KR20110019972A - A method for preparing eucommia ulmoides vinegar composition - Google Patents

Abstract

PURPOSE: An Eucommia ulmoides Oliver vinegar composition is provided to develop functional fermentation beverage for reducing body fat. CONSTITUTION: A method for preparing an Eucommia ulmoides Oliver vinegar composition comprises: a step of extracting Eucommia ulmoides; a step of mixing Eucommia ulmoides Oliver extract and apple concentrate and inoculating alcohol fermentation yeast for primary alcohol fermentation; and a step for inoculating acetic acid fermentation strain to the extract for secondary acetic acid fermentation. An Eucommia ulmoides Oliver vinegar drink contains the composition as an active ingredient.

Description

A method for preparing Eucommia ulmoides vinegar composition

The present invention relates to a method for producing the larvae vinegar composition, more specifically, to extract the larvae to prepare a larvae extract, and to prepare a primary alcohol fermentation by mixing the prepared larvae extract and apple concentrate, acetic acid alcohol-fermented larvae extract The present invention relates to a method for producing a worm vinegar composition comprising fermentation to obtain a worm vinegar composition, and a worm vinegar composition prepared therefrom.

As national income improves and consumers' interest in health-oriented functional foods increases, there is a continuous demand for products that add health functionalities to favorite foods. Therefore, processing using food materials with unique taste, aroma and pharmacological effect The development and commercialization of food will be promoted and its consumption will continue to increase.

In addition, the Korea-US FTA is expected to cause considerable damage to domestic farmers, and to overcome it wisely, it is time to focus on strengthening international competitiveness by developing new products or new food materials. In particular, to protect domestically grown farmers, imports have been banned since 1993, which has been classified as a supply / demand control item. However, domestic consumption of tofu is rapidly decreasing due to economic recession and limited use. Compared with the domestic demand and annual output of the two layers, the domestic consumption is still satisfied. Therefore, in order to improve the competitiveness and productivity of domestic production, the development of processing technology for creating value added is not necessary. It is becoming.

On the other hand, the development of functional food has the advantage that the development cost is cheaper than the drug development and can be commercialized in a relatively short time. In order to utilize tofu as a functional food material, an experimental study on how to use it as a raw material for purpose-oriented functional food is needed.

Vinegar is a typical alkaline food that is known to be effective in breaking down lipid peroxide causing atherosclerosis or thrombosis, preventing atherosclerosis, and releasing corticosteroids that manage stress, digestion and appetite. It contains various ingredients that revitalize the human body and is recognized for its function as a well-being drink. Vinegar has been used mostly for seasoning rather than drinking directly because of its strong sourness. However, some drinking vinegar beverages have been developed, and the company's lectures on the vinegar health theory and the media's health function on the vinegar have attracted the attention of consumers.

In addition, with the fever of well-being of modern consumers, interest in the health functionalities of drinking beverages in the domestic and overseas beverage markets has increased, leading to a change in consumers' purchasing patterns, and the domestic vinegar market generated sales of about 45 billion won in 2007. The industry is analyzing. Moreover, seasoned vinegars, which accounted for about 77% of the total market as of 2005, posted about 30% growth compared to the previous year, and drinking vinegar recorded 67% growth compared to 2004.

On the other hand, Eucommia ulmoides Oliver is a deciduous broad-leaved arboreous tree of the dicotyledonous rosemary locust and grows up to 40 cm in diameter and 15 m in height. Growing in mountains and fields, it is one of China's specialty plants and is cultivated in large quantities in southern China. It is well known for its effects of lowering cholesterol, improving hyperlipidemia and lowering blood pressure, inhibiting cancer cell proliferation, tonic, sedation, and diuresis. It is known. Folks in the leaves decoctions are used for neuralgia, hypertension, or even tea.

Republic of Korea Patent Application No. 10-2001-0003645 "Method for producing chopped vinegar" includes the steps of washing the head worms and extracting the essence for more than 24 hours on low heat, after the step, wash the glutinous or non-glutinous rice in clear water 10 After immersing for more than an hour and cooling the water at room temperature by making a gourd rice, after the step, crush the traditional yeast into a grain of rice, mix all of the tofu extract, cooled soybean rice, and put it in a fermentation vessel 3 at a temperature of 25 degrees to 30 degrees Celsius. It is disclosed a method for producing two vinegar vinegar characterized in that the fermented water for a week to 4 weeks, after the step, by filtration of the fermentation broth at a temperature of 10 to 15 degrees Celsius for a period of 3 to 12 months.

However, the above method is a method of preparing tofu vinegar in the traditional method by using tofu as a main raw material and using rice and traditional yeast, in terms of yield or functionality in order to use as a functional food material for the purpose of using tofu as a functional food material. Very poor situation.

Therefore, the present inventors extract the active ingredient of the untreated soybean, and mix it with apple concentrate as a natural material to prepare a vinegar composition in a two-step fermentation process through primary alcohol fermentation and secondary acetic acid fermentation, based on this As a base, the body fat reduction functional fermented beverage was developed, the quality characteristics thereof, and the present invention was achieved.

Therefore, the object of the present invention

The present invention provides a method for preparing the larvae vinegar composition.

The method according to the present invention comprises the steps of extracting the larvae to prepare a larvae extract; Mixing the Tofu extract and apple concentrate, and then inoculating alcohol fermentation yeast with primary alcohol fermentation; Inoculating acetic acid fermentation strain to the primary alcohol fermented soybean extract, followed by secondary acetic acid fermentation; And obtaining the secondary acetic acid fermented bean vinegar composition.

In the method according to the present invention, the optimum extraction conditions for preparing the larvae extract is preferably 90 to 100 ℃ extraction temperature, 5 to 10 hours extraction time and 40 to 70% (v / w) solvent ratio.

In the method according to the present invention, the primary alcohol fermentation step of the larvae extract is preferably carried out with a fermentation temperature of 25 to 30 ℃, an initial sugar content of 12 to 16 ^o Brix and a fermentation time of 2-3 days.

As the strain used in the primary alcohol fermentation step, a conventional yeast used for alcohol fermentation, for example, Saccharomyces cerevisiae may be used. Preferably, the alcohol fermentation strains DUJ-98, DUK-99, DUL isolated from larvae Any one of -11, DUM-12 and DUN-3 can be used, and more preferably, S. cerevisiae DUJ 98 can be used.

In the method according to the present invention, the second acetic acid fermentation step of the alcohol fermented tofu extract extract inoculated acetic acid fermentation strain in the alcohol fermented tofu extract and the initial acidity 1 to 25%, shaking speed 200 to 250 rpm, fermentation time It is preferably carried out in 6 to 7 days.

As the strain used in the second acetic acid fermentation step, a conventional strain used for acetic acid fermentation may be used, and preferably, the genus Acetobacter sp.

Using the worm extract as described above can be obtained chungbu vinegar composition through the primary alcohol fermentation and secondary acetic acid fermentation step.

The two-step fermentation method through the primary alcohol fermentation and secondary acetic acid fermentation according to the present invention as described above is low in yield and takes a long period of more than 6 months, high content of lactic acid is a conventional parallel combined fermentation that causes off-flavor occurrence The problem of the method can be improved.

In particular, the primary alcohol fermentation is possible to shorten the fermentation time and produce a high yield of drunk, it is expected to contribute to the quality of fermented products. For example, in the case of persimmon vinegar, the persimmon vinegar produced by two-stage fermentation showed higher total tannin content than the commercial persimmon vinegar. In the case of grape vinegar and apple vinegar, the quality of the product produced by the two-step fermentation method is generally It is reported to be excellent.

In the wormwood vinegar composition according to the present invention, the content of nipposide was 0.60 ㎍ / mL, the total acid content was relatively high at 6.36%, and the acetic acid content of each organic vinegar was 5395.56 mg%. In the order of malic acid, succinic acid, tartaric acid and citric acid, the contents were high. Fructose was found to have the highest content of sugar, and dietary fiber content of vinegar was 0.5%. Inorganic components were present in a large amount of K, and then the content of Na, Ca, Mg was high. The total amino acid content of each vinegar was 27.64 mg%, showing the highest aspartic acid content.

The present invention provides a Tobacco vinegar beverage prepared based on the Tobacco vinegar composition.

Tofu vinegar beverage according to the present invention, preferably 3 to 12% by weight of the worm vinegar composition, 3 to 7% by weight apple concentrate, 8 to 15% by weight sucrose, 0.2 to 0.7% by weight honey, 2 to 5% by weight of polydextrose, 0.01 to 0.03 weight percent glutamic acid, 0.01 to 0.05 weight percent ascorbic acid and 70 to 74 weight percent purified water.

As described above, in the present invention, the worm extract extract containing the functional ingredient of nipposide was successfully extracted from the worm, and the optimum extraction conditions according to the extraction temperature, the extraction time, and the solvent ratio were established using the reaction surface analysis method. In addition, the optimum alcohol fermentation conditions were derived by examining the culture conditions for alcohol production, and the fermentation characteristics of acetic acid were successfully monitored and the optimum vinegar fermentation conditions were derived.

According to the method of preparing the chungbu vinegar composition through the two-stage fermentation method of the present invention, chungbu vinegar composition was successfully prepared and functional quality characteristics were analyzed. In addition, through the quality characteristics analysis and consumer inspection, the formulation ratio preferred by consumers was derived, and the development of two-stage vinegared beverage was succeeded.

The wormwood vinegar composition prepared according to the method of the present invention as described above may contribute to the activation and development of the research and use of various types of healthy food or excellent functional dietary foods, economic impact of the commercialization of the wormwood vinegar beverage It is expected to be large.

In addition, considering the current price of tofu, there is sufficient price competitiveness compared to other drinking vinegar beverages on the market, and as a result, the cost reduction effect is expected, and contributing to the activation of the vinegar beverage market by developing new drinking vinegar beverages. It is thought that it can contribute to the increase of farm household income through collective cultivation. In addition, it can be applied to the development of new functional food materials, and it is helpful to revitalize the local economy, and enhance the competitiveness of SMEs through the launch of functional-related products utilizing various bioactive substances of Doochung, and anticipate the export of processed products, create new demand and It is expected to contribute to product development.

In addition, it is possible to apply to the development of a variety of fermentation products using the strain selected in the present invention, it is possible to develop a new functional food using the physiological activity of the fermentation by-products, the strain is applied to the development of fermentation products using similar herbs in domestic It is applicable to the development of fermented products using medicines, and it will be possible to develop high value-added products incorporating the latest science and technology into the food processing process.

Therefore, the successful implementation of the present invention is expected to contribute to the development of new products through technological innovation by not only contributing economically and industrially to domestic farmers, but also improving the shortcomings of the parallel fermentation process.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example 1 Establishment of Optimal Extraction Conditions

In this embodiment, in order to establish the optimum extraction condition of the two worms, the response surface analysis method was used to analyze the indicator effective ingredient according to the extraction conditions and to set the optimum extraction conditions.

The extraction range ( X ₁ ), extraction time ( X ₂ ) and extraction solvent ratio ( X ₃ ), which are important independent variables ( X _i ) in the extraction process, were set in five steps. Extraction experiments were carried out in 16 sections according to the central synthesis plan, and the nipposide content ( Y ₁ ), which is a functional substance of the two insects, was measured as the dependent variable ( Y _n ) affected by these independent variables, and the reaction was performed based on a predictive model equation. The surface was analyzed for optimal conditions. The geniposide content of the extract is The extract was filtered with 0.45 μm filter paper and analyzed using HPLC under the conditions given in Table 1.

TABLE 1

Geniposide Assay Conditions Using HPLC

HPLC specification table Condition equipment  Shimadzu LC 10A column Symmetry C ₁₈ (4.6 X 150 mm, 5 m) Fluidized bed H ₂ O: Acetonitrile = 91: 9 Flow rate  1.0 mL / min Column temperature  40 ℃ Injection volume  20 L Detector  UV 240 nm

The reaction surface for the change of the geniposide content according to the extraction conditions as described above is shown in FIG. The geniposide content was found to be most affected by the extraction temperature. The peak point was the highest point and the maximum value was expected to be 6.18 mg%. The extraction conditions were extraction temperature of 93.98 ° C, extraction time of 6.99 hr, and solvent ratio of the sample to 60.59% (v / w).

From this, the optimum extraction conditions for preparing the Tofu extract were set to 95 ° C., 7 hr, and 60% (v / w) of extraction temperature, extraction time, and solvent ratio, respectively.

Example 2 Establishment of Optimal Alcohol Fermentation Conditions of Tofu Extract

Example 2-1 Preparation of Tofu Extract

Tofu extract used in this example was prepared in March 2007 to purchase tofu (shell) produced in Gyeongbuk Yeongcheon, apple concentrate (70 ^o Brix) was produced in 2007 Was prepared by concentration under reduced pressure.

Using the optimum extraction conditions for the preparation of the larvae extract prepared in Example 1, the chopped worms were reflux-cooled under the conditions of extraction temperature of 95 ° C., extraction time of 7 hours, and extraction hot water solvent ratio of 60 mL / g. An extract was prepared.

Example 2-2: Alcohol fermentation strain selection

In this embodiment, in order to select a yeast suitable for fermentation of the larvae extract, several kinds of yeasts are first separated using YPD solid medium (1% yeast extract, 2% peptone, 2% glucose, pH 6.0), and finally by alcoholic fermentation strain S saccharide five species of my process three Levy jiae (S. cerevisiae) was isolated DUJ-98, DUK-99, DUL-11, DUM-12 and DUN-3. In order to determine the alcohol fermentation ability of the isolated alcohol fermentation strains, the results of the fermentation test were shown in Table 2.

TABLE 2

Fermentation Test of Isolated Yeast for Alcohol Fermentation

Isolate Alcohol concentration (%) Acidity (%) Brix Alcohol resistance
(O.D) DUJ-98 7.6 1.40 5.5 0.683 DUJ-99 7.8 1.44 5.0 0.705 DUL-11 7.4 1.39 5.5 0.673 DUL-12 8.0 1.38 5.0 0.697 DUN-3 6.8 1.42 5.0 0.384

Subsequently, an apple concentrate was added to the above-mentioned worm extract extract up to 15 ^o Brix, and each of the five yeasts was inoculated, and then fermented at a culture temperature of 30 ° C., shaking speed of 120 rpm, and culture time for 3 days. In order to determine the alcohol fermentation ability of the five alcohol fermentation strains, the alcohol content and the alcohol yield were investigated, and the results are shown in Table 3.

TABLE 3

Comparison of Alcohol Content, Alcohol Yield, pH, Acidity and Residue Concentration from Various Yeast Strains

Strain Alcohol content
(%) Alcohol yield ¹
(%) pH Titration
(%) Residue concentration
( ^o Brix) DUJ-98 7.6 99.34 3.95 0.33 5.4 DUK-99 7.0 91.52 4.02 0.30 5.2 DUL-11 7.0 91.52 4.12 0.27 4.8 DUM-12 7.4 96.73 4.17 0.21 4.6 DUN-3 7.2 94.11 4.15 0.22 5.0

                       Final Ethanol Concentration (g / L)

¹ Alcohol Yield (%) = ------------------------------ X 100

                     Initial Glucose Concentration (g / L) X 0.51

As shown in Table 3 above, fermentation at 30 ° C. for 3 days showed similar results in all five strains. The alcohol content was 7.0-7.6%, the titratable acidity was 0.21-0.33, and the pH was 3.95-4.17. The experimental values of the alcohol fermentation broth were not significantly different among the strains. It was confirmed that sufficient alcohol fermentation occurred during the primary alcohol fermentation, which is a preliminary step to produce vinegar, and YPD extract (Yeast extract ) was selected by selecting S. cerevisiae DUJ-98, which has the highest alcohol content. 1%, Peptone 2%, Glucose 2%, pH 6.0) was incubated for 30 hours, incubated for 24 hours, 5% (v / v) was used as the main hair for alcohol fermentation of two worms.

Example 2-3 Establishment of Optimal Alcohol Fermentation Conditions for Tofu Extract

In this example, the optimum conditions for primary alcohol fermentation were established using the prepared soybean extract and selected strains. In other words, S. cerevisia DUJ-98 was used to examine the quality change according to the initial sugar (10 to 18 ^o Brix), fermentation time (1 to 5 days) and fermentation temperature (20 to 36 ℃).

In the alcohol fermentation process, the experimental ranges for initial sugar concentration ( X ₁ ), fermentation time ( X ₂ ) and fermentation temperature ( X ₃ ), which are important independent variables ( X _i ), were set in five steps. The fermentation experiment was conducted in 16 sections according to the central synthesis plan, and the alcohol content ( Y ₁ ), acidity ( Y ₂ ) and sugar ( Y ₃ ) were measured and predicted as the dependent variable ( Y _n ) affected by these independent variables. Based on the model equation, the reaction surface was analyzed to find the optimal condition.

After centrifuging the culture fermented according to the fermentation conditions, 100 mL of the supernatant was taken and alcohol was distilled using an alcohol distillation apparatus (Alcodest 0362392, J. P. Select Co., Spain), and alcohol content was measured using a spirit meter. Total acid was neutralized and titrated with 0.1 N NaOH solution to obtain acetic acid content (%). The sugar content of the fermentation broth was measured three times using a refractometer (model 507-1, Nippon Optical Works Co., Japan).

The regression equation according to the response surface regression analysis on the alcohol content, acidity and residual content is shown in Table 4.

TABLE 4

Polynomial Equation from RSM Analysis on Alcohol Fermentation Process

reaction 2nd polynomial R ² Significance Alcohol content Y _AC = 1.3375-2.38 125X ₁ + 4.8875X ₂ +0.753 125X ₃ + 0.0125X ₁ ² + 0.2375X ₁ X ₂
-0.775X ₂ ² + 0.059375X ₁ X ₃ -0.09375X ₂ X ₃ -0.021875X ₃ ² 0.9016 0.0196 Acidity Y _A = 2.223125-0.092500X ₁ -0.101250X ₂ -0.093125X ₃ + 0.003125X ₁ ²
-0.001 250X ₁ X ₂ +0.016 250X ₂ ² + 0.000937X ₁ X ₃ + 0.000625X ₂ X ₃
+ 0.001406X ₃ ² 0.8284 0.0842 Remnant Y _RS = 35.550-1.096875X ₁ -4.718750X ₂ -1.289063X ₃ + 0.050000X ₁ ²
-0.093750X ₁ X ₂ + 0.675000X ₂ ² + 0.010938X ₁ X ₃ + 0.046875X ₂ X ₃
+ 0.017188X ₃ ² 0.9185 0.0116

The alcohol content showed a value of 1.4-9.2%, and the response surface regression analysis showed that R ² of the regression equation was 0.9016, and significance was recognized within 5% (see Table 4). The effect of fermentation conditions on alcohol content was found to be more affected by fermentation time and initial sugar content than by fermentation temperature (see Table 5). As a result of analyzing the change of alcohol content according to alcohol fermentation by the four-dimensional reaction surface under the experimental conditions designed by the central synthesis plan, the ridge analysis resulted in the shape of saddle point as shown in FIG. As the initial sugar content increased, it increased as the fermentation time increased. The optimum alcohol production conditions were found to be 17.57 ^o Brix initial sugar content, 3.65 day fermentation time and 30.41 ℃ fermentation temperature.

TABLE 5

Variable Analysis of Regression Model Variables in Alcohol Fermentation Conditions

reaction F-B Initial sugar concentration
(° Brix) time
(day) Temperature
(℃) Alcohol content 3.38 ^* 8.73 ^** 2.12 Acidity 5.62 ^** 1.13 2.16 Remnant 5.88 ^** 11.21 ^*** 1.37

^* Significance at 10% level; ^** significance at the 5% level; ^*** Significance at 1% level.

TABLE 6

Estimated Fermentation Conditions for Alcohol Fermentation Process by Ridge Analysis

reaction Fermentation Conditions ¹⁾ X ₁ X ₂ X ₃ Expected value shape Alcohol content 14.45 1.14 25.13 2.03 (min) Saddle point 17.57 3.65 30.41 10.10 (max) Acidity 11.22 2.99 28.68 0.22 (min) Min 17.00 2.81 33.23 0.38 (max) Remnant 10.16 3.15 30.15 2.96 (min) Min 15.37 1.15 26.67 8.07 (max)

¹⁾ X ₁ : initial sugar concentration (° Brix), X ₂ : time (day), X ₃ : temperature (℃)

R ² of the response surface regression equation for the change of total acid according to the primary alcohol fermentation conditions of Tofu extract was 0.8284, and the significance was recognized at the significance level within 10% (see Table 4). The change in total acidity was most affected by the initial sugar content, and there was little effect of fermentation temperature and fermentation time.The predicted peak point was the lowest point (see Fig. 3), and the condition showing the minimum acidity was the initial sugar content. Content was 11.22 ^o Brix, fermentation time was 2.99 days and fermentation temperature was 28.68 ℃ (see Table 6), which indicates that the total acid change in alcohol fermentation increased in proportion to the initial sugar content in the production of vinegar using concentrated apple juice. It was consistent with the report.

R ² of the response surface regression equation for the residual content was 0.9185, which was significant within 5% (see Table 4). The predicted normal point was the lowest point and the lowest value was predicted as 2.96 ^o Brix, where fermentation conditions were initial sugar content of 10.16 ^o Brix, fermentation time 3.15 days and fermentation temperature 30.15 ° C. (see FIG. 4 and Table 6). As shown in the reaction surface of Figure 4 it can be seen that the initial sugar is low, the fermentation time exhibits the lowest content in the range of 2.5 to 3.5 hours. In addition, as shown in Table 6, the effect of fermentation conditions on the residual content was mainly influenced by the fermentation time, and was affected little by the fermentation temperature.

As a result of this, more than 7% of alcohol content is produced at the optimum condition with low alcohol content and total acid content suitable for vinegar production using worm extract, that is, fermentation temperature 28 ℃, initial sugar content 14 ^o Brix and fermentation time 2.5 day. It became. That is, it was found that alcohol fermentation was sufficient to produce Pleurotus vinegar by alcohol fermentation using the Pleurophyllum extract as the optimum alcohol fermentation conditions.

Example 3 Establishment of Optimum Acetic Acid Fermentation Conditions of Tofu Extract

Example 3-1 separation of vinegar fermentation bacteria

Acetic acid bacteria used for acetic acid fermentation were activated for 30 to 24 hours in each sample such as fruit or seed, and then diluted appropriately with physiological saline, and inoculated with 0.2 mL of each sample to smear plate for separating acetic acid bacteria. After culturing for 2 days at, the bacteria forming a transparent ring around the colony were purely separated. The bacteria isolated from the solid medium were again inoculated into acetic acid-producing bacteria medium containing the larvae, shaken and cultured at 30, and finally, Acetobacter sp. DU-4 strain having excellent acetic acid-producing ability was finally selected.

Example 3-2 Preparation of Vinegar Using Tofu Extract

Inoculated with the Acetobacter sp. DU-4 strain stored in the primary alcohol fermentation broth of the larvae extract prepared under the optimum conditions according to the Examples 1 and 2, and then stored at 30 ° C. and 200 in a shaker incubator. Fermentation at rpm was used as a seed when the acid was 4.5%.

Example 3-3: Establishment of Optimum Acetic Acid Fermentation Conditions of Tofu Extract

In order to optimize acetic acid fermentation conditions using Tofu extract, experiments were conducted according to the central synthesis plan. Acetic acid fermentation was inoculated in 200 mL alcohol fermentation broth 10% (v / v) in 500 mL Erlenmeyer flasks, respectively, and fermented in a shaker at 30 ℃. Acetic acid fermentation conditions are the initial acidity (0.5-2.5%), stirring speed (50-250 rpm) and fermentation time (4-8 day) set as factor variables for each of five levels (-2, -1, 0, 1, 2). ), The central synthesis plan was established, and experiments were carried out under 16 established conditions, and the optimum characteristics were determined by measuring the acidity, geniposide content, chromaticity and organic acid content such as quality characteristics of fermented products.

pH was measured using a pH meter (691 pH meter, Metrohm Co., Swiss), the total acid was neutralized titration with 0.1 N NaOH solution and converted to acetic acid content (%). The content of nipposide was measured by the same method used in Example 1.

Chromaticity is measured three times using Hunter's color L value (brightness), a value (red), and b value (yellow) using a color meter (Chromameter, model CR-300, Minolta Co., Japan). Repeated measurements were taken and distilled water (L = 100.00, a = 0.00 and b = + 0.02) was used as a control. Organic acid analysis was performed by diluting each vinegar stock at an appropriate magnification, filtering with 0.45 m membrane filter, and analyzing by HPLC.

<Table 7>

details Condition equipment  Shimadzu LC-10AT (Japan) column  Symmetry C₁₈ (4.6 x 150 mm, 5 m, Waters Co., USA) Mobile phase 0.01 NH ₂ SO ₄ (pH 2.4) Detector  UV 210 nm Flow rate  0.5 mL / min

1) Total Acid Content of Tobacco Vinegar

As a result of analyzing the total acidity of the larvae extract vinegar as described above, the value of 2.67∼6.61% was found, and the R ² of the response surface regression equation was high as 0.9133, and the significance was recognized within 5% (Table 8). Reference). The predicted steady point was the saddle point, and the ridge analysis showed a maximum value of 7.15%. The fermentation conditions that can be obtained were 1.71% of initial acidity, 234.82 rpm, and 6.96 days of fermentation time (see Table 9). The total acid content during the fermentation of acetic acid was affected by the fermentation time and the stirring speed, and there was little effect on the initial acidity within the set range (see Table 10). The fermentation characteristics of Tofu extract vinegar for the total acid showed a tendency to increase as the fermentation time is long, the stirring speed increases as shown in FIG.

<Table 8>

Response surface regression from RSM analysis of acetic acid fermentation process

reaction 2nd polynomial R ² Significance Acidity Y _AC = 13.370000-1.978750X ₁ -0.060838X ₂ -2.223125X ₃ -0.125X ₁ ²
+ 0.000154X ₁ X ₂ + 0.000193X ₂ ² + 0.4975X ₁ X ₃ + 0.001925X ₂ X ₃
+ 0.155000X ₃ ² 0.9133 0.0138 Zeniposide content Y _GC = -0.417500-0.067500X ₁ -0.001650X ₂ + 0.343750X ₃ -0.060000X ₁ ²
+ 0.001000X ₁ X ₂ -0.0000055X ₂ ² + 0.015000X ₁ X ₃ + 0.000350X ₂ X ₃
-0.031 250X ₃ ² 0.9288 0.0080 L Y _L = 49.422500 + 2.070000X ₁ + 0.093700X ₂ + 10.650000X ₃ -1.815000X ₁ ²
+ 0.001300X ₁ X ₂ -0.000336X ₂ ² + 0.590000X ₁ X ₃ + 0.001000X ₂ X ₃
-0.970000X ₃ ² 0.8347 0.0767 a Y _a = -2.235000 + 2.933750X ₁ -0.029938X ₂ + 0.303125X ₃ -0.455000X ₁ ²
+ 0.006650X ₁ X ₂ + 0.0000275X ₂ ² -0.40250X ₁ X ₃ + 0.001725X ₂ X ₃
+ 0.018750X ₃ ² 0.9087 0.0159 b Y _b = 56.983750-5.735000X ₁ -0.044950X ₂ -6.502500X ₃ + 1.670000X ₁ ²
-0.020 600X ₁ X ₂ + 0.000201X ₂ ² + 0.540000X ₁ X ₃ + 0.005000X ₂ X ₃
+0.447 500X ₃ ² 0.9041 0.0182

<Table 9>

Estimated level of fermentation conditions for acetic acid fermentation process by ridge analysis

reaction Fermentation Conditions ¹⁾ X ₁ X ₂ X ₃ Predictive response shape Acidity (%) 0.52 132.47 5.69 2.94 (min) Saddle 1.71 234.82 6.96 7.15 (max) Zeniposide content (ug / mL) 1.61 161.67 4.02 0.37 (min) Saddle 1.97 221.73 7.02 0.64 (max) L 1.17 132.50 7.86 86.35 (min) maximum 2.46 157.32 6.51 89.15 (max) a 0.72 183.60 4.93 -1.95 (min) Saddle 1.06 97.99 7.46 -0.23 (max) b 1.55 125.10 5.68 31.44 (min) at least 1.01 232.12 6.61 36.28 (max)

¹⁾ X ₁ : initial acidity (%), X ₂ : shaking speed (rpm), X ₃ : time (day)

<Table 10>

Variable Analysis for Regression Models of Variables in Acetic Acid Fermentation Conditions

Extraction conditions F-B Initial pH (%) Shaking speed (rpm) Time (day) Acidity 1.96 6.10 ^** 7.21 ^** Zeniposide content 3.13 ^* 4.74 ^** 16.57 ^*** L 1.75 4.38 ^* 6.11 ^** a 6.71 ^** 3.42 ^* 7.36 ^** b 4.61 ^** 10.15 ^*** 4.88 ^**

^* Significance at 10% level; ^** significance at the 5% level; ^*** Significance at 1% level.

2) Geniposide Content of Tofu Extract Vinegar

R ² of the regression equation for the geniposide content of Tofu extract vinegar was 0.9288, and significance was recognized within 1% (see Table 8). As a result of the ridge analysis using the saddle point, the maximum value of the geniposide content was estimated to be 0.64 g / mL, and the optimum fermentation condition was 1.97%, the stirring speed was 221.3 rpm, and the fermentation time was 7.02. It was time (see Table 9). Looking at the reaction surface for the change in the content of the geniposide by fermentation conditions, it appears that the content of the geniposide increases as the fermentation time is longer (see Fig. 6). In other words, the effect of the extraction conditions on the nipposide content was shown to have the largest effect on the fermentation time, as shown in Table 10, and then affected by the stirring speed and initial acidity.

3) Changes in chromaticity of tofu extract vinegar

As a result of measuring the chromaticity of Tofu extract vinegar prepared by central synthesis plan, L value was 86.85 ~ 90.94, a value was -1.70 ~ -0.26, and b value was 31.30 ~ 35.60. The response surface regression equation for chromaticity is summarized in Table 8, the L value was found to be significant within 10%, and the a and b values were within 5%. The color L value showed the highest 89.15 when the initial acidity was 2.46%, the stirring speed was 157.82 rpm, and the fermentation time was 6.51 days. It was. In addition, the value of chromaticity b showed the highest value of 6.61 at the initial acidity of 1.01%, the stirring speed of 232.12 rpm, and the fermentation time of 6.61 day (see Table 9).

As a result of analyzing the influence of the initial acidity, the stirring speed and the fermentation time on the chromaticity (see Table 10), it was found that the L value was affected by the stirring speed and the fermentation time. Was found to be almost absent. In the case of a value, fermentation time was found to have more influence than initial acidity and stirring speed. On the other hand, the b value was affected by all three variables such as initial acidity, stirring speed and fermentation time. As for the reaction surface of color change according to fermentation conditions, as shown in FIG. The longer it was, the higher the value was. The fermentation characteristics of b value according to the fermentation conditions of Tofu extract vinegar increased b value as the stirring speed increased.

4) Changes in Organic Acid Content of Vinegar Extract

The regression equation for acetic acid content of Tofu extract vinegar was found to be significant within 5% (see Table 11), and the predicted peak point was saddle point. A maximum of 6.30% was predicted (see Table 12). As shown in FIG. 8, the reaction surface of acetic acid content change showed a higher content as the fermentation time was longer and the stirring speed was faster. In addition, as shown in Table 13, the effects of fermentation time and stirring speed were significant.

R ² of the regression equation for the citric acid content of the fermentation broth was 0.8909, and significance was recognized within 5% (see Table 11). The estimated peak point was the saddle point, and the maximum value was 6.48 mg%. It was predicted (see Table 12), and the fermentation conditions from which the predicted value was obtained were found to have an initial acidity of 0.57%, a stirring speed of 117.43 rpm and a fermentation time of 6.43 days. As shown in the reaction surface of FIG. 8, the citric acid content increased as the initial acidity decreased, and the effect was significant (see Table 13).

R ² of the regression equation for the maleic acid content of soybean vinegar was 0.8629 and significance was recognized within 5% (see Table 11). The predicted normal point was the maximum and the maximum value was predicted to be 315.34 mg% (see Table 12). ). The maximum fermentation conditions were 1.56% of initial acidity, 161.93 rpm, and 6.29 days of fermentation time. As a result of the reaction surface analysis, the highest amount of malic acid was detected in the range of initial acidity of 1.0 to 2.0%, stirring speed of 100 to 200 rpm, and fermentation time of 5.5 to 7.5 days.

<Table 11>

Polynomials from RSM Analysis for Acetic Acid Fermentation Process

reaction 2nd polynomial R ² Significance Acetic acid Y _AC = 12.674375-0.741250X ₁ -0.060287X ₂ -2.323 125X ₃ -0.250000X ₁ ²
-0.000750X ₁ X ₂ + 0.000163X ₂ ² +0.342 500X ₁ X ₃ + 0.003325X ₂ X ₃
+ 0.158750X ₃ ² 0.9114 0.0147 Citric acid Y _CA = -6.861250-1.258750X ₁ -0.024112X ₂ + 4.831875X ₃ + 1.110000X ₁ ²
+ 0.008450X ₁ X ₂ + 0.000035X ₂ ² -6.77500X ₁ X ₃ -0.000125X ₂ X ₃
-0.313750X ₃ ² 0.8909 0.0259 Malian Y _MA = -452.223125 + 197.032500X ₁ + 1.693025X ₂ + 151.607500X ₃ -37.04500X ₁ ²
-0.040500X ₁ X ₂ -0.005057X ₂ ² -11.86000X ₁ X ₃ + 0.000900X ₂ X ₃
-10.593750X ₃ ² 0.8629 0.0475 Tartaric acid Y _TA = -29.613125 + 13.798750X ₁ + 0.185238X ₂ + 10.583125X ₃ -2.145000X ₁ ²
-0.024450X ₁ X ₂ -0.000276X ₂ ² -0.597 500X ₁ X ₃ -0.010475X ₂ X ₃
-0.615000X ₃ ² 0.8391 0.0716 Suche mountain Y _SA = -15.883750-2.127500X ₁ + 0.412025X ₂ + 8.795000X ₃ + 2.830000X ₁ ²
-0.079000X ₁ X ₂ + 0.000308X ₂ ² + 1.145000X ₁ X ₃ -0.067 100X ₂ X ₃
+ 0.048750X ₃ ² 0.8454 0.0647 Total organic acid Y _TOA = 11.422 500-0.310000X ₁ -0.057475X ₂ -1.955000X ₃ -0.290000X ₁ ²
+ 0.000300X ₁ X ₂ + 0.000159X ₂ ² + 0.270000X ₁ X ₃ + 0.002850X ₂ X ₃
+0.142 500X ₃ ² 0.9053 0.0176

<Table 12>

Estimated Levels of Fermentation Conditions for Acetic Acid Fermentation Process by Analysis

reaction Fermentation Conditions ¹⁾ X ₁ X ₂ X ₃ Predictive response shape Acetic acid (%) 0.53 132.92 5.60 2.49 (min) Saddle 1.64 232.27 7.09 6.30 (max) Citric Acid (mg%) 1.88 146.41 7.84 2.69 (min) Saddle 0.57 117.43 6.43 6.48 (max) Malic acid (mg%) 0.99 91.77 4.24 240.73 (min) maximum 1.56 161.93 6.29 315.34 (max) Tartaric Acid (mg%) 1.07 88.15 4.68 23.33 (min) maximum 1.47 145.41 6.59 29.10 (max) Succinic Acid (mg%) 1.25 91.06 4.45 36.30 (min) Saddle 1.95 79.09 7.07 56.04 (max) Total organic acid (%) 0.55 135.66 5.43 2.80 (min) Saddle 1.65 233.83 7.04 6.62 (max)

¹⁾ X ₁ : initial acidity (%), X ₂ : shaking speed (rpm), X ₃ : time (day)

TABLE 13

Variable Analysis for Regression Models of Variables in Acetic Acid Fermentation Conditions

Extraction conditions F-B Initial pH (%) Shaking speed (rpm) Time (day) Acetic acid 1.28 5.96 ^** 7.46 ^** Citric acid 6.05 ^** 0.78 3.29 ^* Malian 3.76 ^* 6.04 ^** 5.40 ^** Tartaric acid 2.68 4.22 ^* 5.66 ^** Suche mountain 2.31 6.60 ^** 5.11 ^** Total organic acid 1.26 5.50 ^** 6.63 ^**

^* Significance at 10% level; ^** significance at the 5% level; ^*** Significance at 1% level.

R ² of the regression equation for tartaric acid content is 0.8391, and the significance was accepted within 10% (see Table 11), the predicted steady point is maximum jeomyigo maximum value was estimated to be 29.10 mg%, The fermentation conditions were initial pH 1.47 %, Stirring speed 145.41 rpm and fermentation time was 6.59 days (see Table 12). As shown in the reaction surface of FIG. 9, the content of tartaric acid was increased as the stirring speed was faster and the fermentation time was longer, which was similar to the result of malic acid. On the other hand, the tartaric acid content was most affected by the fermentation time, and the influence on the stirring speed and initial acidity was small (see Table 13).

R ² of the regression equation for succinic acid content was 0.8454 and significance was recognized at levels within 10% (see Table 11). The optimum fermentation conditions for succinic acid were 1.95% of initial acidity, 79.09 rpm of agitation speed, and 7.07 days of fermentation time, with a maximum value of 56.04 mg% (see Table 12). Fermentation characteristics for succinic acid according to the fermentation conditions, as shown in Figure 9 it was confirmed that the succinic acid content increases as the stirring speed is slow and the fermentation time is longer. The effect of extraction conditions on succinic acid was shown to be affected by the stirring speed and fermentation time, as shown in Table 13, and had little effect on the initial acidity within the set range.

Acetic acid, citric acid, malic acid, tartaric acid, and succinic acid were all analyzed to calculate the total organic acid content of the fermentation broth. As shown in Table 10, R ² of the regression equation was 0.9053 and significance was recognized within 5%, and the predicted normal point was the saddle point, and the maximum value was 6620 mg% (see Table 12). ).

The fermentation conditions for this predicted value were 1.65% of initial acidity, 233.83 rpm of stirring speed, and 7.04 days of fermentation time. Looking at the reaction surface for the total organic acid content of Figure 9 it can be seen that the higher the stirring speed and the longer the fermentation time, the higher the content. The change in total organic acid content was greatly affected by the stirring speed and fermentation time, and the effect on the initial acidity was slightly low (see Table 13).

5) Establishment of Optimum Acetic Acid Fermentation Conditions of Tofu Extract

The results of the above results were superimposed on the above result values (see Table 14 and FIG. 10). For the preparation of larvae extract vinegar, the larvae extract was collected at 14 ^° C Brix using a strain isolated from larvae at 28 ° C. After fermentation for 2.5 days to obtain an alcoholic fermentation broth, Acetobacter sp. By using DU4, the initial acidity was 1.5%, the shaking speed was 225 rpm, and the fermentation time was adjusted to 6.5 days.

TABLE 14

Optimum fermentation range of conditions for maximum response variables obtained by overlapping reaction surfaces

Fermentation condition Expected range of conditions
(Optimal point) Initial pH (%) 1.0 ~ 2.0 (1.5) Shaking speed (rpm) 200-250 (225) Time (day) 6 ~ 7 (6.5)

Example 4 Analysis of Functional Quality Characteristics of Tobacco Vinegar

In the present embodiment, the chopped vinegar base was prepared under the optimum conditions set in Examples 1 to 3, and the functional quality characteristics of the chopped vinegar were analyzed using the same.

The total acidity of two vinegar vinegar was neutralized and titrated with 0.1 N NaOH solution and converted into acetic acid content (%). In addition, the content of the geniposide of the two vinegar vinegar was measured by the same method used in Example 1.

Chromaticity was measured by the same method as used in Example 3 using a color difference meter (Chromameter, model CR-300, Minolta Co., Japan). The organic acid was analyzed by HPLC after diluting each vinegar stock in an appropriate ratio and filtering with 0.45 μm membrane filter. The analytical conditions used were as follows (column: Sunfire C ₁₈ , mobile phase: 0.01 NH ₂ SO ₄ (pH 2.4), flow rate: 0.5 mL / min, detector: UV 210 nm) and the calibration curve of the standard under the same analysis conditions The organic acid was quantified.

Tobacco vinegar was filtered through a 0.45 μm membrane filter and analyzed using HPLC, respectively. Analytical conditions for free sugar: ULTRON PS-80N column, mobile phase = distilled water, flow rate = 0.6 mL / min, detector (RI detector) were used, and the calibration curve of the standard product was prepared under the same analysis conditions to quantify free sugar. The dietary fiber was first digested with two vinegar vinegar by α-amylase, protease and β-glucosidase, followed by 0.45 μm membrane filter and analyzed by HPLC under the same conditions as free sugar analysis.

TABLE 15

Conditions Used for the Analysis of Free Sugars and Dietary Fiber by HPLC

details Condition equipment  Shimadzu LC-10AT (Japan) column Symmetry C ₁₈ (4.6 X 150 mm, 5 m, Waters Co., USA) Fluidized bed 0.01 NH ₂ SO ₄ (pH 2.4) Detector  UV 210 nm Flow rate  0.5 mL / min

Inorganic Components and Amino Acid Analysis After the dry decomposition of the sample, a certain amount was taken and filtered with a 0.45 μm membrane filter, followed by analysis of the desired inorganic component using an atomic absorbance spectrometer (Analyst 100/300 Atomic absorption spectrometer, Perkin-Elmer Co., Norwalk, USA). According to the test curve, the test solution was analyzed. The amino acid of the sample was added to 2 mL of 6 N HCl, hydrolyzed at 110 ° C. for 24 hours, filtered, concentrated to dryness under reduced pressure, dissolved in sodium citrate buffer (pH 2.2), and filtered after analysis with a 0.45 μm membrane filter.

TABLE 16

Operating Conditions of Amino Acid Analyzer for Amino Acid Analysis

details    Condition equipment    S7130 amino acid analyzer (Sykam Co., Germany) column    LCA K02 / Na Cation separation column Buffer A: 0.12 N Sodium citrate buffer (pH 3.45)
B: 0.20 N Sodium citrate buffer (pH 10.85) Buffer flow rate    0.45 mL / min Ninhydrin flow rate    0.25 mL / min Injection volume    20 uL

Physical quality characteristics and amino acid contents of the two vinegar vinegar prepared under the optimum fermentation conditions (two-step fermentation method of primary alcohol fermentation and secondary acetic acid fermentation) are summarized in Table 17 and 18, respectively. The geniposide content in soybean vinegar was 0.60 ㎍ / mL, the total acid content was 6.36%, and the acetic acid content in each vinegar was 5395.56 mg%, which was the main organic acid component. The contents were higher in order of malic acid, succinic acid, tartaric acid, and citric acid. Fructose was found to have the highest content of sugar, and dietary fiber content of vinegar was 0.5%. Inorganic components were present in a large amount of K, followed by Na, Ca and Mg. The total amino acid content of each vinegar was 27.64 mg%, showing the highest aspartic acid content.

TABLE 17

Physiological and Chemical Properties of Tobacco Vinegar

characteristic value Zeniposide content (/ mL) 0.60 ± 0.01 PH (%) 6.36 ± 0.05 Hunter color index L-value 87.46 ± 0.00 a-value -0.92 ± 0.01 b-value 34.32 ± 0.00 Organic acid (mg%) Acetic acid 5395.56 ± 07.16 Citric acid 2.79 ± 0.06 L-malic acid 248.65 ± 4.79 Tartaric acid 25.51 ± 0.12 Suche mountain 42.74 ± 0.54 synthesis 5715.25 ± 31.65 Sugar content (mg%) glucose 37.72 ± 0.50 Exaggeration 344.90 ± 4.59 saccharose 28.28 ± 0.49 synthesis 410.90 ± 4.59 Dietary heart oil (%) 0.5 ± 0.00 Mineral content (mg%) Ca 5.56 ± 0.04 Fe 1.65 ± 0.01 K 144.75 ± 2.82 Mg 4.73 ± 0.15 Na 10.30 ± 0.22 Zn 0.51 ± 0.01 Total 167.50 ± 2.88

TABLE 18

Amino Acid Content of Tobacco Vinegar

amino acid Content (mg%) Aspartic acid 10.93 ± 0.30 Serine 0.80 ± 0.01 Seronine 1.18 ± 0.07 Glutamic acid 2.14 ± 0.09 Proline 0.00 ± 0.00 Glycine 0.86 ± 0.01 Alanine 1.90 ± 0.13 Cystine 0.00 ± 0.00 Balin 0.57 ± 0.01 Methionine 0.00 ± 0.00 Isoleucine 0.29 ± 0.04 Leucine 0.44 ± 0.01 Tyrosine 0.00 ± 0.00 Phenylalanine 0.00 ± 0.00 Histidine 1.10 ± 0.06 Lee Sin 0.77 ± 0.01 ammonia 6.41 ± 0.37 Arginine 0.25 ± 0.02 Total amino acids 27.64 ± 0.39

Example 5 Preparation, Quality Characteristic Analysis and Consumer Inspection of Tobacco Vinegar Beverage

In order to prepare a functional two vinegar vinegar beverage containing fructooligosaccharide and vitamin C using the two vinegar vinegar (vinegar base) prepared in the optimum conditions in Example 4 according to the combination ratio given in Table 18 below It prepared.

Each two-packed vinegared beverage was heated for 10 minutes at 50 ℃, injected into a bottle or pack (120 mL), sealed and sterilized again at 80 ℃ for 15 minutes, used as a sample after cooling with cooling water.

A consumer test was conducted on 60 students in the food engineering department at Daegu University. Randomly placed 20 mL samples in randomly attached paper containers and presented to the consumer, and scored from 1 (very bad) to 9 (very good) for color, aroma, taste, aftertaste and overall preference. It was evaluated using a 9-point scale method that gives.

The results of each item were analyzed by variance analysis using Statistical Analysis System program, and the items with significant differences were verified at p <0.05 level by Duncan's multi-range test.

TABLE 19

Mixing ratio of Tofu vinegar beverage used in this embodiment

ingredient (%) sample 3% 6% 9% 12% Vinegar Base 3 6 9 12 Apple Juice Concentrate 5 5 5 5 Exaggeration 10 10 10 10 honey 0.5 0.5 0.5 0.5 Polydextrose 3 3 3 3 Glutamic acid 0.02 0.02 0.02 0.02 Ascorbic acid 0.04 0.04 0.04 0.04 Purified water 78.44 75.44 72.44 69.44 Total (%) 100 100 100 100

Physical quality characteristics of each vinegar beverage prepared according to the compounding ratio given in Table 19 are summarized in Table 20. Consumer test results (see Table 21) showed that there was no significant difference in color preference among the samples, and the flavor and taste showed higher preference than other samples with 3 or 6% of the content of Pulverum vinegar. The vinegar beverage containing 3% of chopped vinegar showed a significant difference in preference from other samples ( p <0.05).

TABLE 20

Physiological and Chemical Properties of Vinegar Beverage Based on Tobacco Vinegar

characteristic Vinegar Base Levels of Mixed Drinks (%) 3 6 9 12 pH 3.37 3.33 3.29 3.26 Soluble Solids Content ( ^o Brix) 15.1 15.4 15.5 15.8 Turbidity 0.120 0.135 0.170 0.189 L * 34.44 34.07 33.91 32.65 a * -0.527 -0.493 -0.463 -0.373 b * 3.01 3.43 3.88 4.02

TABLE 21

Consumer preference of vinegar drink based on tofu vinegar

Symbol Vinegar Base Levels in Mixed Drinks (%) 3 6 9 12 color 5.85 ^a 5.56 ^a 5.42 ^a 5.43 ^a Flavor 5.62 ^a 5.02 ^a 3.77 ^b 3.75 ^b flavor 6.67 ^a 6.12 ^a 5.40 ^b 4.13 ^c aftertaste 6.65 ^a 5.98 ^b 5.08 ^c 4.08 ^d Overall satisfaction 6.68 ^a 6.27 ^a 5.12 ^b 4.25 ^c

Means in the same column without ^ad common characters are significantly different ( p <0.05).

Figures 1a to 1c is a graph showing the reaction surface according to the change in the content of nipposide by the extraction conditions of the two worms.

2 is a graph showing the response surface to alcohol content as a function of fermentation temperature, time and initial sugar concentration (alcohol content = 6.0%, 7.0%, 8.0%).

3 is a graph showing the response surface to acidity as a function of fermentation temperature, time and initial sugar concentration (acidity = 0.25, 0.30, 0.35%).

FIG. 4 is a graph showing the response surface to the residual sugar content as a function of initial acidity, shaking speed and time (residue = 4.0, 5.0, 6.0 ^o Brix).

5 is a graph showing the response surface to acidity as a function of initial acidity, shaking speed and time (acidity = 3.5%, 4.0%, 4.5%).

FIG. 6 is a graph showing the reaction surface with respect to the geniposide content as a function of initial acidity, shaking speed and time (geniposide content = 0.45%, 0.50%, 0.55%).

7A-7C are graphs showing response surfaces for Hunter color index values as a function of initial acidity, shaking speed, and time (L = 87, 88, 89; a = −1.5, −1.0, −0.5; b = 32 , 33, 34).

8A-8C are graphs showing the response surface for acetic acid, citric acid and malic acid content as a function of initial acidity, shaking speed and time (acetic acid content = 3.0, 3.5, 4.0%; citric acid content = 3.5, 4.0, 4.5 mg% Malic acid content = 250, 270, 290 mg%).

9A-9C are graphs showing the response surface for tartaric acid, succinic acid and total organic acid content as a function of initial acidity, shaking speed and time (tartaric acid content = 25, 27, 29 mg%; succinic acid content = 44, 46 , 48 mg%; malic acid content = 3.5, 4.0, 4.5 mg%).

FIG. 10 is a graph showing overlapping reaction surfaces for optimization of total organic acid and geniposide content as a function of fermentation conditions.

Claims (7)

Extracting the larvae to prepare a larvae extract; Mixing the Tofu extract and apple concentrate, and then inoculating alcohol fermentation yeast with primary alcohol fermentation; Inoculating acetic acid fermentation strain to the primary alcohol fermented soybean extract, followed by secondary acetic acid fermentation; And Obtaining the second acetic acid fermented Tofu vinegar composition Method for producing a worm vinegar composition consisting of. The method of claim 1, wherein the preparation step of the larvae extract is prepared at 90 to 100 ℃, extraction time 5 to 10 hours and solvent ratio 40 to 70% (v / w) of the larvae vinegar composition Way. The method of claim 1, wherein the primary alcohol fermentation step of the larvae extract is a fermentation temperature of 25 to 30 ℃, an initial sugar content of 12 to 16 ^o Brix and a fermentation time of 2 to 3 days. . The method of claim 1, wherein the second acetic acid fermentation step of the alcohol-fermented soybean curd extract is an initial acidity of 1 to 25%, shaking speed of 200 to 250 rpm, fermentation time of 6 to 7 days Manufacturing method. Tobacco vinegar composition prepared by fermenting tofu extracts according to the method of claim 1 to primary alcohol fermentation, secondary acetic acid. Tobacco vinegar beverage containing the worm vinegar composition according to claim 5 as an active ingredient. According to claim 6, wherein the larvae vinegar beverage is 3 to 12% by weight of vinegar vinegar composition, 3 to 7% by weight apple concentrate, 8 to 15% by weight sucrose, 0.2 to 0.7% by weight honey, 2 to 5% by weight of polydextrose, glutamic acid Tobacco vinegar drink, characterized in that consisting of 0.01 to 0.03% by weight, ascorbic acid 0.01 to 0.05% by weight and purified water 70 to 74% by weight.

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