000 ppm and the content of hesperetin is 200 ppm to 600 ppm. 11. The method of claim 10,
Further comprising the step of mixing the acetic acid fermentation broth with the carbonated beverage. delete delete 11. The method of claim 10, wherein the alcohol fermentation
Wherein the citrus vinegar is fermented with CMY (Citrus-malomelo yeast). delete delete delete

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