KR20220069554A - Method for preparing vinegar using black raspberry pomace fermented by Acetobacter pasteurianus - Google Patents

Abstract

By acetic acid fermenting raspberry wine juice extraction cake, which is discarded as a byproduct of a raspberry wine process, with a vineger starter comprising Acetobacter pasteurianus deposited under accession number KCCM12808P, the present invention relates to a producing method of a functional fermentation vinegar having excellence in acetic acid fermentation results and having high antioxidant activity.

Description

Method for preparing vinegar using black raspberry pomace fermented by Acetobacter pasteurianus}

The present invention is by acetic acid fermentation of bokbunja juiced gourd, which is discarded as a by-product of processing bokbunja, with a vineger starter containing Acetobacter pasteurianus deposited with accession number KCCM12808P, so that the acetic acid fermentation result is excellent and antioxidant It relates to a method for producing a functional fermented vinegar with high activity.

Vinegar is a fermented food with a long history in both the East and the West. Vinegar contains various volatile and non-volatile organic acids, amino acids, sugars, esters, etc., which are derived from raw materials used or generated by the metabolism of microorganisms involved in fermentation. It is a food with

In addition to antioxidant activity that destroys free radicals that are harmful to the human body, vinegar has antibacterial activity, anti-obesity, anti-diabetes, fatigue recovery, promotion of digestive juice secretion, improvement of lipid metabolism, suppression of increase in blood pressure, prevention of arteriosclerosis, high blood pressure, and heart disease, etc. As functionalities have been revealed, it is attracting attention as a functional food (Kim et al., 2020).

Recently, as the demand for natural fermented vinegar manufactured with 100% fruit juice raw materials and high grain content without adding auxiliary materials is increasing, research on the development of vinegar using various natural materials and enhancement of functionality is being conducted.

Despite this interest, traditional fermented vinegar manufactured from fruits such as bokbunja, blueberry, and cucurbiton is not competitive due to the high price of raw materials, so consumption is insufficient despite its high functionality.

Bokbunja (Rubus occidentalis) is characterized by low sugar content and strong sour taste.

In addition, functional compounds such as ellagic acid, gallic acid, caffeic acid, ferulic acid, rutin, myriecetin, luteolin, and kaempfrol are present in large amounts in bokbunja (Shin et al., 2018), and these various components are antioxidants (Wang and Lin). , 2000), immune enhancing activity (Seeram et al., 2006), and biologic activity such as anticancer effect (Eu et al., 2008), the demand for it is increasing.

The amount of bokbunja processed in 2018 was a total of 232 tons, of which beverages accounted for the highest at 53.5%, followed by alcohol 30.3% and juice 9.5% (Fruit Processing Status 2018, Ministry of Agriculture, Food and Rural Affairs).

In order to produce such processed bokbunja processed food, a squeezing process is performed, and the juice yield is about 60 to 70%, resulting in by-products. It is insufficient to the extent that only studies for feed use (Lee et al., 2019) are conducted.

Therefore, in the present invention, in order to increase the utility of by-products discarded after processing bokbunja, vinegar using bokbunja juice was prepared, and physicochemical properties including fermentation characteristics and antioxidant activity were analyzed, and the possibility of use as a material for traditional fermented vinegar was investigated. Let's take a look.

Republic of Korea Patent Registration 10-1428692 (Registration Date August 04, 2014) Republic of Korea Patent Registration 10-0943024 (Registration Date February 10, 2010) Republic of Korea Patent Registration 10-1797464 (Registration Date November 08, 2017)

The present invention provides excellent physicochemical properties and antioxidant activity by acetic acid fermentation of raspberry juice, which is discarded as a by-product of processing bokbunja, with a vineger starter containing Acetobacter pasteurianus deposited with accession number KCCM12808P. It is an object of the invention to provide a method for producing a functional fermented vinegar.

In order to achieve the above object,

The present invention is characterized in that vinegar is prepared by acetic acid fermentation of bokbunja extract with a vinegar starter containing Acetobacter pasteurianus deposited with accession number KCCM12808P. It provides a method for preparing vinegar made by acetic acid fermentation with Pasteurian GBA5.

Fermented vinegar using bokbunja juice prepared according to the present invention has the following effects.

first. Total acidity and alcohol content were 0.49% and 1.32% lower in Pomace than in bokbunja juice, respectively. After acetic acid fermentation, alcohol content and sugar content were 0.72% and 0.6 ° in juice, respectively. appeared as high as Brix.

second. In Pomace, the pH was 0.32 lower and the total acidity was 0.38% higher. From these results, it can be seen that the smooth fermentation proceeded in the juiced gourd. Also, as a result of organic acid analysis, acetic acid was higher in Pomace, and citric acid was higher in Pomace.

third. As a result of measuring the total polyphenol, total flavonoid, and total anthocyanin contents of vinegar using bokbunja extract, it was 51.58 TAE mg/mL, 9.55 RUE mg/mL, and 6.05 CYE mg/mL, respectively, compared to the control, bokbunja extract vinegar. showed high content.

fourth. As a result of measuring the IC ₅₀ values of DPPH and ABTS radical scavenging ability to measure the antioxidant activity of bokbunja extract vinegar, they were 10.23 and 18.30 μL/mL, respectively, which were higher than those of bokbunja extract vinegar. It was confirmed that it appeared high in vinegar.

fifth. Through the results of the first to fourth, when using bokbunja juice that is discarded as a by-product of processing bokbunja according to the present invention, the result of acetic acid fermentation is excellent, and it exhibits high antioxidant activity, so it has a very excellent effect as a functional fermented vinegar. know that you have

1 is a graph showing a comparison of changes in alcohol content of black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.
2 is a graph showing a comparison of changes in sugar concentration between black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.
3 is a graph showing a comparison of the pH change of black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.
4 is a graph showing a comparison of changes in total acidity of black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.
5 is a graph showing a comparison of reducing power according to the concentration of black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.
6 is a graph showing the comparison of FRAP activity according to the concentration of black raspberry pomace vinegar according to the present invention and black raspberry juice vinegar as a control.

Hereinafter, let's take a look at the specific technical contents of the vinegar manufacturing method made by acetic acid fermentation of bokbunja juiced gourd according to the present invention with Acetobacter Pasteurian GBA5.

As mentioned above,

Bokbunja juiced gourd vinegar according to the present invention is produced by acetic acid fermentation with a vineger starter containing Acetobacter pasteurianus deposited under the accession number KCCM12808P of bokbunja juiced meal, which is discarded as a by-product of processing bokbunja.

Looking at the manufacturing process of the bokbunja juiced gourd vinegar in more detail,

Bokbunja juiced gourd was put in 8 to 12 times the weight of water, extracted at 100 ° C for 5 to 7 hours, and concentrated to 15 to 20 ° Brix with a reduced pressure concentrator to prepare a concentrate, and dried yeast S. cerevisiae in the concentrate Preparing an alcoholic fermentation solution by inoculating fermivin to 1×10 ⁹ CFU/kg, stirring, and fermenting at 24 to 26 ° C. for 4 to 7 days;

Acetobacter pasteurianus deposited with the accession number KCCM12808P in the alcoholic fermentation liquid is inoculated with a vineger starter containing acetic acid fermentation.

The vinegar used in the manufacture of the vinegar (vineger starter),

A liquid medium containing 93.5 to 97.2 wt% of distilled water, 0.3 to 0.7 wt% of yeast extract, 0.2 to 0.5 wt% of peptone, and 2.1 to 3.2 wt% of mannitol It was prepared by inoculating Acetobacter pasteurianus deposited under the accession number KCCM12808P in the , and culturing with shaking at 29-31 ℃ for 70-74 hours.

In the acetic acid fermentation step using such a seed seed, the vinegar starter is inoculated into the alcoholic fermentation solution, inoculated at 9 to 12 vol% with respect to the total volume of the alcoholic fermentation solution, and cultured at 29 to 31 ° C. for 20 to 28 days. .

At this time, if the inoculation amount of the seedling is less than 9 vol% or exceeds 12 vol%, acetic acid fermentation may not occur well, so the inoculation amount of the seedling is within the range of 9 to 12 vol% with respect to the total volume of the alcoholic fermentation broth. It is preferable to limit

If the shaking culture exceeds 31 ℃, the acetic acid bacteria may not proliferate well and the acetic acid fermentation may not be successful. Therefore, the temperature of the shaking culture is preferably limited to within the range of 29 ~ 31 ℃.

Hereinafter, let's look at specific examples and test examples of fermented vinegar prepared by acetic acid fermentation of bokbunja juice meal with Acetobacter pasteurianus GBA5.

The present invention relates to bokbunja extract vinegar, and as a control, bokbunja extract vinegar is presented and compared.

[Example]

1. Experimental materials and reagents

The bokbunja used in the experiment according to the present invention was harvested in 2020 in Buan-myeon, Gochang-gun, Jeollabuk-do, thawed at room temperature, and then juiced through a hydraulic juicer (Stainless 70L, Tomotech Ltd., Seoul, Korea) and Pomace was used as a sample for the preparation of vinegar.

Folin-Ciocalteu's phenol reagent, DPPH (2,2'-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) used to measure antioxidant activity, and organic acids Standard materials for quantification, lactic acid, citric acid, malic acid, acetic acid, succinic acid, and yeast extract, peptone, and mannitol used for culturing acetic acid bacteria were purchased from Sigma-Aldrich (St. Louis, MO, USA) and used. and ethanol and methanol were J.T. It was purchased from Baker (Boston, MA, USA), and other reagents were used.

2. Composition of strain used and medium

The yeast used for alcoholic fermentation was Saccharomyces cerevisiae Fermivin (DSM Food Specialties, Ma Delft, Netherlands), an imported commercially available dry yeast, which was incubated in 15% sugar solution at 20 °C for 10 minutes and then activated and used for fermentation.

For acetic acid fermentation, the Acetobacter pasteurianus GBA5 strain deposited under the accession number KCCM12808P possessed by the Berry & Bio Food Research Institute was used, and the medium composition of 0.5 wt% yeast extract, 0.3 wt% peptone, and 2.5 wt% mannitol was used at 30 °C. What was cultured with shaking for 72 hours was used as an acetic acid fermentation starter (seeder).

3. Vinegar Manufacturing

After adding 10 times the weight of water and extracting at 100 °C for 6 hours, the extract is concentrated until 18 °Brix with a vacuum concentrator. Inoculate at ⁹ CFU/kg, stir, and ferment at 25° C. for 6 days to prepare an alcoholic fermentation broth.

Next, after alcohol fermentation was finished, the fermentation broth was filtered, seeded seed 10% (v/v) was inoculated, respectively, and inoculated at 30° C. for 24 days, samples were collected at 3-day intervals and used as samples.

As a control of the bokbunja juice prepared in this way, instead of using the extract, the bokbunja juice concentrated until 18 ° Brix with a reduced pressure concentrator (Buchi R210, Flawil, Switzerland) was used to extract the bokbunja juice. Bokbunja juice vinegar is produced through the same process as for gourd.

Next, changes in alcohol, sugar content, pH, total acid and chromaticity of the prepared bokbunja juice vinegar and the control bokbunja juice vinegar; organic acids; total polyphenol, total flavonoid and total anthocyanin content; DPPH radical scavenging ability; ABTS radical scavenging ability; reducing power; Let's compare and look at the measurement results for ferric reducing antioxidant power (FRAP).

[Test Example]

1. Alcohol, sugar content, pH, total acid and color change

Alcohol content was measured using an alcohol analyzer (Alcoholyzer wine, Anton Paar, Graz, Austria), and sugar content was measured at room temperature using a saccharometer (PAL-1 Pocket Refractometer, ATAGO, Tokyo, Japan).

The pH was measured with a pH meter (S20, Mettler Toledo, Schwerzenbach, Switzerland), and the total acid content was calculated as the amount of citric acid or acetic acid consumed until pH 8.3 was reached by adding 0.1 N NaOH to 10 mL of the sample. .

Chroma meter (CM-5, Minolta, Osaka, Japan) was used to measure L (brightness), a (redness), and b (yellowness) values, and the white plate used ( The values of the calibration plate) were L=92.73, a=-0.03 , b=-0.06, and the L value, a value, and b value were measured three times each and divided by the average value to measure chromaticity.

2. Organic acids

For the measurement of organic acids in vinegar prepared from bokbunja juice and juice, each sample was filtered with a 0.45 μm membrane filter (Millipore, Bedford, MA, USA) and diluted by HPLC (Ultimate 3000, Dionex, Sunnyvale, CA, USA). ) was used for analysis.

As analysis conditions, Aminex HPX-87H (300×10 mm, Bio-Rad, Hercules, CA, USA) was used for the column, and 0.01NH ₂ SO ₄ (Sigma-Aldrich) was used as the solvent, the flow rate was 0.5 mL/min, the injection volume. Silver 10 μL, UV detector wavelength 210 nm was used.

3. Total polyphenols, total flavonoids and total anthocyanin content

The total polyphenol content was measured by applying the National Health Functional Foods Method (KFDA, 2012).

That is, 7.5 mL of distilled water, 0.5 mL of Folin-Ciocalteu's phenol reagent, and 1 mL of 35% sodium carbonate were added to 1 mL of vinegar prepared from bokbunja juice and juice, and then left at room temperature in the dark for 1 hour, followed by a UV/VIS spectrophotometer. (UV-2450, Shimadzu Co., Kyoto, Japan) was used for colorimetric quantification at 760 nm.

At this time, a calibration curve was prepared using tannic acid as a standard material, and the total polyphenol content was obtained from this.

Total flavonoid content was measured by applying the method of Chang et al. (2002).

2 mL of diethylene glycol and 20 μL of 1 N sodium hydroxide were added to the solution prepared at a concentration of 200 μL/mL of each sample, and then reacted in a constant temperature water bath at 37°C for 1 hour. Then, the absorbance was measured at 420 nm using a UV/VIS spectrophotometer. measured.

At this time, the total flavonoid content was calculated using rutin as a standard material and a calibration curve was drawn up from this.

The total anthocyanin content was obtained by applying the method of Kim et al. (1998), extracting 39 mL of 80% methanol solution containing 0.1% HCl in 1 mL of each sample in a shaker incubator (Vision Scientific) for 24 hours, followed by UV/VIS spectrophotometer was used to measure the absorbance at 528 nm.

At this time, a calibration curve was prepared using cyanidin as the standard material, and the total anthocyanin content was obtained from this.

4. Measurement of DPPH radical scavenging ability

Apply the antioxidant activity measurement method (Choi et al., 1993) using DPPH, a free radical, and dilute the concentrate in distilled water to a concentration of 5, 10, 20, or 30 μL/mL, and add 200 μL of ethanol and 0.2 mM to 100 μL of each sample. After adding 300 μL of DPPH solution, it was stirred, reacted at room temperature for 30 minutes, and absorbance was measured at 517 nm using an ELISA reader (Synergy HT, Biotec, Washington DC, USA). As a control, ethanol was added instead of the sample. .

5. Measurement of ABTS radical scavenging ability

The method of Art et al. (2004) was applied for ABTS assay. After adding 195 μL of ABTS radical solution to 5 μL of each sample prepared at 5, 10, 20, and 30 μL/mL concentrations, and reacting for 7 minutes, absorbance was measured at 734 nm using an ELISA reader, and the control was used instead of the sample. Ethanol was added to the experiment.

6. reducing power

The reducing power was measured by using the method of Oyaizu (1986) at each concentration of 5, 10, 20, and 30 μL/mL, in 200 μL of each sample, 250 μL of 0.2 M sodium phosphate buffer (pH 6.6) and 250 μL of 1% potassium ferricyanide. was added and reacted for 20 minutes in a water bath at 50°C, and then 250 μL of 10% trichloroacetic acid was added.

After that, 50 μL of 0.1% ferric chloride was added, and the reducing power was measured at 700 nm.

7. Measurement of ferric reducing antioxidant power (FRAP)

FRAP was measured by applying the method of Blois (1958).

That is, 0.3 M sodium acetate buffer solution (pH 3.6), 10 mM TPTZ, and 20 mM FeCl ₃ .6H ₂ O were prepared and mixed in a ratio of 10:1:1 just before the experiment to prepare a FRAP solution.

After adding 750 μL of FRAP solution and 30 μL of each sample prepared by concentration, the reaction was performed in a constant temperature water bath at 37° C. for 15 minutes, and absorbance was measured at 593 nm.

8. Statistical analysis

Experimental results for each test item according to the present invention were analyzed three times and expressed as mean and standard deviation after statistical processing using SPSS program 23.0 (Statistical Package for Social Sciences, SPSS Inc., Chicago, IL, USA). .

Statistical significance between each test group was analyzed by Student's t-test and one-way ANOVA at p<0.05 level, and a post-hoc test was performed using Duncan's multiple range test.

9. Results and Considerations

1) Alcohol fermentation

An alcoholic fermentation broth was prepared using bokbunja juice and juice, and the results of measuring physicochemical properties at the initial stage of fermentation and after the end of fermentation are shown in Table 1 below.

S. cerevisiae fermibin ( S. cerevisiae Fermivin ) and change of fermentation characteristics of black raspberry juice and Pomace wine fermented at 25℃ for 6 days
fermentation time
(Work)
(Fermentation time
(days))
Bokbunja
wine
(Black raspberry wine)
Alcohol
content (%)
(Alcohol
content (%))
sugar concentration
(°Brix)
(Sugar concentration (°Brix))
pH
Total acidity (%)
(Total acidity (%)) Hunter’s color value L a b 0 juice
(Juice) 0.01±0.01 ¹⁾ 18.01±0.06 3.69±0.01 0.94±0.01 15.10±0.02 46.86±0.01 26.03±0.03 juice gourd
(Pomace) 0.01±0.01 18.07±0.06 3.75±0.01* 0.46±0.01* 15.78±1.73 45.75±0.06* 25.34±0.03* 6 juice
(Juice) 10.22±0.03 7.80±0.01 3.63±0.01 1.20±0.01 24.76±0.01 56.25±0.01 41.38±0.06 juice gourd
(Pomace) 8.90±0.01* 7.03±0.06* 3.59±0.01* 0.71±0.01* 17.91±0.01* 49.60±0.53* 30.70±0.02*

¹⁾ Data are mean ± standard deviation.

*p<0.05 indicates a statistically significant difference with wine using bokbunja juice.

The alcohol content of bokbunja juice and pomace was 10.22% and 8.90%, respectively, from 0.01% at the beginning of fermentation to 10.22% and 8.90% on the 6th day at the end of fermentation. (p<0.05).

In the case of sugar content, the initial sugar content of bokbunja juice and juice extract was 7.80 °Brix and 7.03 °Brix at the end of fermentation at 18.01 to 18.07 °Brix, respectively.

The initial pH of bokbunja juice and juiced meal was 3.69~3.75, and decreased after fermentation was completed.

In the case of total acidity, the initial total acidity of bokbunja juice and juice extract was 0.94% and 0.46%, respectively, which was twice higher than that of the juice extract. It is estimated that the initial total acidity of the juice is higher than that of the juice extract.

The total acidity after fermentation was 1.2% of the extract, which was higher than that of the extract (0.71%) (p<0.05).

According to Lee et al. (2002), chromaticity is known to be affected during fermentation by several factors such as polyphenol content, microbial activity, and SO ₂ .

In the present invention, as a result of measuring the chromaticity of the alcoholic fermentation broth of bokbunja juice and juiced gourd, the L value indicating the brightness, the a value indicating the redness, and the b value indicating the yellowness were all increased at the end of fermentation compared to the beginning of fermentation. In all of the L, a, and b values, the juice juice was higher than that of bokbunja juice (p<0.05).

2) Acetic acid fermentation

The results of examining the changes in alcohol, sugar content, pH, and total acidity during the fermentation period of vinegar prepared by using bokbunja juice and alcoholic fermentation broth from juice extract are shown in FIGS. 1 to 4 .

The change in the alcohol content of vinegar using bokbunja juice and juice went down gradually as fermentation progressed. Vinegar was measured from 8.58% at the beginning of fermentation to 0.15% on the 24th day of fermentation.

This showed a result that satisfies the conditions for fruit vinegar (Kim et al., 2008) set by the Korean Agency for Technology and Standards of the Ministry of Commerce, Industry and Energy.

During the acetic acid fermentation period of vinegar using bokbunja juice and juice, the sugar content gradually decreased as the fermentation period elapsed at 7.27~7.47 °Brix at the beginning of fermentation. It was found to be 5.3 °Brix and 4.7 °Brix.

As a result of examining the change in pH during the fermentation period, the initial pH of bokbunja juice vinegar gradually decreased from 3.66 to 3.38 on the 24th day of fermentation, and the vinegar using juice extract decreased from the initial pH of 3.82 to 3.06. It was confirmed that the final pH was lower than that of the juice.

Lee et al. (2012) reported that the pH decreased as acetic acid production increased due to organic acid production during acetic acid fermentation, which coincided with the trend of decreasing the pH of vinegar using bokbunja juice and juice extract of the present invention.

Acetic acid is used as an indicator of the quality of vinegar, and the Food Codex sets the total acid (acetic acid, w/v%) content of vinegar in the range of 4.0 to 20.0 %, and for persimmon vinegar 2.6 % or more (MFDS, 2007).

The total acidity of vinegar using bokbunja juice and juice extract increased gradually as fermentation progressed. As it shows high total acidity, it is judged that it is possible to use bokbunja juice as a fruit vinegar material.

Table 2 below shows the color change during fermentation of vinegar prepared using bokbunja juice and juice. The L value indicating the brightness, the a value indicating the redness, and the b value indicating the yellowness all decreased during the acetic acid fermentation process, and the bokbunja juice was significantly higher than that of the bokbunja extract during the entire fermentation period ( p<0.05).

Color value of vinegar using bokbunja juice and bokbunja extract Color value Fermentation time (days)
Fermentation time
(days) Black raspberry vinegar Juice Pomace L 0 28.46±0.01 ^1)a2) 22.67±0.03 ^a 3 27.09±0.04 ^b 18.52±0.03 ^b 6 25.76±0.02 ^c 18.34±0.08 ^c 9 25.39±0.08 ^d 18.31±0.06 ^c 12 24.46±0.04 ^e 17.67±0.02 ^d 15 24.20±0.16 ^f 17.33±0.05 ^d 18 24.03±0.04 ^f 17.14±0.07 ^e 21 22.98±0.08 ^g 16.77±0.05 ^f 24 22.21±0.02 ^h 14.23±0.04 ^g a 0 58.79±0.03 ^a 54.68±0.03 ^a 3 57.19±0.03 ^b 50.39±0.03 ^b 6 56.69±0.09 ^c 50.25±0.11 ^c 9 55.85±0.03 ^d 50.11±0.07 ^d 12 55.23±0.05 ^e 49.11±0.07 ^e 15 54.69±0.10 ^f 48.92±0.01 ^f 18 54.19±0.04 ^g 48.68±0.03 ^g 21 53.98±0.07 ^h 48.29±0.02 ^h 24 53.44±0.03 ⁱ 45.57±0.02 ⁱ b 0 44.79±0.03 ^a 36.11±0.03 ^a 3 43.19±0.08 ^b 31.56±0.32 ^b 6 42.87±0.13 ^c 31.54±0.05 ^b 9 42.06±0.01 ^d 31.09±0.04 ^b 12 41.07±0.05 ^e 30.32±0.06 ^c 15 40.72±0.05 ^f 29.41±0.02 ^d 18 40.12±0.07 ^g 29.05±1.15 ^d 21 39.10±0.09 ^h 28.27±0.03 ^e 24 39.03±0.02 ^h 24.53±0.04 ^f

¹⁾ Data are mean ± standard deviation.

²⁾ Different letters in one column indicate statistically significant differences by Duncan's multiple-range test (p<0.05).

3) organic acids

Table 3 below shows the results of the organic acid analysis of vinegar prepared using bokbunja juice and juice. In both types of vinegar, only acetic acid and citric acid were detected.

Organic acid content of bokbunja vinegar using bokbunja juice and juice bokbunja vinegar
(Black raspberry vinegar) Organic acid content (mg/mL) Acetic acid Citric acid Juice 2.80±0.72 1.19±0.38 Pomace 3.09±0.64* 0.52±0.20*

¹⁾ Data are mean ± standard deviation.

*p<0.05 indicates a statistically significant difference from the vines using the test raspberry juice.

The acetic acid contents of vinegar prepared from bokbunja juice and juice were 2.80 mg/mL and 3.09 mg/mL, respectively, and the vinegar prepared from juice extract showed a higher content than vinegar prepared from juice extract (p< 0.05).

This was found to be consistent with that at the end of the acetic acid fermentation, the total acid content prepared from the juice was higher than that of the vinegar prepared from the juice juice.

The content of citric acid was 1.19 mg/mL in vinegar prepared from bokbunja juice, which was about 2.2 times higher than that of juice (0.52 mg/mL).

In general, the organic acid content of vinegar is known to show a lot of difference in the organic acid composition depending on the raw material used (Jung et al., 2019).

Citric acid, succinic acid, and fumaric acid are known to exist in bokbunja fruit (Hong et al., 2012), but only acetic acid and citric acid were detected in this test result. .

4) Antioxidant activity

Polyphenolic substances are one of the secondary metabolites widely distributed in the plant world. They have a phenolic hydroxy group, so they show a property of binding to proteins and other macromolecules, and are known to have physiological activities such as antioxidant effects (Fine, 2000). .

Table 4 below shows the results of measuring total polyphenol, total flavonoid and total anthocyanin content, and DPPH and ABTS radical scavenging ability of vinegar prepared from bokbunja juice and juice.

Antioxidant action of vinegar using bokbunja juice and juice bokbunja vinegar polyphenol content
(TAE ¹⁾ mg/mL) Flavonoid content
(RUE ²⁾ mg/mL) anthocyanins
content
(CYE ³⁾ mg/mL)
DPPH
(IC ₅₀ μL/mL) ⁴⁾
ABTS
(IC ₅₀ μL/mL) Juice 29.09±0.29 ⁵⁾ 6.48±0.09 5.13±0.02 20.12±0.08 35.39±0.15 Pomace 51.58±0.44* 9.55±0.09* 6.05±0.03* 10.23±0.08* 18.30±0.03*

¹⁾ Total polyphenol content was expressed in mg tannic acid (TAE) per 100 g.

²⁾ Total flavonoid content was expressed in mg rutin (RUE) per 100 g.

³⁾ Total anthocyanin content was expressed as mg cyanidin (CYE) per 100 g.

⁴⁾ DPPH and ABTS radical scavenging ability was expressed as the average of the 50% inhibitory concentration obtained through interpolation of the concentration inhibition curve.

⁵⁾ Data are mean ± standard deviation.

*p<0.05 indicates a statistically significant difference from vinegar using bokbunja juice.

The total polyphenol content of vinegar prepared from bokbunja extract was 51.58 TAE mg/mL, which was about 1.7 times higher than that of vinegar prepared from juice (29.09 TAE mg/mL) (p<0.05).

This is presumed to affect the total polyphenol content of the final vinegar as polyphenols present in the peel, pulp, and seeds do not elute properly and are present in large amounts in the by-products in the case of the hydraulic juice process for manufacturing bokbunja undiluted solution.

As a result of measuring the total flavonoids of vinegar using bokbunja juice and juice, the vinegar prepared from the juice was 9.55 RUE mg/mL, and the vinegar prepared from the juice was 6.48 RUE mg/mL, and the total flavonoid content was also increased. Vinegar prepared from gourd was higher than vinegar prepared from juice (p<0.05).

Total flavonoids, like total polyphenols, are known to exist in large amounts in the fruit of bokbunja. They have a strong antioxidant effect and are used as health functional food materials. Since they cannot be synthesized in the human body, it is known that they must be ingested through the diet. (Rice-Evans et al., 1996).

Anthocyanin is chrysanthemin with a cyanidin-based color. It is a water-soluble, water-soluble, bioavailable functional substance with high antioxidant activity (Tsuta et al., 1996), anticancer effect and anti-inflammatory effect (Ryu et al., 2000). have.

As a result of measuring the total anthocyanin content, the vinegar prepared from bokbunja extract was 6.05 CYE mg/mL, which was higher than the 5.13 CYE mg/mL of the vinegar prepared from the juice extract. and total anthocyanin were all high. Polyphenols, flavonoids and anthocyanin compounds, collectively referred to as phenolic compounds, are compounds known as powerful antioxidants. do.

The DPPH radical is a relatively stable free radical and has been widely used to measure antioxidants from natural materials (Lee et al., 2008).

As a result of measuring the IC ₅₀ value of the DPPH radical scavenging ability of vinegar prepared with bokbunja juice and juice, the vinegar prepared with bokbunja extract (10.23 μL/mL) was about 2 compared to vinegar (20.12 μL/mL) from bokbunja extract. It exhibited twice as much activity (p<0.05).

As a result of measuring the ABTS radical scavenging ability, similarly to the DPPH radical scavenging ability, vinegar (18.30 μL/mL) using Bokbunja extract showed about twice as much activity as vinegar (35.39 μL/mL) using juice (p<< 0.05).

Kim et al. (2017) reported that there are flavonoids and tannins as representative plant-based polyphenol materials, and that the total phenol content and antioxidant activity are proportional because the antioxidant mechanism of phenolic compounds is due to the radical scavenging action, which shows the same results as the present invention. indicated.

5) reducing power and FRAP

Among the various mechanisms showing antioxidant action, the action of donating electrons to active oxygen and free radicals to stabilize them is called reduction, and since substances with reducing power act as electron donors, they inhibit the production of intermediate products in the lipid peroxidation process and It is known to act as an antioxidant (Yoshino and Murakami, 1998).

5 and 6 show the results of measuring the reducing power and FRAP activity of vinegar prepared from bokbunja juice and juice.

At the same concentration, vinegar prepared from bokbunja extract showed a higher reducing power than vinegar prepared from juice, showing the same trend as the results of studies on DPPH and ABTS radical scavenging activity.

Also, as a result of measuring FRAP activity, vinegar prepared from bokbunja juice showed higher activity than vinegar prepared from bokbunja juice.

These results were consistent with the findings of Park et al. (2008), who reported that DPPH and ABTS radical scavenging activity, reducing power, and FRAP activity were highly correlated with total polyphenol, total flavonoid and total anthocyanin content.

Therefore, it is judged that vinegar using bokbunja extract has excellent antioxidant activity and will show the possibility of manufacturing vinegar with enhanced functionality.

In the case of using bokbunja pomace, which is discarded as a by-product of bokbunja processing according to the present invention, the result of acetic acid fermentation is excellent, and it exhibits high antioxidant activity and has a very excellent effect as a functional fermented vinegar, so it has great industrial applicability .

Korea Microorganism Conservation Center (Overseas) KCCM12808P 20201020

<110> Jeonbuk Institute for Food-Bioindustry BERRY&BIOFOOD RESEARCH INSTITUTE RYU, Yoon Sang <120> Method for preparing vinegar using black raspberry pomace fermented by Acetobacter pasteurianus <130> 2020-378 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1442 <212> RNA <213> Acetobacter pasteurianus <400> 1 gctggcggca tgcttaacac atgcaagtcg cacgaaggtt tcggccttag tggcggacgg 60 gtgagtaacg cgtaggtatc tatccatggg tgggggataa cactgggaaa ctggtgctaa 120 taccgcatga cacctgaggg tcaaaggcgc aagtcgcctg tggaggagcc tgcgtttgat 180 tagctagttg gtggggtaaa ggcctaccaa ggcgatgatc aatagctggt ttgagaggat 240 gatcagccac actgggactg agacacggcc cagactccta cgggaggcag cagtggggaa 300 tattggacaa tgggggcaac cctgatccag caatgccgcg tgtgtgaaga aggtcttcgg 360 attgtaaagc actttcgacg gggacgatga tgacggtacc cgtagaagaa gccccggcta 420 acttcgtgcc agcagccgcg gtaatacgaa gggggctagc gttgctcgga atgactgggc 480 gtaaagggcg tgtaggcggt ttgtacagtc agatgtgaaa tccccgggct taacctggga 540 gctgcatttg atacgtgcag actagagtgt gagagagggt tgtggaattc ccagtgtaga 600 ggtgaaattc gtagatattg ggaagaacac cggtggcgaa ggcggcaacc tggctcatta 660 ctgacgctga ggcgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg 720 ctgtaaacga tgtgtgctag atgttgggtg acttagtcat tcagtgtcgc agttaacgcg 780 ttaagcacac cgcctgggga gtacggccgc aaggttgaaa ctcaaaggaa ttgacggggg 840 cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgcagaac cttaccaggg 900 cttgaatgta gaggctgcag gcagagatgt ctgtttcccg caagggacct ctaacacagg 960 tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg 1020 caacccctat ctttagttgc catcaggttg ggctgggcac tctagagaga ctgccggtga 1080 caagccggag gaaggtgggg atgacgtcaa gtcctcatgg cccttatgtc ctgggctaca 1140 cacgtgctac aatggcggtg acagtgggaa gctaggtggt gacaccatgc tgatctctaa 1200 aagccgtctc agttcggatt gcactctgca actcgagtgc atgaaggtgg aatcgctagt 1260 aatcgcggat cagcatgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1320 caccatggga gttggtttga ccttaagccg gtgagcgaac cgcaaggacg cagccgacca 1380 cggtcgggtc agcgactggg gtgaagtcgt aacaaggtag ccgtagggga acctgcggct 1440 ga 1442

Claims (4)

Acetobacter pasteuria of Acetobacter pasteurian, which is characterized in that the bokbunja juice is fermented with acetic acid to prepare vinegar with a vinegar starter containing Acetobacter pasteurianus deposited with accession number KCCM12808P Vinegar manufacturing method made by acetic acid fermentation with Nous GBA5.
The method according to claim 1,
Bokbunja vinegar is prepared by putting bokbunja juice in 8 to 12 times the weight of water, extracting it at 100 °C for 5 to 7 hours, and then concentrating it to 15-20 °Brix with a reduced pressure concentrator to prepare a concentrate, and dry yeast in the concentrate. S. cerevisiae Fermivin was inoculated to 1×10 ⁹ CFU/kg, stirred, and fermented at 24 to 26 ° C. for 4 to 7 days to prepare an alcoholic fermentation solution;
Acetobacter paste for bokbunja juice, characterized in that it comprises the step of inoculating a vineger starter containing Acetobacter pasteurianus deposited with the accession number KCCM12808P in the alcoholic fermentation liquid and acetic acid fermentation Vinegar manufacturing method made by acetic acid fermentation with Tulianus GBA5.
3. The method according to claim 2,
Vineger starters are
A liquid medium containing 93.5 to 97.2 wt% of distilled water, 0.3 to 0.7 wt% of yeast extract, 0.2 to 0.5 wt% of peptone, and 2.1 to 3.2 wt% of mannitol Acetobacter pasteurianus ( Acetobacter pasteurianus ) deposited under the accession number KCCM12808P in Acetobacter pasteurianus, characterized in that it was prepared by inoculating and culturing with shaking at 29-31 ° C. for 70-74 hours. Vinegar manufacturing method made by acetic acid fermentation with GBA5.
3. The method according to claim 2,
The step of acetic acid fermentation is to inoculate a vineger starter in the alcoholic fermentation broth, inoculated at 9-12 vol% with respect to the total volume of the alcoholic fermentation broth, and cultured at 29-31 ° C. for 20-28 days, bokbunja Vinegar manufacturing method made by acetic acid fermentation of juiced gourd with Acetobacter Pasteurian GBA5.

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