KR101372588B1 - A preparing method of blended type vineger - Google Patents

Abstract

According to the present invention, the brown rice Takju was prepared by varying the ratio of fermenting agent and the quality characteristics of brown rice vinegar using the same were investigated. The sugar content decreased as the alcohol fermentation progressed. The free sugar showed only glucose and maltose at the beginning of fermentation, fructose and sucrose were detected from the first day of fermentation, and decreased with the fermentation period. On the other hand, the titratable acidity of brown rice vinegar was highest in BV (5.1 ~ 5.3%) according to the leaven and coenzyme ratio.According to the organic acid content, the major organic acid, acetic acid, was highest in AV at 5,054 mg% on day 12. BV was highest at 5,005 mg% on day 24, CV, DV and EV at 5,154, 5,243 on day 18, respectively. The highest concentration was 5,062 mg%, and the remaining organic acids were higher in the cultivation of yeast and coenzyme mixture than in the single addition of yeast or coenzyme, and the content of γ-aminobutyric acid (GABA), the major bioactive substance of brown rice, was higher. Although the relative concentration of acetic acid fragrance was slightly decreased, the relative concentration of sensory good fragrance was increased.

Description

Fermentation method of compromised brown rice vinegar {A preparing method of blended type vineger}

The present invention relates to a brown rice vinegar fermentation method characterized by using a main fermentation yeast and coenzyme.

The fermenting agent is divided into soup and basal liquor under the jurisdiction law, and soup is a sugar agent that converts starch into sugar, which is the traditional yeast of Korea, which has been used gradually since independence ( koji ), Coenzyme and purified enzyme have been developed and distributed since 1960. The traditional liquor of our country has various kinds of liquor such as Takju, Yakju, and Soju, which are brewed with rice as raw material and leaven as fermenting agent. Since conventional Nuruk is manufactured in natural fermentation state by using raw starch as it is without sterilization by coarse grinding of grains, it is produced in natural fermentation state, so that various kinds of microorganisms such as mold, yeast, bacteria, etc. Having yeast alcohol fermentation ability by yeast at the same time can be produced by traditional yeast alone, and because it is brewed by the composition of several strains, there is a variety of flavors of Takju. However, due to unnecessary microbial growth, the enzyme power and the number of yeasts are lowered, which makes it difficult to maintain the uniformity of the product. Coenzyme (improved yeast) enhances glycation by artificially culturing saccharifying enzyme-producing bacteria such as Rhyzopus , Usami and 0 ryzae , which are the main strains of conventional yeast, and simultaneously pursues the complex taste and saccharifying power of conventional yeast. This prevents the advantage that the wine is manufactured with a uniform quality. Therefore, the enzyme activity, organic acid production capacity, and alcohol fermentation power by microorganisms vary depending on the type of fermentation agent, which is considered to have a great influence on the quality characteristics of volatile flavor components, taste, color, etc. of Takju.

Vinegar is an acidulant that enhances the taste of food, and is a representative fermented food with unique aroma and sour taste produced during fermentation. Consumption patterns were changed to high-quality fermented vinegars such as fermented vinegar diluted with spirits, fermented vinegar containing inorganic salts and grain vinegar containing more than 4% of grain, followed by increased grain content, and fermented black vinegar rich in organic acids and amino acids. Recently, high-quality fermented vinegars are commercially available using traditional culture and fermentation methods, which are of increasing interest. Political culture method of traditional ripening method is natural fermentation, the characteristics of the raw material remain a lot of nutritional properties, and a lot of sensory excellent volatile fragrances are detected, the market size is expected to grow significantly due to the trend and diversification of vinegar. The studies on the quality characteristics of political vinegar have been reported so far, including the current status and outlook of the domestic vinegar market, the quality characteristics of apple cider vinegar according to agitation and political cultivation, the quality characteristics of tomato vinegar according to the properties and political fermentation, types of yeast and The organic acid and volatile components of brown rice vinegar according to fermentation method, amount of yeast addition and quality characteristics of brown rice vinegar with different culture methods are actively reported.

The present inventors have prepared a rice bran using conventional yeast in the telegram, and then prepared a brown rice vinegar by the property and political culture method to investigate its quality characteristics and applied for a patent (10-2010-0005697). At this time, if you go through the long-term fermentation process according to conventional Nuruk alone, off-flavor could occur. Therefore, the present invention investigated the quality characteristics of brown rice vinegar according to the ratio of the fermenting yeast and coenzyme, and in particular, attempted to improve the flavor component and quality by standardizing the soaking.

Therefore, an object of the present invention is to provide an eclectic brown rice vinegar fermentation method characterized by using a mixture of yeast and coenzyme.

The object of the present invention is to prepare an eclectic brown rice vinegar; It was achieved through the step of evaluating the quality of brown rice vinegar fermented according to the manufacturing method.

The present invention has an excellent effect of increasing the content of various organic acids and amino acids and organoleptic fragrance component by the fermentation method of cultivated brown rice vinegar by mixing koji and coenzyme.

1 is a graph showing the change of alcohol and sugar content in brown rice alcohol fermentation according to the addition amount of Nuruk and coenzyme according to the present invention.
2 is a view showing the change of titratable acidity and pH during alcohol fermentation of brown rice by the difference in content of koji and coenzyme according to the present invention.
Figure 3 is a view showing the change of titratable acidity and pH during stationary vinegar fermentation of brown rice by the difference in content of koji and coenzyme according to the present invention.
Figure 4 is a fragrance component pattern of the chromatogram during stationary fermentation of brown rice vinegar by various yeast and coenzyme ratio according to the present invention.
5 is an analysis graph of the main flavor components of brown rice vinegar by the ratio of various yeast and coenzyme according to the present invention.
6 is a view showing a polar frequency pattern of organic acids of brown rice vinegar at various yeast and coenzyme ratios according to the present invention.

The experimental materials and Used strain

The brown rice used in this experiment was purchased from ordinary brown rice grown in Sangju, Gyeongbuk in 2010, and the liquefied enzyme α-amylase (14,500 unit / g, Daiwa kasei, Japan) used for alcohol fermentation was E & B Biotech Co., Ltd. E & Bio tech Co., Ltd., Korea) was used to store at room temperature. The yeast was purchased from Sangjugoza Co., Ltd. and coenzyme CU210^®(4,000sp,Asp . usami) Was purchased from Korea enzyme Co., Ltd. (Hwaseong, Korea). Yeast used for alcohol fermentation in brown rice is stored in fermentation engineering laboratory.Saccharomyces cerevisiae GRJ was passaged at 30 ° C. for 24 hours in YPD agar medium (yeast extract 1%, peptone 2%, glucose 2%, agar 2%, pH 6.0).Acetobacter pomorum KJY 8 (KCTC 10173BP) was prepared as a solid medium (glucose 3%, yeast extract 0.5%.₃ 1%, ethanol 3%, agar 2%, pH 7.0) was used for 30 hours at 48 ℃, and then refrigerated at 4 ℃.

Grandma and Herb

The primary hair was 1 500 mL of purified water in 500 g of koji water, saccharified at 55 ° C. for 6 hours, filtered through a nonwoven fabric, and adjusted to 10 ^o Brix. The saccharified solution was sterilized at 121 ° C. for 15 minutes and then Saccharomyces cerevisiae GRJ was inoculated and incubated in an incubator (HB-103-2H, Hanbaek Scientific Co., Bucheon , Korea) at 30 ° C for 24 hours, and 5% (v / w) was used for the amount of raw material. The used seeds were diluted with alcohol content of brown rice alcohol fermentation solution to 6% alcohol and then Acetobacter pomorum KJY 8 (KCTC 10173BP) was inoculated and stirred at 250 rpm at 30 ° C. for 10 days.

Ingredient Analysis for Fermented Raw Materials

The contents of general and free amino acids of brown rice, koji and coenzyme used as fermentation raw materials of brown rice vinegar were investigated.

Brown Rice Alcohol Fermentation According to Fermentation Rate

Add 600 g of brown rice powder sample to 5L fermentation tank, add water to 480% (v / w), add 0.03% (v / w) of liquefied enzyme, and add 90 ° C in shaking incubator (HB 205SWM, Hanbeak Scientific Co., Korea). Hydrolysis at 150 rpm for 60 minutes. The yeast and coenzyme ratios in the yeast addition amount of 30% (w / w) compared to the brown rice suspension liquefied as the optimum conditions of the telegram were A ( Nuruk 100: crude enzyme 0), B ( Nuruk 75: crude enzyme 25), and C ( Nuruk 50: crude enzyme 50), D ( Nuruk 25: crude enzyme 75) and E ( Nuruk 0: crude enzyme 100) were added, and then incubated with incubator 5% (v / w) (HB-103-2H) , Hanbaek Scientific Co., Korea) was incubated at 30 ℃ for 3 days, and after the fermentation was finished by primary filtration with a nonwoven fabric and centrifuged at 13,000 rpm for 5 minutes was used as an analytical sample.

Acetic Acid Fermentation According to Fermentation Agent Ratio

The brown rice alcohol fermentation filtrate according to the addition of koji and coenzyme agent was adjusted to alcohol content 6% (v / v), and then inoculated with 10% (v / v) seed incubator (HB-103-2H, Hanbaek Scientific) Co., Korea) was incubated at 30 ℃ for 24 days. Brown fermented brown vinegar AV ( Nuruk 100: crude enzyme 0), BV ( Nuruk 75: crude enzyme 25), CV ( Nuruk 50: crude enzyme 50), DV ( Nuruk 25: crude enzyme 75) and EV ( Nuruk 0: Crude enzyme 100) was used as analytical sample supernatant centrifuged for 5 minutes at 13,000 rpm.

General compositional analysis

For the analysis of general ingredients of fermented raw materials (brown rice, yeast and crude enzyme), moisture was subjected to atmospheric pressure drying method according to AOAC method, micro-Kjeldahl method for crude protein content, and Soxhlet extraction method for crude fat content. It was measured by.

The results of analyzing the general components of brown rice, koji and agonist used as fermented raw materials of brown rice vinegar are shown in [Table 1]. Brown rice was the highest in carbohydrate 77.93%, water content 13.38%, crude protein 5.90%, crude fat 1.64%, crude ash 0.99%, crude fiber 0.16%. Yeast showed high carbohydrate 74.49%, water content 11.26%, crude protein 11.09%, crude ash 1.64%, crude fat 1.50%, crude fiber 0.02%. Coenzyme was the highest in carbohydrate 62.89%, crude protein 22.12%, water content 6.82%, crude ash 6.32%, crude fat 1.81%, crude fiber 0.04%. Therefore, the protein content is higher in the order of coenzyme, yeast, and brown rice, and the free amino acid content is expected to increase as the ratio of coenzyme is increased.

Alcohol content and sugar content

Alcohol content was calculated by distilling 100 mL of sample and converting the value measured using a spirit meter to Gay Luccac Table.,Japan).

As a result of investigating the alcohol content change of brown alcohol fermentation broth according to the ratio of fermentation agent, as shown in Fig. 1, it was the highest as 9.1% in the second day of fermentation (B), (8), (8), (8), The higher the coenzyme ratio, the lower the alcohol content (A) was the lowest (7.7%). The sugar content decreased drastically as alcohol fermentation progressed, and gradually decreased after 3 days of fermentation. As a result of examining the titratable acidity, the fermentation period increased in all the additions as shown in FIG. 2 and appeared to be 0.7∼0.8% after the end of fermentation. pH decreased steadily during the alcohol fermentation period.

Titratable acidity  And pH

The titratable acidity was 1 ~ 3 drops of 1% phenolphthalein indicator in 1 mL of sample, neutralized to 0.1 N NaOH, and converted into acetic acid (%). PH was pH meter (Metrohm 691, Metrohm UK Ltd., Herisau, Switzerland) at room temperature.

The result of examining the change of titratable acidity of brown rice acetic acid fermentation according to the ratio of fermentation agent is shown in FIG. 3. Initially, the titratable acidity increased rapidly in AV compared to the other additions, but the final titratable acidity was highest in BV (5.1 ~ 5.3%). Overall, titratable acidity increased from 15 to 18 days of acetic acid fermentation and decreased after 18 days. The initial pH ranged from 3.6 to 4.0, and it decreased steadily as the fermentation progressed, resulting in 3.3 to 3.5 on the 15th day of fermentation.

Free sugar content

Free sugar was analyzed by high performance liquid chromatography (HPLC, Waters 1515, Waters Co., Milford, USA) by passing the alcohol fermentation broth through a Sep-pak C18 cartridge, followed by filtration through a 0.45 μm membrane filter. Carbohydrate analysis column (3.9 × 300 mm, Waters Co.), mobile phase used 75% acetonitrile (JTbaker Co., Phillipsburg, USA) with flow rate of 1.0 mL / min and injection volume of 20 μL. It was analyzed by RI detector (M410 RI, Waters Co.).

The results of analyzing the change of free sugar content of brown rice alcohol fermentation according to the ratio of fermentation agent are shown in [Table 2]. In the early stage of fermentation, only glucose and maltose were detected, and fructose and sucrose were detected from day 1 of fermentation. Also, there was no big difference according to the ratio of fermentation agent.

Organic acid content

The organic acid was diluted 5 ~ 10 times with acetic acid fermentation solution, passed through Sep-pak C18 cartridge, filtered through 0.45 μm membrane filter, and analyzed by High Performance Liquid Chromatography (HPLC, Waters 1515, Waters Co., USA). AtlantisTM dC18 (3.9 × 150 mm, Waters Co.), 20 mM NaH2PO4 (pH 2.7) for mobile phase, flow rate 1.0 mL / min, injection volume 20 μL, detector UV (Waters) 2487, 210 nm) was used.

The results of examining the change of organic acid content of brown rice acetic acid fermentation according to the ratio of fermentation agent are shown in [Table 3]. The major organic acid, acetic acid, was highest in AV at 5,054 mg% on day 12, BV was highest at 5,005 mg% on day 24, and CV, DV and EV were highest at 5,154, 5,243 and 5,062 mg% on day 18, respectively. In addition, other lactic acid showed a higher content in the yeast and coenzyme-added mixture than the single addition of the yeast or coenzyme.

Free amino acids  analysis

The free amino acid for fermented raw materials was added 100 mL of 60% ethanol solution to 5 g of the sample, and the filtrate was repeatedly extracted at 80 ° C. for 2 hours, and concentrated under reduced pressure at 45 ° C. to give 5 mL of 0.1 N sodium citrate buffer (pH 2.2) solution. In addition, 30 mL of ethanol was added to 10 mL of free amino acid in acetic acid fermentation solution. The solution was left at room temperature overnight to precipitate and remove the protein. The supernatant was centrifuged at 8,000 rpm for 15 minutes, and the upper layer was taken up and dried in a heated bath. Diluted by adding 10 mL of citrate buffer. After pretreatment, the filtrate was filtered with a 0.45 um membrane filter using an amino acid autoanalyzer (L-8800, Hitachi Co., Tokyo, Japan).

The results of analysis of the free amino acid content of brown rice, koji and coenzyme used as fermentation ingredients of brown rice vinegar are shown in [Table 4]. The major free amino acids of brown rice were asparagine, glutamic acid, ammonium chloride, alanine and γ-aminoburyric acid (GABA). The major free amino acids of yeast were proline, ammonium chloride, alanine, tryptophan, glutamic acid, asparagine, and GABA. The major free amino acids of coenzyme were ammonium chloride, glutamic acid, arginine, GABA, alanine, serine, lysine, asparagine, histidine, threonine and tyrosine. The content of GABA, which is known as the major functional enhancer of brown rice, was 10.18 mg% in brown rice, 45.66 mg% of yeast, and 205.68 mg% of coenzyme.The total free amino acid was 189.04 mg% in brown rice and 1,268.56 mg% in yeast, coenzyme. The formulation was found to be 2,627.46 mg%.

The results of analyzing the free amino acid content of brown rice vinegar according to the ratio of fermentation agent are shown in [Table 5]. Threonine, glutamic acid, alanine, valine, leucine, and phenylalanine were more than 10 mg% in brown rice vinegar (BV ～ EV) prepared by adding coenzyme rather than brown rice vinegar (AV). In particular, γ-aminobutyric acid, the main bioactive substance of brown rice, showed the lowest content in AV (2.02 mg%), BV 7.60 mg%, CV 6.92 mg%, DV 8.36 mg%, EV 9.15 mg%. The higher the γ-aminobutyric acid was, the higher the tendency was. The total free amino acid content was the lowest in AV (132.06 mg%), and BV ~ EV showed similar content around 190 mg%. Therefore, the content of essential free amino acid and total free amino acid was higher than that of brown rice vinegar (BV ~ EV) prepared by addition of coenzyme, compared to brown rice vinegar (AV) prepared by adding yeast. There was no difference.

Electronic nose  analysis

As the electronic nose used in the fragrance pattern analysis of the sample, an electronic nose system using a surface acoustic wave (SAW) sensor (zNose 7100, Electronic Sensor Technology, Newbury Park, CA, USA) was used. 2 mL of vinegar was added to 40 mL vial (Supelco, Bellefonte, PA, USA), sealed with Teflon-coated septa (PTFE / silicone septa, Supelco), and left at room temperature for 24 hours. The headspace part was separated into a single material by a DB-5 capillary column (Supelco, Bellefonte PA, USA) by a carrier gas (high purity helium: 99.9995%), and then detected by a SAW sensor. In order to check the reproducibility, the experiment was repeated three times for each sample, and the temperature conditions of the apparatus used were SAW sensor 30 ℃, column 60 ℃, valve 120 ℃, inlet 150 ℃, trap 220 ℃. The measured fragrance pattern was analyzed using VaporPrintTM program (Misrosense 4.88, Electronic Sensor Technology, Newbury Park, CA), and principal component analysis (PCA) was performed using SAS program (version 8.1) on the detected main peak. Analyze and compare the patterns between samples.

4 shows the change of the electronic nasal pattern during acetic acid fermentation of brown rice vinegar according to the ratio of fermentation agent. On day 0 of acetic acid fermentation, peaks were different according to the ratio of yeast and coenzyme at 2.30, 3.20, 3.54, 3.80, 4.96, and 6.70 seconds. As the fermentation progressed, high peaks appeared at 0.60, 0.94, 1.58, and 1.70 seconds, and the peaks between 2 and 4 seconds after fermentation showed significant differences according to the leaven and coenzyme ratios. Therefore, the peaks within 2 seconds are assumed to be the peaks represented by acetic acid produced by acetic acid fermentation, and the peaks after 2 to 4 seconds are estimated to be the peaks produced by the leaven and coenzyme ratio. Analysis of the main components of the electronic nose pattern peaks of Figure 4 shows the difference in the fragrance pattern according to the yeast and the ratio of the agent is shown in FIG. On day 0 of fermentation, differences in fragrance patterns were observed in AV, BV, and CV, but no obvious differences were observed in DV and EV. On day 12 of fermentation, DV and EV were fermented in a similar pattern as on day 0. On the 24th day of fermentation, the AV and EV fermented by using the yeast and coenzyme alone showed distinct odor pattern differences, and BV, CV, and DV fermented with the yeast and coenzyme mixed ratio showed similar fragrance patterns. In addition, as a result of the electronic pattern analysis of brown rice vinegar on the 24th day of fermentation in the frequency pattern, it was found that it appeared in a unique pattern according to the proportion of each yeast and coenzyme (FIG. 6).

Volatile Component Analysis

To collect volatile components of the sample, volatile components were adsorbed using SPME fibers (Supelco, Bellefonte, PA, USA) coated with carboxen / polydimethylsiloxane (CAR / PDMS, 75 μm thickness). Before adsorbing volatile components, the fiber was preheated at 250 ° C for 5 minutes with GC. SPME capture was added to the headspace vial (22.5 × 75 mm, PTFE / silicon septum, aluminum cap) by adding 5 mL of sample to 25% NaCl. Preheated SPME fiber was injected. Samples were preheated for 5 minutes in a 35 ° C. heating block and SPME fibers collected volatiles for 10 minutes, then injected into GC and held for 2 minutes for desorption. For volatile analysis, GC (Agillent GC 6890, Palo Alto, CA, USA) with mass selective detector (MSD) was used, and HP-FFAP capillary column (30 m × 0.25 mm × 0.25 μm) and He were carriers. Used as gas (1 mL / min). At this time, the oven temperature was maintained at 35 ° C. for 10 minutes and was raised to 5 ° C. per minute up to 100 ° C. and 10 ° C. up to 210 ° C. for 10 minutes. The MS source, MS quadrupole, and transfer line were 230, 150, and 280 ° C, respectively. The MS library was Wiley7Nist0.5 (Wiley7Nist0.5 Library, mass spectral search program, version 5.0, USA).

The results of analyzing the volatile flavor components of brown rice vinegar according to the ratio of fermentation agent are shown in [Table 6]. The major volatile components of brown rice vinegar according to yeast and coenzyme ratio were detected in 20 species such as acetic acid, 3-methyl butyl acetate, acetoin, and isomayl alcohol. The main volatile component was acetic acid, which showed the highest relative concentration of AV (67.56%) and low EV (55.53%). Acetoin, the major fragrance component of butter or fermented milk, and the cause of bad smell in brews and juices, was highest at 4.74% in AV and low at around 3% in coenzyme added groups. Ethyl acetate, isobutyl acetate, 3-methyl butyl acetate, and phenethyl acetate, which showed rose, floral, sweet and fruity flavors, were higher in samples containing coenzyme than in AV (BV ~ EV). There was no difference. Therefore, as a result of the separation and identification of the fragrance components using the SPME method, the relative concentration of acetic acid flavor was slightly reduced compared to the brown rice vinegar (AV) prepared by adding some coenzyme (BV ~ EV) to brown rice vinegar (AV). However, since the relative concentration of the sensory good fragrance increased, it would be better to prepare brown rice vinegar by combining yeast and coenzyme.

The present invention is a very useful invention in the vinegar fermentation industry because it has an excellent effect of providing a compromised brown rice vinegar that is new and technologically advanced by admixing yeast and coenzyme in stationary fermentation.

Claims (3)

To the powder of brown rice, 30% (w / w) of a mixture of koji and coenzyme was added to the whole of the brown rice suspension hydrolyzed by adding 0.03% (v / w) of liquefied enzyme α-amylase. After fermentation of 5% (v / w) hair seedlings and 3 days of incubation at 30 ° C., the filtered alcoholic fermentation filtrate was adjusted to 6% (v / v) alcohol content, and then, in the brown rice alcohol fermentation solution, Acetobacter pomorum KJY 8 (KCTC 10173BP) Inoculated with a separate seed vinegar obtained by inoculating 10% (v / v) of the eclectic brown rice vinegar fermentation method characterized in that the stationary culture at 30 ℃ for 24 days.
delete Brown rice vinegar characterized in that prepared according to the method of claim 1.

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