KR101984400B1 - Process for producing antioxidant and anti-fatigue efficacy of mulberry vinegar - Google Patents

Abstract

The present invention relates to a method for producing a plum fermentation vinegar using an alcohol fermentation broth and an acetic acid fermentation broth, and an antioxidant and anti-fatigue effect when the fermented vinegar is consumed.
The method for producing the fermented vinegar of the present invention comprises mixing 0.5 to 8 parts by weight of a plum concentrate and 15 to 40 parts by weight of a concentrate with 100 parts by weight of purified water, adjusting the sugar content to 10 to 20 ° Brix, And fermenting acetic acid to ferment the acetic acid to produce the fermented vinegar of acetic acid. The fermenting alcoholic fermentation strain haromyces cerevisiae is used. Acetobacter aceti, which is inoculated into the alcoholic fermentation broth, is used. Wherein the fermented broth is fermented for 6 to 12 days while inoculating the fermented broth with the fermented broth and incubated at a temperature of 25 to 37 DEG C at a temperature of 150 to 300 rpm to produce an acetic acid fermented broth, And the mixture is heated and sterilized at a temperature of 80 ° C for 5 to 20 minutes, followed by centrifugation and filtration to prepare a fermented vinegar.

Description

[0001] The present invention relates to a method for producing an antioxidative and antipyroflocculant,

More particularly, the present invention relates to a method for producing ginger vinegar with antioxidant and anti-fatigue effect, which comprises culturing Saccharomyces ceresivisiae KCCM 11306 (YM broth culture) as an alcohol strain in an amount of 30 And then cultivating Acetobacter aceti KCCM 12654 (mannitol broth cultivation) at 30 ° C for 48 hours to prepare an antioxidative and anti-fatigue effect.

Plum ( Prunus mume ) is a fruit of plum tree which is widely produced in Korea. It is rich in minerals such as fibrin, free sugar, calcium and iron, and also contains many organic acids such as succinic acid, citric acid, malic acid and tartaric acid It has been used as a medicinal herb for the effects of roots, leaves, flowers and fruits of plum in the oriental medicine and the private sector for the effects of dryness, mud, geography, geomorphism, poisoning, detoxification, vomiting and fever. Recently, it has been proved that it is effective against aging prevention, hangover and constipation, and it is known that there is an antibacterial effect against induced bacteria such as typhoid fever, cholera and food poisoning.

Vinegar is a typical seasoning, both east and west, and is followed by a lot of Korean liquor. The main ingredient of vinegar is acetic acid, which contains volatile or nonvolatile organic acids, sugars, amino acids and esters, and has a unique aroma and sour taste. These vinegar ingredients are known to have effects such as prevention of adult diseases such as arteriosclerosis, hypertension, cholesterol lowering effect, and body fat reduction.

The vinegar is classified into vinegar, vinegar, or synthetic vinegar made by diluting acetic acid with water, mainly made from grains and fruits.

Recently, as the food culture has improved along with the economic growth, the awareness of health has been increasing, and research on natural vinegar has been actively carried out using a variety of natural fruits. However, Patent Publication No. 10-1518391 and Korean Patent Laid-Open Publication No. 2015-0095292.

Korean Patent Publication No. 10-1518391 relates to a method for preparing a fermented starch of mulberry by mixing pectinase for smooth acetic acid fermentation at the beginning of fermentation and replacing the liquid fructose with a natural raw material, Publication No. 2015-0095292 discloses a method for fermenting a fermented food belonging to a fermented food belonging to a fermented food belonging to a fermented food such as a soy sauce, a kimchi, a fermented food, a fermented vinegar, a lactic acid bacteria food and the like by extracting and culturing (including an active culture) Next, by mixing a food raw material with the liquid microorganism and preparing a useful microorganism containing the active ingredient of the food raw material, the sugar microorganism containing the active ingredient of the food raw material is administered with sugar, yeast, etc. to the alcohol fermented product obtained by alcohol fermentation The present invention relates to a method for producing a fermented vinegar obtained by fermenting acetic acid to obtain a fermented acetic acid by administering microorganisms and acetic acid bacteria containing a raw material effective ingredient All.

However, the present invention aims to establish the optimum conditions of the plum vinegar using the main ingredient of the plum, and to provide the antioxidant and anti-fatigue effect thereof. Fatigue can be defined as a condition in which a person can not perform a temporary activity, and the treatment and prognosis are unclear. The accumulation of fatigue induces symptoms of decreased resistance, aggravation of disease, It lowers personal work and productivity, which causes economic losses on a national level.

The above-mentioned prior art is different from producing the fermented vinegar by lowering the intake of the liquid fructose or by using the fermented acetic acid using the useful microorganism, and producing the fermented vinegar for providing the antioxidant and anti-fatigue effect as in the present invention.

Korean Registered Patent No. 10-1508391 (Feb. Korean Patent Laid-Open Publication No. 2015-0095292 (Aug. 21, 2015)

It is an object of the present invention to provide a method for producing a plum fermentation vinegar using an alcohol fermentation broth and a fermented broth as a concentrate of a mussel concentrate and a concentrate.

It is another object of the present invention to provide a method for producing a plum fermentation vinegar by using an alcohol fermentation broth and an acetic acid fermentation broth in the concentration of a mussel concentrate and an apple concentrate.

Another object of the present invention is to provide an antioxidant and anti-fatigue effect when the fermented vinegar is consumed.

In order to accomplish the above object, the present invention provides a process for preparing a fermented vinegar, comprising mixing 0.5 to 8 parts by weight of a plum concentrate and 15 to 40 parts by weight of a concentrate with 100 parts by weight of purified water, adjusting the sugar content to 10 to 20 ° Brix, An alcohol fermentation broth is inoculated and fermented to prepare an alcohol fermentation broth, and acetic acid bacteria are inoculated in the alcohol fermentation broth to ferment the acetic acid.

10 to 30 parts by weight of an apple concentrate may be used instead of 15 to 40 parts by weight of the above concentrate, or 3 to 10 parts by weight of one or more sugar in honey or sugar may be used.

The alcohol fermentation strain of the present invention, haromyces cerevisiae , is inoculated into the mixture of the above concentrate and the concentrate and fermented at 25 ° C to 37 ° C for 6 to 10 days. The alcohol fermentation broth is inoculated with Acetobacter aceti, Fermented for 6 to 12 days while shaking at a temperature of 37 ° C at 150 to 300 rpm to prepare an acetic acid fermentation broth. Instead of the above concentrated concentrate, an apple concentrate, honey, sugar or the like may be mixed and fermented. The acetic acid fermentation broth is heated and sterilized at a temperature of 65 ° C to 80 ° C for 5 to 20 minutes, followed by centrifugation and filtration to prepare a plum vinegar.

The present invention relates to a method for producing a plum fermented vinegar by mixing a concentrated liquid of a mussel or an apple concentrate or a sugar such as honey or sugar, followed by alcohol fermentation and acetic acid fermentation to produce a plum fermented vinegar. Inorganic phosphoric acid, and lactic acid.

In addition, the intake of plum vinegar is metabolized in the body to increase the accumulation of glycogen, which directly plays a role in the energy production through the corresponding process in the body, the amount of glycogen directly affects the exercise capacity.

Malondialdehyde, an indicator of lipid peroxidation, and glutathione peroxidase, an antioxidative enzyme, were found to be inhibited by lipid peroxidation and the activity of antioxidant enzyme, GPx, And the plum fermented vinegar provides antioxidant effect.

FIG. 1 is a graph showing changes in alcohol content and change in sugar content according to alcohol fermentation time after mixing the concentrate of the present invention with the concentrate of the present invention.
FIG. 2 is a graph showing changes in alcohol content and acetic acid content according to the acetic acid fermentation time after mixing the concentrate of the present invention with the concentrate of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings.

In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

FIG. 1 is a graph showing change in alcohol content and change in sugar content according to alcohol fermentation time after mixing of the concentrate of the present invention and the concentrate of the present invention. FIG. And the change of the alcohol content and the acetic acid content according to the amount of water. Acetic acid fermentation using the alcohol fermentation conditions was performed by fermenting Acetobacter aceti (KCCM 12654), which is a host of acetic acid bacteria, at 30 ° C and 200 rpm.

Plum ( Prunus mume ) which is the main material of the present invention uses fruit, and roots, leaves and flowers have been used as traditional Chinese medicine in herbal medicine.

Plum is a round fruit that usually ripens green from the end of May to the middle of June. It is originated in China. It has been used for about 3,000 years from the ancient Chinese Mongolian Materia Medica, In Korea, it has been reported to have been used as a medicinal plant in Korea since the beginning of the Three Kingdoms period. Its typical efficacy has been to lower the heat of the lungs, eliminate thirst, reduce edema, ), Detoxification, and hemostasis (hemostasis).

Especially, plum is a typical alkaline food which has the effect of improving the constitution which prevents the acidification of blood and constitution caused by the excessive consumption of meat and the favorite food of the modern people, and maintains the body to be weakly alkaline when consumed steadily. It is also said that poison of food, poison of blood, poison of water, poison of water is removed, and three poison is removed, and picric acid which is contained in plum acts as a poisonous substance decomposition, It can be said to be effective. It promotes the secretion of digestive juices, improves digestion defects, and treats gastrointestinal disorders. It regulates excessive secretion of gastric acid and is effective for overeating and stomachache.

  1. Preparation of plum concentrate

In the method of manufacturing the mash concentrate of the present invention, the flesh of the flesh is cooked and washed with water before the flesh is discolored, the moisture on the surface of the flesh chamber is removed, and the seeds are separated from the flesh chamber. Amygdalin, a glycosylated glycoside of two molecules of glucose, is contained in the nucleus of the amniotic fluid of the cheonggwamjil, which produces hydrogen cyanide (cleavage-HCN), two molecules of glucose and benzoaldehyde by the enzyme Emulisin.

A primary enzyme digestion step is carried out to extract the juice from the juice obtained by removing the hard seed nucleus and heat the fresh juice produced by the juice to inactivate the enzyme activity. The primary digestion step decomposes the starch and pectin contained in the pulp-free flesh to facilitate juicing. That is, it is heated at a temperature of 55 ° C to 70 ° C for about 8 to 25 hours, and then rapidly heated at a high temperature of 100 ° C for 5 minutes to inactivate the enzyme activity. Heating the fresh broth at a temperature of 55 ° C to 70 ° C for about 8 to 25 hours is intended to inactivate the enzyme while concentrating the optimum concentration.

The fresh juice of the cheongmilm which was heated in the primary enzyme decomposition step was cooled to 30 ° C, and the cooled juice of the cheongmiljang was filtered and heated at 90 ° C to 100 ° C for 5 to 15 hours to obtain plums of 50 ° Brix to 70 ° Brix A concentrate is prepared . In order to maintain the flavor and aroma of the fresh juice juice at optimal temperature for 90 to 100 ° C for 5 to 15 hours in the fresh juice juice heated in the primary enzyme decomposition step, Lt; / RTI > filter.

2. Preparation of concentrate

The pear is a fruit of a deciduous perennial plant belonging to the pears (Pyrus). It is a representative alkaline food which is beneficial to our body. Its moisture content is 85 ~ 88% and its main ingredient is carbohydrate. The sweetness of the sugar is 10 ~ 13% and the protein content is about 0.3%. It is not much different from other fruits, but it has a high dietary fiber content and excellent constipation and dressing effect. Also, the excretion of polycyclic aromatic hydrocarbons . ≪ / RTI > Vitamin B and C are abundant in the abdomen, and it contains antioxidant, anti-cancer and anti-inflammatory effects because it contains a large amount of polyphenols such as chlorogenic acid, catechin, epicatechin and arbutin and flavonoids such as quercetin and leuteolin. In addition, polyphenols in the diet are excellent in blood cholesterol lowering, immune function enhancement, cancer cell growth inhibition and antioxidant activity, and leuteolin in flavonoids has the effect of stopping bronchitis, sputum and cough. Quercetin has excellent effect on thyroid related diseases I have.

The husks and seeds of the ripe pear were removed and the flesh of the pear was heated at a temperature of 50 ° C to 90 ° C for 15 to 30 hours to concentrate at a concentration of 60 ° Brix to 80 ° Brix and filtered to prepare a concentrate do. If the temperature at the time of concentration is 50 DEG C or less, the concentration time is excessively uneconomical, and if the temperature is 90 DEG C or higher, the taste and the smell of the boil disappear.

3. Preparation of apple concentrate

The peel and seed parts of the ripe apple are removed, and the flesh of the apple is heated at a temperature of 50 to 90 DEG C for 10 to 25 hours, concentrated to a concentration of 50 to Brix to 70 Brix , and filtered to prepare an apple concentrate . If the temperature at the time of concentration is 50 DEG C or lower, the concentration time is excessively uneconomical, and if the temperature is 90 DEG C or higher, the fragrance of the apple will be lost.

 4. Alcohol Fermentation Strain

Alcohol fermentation strains are cultured on YM plate medium and cultured in YM liquid medium (Table 1) at 28 ° C to 32 ° C for 40 to 55 hours to be used as a master in Saccharomyces cerevisiae (KCCM 11306). Culturing the alcohol fermentation strain at 28 ° C to 32 ° C is because the activity of the strain Saccharomyces cerevisiae is optimal at the temperature.

[Table 1] Liquid medium composition of alcohol strain

YM liquid medium Composition component ratio(%) Malt extract 0.3 Yeast extract 0.3 Dextrose 1.0 Peptose 0.5

5. Acetic acid bacteria

Acetobacter aceti (KCCM 12654) is cultivated in a mannitol broth liquid medium composition of Table 2 shown below at 29 ° C to 31 ° C for 40 to 55 hours and used as a seed. This is because the activity of the strain Acetobacter aceti is optimal at the above temperature when the cultured acetic acid bacteria is cultured at 29 ° C to 31 ° C.

[Table 2] Liquid medium composition of acetic acid strain

Liquid medium Composition component ratio(%) Yeast Extract 0.5 Mannitol 2.5 Peptose 0.3

6. Preparation of fermented vinegar with antioxidant and anti-fatigue effect

0.5 to 8 parts by weight of the 50% Brix to 70 Brix of the plum concentrate and 15 to 40% by weight of the concentrate of 60% Brix to 80% Brix were mixed with 100 parts by weight of purified water to adjust the sugar content to 10-20 ° Brix , alcohol fermentation strain haromyces cerevisiae (KCCM 11306) when the immunization by fermentation during the 25 ℃ ~ 37 ℃ 6 ~ 10 il alcohol content range of the resulting fermentation broth is a 5 to 10%. When the fermentation is performed at a temperature of 25 ° C or lower when fermenting, the activity of the alcoholic fermentation microorganism is lowered, which makes economical fermentation difficult. When the fermentation is performed at a temperature of 37 ° C or higher, the activity of the microorganism is lowered and the fermentation of alcohol is delayed .

When the plum was mixed with the purified water, 10 to 30 parts by weight of an apple concentrate of 50 ° Brix to 70 ° Brix was mixed in place of the concentrate, and the sugar content was adjusted to 10 to 20 ° Brix. Then, an alcohol fermentation strain, haromyces cerevisiae (KCCM 11306) When inoculated and fermented at 25 ° C to 37 ° C for 6 to 10 days, the alcohol content of the obtained fermentation broth is 5 to 10%.

Instead of the above concentrated concentrate or apple concentrate, 3 to 10 parts by weight of one or more sugar components in oligosaccharide, fructose, glucose, honey or sugar may be mixed for alcohol fermentation.

After filtration of the fermentation broth obtained from the alcohol fermentation, the filtrate is inoculated with Acetobacter aceti (KCCM 12654), fermented for 6 to 12 days while being shaken at a temperature of 25 to 37 ° C at 150 to 300 rpm Acetobacter aceti (KCCM 12654) is the most active in the temperature range of 25 ℃ ~ 37 ℃, so the activity of the strain is lowered at the temperature below 25 ℃. And the strain is not actively growing due to the influence of high temperature at a temperature of 37 캜 or higher.

The acetic acid fermentation broth is heated and sterilized at a temperature of 65 ° C to 80 ° C for 5 to 20 minutes, followed by centrifugation and filtration to prepare a plum vinegar. The centrifugation is preferably performed at 2500 to 4000 rpm, and the filtration is preferably performed with a filter net of 50 to 150 mesh.

[Example 1]

The optimum conditions for the production of the plum fermentation vinegar were as follows. The concentrate of 69 ° Brix was mixed with purified water and adjusted to 16 ° Brix with a stock solution containing 3% (v / v) of mussel concentrate of 59 ° Brix. (IB-600M, JEIO TECH, Korea) at 28 ℃ ~ 32 ℃ after the inoculation. The filtered filtrate is used as acetic acid fermentation substrate.

The acetic acid fermentation was performed by inoculating 10% (v / v) of the herbal composition into the total volume of the fermentation broth of the plum alcohol with the fermentation broth of the plum alcohol in a fermenter (KF-5L, Korea fermentor Co., Korea) Vinegar is produced.

As shown in FIG. 1, when the fermented vinegar was prepared, the sugar fermentation rate was rapidly decreased from 16 ° Brix on the first day to 9 ° Brix on the third day and 6.4 ° Brix on the 9th day. , And the alcohol content increased from 5.8% by volume on the 5th day to 7.0% by volume on the 9th day, which was 0% by volume on the first day of the first day.

In the acetic acid fermentation of FIG. 2, the alcohol content decreased from 7.0% by volume on the first day to 3.5% by volume on the sixth day and almost no alcohol content remained at 0.0% by volume on the 12th day. On the other hand, titratable acidity, that is, the acidity determined by titrimetric method using the alkaline standard solution, increased from 1.0 vol% on the first day to 4.0 vol% on the sixth day and 5.5 vol% on the 12 th day.

<Physicochemical properties of the fermented vinegar of the present invention>

  end. pH

[Table 3] pH change of fermented vinegar of plum

ingredient Early composition of fermentation Solution of fermentation of plum alcohol Plum fermented vinegar pH 2.73 ± 0.01 2.81 ± 0.02 2.53 + - 0.01

Data values were expressed as mean ± SD. of triplicate determinations.

[Table 3] shows the results of measurement of changes in pH and total acidity during the process of making plum fermentation vinegar. The pH of the composition before fermentation was 2.73 ± 0.01 and the pH of plum vinegar was lowered to 2.53 ± 0.01 .

  I. Free sugar

[Table 4] Free sugar content of plum vinegar Unit: mg%

Free sugar component content Glucose 6414.36 Maltose 11.33 Fructose N.D. Sucrose N.D. Total free sugar content 6425.69

N.D. : Not detected

[Table 4] shows the free sugar content of the fermented vinegar of plum by HPLC analysis. As a result, free sugars were identified as glucose and maltose, and glucose content was the highest.

  All. Free amino acid

[Table 5] Free Amino Acid Content of Plum Vinegar Unit: ppm

Free amino acid component content L-Aspartic acid 7.56 L-Threonine 0.21 L-Serine 1.48 L-Glutamic acid 1.05 Glycine 0.09 L-Alanine 0.02 L-cystine 1.54 L-valine 1.12 L-Methionine 0.12 L-Isoleucine 0.31 L-Leucine 1.14 L-Tyrosine 5.46 L-Thenylalanine 4.43 L-Histidine 32.93 L-Lysine 4.11 Ammonia 1.05 L-Arginine 20.76 L-Proline 0.77 Total free amino acid content 83.32

Table 5 shows that the free amino acid content of the fermented vinegar was relatively high in L-Histidine (32.93 ppm) and L-Arginine (20.76).

  la. Organic acid

[Table 6] Contents of organic acid in the fermented vinegar of plum Unit: mg%

Organic acid content                   content Acetic acid 4034.46 Oxalic acid 72.76 Citric acid 1530.65 Succinic acid 1075.51 Malic acid 140.95 Lactic acid 390.87 Total organic acid content 7245.20

Table 6 shows that acetic acid, malic acid, citric acid, succinic acid and oxalic acid were found in the organic acid content of the fermented vinegar, and acetic acid was the highest as 4034.46 mg% in the main organic acid. citric acid and succinic acid were 1530.65 mg% and 1075.51 mg%, respectively. Especially, citric acid and succinic acid were detected in plum and were higher than other organic acids.

  hemp. Inorganic component

[Table 7] Contents of Inorganic Components of Plum Vinegar Unit: ppm

Inorganic component                   content Mg 3.45 Ca 2.75 Na 10.47 K 95.64 Fe 0.24 Mo 0.04 F 0.30

Table 7 shows that the content of potassium (K) was the highest at 95.64 ppm and the contents of sodium (Na), magnesium (Mg) and calcium (Ca) The contents were as high as 10.47 ppm, 3.45 ppm and 2.75 ppm. In addition, the contents of iron (Fe), phosphorus (P) and manganese (Mn) were somewhat low.

&Lt; Comparison of antioxidative activity of the fermented vinegar of the present invention &

  end. DPPH radical scavenging activity

DPPH radical scavenging activity of each sample was measured by UV / Vis-spectrometer (Hitachi, Japan) at 517 nm using the reducing ability of α, α'-diphenyl-β-picrylhydrazine (DPPH) Tokyo, Japan). 1 ml of 0.1% dibutylated hydroxytoluene (BHT) and 0.1% α-tocopherol solution, which were used as a positive control, were mixed with 3 ml of 4M DPPH solution for 5 seconds in a vortex mixer. The absorbance was measured by reacting in a dark place. Control was expressed as the ratio of decrease in absorbance to control by adding 1 ml of ethanol instead of the sample.

  I. Reducing power

The reducing power of the sample was measured by modifying the method of Yildirim et al. (Yildirim, A., Mavi, A. and Kara, A. A. 2001).

That is, 2.5 ml of potassium phosphate buffer solution (0.2 M, pH 6.6) and 2.5 ml of potassium ferricyanide (1%, w / v) were added to 1 ml of each sample and the positive control group and then mixed for 30 minutes Lt; / RTI &gt;

2.5 ml of trichloroacetic acid (10%, w / v) was added to the reaction solution, and the mixture was centrifuged at 3,000 rpm for 10 minutes.

1 ml of the supernatant was taken into a test tube and 1 ml of distilled water and 0.2 ml of FeCl ₃ (0.1%, w / v) were added and the absorbance was measured at 700 nm with a UV / Vis-spectrophotometer (Hitachi).

All. ABTS ^· radical scavenging activity

The radical scavenging ability of ABTS for the samples was measured by modifying the method of Biglari et al. (Biglari, F., AIKarkhi, A. M. F. 2008). 7 mM 2,2-azobis (2-aminopropane) dihydrochloride (AAPH) was mixed with 2.45 mM ABTS and reacted in a dark place at 23 ° C for 16 hours. The concentration of the ABTS solution was adjusted so that the absorbance was about 0.700 at 734 nm. Absorbance was measured with a UV / Vis-spectrophotometer (Hitachi) at 734 nm while mixing 0.1 ml of each sample and positive control group and 3.9 ml of ABTS solution and reacting at 23 ° C for 6 minutes. The positive colonies used 0.1% BHT and 0.1% α-tocopherol. ABTS radical scavenging ability was calculated according to the following equation.

Scavenging activity (%) = (Control O.D.-Sample O.D.) / Control O.D.

  la. Total polyphenol content

Total polyphenol content was determined by the Folin-Ciocalteu method (Slinkard K, Singleton VL. 1977). To the 0.1 mL of the sample, 8.4 mL of distilled water and 0.5 mL of a 2N Folin-Ciocalteu reagent (Sigma-Aldrich, Co., St. Louis, Mo., USA) were added, and 20% Na ₂ CO ₃ (Junsei Chemical Co., Japan) was added and left for 2 hours. The absorbance of the reaction was measured using a spectrophotometer (U-1800, Hitachi Co., Ltd., Tokyo, Japan) at 725 nm and a standard curve using gallic acid (Sigma-Aldrich, Co., Respectively.

  hemp. β-carotene bleaching assay

The antioxidative effect of β-carotene linoleate system was measured by the method of Mattaus (Mattaus B. 2002). Carotene solution was prepared by dissolving 1 mg of β-carotene (Sigma-Aldrich Co.) in 10 mL of chloroform. 10 mL of β-carotene solution was taken in a 100 mL round-bottomed flask, and 20 mg of linoleic acid (Sigma-Aldrich Co.) (Sigma-Aldrich Co.) was added thereto. The chloroform was removed with a rotary evaporator at 40 ° C, and then 100 mL of distilled water was added thereto, followed by shaking to prepare an emulsion solution. To 0.2 mL of the emulsion solution, 0.1% α-tocopherol (0.1%) and 8 μL of 0.1% BHT solution were added to the sample addition group, ethanol (control) and a positive control, and stored in an incubator (HB-103MP, Hanbaek Scientific Co.) Respectively. Absorbance was measured at 490 nm every 15 minutes for 0 min to 180 min during the storage period.

  bar. SOD-like activity

SOD (superoxide dismutase) -like activity was measured by pyrogallol (Sigma-Aldrich, Co.), which catalyzes the conversion of active oxygen species to hydrogen peroxide (H ₂ O ₂ ) according to the method of Marklund and Marklund (Marklund S, Marklund G. 1975). , St. Louis, Mo., USA). After adding 10 μL of each sample to a 96-well plate, 150 μL of Tris-HCl buffer solution (50 mM Tris aminomethane, 10 mM EDTA, pH 8.0) and 10 μL of 7.2 mM pyrogallol were added. Then, the reaction was carried out at room temperature for 10 minutes, and 50 μL of 1 N HCl was added to stop the reaction, and the absorbance was measured at 420 nm using a microplate reader. The SOD-like activity was expressed as a percentage (%) of the difference in absorbance between the sample added and the non-added sample.

)Superoxidedismutase-likeactivity (%) = (1-

)

A: absorbance of the sample-added sphere, B: absorbance of the sample-free sphere

  four. Hydrogen peroxide scavenging activity

The H ₂ O ₂ scavenging activity was determined by adding 100 μL of each sample to a 96-well plate according to the method of M (MHE, 1985). 20 μL of hydrogen peroxide (Junsei Chemical Co., Ltd., Japan) was added thereto, followed by incubation at 37 ° C for 5 minutes. After the reaction was completed, 30 μL of peroxidase (1 unit / mL; Sigma-Aldrich, Co., St. Louis, MO, USA) was added to 1.25 mM ABTS (Sigma-Aldrich, Co., After incubation for 10 min in an incubator at 37 °, the absorbance was measured at 405 nm with a microplate reader.

Ah. result

  - DPPH radical scavenging activity

Fig. 1. DPPH radical scavenging activity of PPB and PPV. Data values were expressed as mean ± SD, values with different letters are significantly different at p < 0.05. PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: prunus mume  and pear vinegar

     The hydrogen donating ability of PAB and PPV was measured as shown in Fig. Positive controls, butylated hydroxytoluene (BHT; 0.1%) and α-tocopherol (0.1%), were 87%. PPB and PPV were 40% and 31%, respectively.

- Reduction power measurement

Fig. 2. Reducing power effects of PPB and PPV. Data values were expressed as mean ± SD, values with different letters are significantly different at p < 0.05. PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: prunus mume  and pear vinegar

     The reducing power of PPB and PPV is shown in Fig.2. Positive control 0.1% BHT and 0.1% α-tocopherol were 2.5 and 2.6, respectively. PPB and PPV were 1.96 and 2.21, respectively. Therefore, it was confirmed that the fermentation vinegar PPV had better reducing power than PPB as the fermentation stock solution.

- ABTS ^· radical scavenging activity

Fig. 3. ABTS ^{. +}  radical scavenging activity of PPB and PPV. Data values were expressed as mean ± SD, values with different letters are significantly different at p < 0.05. PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: prunus mume  and pear vinegar

The results of measuring ABTS ⁺ radical scavenging activity of PPB and PPV are shown in Fig. Both groups showed lower tendency than the positive control, 0.1% BHT and 0.1% α-tocopherol. PPB showed 42% and PPV had 55% radical scavenging activity. It was confirmed that the ABTS ^· radical scavenging activity of PPV, which is plum vinegar, was superior to that of the PPB, which is the raw material of the fermentation.

 [Table 8] Measurement of β-carotene bleaching Unit:%

0.1% alpha-Tocopherol 0.1% BHT PPB PPv 0 h 100.86 + - 2.21 101.45 + 0.79 98.24 + 1.24 103.47 + 1.83 180 h 85.21 + - 12.32 87.57 ± 5.89 68.97 + - 5.58 79.22 + - 6.53

[Table 8] ß Carotene bleachimg assay of PPB and PPV. Data values were expressed as mean ± SD of triplicate determinations . PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: prunus mume  and pear vinegar

    [Table 8] shows the result of β-Carotene bleaching assay to investigate lipid peroxidation inhibitory activity of PPB and PPV. After 3 hours of reaction time, the antioxidative activity of PPV was 79% higher than that of PPB, which was similar to the positive control of 0.1% BHT and 0.1% of α-tocopherol. Lipid peroxyl radical scavenging activity.

  - Measurement of SOD-like activity

Fig. 4. SOD-like activity of PPB and PPV. Data values were expressed as mean ± SD, values with different letters are significantly different at p < 0.05. PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: prunus mume  and pear vinegar

     The results of SOD-like activity changes of PPB and PPV are shown in Fig. PPV was 17.9% and PPV was 69.8%. These values were similar to those of 0.1% ascorbic acid, which is a positive control with 71.8% of activity, suggesting that SOD - like activity was the best among the antioxidant activities of.

  -Hydrogen peroxide scavenging activity measurement

Fig. 5. Hydrogen peroxide scavenging activity of PPB and PPV. Data values were expressed as mean ± SD, values with different letters are significantly different at p < 0.05. PAB: prunus mume  and apple initial broth, PPB: prunus mume  and pear initial broth, PPV: Prunus mume  and pear vinegar

The changes in H ₂ O ₂ scavenging ability of PPB and PPV are shown in Fig. Positive control 0.1% BHT and 0.1% α-tocopherol showed 88.9% and 85.9% hydrogen peroxide elimination, respectively. PPB and PPV were 18.8% and 34.1%, respectively. However, PPV showed a similar activity to the ABTS ⁺ radical scavenging activity by showing the hydrogen peroxide scavenging ability higher than that of PPB.

  - Total polyphenol content measurement

[Table 9] Changes in total polyphenol content after acetic acid fermentation Prunus mume  initial broth.

Unit: mg%

Raw materials Fermented products PPb PPV 40.83 + - 1.02 25.86 + - 0.33

¹⁾ PPB: prunus mume and pear broth

²⁾ PAB: prunus mume and apple broth

³⁾ PPV: prunus mume and pear vinegar

     The total polyphenol content of the fermentation broth (PAB) and the fermented vinegar (PPV) during fermentation was measured as shown in [Table 9]. The total polyphenol content was decreased to 25.86 mg% in the case of plum vinegar (PPV) in the initial fermentation broth, indicating that the polyphenol component was degraded or oxidized by autoxidation during the fermentation period do.

< Anti Fatigue Effect of Plum Vinegar>

  end. Experimental animal breeding and diet

In the experimental animals, 36 male SD rats of 4 weeks old were purchased from Hyochang Science (Busan). After 1 week of adaptation period, rats were divided into two groups by sedimentary control (SC), exercised control (EC) , Prunus mume juice (PJ), 5% Prunus mume vinegar, 5% PV, and 7.5% prunus mume vinegar. The animals were kept in constant temperature at constant temperature (23 ℃), humidity (50%), 12 hours interval (08: 00 ~ 20: 00) and animals were separated into two cages in an acrylic cage.

The basic diet used in this experiment was a solid diet, free diet and water intake, and body weight change, water intake, and dietary intake were measured every two days.

  I. Exercise experiment design

Except the SC group, the rest of the subjects were orally administered the sample at a constant time of 7 ml / kg during the 4 weeks of experimental period, and then performed a high intensity progressive exercise using a trade mill.

The exercise program was performed at 20m / min (10 min) for the first week, 25m / min (20min) for the second week, 25m / min (30min) for the third week and 30m / The experiment was carried out at 40 m / min to set the exhaustion time for 10 seconds on the electric impact plate during the exercise.

The rats were sacrificed using ethyl ether, blood samples were collected from the inferior vena cava, liver and gastrocnemius muscle were collected, washed with physiological saline, and stored in a deep freezer at 80 ℃.

The rats used in all experiments complied with the guidelines of animal experiments conducted by Dong-A University Animal Ethics Committee (DIACUC-17-1)

  All. Biochemical Indicator Analysis

Serum inorganic phosphorus, ammonia and lactate were purchased from Biovision (USA) and Bioassay (USA), respectively.

Liver and muscle glycogen contents were measured by mixing 400 μL of 30% KOH in 0.25 g of tissue, heating at 100 ° C. for 30 minutes, cooling, cooling 1 mL of ethanol, centrifuging at 6000 rpm, 15 minutes, 4 ° C. and adding 500 μL of distilled water Mixed samples were measured using anthrone reagent.

Lactate dehydrogenase (LDH) and creatine kinase (CK) activities were measured by using Bioassay (USA) kit.

Malondialdehyde (MDA) and glutathione peroxidase (GPx) contents were measured by purchasing Biovision (USA) kit.

  la. result

  ① Dietary intake and dietary efficiency changes

[Table 10] Effects of Prunus mume  vinegar on weight gain, food intake and FER

Group SC ^One) EC ²⁾ PJ ³⁾ PV5 ⁴⁾ PV7.5 ⁵⁾ Body weight (g) Initial 227.21 ^a 226.14 ^a 223.16 ^a 221.47 ^a 214.27 ^a Final 362.42 ^a 347.86 ^ab 341.09 ^ab 332.08 ^ab 327.65 ^b Weight gain (g / day) 6.76 ^a 6.09 ^a 5.89 ^a 5.53 ^a 5.35 ^a Food intake (g / day) 49.89 ^a 50.58 ^a 51.16 ^a 48.81 ^a 50.48 ^a FER ⁶⁾  (%) 13.74 ^a 12.05 ^b 11.56 ^b 11.35 ^b 10.62 ^b

¹⁾ SC: sedentary control

²⁾ EC: exercised control

³⁾ PJ: Prunus mume juice

⁴⁾ PV5: Prunus mume vinegar 5% diluted drink

⁵⁾ PV7.5: Prunus mume vinegar 7.5% diluted drink

⁶⁾ FER (food efficiency ratio): weight gain (g / day) / food intake (g / day)

Data values were expressed as mean ± SD. Values with different letters are significantly different at p <0.05.

[Table 10] recorded changes in body weight, dietary intake and dietary efficiency among experimental animals. The weight difference between the groups at the end of the experiment showed that the weight gain was suppressed in a concentration dependent manner especially when the plum vinegar was consumed. The weight gain, the dietary intake and the diet Efficiency was also affected.

② Changes in long-term weight

[Table 11] Effects of Prunus mume  vinegar on organ weights in rats with exhaustive exercise

Organs
SC ^One) EC ²⁾ PJ ³⁾ PV5 ⁴⁾ PV7.5 ⁵⁾ g / body weight 100g Liver 3.33 ^a 3.06 ^a 2.97 ^a 3.09 ^a 3.01 ^a Heart 0.3 ^a 0.31 ^a 0.31 ^a 0.31 ^a 0.30 ^a Kidney 0.74 ^a 0.72 ^a 0.74 ^a 0.74 ^a 0.72 ^a Abdominal fat 0.79 ^ab 0.82 ^a 0.65 ^ab 0.51 ^b 0.48 ^b Perirenal fat 0.34 ^a 0.34 ^a 0.29 ^a 0.28 ^a 0.28 ^a Epodidymal fat 1.32 ^a 1.27 ^a 1.12 ^ab 1.06 ^ab 1.02 ^b Gastrocnemius
muscle 0.48 ^a 0.43 ^a 0.47 ^a 0.48 ^a 0.48 ^a Spleen 0.20 ^a 0.18 ^a 0.19 ^a 0.18 ^a 0.19 ^a Lung 0.53 ^a 0.46 ^a 0.46 ^a 0.41 ^a 0.41 ^a

¹⁾ SC: sedentary control

²⁾ EC: exercised control

³⁾ PJ: Prunus mume juice

⁴⁾ PV5: 5% diluted drink for Prunus mume vinegar

⁵⁾ PV7.5: 7.5% diluted drink for Prunus mume vinegar

Data values were expressed as mean ± SD. Values with different letters are significantly different at p <0.05.

[Table 11] shows the result of measuring the organ weights of each group. In liver, abdominal fat, epididymal fat, and lung, there was a slight difference when the vinegar group and control group were compared. Particularly, in the abdominal fat, PV7.5 group showed a difference of 40% compared to the control group, which was also observed in epididymis.

③ Endurance measurement

Fig. 6. Effect of PV on running endurance time in rats exhausted by exercise. Data values are expressed as means ± Values with different letters are significantly different at p &Lt; 0.05. SC: sedentary control, EC: exercised control, PJ: Prunus mume  juice, PV5: 5% diluted drink for Prunus mume  vinegar, PV7.5: 7.5% diluted drink for Prunus mume  vinegar

Fig. 6 was recorded as one hour of exercising until exhaustion in order to measure the endurance of the experimental animals. The time taken until exhaustion was about 30 minutes in the non-exercise group, while the exercise group and the plum concentrate group were 63 Min, and 62 minutes, respectively. In the group fed with 5% and 7.5% diluted acetic acid vinegar, exercise time was more than 70 minutes, and endurance was improved.

④ Measurement of blood fatigue substance

Fig. 7. Effect of PV on (A) serum ammonia (B) inorganic phosphate and (C) lactate in rats exhausted by exercise. Data values are expressed as means ± Values with different letters are significantly different at p &Lt; 0.05. SC: sedentary control, EC: exercised control, PJ: Prunus mume  juice, PV5: 5% diluted drink for Prunus mume  vinegar, PV7.5: 7.5% diluted drink for Prunus mume  vinegar

Fig. 7 is the measurement value of ammonia, inorganic phosphoric acid, and lactic acid in plasma, and this item is a fatigue index substance, which is measured in high fat content. In the SC, EC, and PJ groups, there was no significant difference in contents of ammonia, inorganic phosphoric acid, and lactic acid as fatigue substances, but in the PV5 and PV7.5 groups, Of the total fatigue life.

  ⑤ Measurement of muscle and liver glycogen content

Fig. 8. Effect of PV on (A) liver and (B) muscle glycogen accumulation in rats exhausted by exercise. Data values are expressed as means ± Values with different letters are significantly different at p &Lt; 0.05. SC: sedentary control, EC: exercised control, PJ: Prunus mume  juice, PV5: 5% diluted drink for Prunus mume  vinegar, PV7.5: 7.5% diluted drink for Prunus mume  vinegar

Fig. 8 showed that the content of glycogen stored in the liver and muscle was lower in the SC and EC groups than in the PJ, PV5 and PV7.5 groups, as a result of measurement of the glycogen content of the liver and muscles. Thus, Which is an increase in the accumulation of oxygen. Physical fatigue is associated with a lack of energy during exercise, and glycogen is an energy storage form that plays a direct role in energy production through the body's processes. The amount of glycogen affects direct exercise capacity.

  ⑥ LDH and CK content measurement

Fig. 9. Effects of PV on (A) lactate dehydrogenase and (B) creatine kinase activities in rats exhausted by exercise. Data values are expressed as means ± Values with different letters are significantly different at p <0.05. SC: sedentary control, EC: exercised control, PJ: Prunus mume  juice, PV5: 5% diluted drink for Prunus mume  vinegar, PV7.5: 7.5% diluted drink for Prunus mume  vinegar

Fig. 9 showed LDH and serum CK content in muscle tissue. LDH content was lowest in SC group, PV7.5 group was highest and CK content was decreased according to consumption of plum vinegar Respectively. LDH is an enzyme involved in the conversion of lactic acid to pyruvic acid by the side effects of NADH and NAD ⁺ , and CK is known as an indicator of enzyme or muscle damage catalyzing the energy production process of converting ADP to ATP .

  ⑦ Antioxidant activity measurement

Fig. 10. Effect of PV on (A) malondialdehyde, (B) glutathione peroxidase and (C) superoxide dismutase activities in rats exhausted by exercise. Data values are expressed as means ± Values with different letters are significantly different at p <0.05. SC: sedentary control, EC: exercised control, PJ: Prunus mume  juice, PV5: 5% diluted drink for Prunus mume  vinegar, PV7.5: 7.5% diluted drink for Prunus mume  vinegar

Fig. 10 showed that malondialdehyde, an indicator of lipid peroxidation, and glutathione peroxidase, an antioxidative enzyme, showed low malondialdehyde levels in the PV5 and PV7.5 groups and suppressed lipid peroxidation in (A) In (B), the activity of antioxidant enzyme GPx was high in PV5 and PV7.5. Therefore, lipid peroxidation seems to be inhibited by the activity of these antioxidant enzymes.

Claims (7)

A method for producing a plum fermentation vinegar,
The fresh juice produced by juicing the browning room was heated at a temperature of 55 ° C to 70 ° C for 8 to 25 hours and rapidly heated at a high temperature of 100 ° C for 5 minutes to inactivate the enzyme activity. Filtered through a mesh screen mesh, and heated at 90 to 100 ° C for 5 to 15 hours to prepare a 50% Brix to 70 Brix plum concentrate,
The peel and seed parts of the ripe pear are removed and the flesh of the pear is heated at a temperature of 50 ° C to 90 ° C for 15 to 30 hours to concentrate to a concentration of 60 ° Brix to 80 ° Brix and filtered to prepare a concentrate ,
Saccharomyces cerevisiae (KCCM 11306) was subcultured on a YM plate medium and cultured in a YM liquid medium at a temperature of 28 ° C ~ 32 ° C for 40 ~ 55 hours.
Acetobacter aceti (KCCM 12654) was cultivated in mannitol broth liquid medium at 29 ℃ ~ 31 ℃ for 40 ~ 55 hours,
0.5 to 8 parts by weight of the plum concentrate and 15 to 40 parts by weight of the concentrate are mixed with 100 parts by weight of purified water to adjust the sugar content of the fermented vinegar to 10 to 20 ° Brix and then inoculated with the alcohol fermentation broth, &Lt; / RTI &gt;
Wherein the acetic acid bacteria are inoculated in the alcoholic fermentation broth and fermented with acetic acid, thereby producing an antioxidant and anti-fatigue effect.
delete delete delete delete delete The fermented vinegar of plum prepared by the method of claim 1

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