JP3195592B2 - Persimmon vinegar and its production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to persimmon vinegar and a method for producing the same. More specifically, the present invention relates to a persimmon vinegar produced by inoculating a novel strain suitable for producing persimmon vinegar on astringent persimmon and fermenting the same, and a method for producing the same.

[0002]

2. Description of the Related Art Vinegar is classified into brewed vinegar produced by acetic acid fermentation of starch and alcohol, and synthetic vinegar produced using glacial acetic acid, water, spices and coloring agents. Brewed vinegar includes rice, sake lees, lemon, persimmon, apples, grape vinegar, etc. depending on the raw material, and mainly contains acetic acid produced by acetic acid fermentation, and a small amount of volatile or nonvolatile organic acids, sugars, amino acids and It is a typical fermented food with unique aroma and acidity because it contains esters and the like.

[0003] Persimmons are sweet persimmon (Diospyros kaki, L.) and astringent persimmon (Dio
spyros kaki, T.), with less fertilization and use of pesticides and higher palatability than other fruits.
Production and consumption are increasing every year, reaching M / T, 110,000M / T in 1991 and 155,000M / T in 1992.

[0004] Sweet persimmons have high palatability and are mostly used as fresh fruits. However, due to the softening phenomenon during distribution and storage and physiological obstacles due to environmental changes, the quality of the persimmon easily deteriorates. Many inferior fruits that cannot be converted easily occur. In addition, in the case of astringent persimmon, not only must the astringent or softening process be performed, but also preference is lower in preference than sweet persimmon. In addition, over-production has caused prices to plunge, and some have been thrown into trees without being able to be harvested, causing many economic losses.

[0005] Persimmon is rich in carbohydrates and vitamins A and C, and is known to be effective in contracting the large intestine, promoting intestinal juice secretion, and antitussive. However, despite its nutritional benefits, persimmons are limited in their use compared to other fruits due to the above-mentioned problems, and skewer persimmons, soft persimmons, mizumasa, dried persimmons, persimmon powder, persimmons Utilization is improved by providing processed foods such as vinegar.

[0006] Above all, persimmon vinegar is a traditional fermented food that has been produced and used by ordinary farmers for a long time, and has been favored as a folk remedy for removing hangovers, relieving fatigue and intestinal action. Recently, rapid economic growth and a shift in awareness of health have increased interest in traditional vinegar made from natural ingredients. Conventional persimmon vinegar that has been marketed and distributed until now is shipped as a product around July of the following year by natural fermentation of parallel persimmon fermentation of persimmons harvested in mid-November. However, the tannin component, which is abundant in persimmons, becomes a substrate for astringency and polyphenol oxidase, causing browning, inhibiting alcohol fermentation, and forming a complex with enzyme proteins, resulting in the formation of colored precipitates. May be done. Moreover, it is susceptible to contamination by various bacteria and is unsanitary.
Furthermore, persimmon vinegar has a different hue depending on the product, so that not only is the product value low, but also economical efficiency is low because long-term fermentation for 5 to 6 months makes mass production and storage difficult.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide a persimmon vinegar with improved product value and economic efficiency. A further object of the present invention is to provide a method for producing the persimmon vinegar.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above problems, and as a result, inoculated with a new strain suitable for the production of persimmon vinegar, using astringent persimmon as a raw material, and performing alcohol fermentation and acetic acid production. By producing the persimmon vinegar by fermentation, we succeeded in producing a persimmon vinegar without the above problems.

That is, the present invention is a persimmon vinegar having a total acid content of 4% or more, which is obtained by fermenting astringent persimmon. The present invention also crushes astringent persimmons and inoculates a sake mother,
Next, alcohol fermentation strain Saccharomyces kluyveri DJ
After inoculating 97 (KCTC 8842P) and performing alcohol fermentation at a temperature of 20 to 30 ° C, 150 to 1 under the same temperature as alcohol fermentation
A method for producing persimmon vinegar, comprising inoculating acetic acid fermented seed vinegar with stirring at 220 to 250 rpm for 70 hours to effect acetic acid fermentation. Hereinafter, the present invention will be described in detail.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION As will be specifically shown in the following Examples and Experimental Examples, the persimmon vinegar-containing beverage of the present invention is produced through the following steps. (1) Establishing the optimum conditions for alcohol fermentation and acetic acid fermentation in the production of persimmon vinegar by reaction surface analysis; (2) Using the fermentation strain corresponding to the crushed astringent persimmon under the established optimum conditions To produce persimmon vinegar by alcohol fermentation and acetic acid fermentation; (3) Measure the pH of the produced persimmon vinegar with a pH meter and determine the total acid to 0.
(4) measuring the hue and brownness of the produced persimmon vinegar with a color difference meter and indicating the L, a, b values of the Hunter; (5) ) Step of examining the turbidity change by measuring the absorbance using a certain amount of the produced persimmon vinegar; (6) Standard curve prepared with glucose after measuring the reducing sugar of the produced persimmon vinegar by DNS method (7) filtering the manufactured persimmon vinegar stock solution and analyzing it by HPLC for free sugars and organic acid components; (8) free persimmon vinegar contained in the manufactured persimmon vinegar Analyzing amino acids using an automatic amino acid analyzer; (9) calculating the total amount of tannin contained in the manufactured persimmon vinegar by measuring absorbance; (10) atomic absorption of the inorganic components of the manufactured persimmon vinegar Spectrophotometer and
(11) Analyzing the alcohol content of the produced persimmon vinegar by gas chromatography; (12) Volatile components of the produced persimmon vinegar were compared with Liquids according to the method of Maarse and Kepner. Continuous distillation extraction (SDE) devised by Nilerson
(13) mixing persimmon vinegar with food additives such as polydextrose, liquid fructose, pear concentrate, honey, citric acid, etc. The step of producing a beverage containing.

In the present invention, first, a persimmon vinegar is produced by inoculating a strain into crushed astringent persimmon and subjecting it to alcohol fermentation and acetic acid fermentation. In addition, each analysis method and the result of the manufactured persimmon vinegar are shown in the experimental example described later.

In the present invention, as a raw material of persimmon vinegar, 19
Shibu persimmon (D) produced in Cheongdo-gun, Gyeongsangbuk-do, Korea in November 1995
iospyros kaki, L.) was used after degassing at 30 ° C. with a carbon dioxide concentration of 80% or more.

In the alcohol fermentation of the present invention, Saccharomy isolated and identified from persimmon as an alcohol fermentation strain.
ces kluyveri DJ 97 with YPD (yeast extract 1%, pepto
ne 2%, glucose 2%, agar 2%) The cells were cultured in a slant medium at 25 ° C. for 24 hours and used while refrigerated at 4 ° C. In the acetic acid fermentation, a normal strain used in conventional persimmon vinegar production as an acetic acid bacterium was cultured at 30 ° C for 72 hours in the medium shown in Table 1 below at 4 ° C.
Used while refrigerated. The alcohol-fermenting strain was deposited with the Institute of Biotechnology on October 31, 1997, under accession number KCTC 884.
Deposited at 20.

[0014]

[Table 1]

The isolation of the alcohol-fermenting strain was performed using YPD YM (yea
st extract 0.3%, malt extract 0.3%, glucose 0.5%,
(Bacto peptone 0.5%, pH 6.0) medium was used, and the acetic acid bacteria were separated using the media shown in Table 2 below.

[0016]

[Table 2]

[0017] To prepare alcoholic mothers for alcohol fermentation, persimmons from which astringency has been removed are crushed and squeezed to give persimmon juice having a sugar concentration of 10%, which is sterilized with 15 lb for 15 minutes. Inoculate the sterilized medium with the bacteria and shake at 150 rpm for 25 minutes.
Incubation is performed at 38 ° C for 38 hours. The obtained culture was used as a mother for alcohol fermentation. As seed vinegar, persimmon juice is subjected to alcohol fermentation, the persimmon juice is sterilized, and acetic acid bacteria are shaken at 30 ° C., 200 rpm for 72 hours in the persimmon juice to which sugar, alcohol and acetic acid are added. It was prepared by culturing.

In the present invention, the yield of vinegar is represented by the amount of vinegar produced per 100 g of persimmon in% (w / v), and the alcohol fermentation yield is expressed as a theoretical yield relative to the amount of alcohol produced by the initial sugar concentration. Calculated as follows.

[0019]

(Equation 1)

In the present invention, a persimmon vinegar-containing beverage is produced by blending a food additive and purified water with the persimmon vinegar obtained above. The above food additive, polydextrose, liquid fructose, pear concentrate, a honey, additionally Sutebion 100S, vitamin B _2, vitamin C, nicotinamide,
Citric acid, sodium citrate, sodium L-glutamate, L-menthol, red L-500, drink flavor and the like are used.

Specific examples of the composition of the persimmon vinegar-containing beverage include:
100% natural persimmon vinegar 1-10%, polydextrose 0.7-1.7
%, Liquid fructose 7-13%, pear concentrate (68 Brix) 1.0-2.0%, honey 0.3-0.6%, Stebion 100S 0.015-0.025%, vitamin B ₂ 0.0004-0.0008%, vitamin C 0.02-0.05%, nicotine Acid amide 0.005-0.015%, citric acid 0.02-0.08%,
Sodium citrate 0.015-0.025%, sodium L-glutamate 0.01-0.02%, L-menthol 0.0001-0.0002%
, Red L-500 0.025-0.035%, Drink flavor 0.
15-0.25%, the rest contains purified water, most preferably 100
4% natural persimmon vinegar, 1.4% polydextrose, liquid fructose 10.
5%, pear concentrate (68 Brix) 1.5%, honey 0.47%, Stebion 1
00S 0.019%, Vitamin B ₂ 0.0006%, Vitamin C 0.04%,
Nicotinamide 0.009%, citric acid 0.05%, sodium citrate 0.019%, sodium L-glutamate 0.014%, L-
Menthol 0.00014%, Red L-500 0.03%, Drink flavor 0.19%, balance includes purified water.

[0022]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to Examples and Experimental Examples, but the scope of the present invention is not limited only to these Examples and Experimental Examples.

[Example 1] (Survey of alcohol fermentation conditions and acetic acid fermentation conditions by reaction surface analysis) In the reaction surface analysis method, when two or more regression variables affect the reaction Statistical analysis of the surface formed by variables and design related to this.

In the reaction surface analysis, as in the case of the regression analysis, the reaction amount due to the change of the regression variable is predicted from the functional relationship between the regression variable and the reaction variable, and at the same time, the reaction amount becomes maximum, minimum, or optimal. Estimate the value of the regression variables and design an experiment that can perform such a procedure most rationally.

In the reaction surface analysis, a first-order model and a second-order model represented by the following equations are used to estimate an unknown functional relationship between a regression variable and a response variable:

[0026]

[Equation 2] Primary model: Y = β ₀ + β ₁ X ₁ + β ₂ X ₂ +... + Β _k X _k + ε

[0027]

(Equation 3)

The multiple regression model exemplified in [1] is used. To estimate the parameter of this function, the least squares method is used as in the regression analysis. It is effective to collect the data for estimating the parameters in an experiment designed for reaction surface analysis. At this time, alcohol and acetic acid fermentation conditions were established by the reaction surface experiment design used.

<First Step: Alcohol Fermentation Conditions> A central synthesis design was used to optimize the reaction conditions of alcohol fermentation, and SAS (statistical analysis s) was used for the reaction surface analysis.
ystem) package was used. Experimental variables during alcohol fermentation include sugar content ( ⁰ Brix) (X ₁ ), shaking speed (X ₂ ), and fermentation time (X ₃ ).
-1,0,1,2) and shown in Table 3 below.

[0030]

[Table 3]

The reaction variables (Y _n ) relating to the quality characteristics of alcohol fermentation include alcohol content (Y ₁ ), residual sugar content (Y ₂ ), total tannin content (Y ₃ ), pH (Y ₄ ), and acidity (Y ₅ ), hue (Y ₆ ), etc. A central synthesis design method shown in Table 4 was established so that the above three factor variables (X ₁ , X ₂ , X ₃ ) have the above five levels (−2, −1, 0, 1, 2). The experiment was performed under 16 setting conditions in the table. At this time, the fermentation temperature was fixed at 25 ° C. in consideration of the characteristics of the strain, and the fermented solution was filtered with a filter (Whatman No. 1), and the whole amount was kept constant and used as an analysis sample.

[0032]

[Table 4]

Since the reaction surface analysis method is a method for finding an optimum reaction condition by using a graph, the optimum condition can be determined by using contour lines. In alcohol fermentation using persimmons, the alcohol content in the fermented liquid after fermentation is important as a substrate for acetic acid fermentation, which is the next operation.
Also, the color difference (ΔE) can be a good quality index for producing vinegar having a persimmon hue.

Accordingly, the degree of change with respect to the fermentation time, the shaking speed and the sugar content, which are the respective experimental variables, are shown on a contour map, using the alcohol content and the color difference as limiting functions.

[0035] Reaction surfaces due to various variables (fermentation time, shaking speed, and sugar content) during alcohol fermentation do not always coincide with each other, so that appropriate restrictive conditions are required. Within the given experimental conditions, the appropriate alcohol content for acetic acid fermentation 6.0-7.0%, the total color difference of the acceptable quality change index range 30.00-41.00 set as limiting conditions,
The experimental conditions satisfying this restriction condition were displayed on an overlapping contour map.

First, FIG. 1 shows a case where the relationship between the shaking speed and the sugar content is made a function. It was shown that the sugar content and the shaking speed satisfying the above restriction conditions were 13 to 16% and 110 to 200 rpm, respectively.

FIG. 2 shows a case where the sugar content and the fermentation time are used as variables under the same restriction conditions as described above. The sugar content and the fermentation time satisfying the above-mentioned restriction conditions are 13 to 15%, 70 to 1 respectively.
It was shown to be 40 hours.

FIG. 3 shows a case where the fermentation time and the shaking speed are used as variables. It was shown that the fermentation time and the shaking speed satisfying the above restriction conditions were 80 to 100 hours and 50 to 150 rpm, respectively.

FIG. 4 shows the results of a reaction surface analysis with the shaking speed fixed at 100 rpm and only the fermentation time and sugar content as variables. 6 A suitable alcohol content as a substrate for acetic acid fermentation
~ 7%, the color difference is 30.00 ~ 41.00, and the limit condition that can satisfy the efficiency of persimmon vinegar production due to the characteristics of the strain, etc. are determined to be 13 ~ 16% sugar content and 96 ~ 120 hours fermentation time. Was.

FIGS. 5 and 6 show reaction surfaces prepared for observing the degree of change in alcohol content and hue (= color difference: ΔE) depending on sugar content and fermentation time. FIG. 5 shows that although the alcohol content is hardly affected by the fermentation time, it is strongly affected by the sugar content, and that the alcohol content is greatly increased by increasing the sugar content. FIG. 6 also shows that the color difference takes a larger value as the fermentation time increases in a low sugar content range, and takes a lower value as the fermentation time increases in a high sugar content range.

Table 5 shows a regression equation of a secondary model obtained by performing a reaction surface analysis with the shaking speed fixed and only the sugar content and time as independent variables. This indicates that the model formula was established with a relatively high coefficient of determination for the alcohol content and the color difference.

[0042]

[Table 5]

As a result of the reaction surface analysis, the conditions obtained to optimize the alcohol fermentation were as follows: the fermentation time was 96 to 120 hours,
It was found that the sugar content was 13.0-16.0% and the shaking speed was 100 rpm. Therefore, the optimal conditions in alcohol fermentation with the alcohol content indicating the yield of alcohol fermentation being 6.0 to 7.0% and the color difference that can be made as a quality index being 30.00 to 41.00, the fermentation time is 90 to 120 hours and the sugar content is 13.0 ~ 16.
0%, and the shaking speed was shown to be 100 rpm.

<Second stage: acetic acid fermentation conditions> In order to optimize the acetic acid fermentation reaction conditions, the central synthesis design method was used as in the case of the alcohol fermentation, and the reaction surface regression analysis was performed.
SAS was used. Shaking speed as an independent variable during acetic acid fermentation
(X ₁ ) and fermentation time (X ₂ ) were set, and these factor variables were coded into five levels (-2, -1,0,1,2) and are shown in Table 6 below.

[0045]

[Table 6]

The reaction variable (Y_n)
As total acid (Y₁), pH (Y_Two), Brownness (Y _Three), Turbidity (Y_Four), Hue
(Y_Five) Etc. were used.

The central synthesis method shown in Table 7 is established so that the two factor variables (X ₁ , X ₂ ) have the above five levels (-2, -1,0,1,2). The experiment was performed under the ten conditions set in the table. At this time, the fermentation temperature and the initial acidity were adjusted to a fermentation temperature of 30 ° C. and an appropriate acidity of 1.0 in consideration of the characteristics of the strain, etc., and after fermentation, the mixture was centrifuged and the supernatant was used as analytical data.

[0048]

[Table 7]

In the acetic acid fermentation, the total acidity (indicating the yield of acetic acid fermentation) and the color difference (a quality index for producing vinegar close to the hue of persimmon) among the various reaction variables, as in the case of the alcohol fermentation described above. Was used as the restriction function.

Therefore, in the case of acetic acid fermentation, regions where the limiting conditions match when the dependent variable is the total acidity and the color difference and the independent variable is the fermentation time and the shaking speed are shown by overlapping contour maps.

As shown in FIG. 7, the total acidity was 6.8-8.3%, and the color difference was 4
The fermentation time and the shaking speed satisfying the condition of 7.34 to 47.70 were respectively 150 to 170 hours for the fermentation time and 220 to 250 rpm for the shaking speed.

FIGS. 8 and 9 show reaction surfaces prepared to measure the degree of change in the total acidity and color difference with changes in the shaking speed and fermentation time, which are independent variables. The effect of each independent variable on the obtained reaction surface can be determined by the magnitude of the gradient. FIG. 8 shows that the effects of shaking speed and fermentation time on the change in total acidity are similar, while FIG. 9 shows that the color difference has a lower value as the fermentation time is shorter and the shaking speed is faster. It was shown to take.

As a result of the reaction surface analysis, it was found that the optimal conditions for acetic acid fermentation satisfying the control function were a fermentation time of 150 to 170 hours and a shaking speed of 220 to 250 rpm. Therefore, the total acidity indicating the yield of acetic acid fermentation is 6.8-7.8%, and the color difference is 40.00-
The optimum conditions for acetic acid fermentation satisfying the condition of 40.30 were shown to be a fermentation time of 150 to 170 hours and a shaking speed of 220 to 250 rpm.

Example 2 Production of Persimmon Vinegar Persimmon vinegar was produced according to the optimum conditions established by the reaction surface analysis for alcohol fermentation and acetic acid fermentation in Example 1 above. That is, crushed astringent persimmon, 0.1% pectinase (V
ision Co., Sumyzyme MC) at 40 ° C. for 12 hours.
Thereafter, 5% of sake brewer was inoculated, and pectinized astringent persimmon was subjected to alcohol fermentation at 25 ° C. under the established optimum conditions. The fermentation product was filtered through a cheese cloth and the filtrate was used as a substrate for acetic acid fermentation. In acetic acid fermentation, 5.0% seed vinegar is inoculated into the filtered alcohol fermentation liquor, and wo
rking volume 4L fermenter (Korea Fermentor Co., Ltd.
KF-5L). After fermentation, it was used as an analytical sample.

Under the conditions established by the central synthesis design method, R ² of the regression equation for the alcohol content was 0.9386, the residual sugar was 0.9479, and the color difference was 0.8576, which was significant. Based on this fact, the influence of each independent variable was examined by the magnitude of the gradient for the non-independent variable on the surface. As a result, the alcohol content was hardly affected by the fermentation time, but increased greatly with the sugar content. As for the color difference, as the fermentation time became longer, a large value was obtained at a low sugar content, but a small value was obtained at a high sugar content.

In alcohol fermentation, as described above, changes in alcohol content and color difference with the sugar content and fermentation time can be expected. As a result of examining the optimal conditions by reaction surface analysis, the fermentation time was 96 to 130 hours, and the initial sugar content was 13.0
116.0%, shaking speed was established at 100rpm. Taking into account these optimum conditions, as shown in FIG. 10, crushed pears were treated with 0.1% pectinase (Vision (Co) Sumyzyme MC) for 40%.
Enzyme treatment at 12 ° C for 12 hours, SaccharomycesKluyveri DJ 9
7 (KCTC 8842P) was inoculated with 5% of the sake mother obtained by culturing in astringent persimmon extract medium, and shaken at 25 ° C, 100 rpm, 11
It was subjected to alcohol fermentation for 3 hours.

Under the conditions established by the central synthesis scheme for acetic acid fermentation, R ² of the regression equation for total acid is 0.9747,
The color difference was 0.7057, which was significant.
Shaking speed and fermentation time had a similar effect on total acid, lower color difference values were obtained with shorter fermentation time and higher shaking speed.

As a result of examining the optimum conditions for acetic acid fermentation by reaction surface analysis, the fermentation time was established at 150 to 170 hours, and the shaking speed was established at 220 to 250 rpm. As shown in FIG. 10, put the alcohol fermentation broth into a fermenter of working volume 4L, inoculate 5.0% seed vinegar, 30 ℃, 230rpm,
It was subjected to acetic acid fermentation at 0.1 vvm for 216 hours.

[Experimental Example 1] (Changes in pH and Total Acid of Persimmon Vinegar) In this experimental example, the pH of the persimmon vinegar produced in Example 2 was measured with a pH meter, and the total acid was measured with a 0.1N NaOH solution. Neutralized and converted to acetic acid. PH and total acid of astringent persimmon during alcohol fermentation,
As shown in FIG. That is, the pH was initially 5.65,
It gradually decreased as the fermentation time elapsed, and reached 4.27 after 5 days of fermentation. Total acid was initially 0.15%,
It gradually increased as the fermentation time passed, and was 0.29% after 5 days of fermentation. FIG. 12 shows the pH and total acid of astringent persimmon during acetic acid fermentation. The pH was initially 4.25, but gradually decreased over the course of the fermentation time to 2.98 after 8 days of fermentation, and this pH decrease was slower than that of alcohol fermentation. Total acid is 1.
It was 68%, but gradually increased as the fermentation time elapsed, and reached 5.81% after 8 days of fermentation, indicating that the total acid increased sharply compared to alcohol fermentation.

[Experimental Example 2] (Changes in hue and brownness of persimmon vinegar) A test for the color of the persimmon vinegar produced in Example 2 was carried out using a color difference meter (C
hromometer, Model CR-200, CT-210, Minolta Co., Jap
An) by measuring Hunter's L, a, b values,
Brownness (a / b) was also determined. The color difference (ΔE) of persimmon vinegar was based on the color of the crushed raw material persimmon. Changes in the hue and brownness of the astringent persimmon during alcohol fermentation are as shown in Table 8 below.

[0061]

[Table 8]

The L value indicating lightness and the b value indicating yellowness were each higher than that of sweet persimmon, and the a value indicating redness was lower. Brownness
(a / b) increased gradually as alcohol fermentation proceeded, but was lower than that of sweet persimmon.

The color change of astringent persimmon during acetic acid fermentation is shown in Table 9 below.
It is as shown in FIG. The L and b values were high and the a value was low, but the brownness tended to decrease gradually.

[0064]

[Table 9]

[Experimental Example 3] (Change in turbidity of persimmon vinegar) The turbidity of the persimmon vinegar produced in Example 2 was determined by taking a certain amount of a sample.
The absorbance at 660 nm was measured and shown. The turbidity of the astringent persimmon was 2.85 at the beginning, but gradually increased as the fermentation time elapsed, and reached 3.70 on the fifth day of alcohol fermentation. The results are as shown in Table 10 below.

[0066]

[Table 10]

[Experimental Example 4] (Changes in reducing sugars and total sugars in persimmon vinegar) The reducing sugars in the persimmon vinegar produced in Example 2 were measured using the DNS method and compared with a standard curve prepared with glucose. Its content was determined. For total sugar, sample 20mL and distilled water 180m
After diluting to 25 L and adding 20 mL of 25% HCl, the mixture was hydrolyzed on a boiling bath for 3 hours, and the hydrolyzed solution was neutralized with a 10% NaOH solution to give 25%.
The volume was adjusted to 0 mL. The quantification was performed in the same manner as for the reducing sugar.

As shown in Table 11 below, the reducing sugars and total sugars of astringent persimmon during alcohol fermentation were initially 12 respectively.
5.4, 132.0 mg / mL, but decreased significantly as the fermentation time elapsed, and on the 5th day of fermentation, 4.1 and 4.2 m, respectively.
g / mL.

[0069]

[Table 11]

The contents of reducing sugars and total sugars during acetic acid fermentation were initially 3.2 and 3.5, respectively, as shown in Table 12 below.
mg / mL, but gradually decreased as the fermentation time elapsed, and reached 2.8 and 2.9 mg / mL on the fifth day of fermentation. No major changes have appeared since then.

[0071]

[Table 12]

[Experimental Example 5] (Analysis of free sugars and organic acids of persimmon vinegar) In order to analyze free sugars and organic acids of persimmon vinegar produced in Example 2, fats and oils were extracted from crude persimmon vinegar with hexane. The dye and protein components were removed from the components using a 0.45 μm membrane filter and Sep-pak _C18 . To analyze ascorbic acid in organic acids, dilute 5 mL of distilled water to 1 mL of sample,
Treat with sonication for 5 minutes, filter and save the filtrate. 5 mL of distilled water was further added to the residue, and the same operation as above was performed. After this operation was repeated three times, the total amount of the filtrate was adjusted to 20 mL, the mixture was filtered with HPLC filter paper, and analyzed under the conditions shown in Table 13 below.

[0073]

[Table 13]

During alcohol fermentation, free sugars including glucose, fructose and sucrose were detected. As shown in FIG. 13, the sweet persimmon liquid contains 0.01% glucose, 0.15%
Fructose and 0.01% sucrose. For the astringent persimmon, its free sugar content is shown in Table 14 below.
Is shown in

[0075]

[Table 14]

As shown in Table 14, immediately after fermentation, glucose was the most abundant of free sugars. One day after fermentation, the glucose content sharply decreased from 5.63% to 2.64%, and the fructose content gradually changed from 5.21% to 4.30%. For sucrose, only a trace amount remained at 0.62%. Then, as alcohol fermentation progresses,
Free sugars decreased, and after 5 days of fermentation only fructose was reduced to 0.
10% appeared and glucose and sucrose were traces.

Experimental Example 6 (Quantification of Free Amino Acids in Persimmon Vinegar) To 10 mL of persimmon vinegar produced in Example 2, 30 mL of ethanol was added.
The protein was precipitated and removed overnight at room temperature. The supernatant was centrifuged at 3,000 rpm for 10 minutes, and only the supernatant was taken and dried in a boiling water bath. Add pH 2.2 citrate buffer
After adding and diluting 10 mL, 0.45 μm membrane filter
And filtered. The filtrate was analyzed with an automatic amino acid analyzer under the conditions shown in Table 15 below.

[0078]

[Table 15]

Experimental Example 7 (Quantification of Total Tannin of Persimmon Vinegar) The total tannin of persimmon vinegar produced in Experimental Example 2 was quantified according to AOAC. That is, after mixing 1 mL of the sample solution with 5 mL of the Folin-Denis reagent, 5 mL of a saturated Na ₂ CO ₃ solution was mixed with stirring, and the resulting solution was allowed to stand at room temperature for 30 minutes. Its 760n
The absorbance at m was measured and compared to the absorbance value of a previously prepared standard curve to estimate total tannin. At the beginning of the alcohol fermentation, the total tannin of the astringent persimmon was 6.44 mg / mL, but gradually decreased as the fermentation time elapsed.
On the day, it was 4.12 mg / mL. Changes in total tannin during acetic acid fermentation are as shown in Table 16 below.

[0080]

[Table 16]

The change in total tannin was initially 4.10 mg / mL
But gradually decreased as the fermentation progressed
On the day, it was 3.82 mg / mL. This decrease was less than in alcoholic fermentation, and no inhibition of acetic acid fermentation by total tannin was observed.

[0082] Experimental Example 8] (persimmon vinegar inorganic component analysis) decomposing agent persimmon vinegar 100mL prepared in Example _{2 (HClO 4: H 2 SO} 4: H 2
(O ₂ = 9: 2: 5, v / v) Add 25 mL and slowly heat on a hot plate from a low temperature until the solution is completely decolorized so as to decompose the compounds contained in the persimmon vinegar Then, filtration (Whatman No. 2) was performed and the filtrate 100 mL
I got The filtered sample was analyzed for inorganic components using an atomic absorption spectrophotometer under the conditions shown in Table 17 below. molybd
The es blue colorimetric method was used to determine the phosphorus content in persimmon vinegar.

[0083]

[Table 17]

[Experimental Example 9] (Change in alcohol content of persimmon vinegar) After adding 1 mL of n-amyl alcohol as an internal standard substance to 100 mL of the clear sample solution prepared in Example 2, 100 mL of deionized water was added. Followed by heat distillation. 20 mL of the distillate was subjected to gas chromatography under the conditions shown in Table 18 below.

[0085]

[Table 18]

The results of analysis of the alcohol components during alcohol fermentation are shown in FIG. 14 and Table 19 below.

[0087]

[Table 19]

From the astringent persimmon, six kinds of alcohols including methanol, ethanol, isopropyl alcohol, n-propyl alcohol, isobutyl alcohol and isoamyl alcohol were detected together with acetaldehyde. As alcohol fermentation progressed, the content of all of the above-mentioned alcohols tended to increase until day 4 of fermentation, but from day 5 of fermentation, all of them except ethanol tended to decrease gradually. there were. The content of ethanol was 346 ppm, but increased to 75,673 ppm on the fifth day of fermentation.

[Experimental example 10] (Change in volatile components of persimmon vinegar) Like persimmon juice and persimmon vinegar, like method according to Maarse and Kepner
Volatile components were collected using a continuous distillation extraction (SDE) method devised by ns and Nilerson and identified by GC and GC-MS. That is, 500 mL of a sample and 25 mL of distilled water were placed in a flask maintained at 100 ° C.
0 mL was added together with 4-decanol (10 ppm), which is an internal standard substance, and mixed and stirred. Another flask maintained at 40 ° C. was charged with ether. Volatile components were collected from these two flasks for 1 hour, and the conditions shown in Table 20 below were satisfied.
Analysis was performed using GC and GC-MS.

[0090]

[Table 20]

As shown in FIGS. 15 and 16 and Table 21 below, eight kinds of volatile components were detected after alcohol fermentation and ten kinds of volatile components were detected after acetic acid fermentation.

[0092]

[Table 21]

The ethanol content of each sample decreased most greatly after acetic acid fermentation, whereas acetic acid was not detected after alcoholic fermentation, but reached 46,982.72 ppm after acetic acid fermentation. In addition, the ethyl acetate of astringent persimmon after alcohol fermentation
184.43 ppm detected, but after completion of acetic acid fermentation 65.90 ppm
Greatly reduced until. 2-hydroxy-2-butanone and 3-
Other volatile components except methyl-butanoic acid had lower contents in the sample after acetic acid fermentation than in the sample after alcohol fermentation.

Example 3 (Production of Persimmon Vinegar Beverage Composition) A persimmon vinegar-containing beverage composition was produced using the persimmon vinegar produced in Example 2 in combination with the food additives shown in Table 22 below.
All ingredients used have passed official standards for foods and food additives. At the time of use, groundwater was filtered with a demineralization device to obtain purified water. According to the mixing ratio shown in Table 22, persimmon vinegar and an additive were mixed and formed together with purified water. Until the mixture is completely dissolved
Stirred at 50 ° C. for 30 minutes. The mixture was instantaneously sterilized at 90 ° C. for 30 seconds with a heat exchanger, and then passed through a 0.5-micron filter. The sterilized and filtered liquid is filled into a bottle by an automatic filling device in a fixed amount, and after sealing, the bottle is heated to 90 ° C.
For 15 minutes, cooled to 40 ° C., and air bubbles were removed. After that, a trademark was applied by an automatic labeling machine, and the product was put into an inner box and an outer box by predetermined quantity, packaged, inspected item by item, and shipped as a finished product only to a product conforming to the specification of the product standard.

[0095]

[Table 22]

[0096]

According to the present invention, astringent persimmon is transformed into a novel strain Sacc
Fermentation using haromyces kluyveri DJ 97 (KCTC 8842P) provides persimmon vinegar with improved product value and economic efficiency.

[Brief description of the drawings]

FIG. 1 shows the degree of change in alcohol content as a function of shaking speed and sugar content.

FIG. 2 shows the degree of change in alcohol content when sugar content and fermentation time are used as variables.

FIG. 3 shows the degree of change in alcohol content when the fermentation time and the shaking speed are used as variables.

FIG. 4 shows the results of a reaction surface analysis with the shaking speed fixed and only the fermentation time and sugar content as variables.

FIG. 5 shows the degree of change in alcohol content depending on the sugar content and the fermentation time.

FIG. 6 shows the degree of change in hue depending on the sugar content and the fermentation time.

FIG. 7: When the independent variables are fermentation time and shaking speed,
Indicates an area where the restriction condition matches.

FIG. 8 shows a reaction surface formed to determine the degree of change in total acid with the shaking speed and time, which are independent variables.

FIG. 9 shows a reaction surface formed to determine a change in color difference depending on shaking speed and time, which are independent variables.

FIG. 10 is a schematic diagram of a method for producing astringent persimmon vinegar.

FIG. 11 shows pH and total acid during alcoholic fermentation.

FIG. 12 shows pH and total acid of astringent persimmon during acetic acid fermentation.

FIG. 13 shows free sugar content during alcoholic fermentation.

FIG. 14 shows the results of analysis of alcohol components during alcohol fermentation.

FIG. 15 shows a gas chromatogram obtained by analyzing volatile components after alcohol fermentation.

FIG. 16 shows a gas chromatogram obtained by analyzing volatile components of persimmon vinegar.

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-61-202679 (JP, A) "Science of series foods Science of vinegar" edited by Shigeru Otsuka et al., Asakura Shoten, p. 100, 1996 (58) Field surveyed (Int. Cl. ⁷ , DB name) C12J 1/00-1/10 JICST / JAFIC (JOIS) WPI (DIALOG)

Claims (1)

(57) [Claims] [Claim 1] The astringent persimmon is crushed and inoculated with a sake mother, followed by alcohol fermentation strain Saccharomyces kluyveri DJ 97 (KCTC
8842P) and fermented with alcohol at a temperature of 20-30 ° C, and then at the same temperature as alcohol fermentation for 150-170 hours22
A method for producing persimmon vinegar, comprising inoculating acetic acid fermented seed vinegar while shaking at 0 to 250 rpm to effect acetic acid fermentation.