KR100330341B1 - Grape vinegar produced by two stage-fermentation - Google Patents

Abstract

PURPOSE: A process for preparing grape vinegar by two-stage fermentation is provided, thereby preparing the grape vinegar having a large quantity of organic acids, free amino acids and copper, iron, potassium and sodium and a small quantity of lactic acid causing a bad smell. CONSTITUTION: A process for preparing grape vinegar by two-stage fermentation comprises the steps of: removing seeds from grapes; pulverizing the grapes; alcohol fermenting the pulverized grapes using Saccharomyces cerevisiae; and acetic acid fermenting the alcohol fermented grapes using acetic acid fermenting bacteria, wherein the prepared grape vinegar has pH 3.2, and a large quantity of organic acids, particularly, 340 mg% of tartaric acid and 5305 to 5306 mg% of acetic acid.

Description

Grape vinegar produced by two stage-fermentation}

The present invention relates to grape vinegar by two stage fermentation. More specifically, the present invention relates to grape vinegar containing alcohol and fermentation of acetic acid, and then fermenting acetic acid again, which is free of off-flavor and salt, and contains a large amount of organic acid, free amino acid and trace components.

Vinegar is a fermented food that has been handed down for a long time in both East and West. Since ancient times, our nation has manufactured and used various kinds of vinegar at home. Vinegar is used not only as a seasoning for all foods with salt, but also as a raw material for pickled foods, mayonnaise, ketchup, sauces, etc. Vinegar is largely classified into synthetic vinegar and brewed vinegar. In Korea, synthetic vinegar was produced and consumed by the 1960s. Since then, the production of brewed vinegar using grains, fruits, and alcohol has been in full swing due to the improvement of the economic level. Recently, instead of the synthetic vinegar, which is reported to be harmful, it is consumed as an important seasoning in the diet. In addition, as the metabolic control function of vinegar is reported, the use of vinegar is also diversified, vinegar beverages are being developed in addition to seasonings. In addition, in terms of quality, dietary trends have been advanced to satisfy consumers' tastes and flavors.

In recent years, the consumption of vinegar has steadily increased as meat consumption has increased due to the westernization of diet. In the western world, where a large amount of meat is consumed, vinegar consumption is four times higher than in our country, and grape vinegar has a long history with wine in France, the region where grapes are produced. . In Korea, grape vinegar forms the fruit vinegar market together with apple vinegar and persimmon vinegar, and consumption of grape vinegar with unique colors and flavors is due to the nutritional constituents of grapes and the anthocyanin pigment of grape skins compared to general fruit vinegars. Is expected to continue to grow in the future and to diversify its use. As the prevention of geriatric diseases such as cholesterol lowering effect is known, many conventional grape vinegars made from 100% grape fruit are commercially available, but little research has been made according to the method and characteristics of grape vinegar.

The present inventors divided the grape vinegar production method into two stages of alcohol fermentation and acetic acid fermentation to produce grape vinegar in a short period of time and grape onion vinegar added onion grape juice reported to have a serum cholesterol lowering effect to the prepared grape vinegar The present invention was completed by comparing and analyzing the quality of grape vinegar and conventional grapevine vinegar of the present invention after manufacturing and confirming that the grape vinegar and grape onion vinegar of the present invention are of high quality.

Accordingly, an object of the present invention is to give a grape vinegar rich in organic acids, free amino acids, trace components without the odor and odor by acetic acid fermentation again after alcohol fermentation. Another object of the present invention is to provide a grape onion vinegar added onion juice having a serum cholesterol lowering effect to the grape vinegar.

The above object of the present invention is to remove the seed and the seedling and the vinegar prepared from the juice of the grapes from which the seeds are removed, added to the crushed grapes and fermented acetic acid again after alcoholic fermentation to obtain the present invention vinegar vinegar and add onion juice to this grape vinegar Onion vinegar was prepared, and then the physicochemical properties of grapes and grape onion vinegar were investigated, and their quality was compared with those of commercial grape vinegar.

Hereinafter, the configuration and operation of the present invention.

Figure 1 shows the changes in sugar and alcohol content during the fermentation of the grapevine vinegar of the present invention.

Figure 2 shows the change in pH and total acid during the fermentation of grape vinegar of the present invention.

In the present invention, after sterilizing the grape juice filtrate from which the seed was removed, inoculated and cultured with Sacchromyces cerevisiae YJK20 (KCTC 8841P) in Sacchromyces cerevisiae YJK20 (KCTC 8841P), the seed was prepared in the alcoholic fermented grape juice PA97 in PA97 (Acetobacter sp. PA97) inoculated and cultured to prepare; Seeding the crushed grapes by removing the seed and inoculated with alcohol prepared in one step, inoculated with seed vinegar and acetic acid fermentation in two steps to prepare the grape vinegar of the present invention; Preparing onion grape vinegar by adding onion juice to the prepared grape vinegar; The grape grape vinegar and grape vinegar prepared above were centrifuged to measure the alcohol content of the supernatant from which the precipitate was removed, microalcohol analysis, pH measurement, total acidity measurement, and color and turbidity of the present invention. Examining the peculiarities; Investigating the physicochemical properties of commercially available grape vinegar in the same manner as described above and comparing the physicochemical properties of the investigated grape vinegar and grape onion vinegar; The present invention removes the fat, pigment and protein components from the grape vinegar and grape onion vinegar, and then analyzes the organic acids, oxalic acid, malic acid, citric acid, tartaric acid, succinic acid, lactic acid and acetic acid contained in grape vinegar and grape onion vinegar. Comparing with organic acids contained in commercial grape vinegar; After removing the protein in the grape vinegar and grape vinegar vinegar solution of the present invention, the membrane filtrate is analyzed by amino acid automatic analysis of the free amino acid and compared with the free amino acid contained in the commercial grape vinegar; The present invention is composed of the steps of adding a disintegrating agent to the grape vinegar and grape onion vinegar solution to completely decompose and analyze the trace components with atomic absorption spectrophotometer and compare with the trace components contained in the commercial grape vinegar.

The grapes used in the present invention were grown in Gyeongsangbuk-do and purchased directly from the farmhouse, and two commercial grape vinegars used in the comparative analysis were prepared from Chaechaesan grape vinegar and wheat grain food prepared by Daechang Brewing Cooperative Corporation, Geobong grape vinegar was sold and used as a sample.

Saccharomyces cerevisiae YJK20 (KCTC 8841P) in Saccharomyces cerevisiae used in the present invention was distributed from the Biotechnology Research Institute Gene Bank, and the strain stored in the Traditional Food Research Institute of Kyungpook National University of Science was used. Acetobacter sp. PA97 ( Acetobacter sp. PA97), a common fermentation strain of acetic acid, was used by receiving a strain stored in the Institute of Traditional Food Research at Kyungpook National University of Science. YJK 20 (KCTC 8841P) was used as the YPD in Saccharomyces cerevisiae. (Yeast extract 1%, peptone 2%, glucose 2%, agar 2%) incubated in a slope medium at 25 ℃, 24 hours and used for 4 hours refrigerated storage. The acetobacter genus PA97 is a medium for acetic acid bacteria storage (20% potato Juice 1L, meat extract 3g, yeast extract 3g, glycerin 15g, glucose 5g, CaCO ₃ 10g, agar (agar) 20g, pH 7) incubated for 30 hours, 72 hours and used while refrigerated at 4 ℃.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Grape Vinegar Preparation

1st process: Main hair and seed

After removing the seeds of grapes, sterilized the filtrate and inoculated YJK20 ( Saccharomyces cerevisiae YJK20) (KCTC 8841P) to Saccharomyces cerevisiae , which is stored at the Institute of Traditional Food Research, Kyungpook National University of Science and incubated for 38 hours at 25 ℃ and 150rpm. 5% (w / v) was used as the main hair, and the seed was inoculated with grape juice to inoculate PA97 ( Acetobacter sp. PA97) in acetobacter, and incubated at 30 ° C and 200 rpm for 72 hours for 10% (v / v). ) Was used as the seed.

Second Process: Grape Vinegar Production by Two Stage Fermentation

In this step, the vinegar was prepared by removing the decayed portions of the grapes and washing the washed 10kg grapes with alcohol and acetic acid in two stages. That is, in the first stage of alcohol fermentation, seed was inoculated with 500 mL prepared in the first step in the state of being crushed with a blender mixer, and then fermented with alcohol for 5 days while stirring intermittently in a 25 ° C incubator. Removed to obtain a filtrate of 5.9% alcohol content. Two-stage acetic acid fermentation was inoculated with 5 ml of grape alcohol fermentation broth and inoculated with 500 mL of seed, followed by adjusting the aeration rate according to the growth of acetic acid bacteria at 25 ° C in a fermenter (Korea Fermentor Co., Ltd KF-51) at 25 ° C. The supernatant from which the precipitate was removed after centrifugation by acetic acid fermentation was used as analytical sample.

Example 2 Preparation of Grape Onion Vinegar

In this embodiment, grape onion vinegar is prepared by adding 3% (v / v) of onion juice to grape vinegar obtained by two-step fermentation by alcoholic and acetic acid fermentation in the first and second steps of Example 1. It was.

Example 3 Physicochemical Properties of Grape Vinegar of the Invention

The alcohol content of the grape vinegar of the present invention was calculated by distilling the supernatant after centrifugation of the culture medium, and the value measured by alcohol hydrometer was converted into Gay Lussac Table, and the microalcohol analysis was measured using an oxidation method. The pH was measured using a pH meter (Metrohm 691, Swiss), and the total acid was 10 mL of the sample, added to distilled water, and then adjusted to 100 mL. Converted to (%). For color, 20 ml of sample was taken and centrifuged at 3,000 rpm for 15 minutes using a colorimeter (Chrometer, Moder CR-300, CT310, Minolta Co., Japan) using L (whiteness), a (redness), b (yellowness). ) And ΔE (color difference). Turbidity was measured as absorbance at 660 nm with a certain amount of samples taken. The sugar content was expressed as .Brix using a refractometer. As a result, in Fig. 1, the grape juice (12 ° Brix) as a raw material after the first step of alcohol fermentation showed a change in sugar content and alcohol content. The sugar content was significantly reduced at 11.9 ° Brix at initial sugar content at 1 and 2 days of fermentation, and showed 5 ° Brix at 3 days of fermentation. The alcohol content increased sharply on the second day of fermentation to 5.41% and gradually decreased after peaking at 5.93% on the third day of fermentation. Figure 2 shows that after the first alcoholic fermentation of grape juice, seed inoculation was carried out in two stages of acetic acid fermentation, and as a result, there was no significant change from 0.78% of the total acidity to the third day of fermentation. The total acid content is shown. The pH gradually decreased from the initial pH of 3.59 to pH 3.21 on the 6th day of fermentation and there was no change of total acid and pH after 6 days of fermentation.

Comparative Example 1 Comparison of Physicochemical Properties of Invention Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar

In this comparative example, the catabolism of grape vinegar A product of the present invention prepared by fermentation by alcohol fermentation and acetic acid fermentation in Example 1, grape grape vinegar B product prepared in Example 2, and commercial grape vinegar C and D products The mechanical properties were examined in the same manner as in Example 3, and compared. As a result, as shown in Table 1, the sugar content of commercially available grape vinegar D products was slightly higher than that of other products as 5.41。 Brix, and the grape vinegar A products and grape onion vinegar B products of the present invention prepared by two-step fermentation were respectively 5.13 and 4.98。 Brix. Residual alcohol content was relatively high as 0.23% in the commercial grape vinegar C products, residual alcohol was not detected in the grape vinegar A product of the present invention and grape grape vinegar B product of the present invention. The pH level was similar, but the commercial grape vinegar D product was the lowest at pH 2.93. Total acid showed a lot of difference according to the sample. The grape vinegar A product of the present invention prepared by the two-step fermentation method was 5.37% of total acid content, 5.16% of grape onion vinegar B product prepared by adding onion juice and 6.36 of commercial grape vinegar D product. % Higher than other products. The color of grape vinegar A product, grape onion vinegar B product, and commercial grape vinegar C product of the present invention showed similar levels of L, a, and b value, whereas the commercial grape vinegar D product had significantly higher L and b values than other products. High (Table 2). In addition, turbidity was significantly low in the commercial grape vinegar C product was 0.095, the highest in the commercial grape vinegar D product of the present invention (Table 2). The general vinegar vinegar is added to about 30% grape juice, the final product is diatomaceous earth and ultra-filtration, while the grape vinegar A product of the present invention and grape vinegar vinegar B as a filtration method by centrifugation It is believed to be due to the production.

Comparison of the Contents and pH of Sugars, Alcohols, Acids in Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar Physicochemical Properties Sample A B C D Sugar concentration (。 Brix) 5.13 4.98 4.32 5.41 Alcohol(%) - - 0.23 0.09 pH 3.20 3.24 3.06 2.93 Acidity (%) 5.37 5.16 4.65 6.36 A: Invention Grape Vinegar B: Invention Grape Onion Vinegar C: Herbal Acid Vinegar Vinegar of Daechang Brewing Farming Corporation D: Geobong Grape Vinegar of Wheat Grain Food

Comparison of Turbidity and Color in Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar Physicochemical Properties Sample A B C D L 25.53 30.97 57.17 18.34 a +27.81 +34.91 +28.67 +18.49 b +39.94 +49.89 +64.61 +29.69 ΔE 88.94 91.99 82.63 88.84 Turbidity 0.599 0.368 0.095 0.826 A: Invention Grape Vinegar B: Invention Grape Onion Vinegar C: Herbal Acid Vinegar Vinegar of Daechang Brewing Farming Corporation D: Geobong Grape Vinegar of Wheat Grain Food

Comparative Example 2 Analysis of Organic Acid in Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar

The organic acid analysis of the grape vinegar, grape onion vinegar and commercial grape vinegar of the present invention is to remove the fat and oil components of the grape vinegar solution with hexane and then remove the pigment and protein components by 0.45μm membrane filter and Sep-pak C ₁₈ filtration. And then analyzed. Instruments and conditions used were HPLC (Waters Co.), μ-Bondapak C ₁₈ column, column oven temperature 40 ° C., distilled water in mobile phase, flow rate 0.6 mL / min, injection volume 5 μl, RI detector was used. In addition, calibration curves of oxalic acid, citric acid, tartaric acid, succinic acid, lactic acid, and acetic acid standards were prepared under the same analysis conditions to quantify each organic acid. As a result, oxalic acid, tartaric acid, lactic acid, acetic acid, citric acid and succinic acid were detected as organic acids of grape vinegar, and the analysis results are shown in Table 3. Acetic acid content of the organic acid contained in each vinegar was 4.3 ~ 5.3% of the main organic acid components and the organic acid content is different depending on the product, citric acid in the present invention grape vinegar A product and commercial grape vinegar C product It was a relatively high content. Organic acids, including acetic acid, formed the acidity and sweetness of vinegar and had a significant impact on vinegar quality. The tartaric acid content was 340.0 in the grapevine vinegar A product of the present invention, 316.7 in the grape grape vinegar B product of the present invention, 322.6 in the commercial grape vinegar C product, and 391.7 mg% in the commercial grape vinegar D product. The lactic acid content reported as the odor of vinegar is 277.4mg% of commercial grape vinegar D products, which is markedly higher than other products, which may affect overall palatability and result in deterioration of product quality.

Comparison of Organic Acid Analysis in Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar Organic acid Sample A B C D Oxalic acid 28.2 29.1 27.3 47.2 Tartaric acid 340.0 316.7 322.6 391.7 Lactic acid 186.3 186.4 218.4 277.4 Acetic acid 5305.6 4634.1 4347.8 4949.4 Citric acid 141.4 19.6 242.6 5.5 Succinic acid 21.9 55.1 59.1 46.8 A: Invention Grape Vinegar B: Invention Grape Onion Vinegar C: Herbal Acid Grape Vinegar D of Daechang Brewing Co., Ltd.

Comparative Example 3 Free Amino Acids of Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar of the Present Invention

In order to quantify the free amino acid of grape vinegar, grape onion vinegar and commercial grape vinegar of the present invention, 30 mL of ethanol was added to 10 mL of sample, and then allowed to stand at room temperature overnight to precipitate and remove the protein, followed by centrifugation of the supernatant at 3,000 rpm for 10 minutes. It was taken, heated in a hot water and dried. Dilute this solution by adding 10 mL of citrate buffer at pH 2.2, and filter the filtrate with 0.45 μm membrane filter using amino acid autoanalyzer (LKB 4150, alpha autoanalyzer, Ultrapac 11 cation) by ninhydrin method. exchange resin). As a result, in Table 4, the total free amino acid content was 9.21-15.59 mg%, which was different depending on the product. Commercial grape vinegar D product was the highest at 15.59 mg% and grape onion vinegar B product was the highest at 12.00 mg%. Since the composition of the amino acids contained in each vinegar is different, the grape vinegar A product of the present invention had a high asparagine content of 6.72 mg%, while the grape onion vinegar B product prepared by the same two-step fermentation had a high alanine of 7.13 mg%. Commercial grape vinegar D, which had a high total free amino acid content, was high in glutamic acid, threonine, and glycine.

Comparison of Free Amino Acid Contents of Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar amino acid A B C D Aspartic acid 5.0 5.5 3.7 - Threonine 1.6 7.8 6.7 21.0 Serine 1.4 4.3 3.7 12.5 Asparagine 67.2 - - 8.3 Glutamic acid - 15.1 9.7 34.7 Glycine 1.3 5.7 4.9 21.3 Alanine 11.5 71.3 47.0 1.5 Valine 1.1 5.6 4.8 13.8 Cystine 1.8 - - - Methionine 0.8 0.2 0.7 4.3 Leucine 0.4 1.3 9.0 19.0 Tyrosine - 3.2 4.7 19.5 Sum 92.1 120.0 94.9 155.9 A: Invention Grape Vinegar B: Invention Grape Onion Vinegar C: Herbal Acid Grape Vinegar D of Daechang Brewing Co., Ltd. D: Giant grape vinegar unit of wheat grain food: μg / mL

Comparative Example 4 Analysis of Trace Components of Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar of the Present Invention

25 mL of a disintegrating agent (HClO ₄ : H ₂ SO ₄ : H ₂ O ₂ = 9: 2: 5, v / v) was added to 100 mL of the solution of grape vinegar, grape onion vinegar and commercial grape vinegar of the present invention and slowly heated at low temperature. After complete decomposition, the resulting mixture was filtered and purified to 100 mL. The sample was analyzed under the conditions of Table 5 using an atomic absorption spectrophotometer (spectra A-800, Varian Co.). As a result, Cu in each vinegar was below 0.25 ppm and Fe content was below 5.6 ppm (Table 6). The content of K was considerably high in all vinegars, especially 871.39 ppm in product A of the present invention, which was much higher than other vinegars. In the present invention, grape vinegar A product prepared by two-step fermentation showed higher K, Na, Cu content than grape onion vinegar B product, and Na content was particularly high in commercial grape vinegar C products.

Conditions for Trace Component Analysis by Atomic Absorption Spectrophotometer of Grape Vinegar Trace ingredient λ _max (nm) Atomization method fuel Cu 324.8 Absorption Acetylene gas Fe 248.3 Absorption Acetylene gas K 766.5 Absorption Acetylene gas Na 589.0 Absorption Acetylene gas

Comparative Analysis of Trace Components of Grape Vinegar, Grape Onion Vinegar and Commercial Grape Vinegar Trace ingredient Sample A B C D Cu 0.06 0.13 0.25 0.24 Fe 5.66 1.98 5.51 2.14 K 871.39 449.65 377.72 382.07 Na 11.62 2.45 94.63 6.82 A: Invention Grape Vinegar B: Invention Grape Onion Vinegar C: Herbal Acid Grape Vinegar D of Daechang Brewing Co., Ltd.

As described above through the above Examples and Comparative Examples, the grape vinegar of the present invention obtained in a short time by carrying out a two-step fermentation process by removing the seeds and alcohol-fermented and then acetic acid-fermented again grapes and onions in the grape vinegar Grape onion vinegar obtained by the addition of juice contains organic acids, especially tartaric acid and citric acid, free amino acids and a large amount of components such as copper, iron, potassium, sodium, etc. It is a useful invention.

Claims (2)

Saccharomyces cerevisiae, an alcoholic fermentation strain, is cultured by alcohol fermentation followed by acetic acid fermentation using a conventional acetic acid fermentation strain. Grape vinegar, characterized in that the tartaric acid (tartaric acid) 340 mg%, acetic acid (acetic acid) contained 5305 to 5306 mg%. delete