KR20020034718A - Fermentation apparatus for soy sauce and liquid food and fermentation method using thereof - Google Patents

Abstract

PURPOSE: A fermentation apparatus for soy sauce and liquid foods and a fermentation method using the same are provided, therefore the oxygen dissolving rate can be increased and the fermented solution can be easily recovered. CONSTITUTION: The fermentation apparatus for soy sauce and liquid foods comprises a fermenter(10), a raw material storing tank(20), an air supplying device(30), a bubble inhibiting device(40), and a fermented solution recovering tank(50). Particularly, the fermenter(10) comprises a cover(11), a cooling pipe(13) which is positioned at the top of the cover(11) and inhibits the loss of alcohol and volatile compounds, and a multipore plate(14) which is positioned at the bottom of the fermenter(10) and generates micro-sized air bubbles to increase the oxygen dissolving rate into the fermented solution.

Description

Fermentation apparatus for soy sauce and liquid food and fermentation method using the same

The present invention relates to a fermentation apparatus for soy sauce and liquid food, and a fermentation method using the apparatus. More specifically, it contains a large amount of protein or amino acid, such as fermentation tank (10) for receiving and fermenting soy sauce or liquid food that becomes a fermentation substrate, raw material storage tank (20) for storing raw liquid food, and air supply device (30) and soy sauce. Bubble discharge prevention device (40), which is a special flow pipe that prevents the mixing of bubbles caused by carbon dioxide, which is generated in large quantities as injection air and metabolites during fermentation into fermentation broth that is continuously discharged through the discharge port during continuous fermentation of a liquid food. And it relates to a fermentation broth recovery tank (50) for storing the produced fermentation broth and a method of fermenting soy sauce or liquid foods in batch, semi-continuous and continuous alcohol fermentation or organic acid fermentation.

Korean traditional soy sauce is prepared by soaking soy sauce with meju and brine made by natural inoculation bacteria, aging for 2 to 3 months, and then separating it into soy sauce and miso. The traditional soy sauce is a little lactic acid is produced depending on the aging temperature, but alcohol fermentation rarely occurs, it lacks flavor. Therefore, there is a need to prepare soy sauce with excellent flavor and quality through the fermentation process as a ripening process.

Korean Patent Laid-Open Publication No. 2000-58292 discloses a device equipped with an air pipe that can supply oxygen to a tank body, a temperature sensor, an air aeration device, and a hot / cold water pipe in a soy sauce fermentation tank. The known fermentation device is easy to control the temperature, but after the fermentation is finished, the recovery of the fermentation broth is not easy and the air supply is also not easy, there is a disadvantage that requires a plurality of air pipes.

In addition, Korean Patent Publication No. 1993-36 discloses continuous rapid fermentation of liver in a bioreactor filled with immobilized yeast cells. However, the known method also has a limitation in the recovery method and the air supply of the fermentation broth.

In view of the above, the inventor mounted a porous plate 14 at the bottom of the fermentation tank to increase the oxygen dissolution rate by dispersing the sterilized air into fine bubbles, and bubbles were mixed in the discharge fermentation solution during continuous fermentation. The present invention has been completed by producing fermented soy sauce with increased flavor by fermenting soy sauce by batch, semi-continuous and continuous alcohol fermentation using the present invention fermentation apparatus equipped with an anti-foam discharge preventing device 40.

Accordingly, an object of the present invention is to provide a fermentation apparatus which can increase the oxygen dissolution rate in the fermentation broth and prevents bubbles from mixing into the fermentation broth discharged during continuous fermentation.

Another object of the present invention to provide a method for producing soy sauce and liquid fermented food using the fermentation device.

The object of the present invention is a fermentation tank (10) for accommodating soy sauce or liquid food fermentation substrate and the raw material storage tank 20 for supplying the raw material for fermentation and the air supply device for mixing the air supply and fermentation substrate (30) And a porous plate 14 at the bottom of the fermentation tank 10 in a fermentation system consisting of a bubble discharge preventing device 40 and a fermentation liquid recovery tank 50 to prevent bubbles from being mixed in the discharged fermentation liquid continuously discharged during continuous fermentation. By using the present soy sauce and liquid food fermentation apparatus prepared to increase the concentration of dissolved oxygen in the fermentation broth and to prevent the mixing of bubbles into the fermentation broth discharged by mounting the foam discharge prevention device 40 on the upper side, the soy sauce is batch, half Achieved by producing alcoholic fermented soy sauce by continuous and continuous fermentation.

1 is a perspective view of a fermentation apparatus filled with an immobilized enzyme according to an embodiment of the present invention.

Figure 2 is a perspective view of a fermentation apparatus of soy sauce and liquid food using a yeast seed culture medium according to an embodiment of the present invention.

Figure 3 is an enlarged perspective view of the bubble discharge prevention device mounted on the fermentation apparatus of the present invention.

Figure 4 shows the alcohol concentration produced during batch fermentation using the fermentation apparatus of the present invention filled with immobilized Zygosaccharomyces.rouxii and Candida.verstilis according to the fermentation time.

Figure 5 shows the change in alcohol concentration and sugar concentration during batch fermentation of soy sauce by immobilized yeast.

Figure 6 shows the change of sugar and alcohol concentration during semi-continuous alcoholic fermentation of soy sauce in the fermentation apparatus of the present invention filled with immobilized Z.rouxii yeast cells.

Figure 7 shows the change of sugar and alcohol concentration during semi-continuous alcoholic fermentation of soy sauce in the fermentation apparatus of the present invention filled with immobilized C.versitilis yeast cells.

Figure 8 shows the change in alcohol concentration during continuous fermentation of soy sauce by the fermentation apparatus of the present invention filled with immobilized Z.rouxii and C.versitilis .

Explanation of symbols on the main parts of the drawings

10 fermenter 11 fermenter lid

12: perforated plate 13, 52: cooling tube

14: porous plate 20: raw material storage tank

21: Peristaltic Pump 22: Raw Material Transfer Line

30: air supply device 31: air filter

32: air transfer line 40: bubble discharge prevention device

41: residual fermentation liquid discharge pipe 42: upstream pipe

43: siphon phenomenon prevention tube 44, 45: locking device

50 fermentation broth recovery tank

The present invention is to prepare a meju with soybeans, and then mixed with 20% brine to prepare soy sauce, aged, filtered and sterilized at high temperature to prepare a time zone; Immobilizing the yeast cells in alumina ceramic beads; Batch fermentation in a fermentation chamber air-conditioned at 28 ℃ while filling the fermentation apparatus of the present invention hot-pasteurized in the step and the yeast immobilized in the step and injecting air at a rate of 0.5vvm; Measuring the non-alcohol production rate and the non-substrate utilization rate during the batch fermentation of the step; When the average residence time required for complete alcoholic fermentation of soy sauce in batch fermentation in the above step was constant (4 days), fermented soy sauce was recovered and recovered so that the dilution rate was 0.25day ^-1 by dividing the fermentation method into semi-continuous type. Supplying the substrate soy sauce semi-continuous fermentation; A continuous fermentation step of converting the semi-continuous fermentation into a continuous fermentation method so that the daily dilution rate is 0.25day ^-1 like a semi-continuous fermentation, subdividing the substrate soy continuously and feeding it at a constant rate and continuously recovering it; Continuous fermentation of soy sauce while inoculating yeast seed culture broth after inoculating yeast seed culture medium without filling immobilized enzyme in the fermentation apparatus of the present invention; Analyzing lactic acid content, glucose content, total nitrogen, color, salt, alcohol content and free amino acid content of the fermented soy sauce of the present invention; The present invention consists of a step of conducting aroma component and taste test of fermented soy sauce.

In this step, the filling amount of the immobilized yeast (specific gravity = 2.1739) is appropriately 25 to 35% (v / v) of the effective capacity of the fermentation tank, and preferably 20 to 30% (v / v) of the fermentation tank capacity.

Dilution of the semi-continuous or continuous manner during fermentation substrate in the above step is 0.15~0.35 day ^-1 and preferably 0.20~0.30day ^-1.

In the step, the fermentation temperature in batch, semi-continuous or continuous fermentation was adjusted to 20 ~ 30 ℃ and preferably adjusted to 25 ~ 30 ℃.

In this step, the injection rate of air in a batch, semi-continuous or continuous fermentation was 0.05 to 0.1 vvm, preferably aerated at 0.05vvm.

Hereinafter, the configuration and operation of the fermentation apparatus of the present invention will be described in detail with reference to the accompanying drawings.

1 is a perspective view of a fermentation apparatus filled with an immobilized enzyme according to an embodiment of the present invention. The fermentation apparatus of the present invention is a fermentation apparatus filled with microorganisms or enzymes immobilized on an immobilized carrier, and a raw material storage tank 20 for supplying a fermentation substrate soy sauce or a liquid food and a substrate and an air supply and a fermenter. It consists of an air supply device for mixing the vaginal 30 and the bubble discharge prevention device 40 and the fermentation liquid recovery tank 50 to prevent the bubbles are mixed in the fermentation broth discharged during continuous fermentation.

The fermentation tank 10 is made of pyrex glass or 316 sus or acid and flame resistant FRP resin having an inner diameter of 18 cm x height 120 cm (capacity 30 L) or an inner diameter of 18 cm x height 60 cm (capacity 15 L). The fermenter lid 11 for sealing the fermenter is covered and the fermenter inside the perforated plate 12 consists of four stages to fill the immobilized enzyme or microorganism 15 and the fermenter lid 11 In order to prevent volatilization of alcohol and volatile aromatic components produced by fermentation, a cooling tube 13 is installed, and a porous plate 14 is mounted at the lower end of the fermentation tank 10 to supply air supplied from the air supply device 30. By making it into fine bubbles, it increases the rate of oxygen dissolution into the fermentation broth.

The raw material storage tank 20 is connected to the peristaltic pump 21 and can supply the fermentation raw material at a constant speed to the fermentation tank 10 through the raw material transfer line 22 and the locking device to the raw material transfer line 22 In the case of batch fermentation (23), the locking device was closed to allow the substrate to be fermented.

The air supply device 30 functions to create compressed air, and the compressed air is sterilized while passing through the air filter 31 and transferred to the fermentation tank 10 through the air transfer line 32.

The foam discharge prevention device 40 is mounted to the fermentation tank upper side portion and the fermentation broth discharge line 53 is connected to the fermentation broth recovery tank 50 from the fermentation tank (10). The anti-foaming device is shown in more detail in FIG.

The fermentation liquor recovery tank 50 has a function of storing the fermentation broth discharged after the fermentation is completed by the foam discharge prevention device 40 and the fermentation broth discharge line 53 and the tank lid for sealing at the top of the fermentation liquor recovery tank 50. There is a 51 and the upper end is equipped with a cooling tube 52 to prevent the volatilization of alcohol and volatile flavor components in the fermentation broth during storage.

2 is a perspective view of a fermentation apparatus according to an embodiment of the present invention. The fermentation apparatus of the present invention is a device for inoculating fermentation of a seedling culture without immobilized microorganisms or immobilized enzymes, and is the same as the fermentation apparatus as shown in FIG. 1, except that the fermentation tank 10 has a perforated plate for filling immobilized microorganisms or enzymes. (12) is not included.

Figure 3 is an enlarged perspective view of the bubble discharge prevention device 40 mounted to the fermentation apparatus of the present invention. In the present invention, the foam discharge prevention device 40 is connected to the residual fermentation liquid discharge pipe 41 and the fruit pipe inlet 42 on the upper side of the fermentation tank 10, and is in communication with the siphon phenomenon prevention tube 43, and fermentation during continuous fermentation. After the fermentation broth is completed, the fermentation broth enters through the remaining fermentation broth discharge pipe 41 and the fruit pipe inlet 42, passes through the siphon phenomenon prevention tube 43, and is collected in the fermentation broth recovery tank 50 via the fermentation broth discharge line 53. The height of the residual fermentation liquid discharge pipe 41 and the inlet pipe 42 are both lower than the end height of the siphon phenomenon prevention tube 43 and the ends of the siphon phenomenon prevention tube 43 are double stranded to prevent the siphon phenomenon. Even if bubbles are generated due to bubbles generated by carbon dioxide generation and air injection during fermentation, the inlet into the fermentation broth solution tank 50 is prevented because the inlet of the fruit pipe 42 is locked in the fermentation broth. In addition, the locking device (44, 45) is mounted, in the case of batch fermentation, during the fermentation, the inlet locking device (23) and the outlet locking device (44) is closed, and when the fermentation is completed, the inlet locking device (23) is opened to release the fermentation broth. In the case of continuous fermentation, the locking devices 23 and 44 were kept open to facilitate the continuous supply and recovery of fermentation broth.

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 Batch Alcohol Fermentation of Traditional Soy Sauce by the Fermentation Apparatus of the Invention Filled with Immobilized Yeast

First Step: Preparation of Alcohol Fermentation Time Zone

Prepared meju by conventional method using Taekwang soybean produced in 1999 (produced ingredient ingredient of Gyeongbuk Agricultural Products Garden), and mix it with 20% brine and 1: 3 ratio to make soy sauce. and soy sauce were separated by filtration twice. The filtered soy sauce was further aged for 2 months at room temperature, and then sterilized at 112 ° C. for 8.48 min to be used as an alcohol fermentation time zone.

Second Step: Immobilization of Yeast Cells

Porous alumina ceramic beads were prepared to immobilize the yeast cells. Using alumina of # 240 (Alumina: Al ₂ O ₃ ) as an aggregate and feldspar ((NaK) AlSi ₃ O ₈ ) as a binder, mix alumina 58%, distilled water 35%, feldspar 5.8% and PVA 1.2% by mass ratio. A 5 mm diameter bead was completed.

In order to immobilize Candida versatilis and Candida versatilis and Zygosaccharomyces rouxii on the carrier, a mixture of conventional soy sauce and soybean extract in a volume ratio of 1: 1 is added to sterilized soy sauce medium. Yeast was inoculated and incubated at 30 ° C. for 5 days. After centrifugation of the cultured yeast seed culture medium, a slurry was prepared with sterile water to immobilize the yeast on the prepared carrier.

Step 3: Batch Fermentation Using Immobilized Yeast

Immobilized yeast of Candida versatilis and Candida versatilis and Zygosaccharomyces rouxii prepared in the second process, respectively, 10 kg (4.6 L), respectively, in 15 L and 30 L of the present invention fermenter 10 Each 20 kg (9.2 L) was divided into two stages and four stages in the fermenter, and the top of the perforated plate 12 was filled. 10 kg and 20 kg of sterilized conventional soy sauce were added to the fermentation tank 10 filled with the immobilized yeast from the raw material storage tank 20 by using a ferritic pump 21 and an air supply device ( Compressed air produced by 30) is passed through the air filter 32 to be sterilized, and then made into fine bubbles by the porous plate 14 mounted on the bottom of the fermentation tank 10, and injected at a rate of 0.5vvm to 28 ± 0.5 ° C. Alcohol fermentation was performed in the air-conditioned fermentation chamber to produce soy sauce containing 1.8% (v / v) alcohol. Upon completion of the batch fermentation, the locking device 23 was opened to recover the produced alcoholic soy sauce.

Experimental Example 1: Measurement of the fermentation constant during the ash fermentation of soy sauce using the fermentation apparatus of the present invention filled with immobilized yeast

Specific rate of product formation and specific rate of substrate utilization in the fermentation of alcohol at the maximum rate during the batch fermentation are based on the concentration of beads that fix the yeast in the substrate (X). , g / L) and the concentration of the resulting product (P, g / L), the concentration of the substrate used (s, g / L) and the fermentation time (t, day) by measuring the following formula (I), Ⅱ) and the unit of non-alcohol production rate and non-substrate utilization rate are day ^-1 . The retention time (RT) is the total fermentation (day) during batch fermentation of the substrate in the fermentation system.

Non-alcohol Production Rate =

Specific Substrates (Speed) =

As a result, the sugar and alcohol concentration change of soy sauce is shown in Fig. 4, and after fermentation, Z. rouxii or C is produced from the soy sauce with 4% sugar concentration until 1.8% (v / v) alcohol is produced. Immobilized yeast of versatilis took 16 days, and 2 fermentations took 9 days, 3 times 8 days, 4 times 5 days, and from 6 fermentations, 3 consecutive times 1.8% after 4 days of fermentation. Soy sauce containing (v / v) alcohol was stably produced, and the retention time was 4 days at the highest alcohol fermentation rate during batch fermentation of soy sauce.

In addition, as shown in FIG. 5, Z.rouxii and C. versatilis temporal (day- ¹ ) alcohol production equations are represented by the following formulas (III) and (IV), respectively, and fermented using Z. rouxii immobilized yeast. Alcohol production rate was high and the production rate of alcohol, which is a product produced in 1L substrate from the non-alcohol production rate, that is, the weight of immobilized yeast unit immobilized on the carrier contained in 1L soy sauce (g, including the carrier weight) (day ^-1 ) were 0.0033day ^-1 and 0.0031day ^-1, respectively.

In addition, the specific substrate consumption rate was -0.0087 day ^{-1 in} both Z.rouxii and C.versatilis immobilized yeast, and the time- consuming sugar consumption equation was as shown in the following equation (V). It was found that consumption of sugar produced 0.36 to 0.37% (w / v) of alcohol.

Example 2: Semi-continuous fermentation of conventional soy sauce using the fermentation apparatus of the present invention filled with immobilized yeast

From day 52 of batch alcohol fermentation in Example 1, the fermentation method was switched to semi-continuous. Semi-continuous fermentation recovers a certain amount of substrate soy sauce in the fermentation tank 10 by opening the locking device 23, and locks the same amount of new soy sauce from the raw material storage tank 20 by using the peristaltic pump 21. Was carried out while replenishing into the fermenter (10) in the open state, and the recovery amount of the soy sauce was filled with immobilized Z.rouxii and a large fermentation apparatus containing 20 kg of substrate soy sauce and substrate soy sauce filled with immobilized C.versatilis. 5 kg and 2.5 kg of fermented soy sauce, each weighing 0.25day ^-1 , were collected at a given time every day in a small fermentation apparatus containing kg, and then supplemented with fresh soy sauce containing 4% sugar as recovered. The air sterilized at 0.5 ° C. was passed through the porous plate 14 at a rate of 0.5 vvm, and semi-continuous fermentation was performed while injecting air, and the change of alcohol and sugar content of the liver was measured over time during fermentation.

As a result, as shown in FIG. 6, Z.rousii -fixed fermentation apparatus of the present invention in the fermentation apparatus of the soybean during the continuous fermentation of soy alcohol has a specific alcohol production rate of 0.0045 day ⁻¹ and a sugar consumption rate of 0.01 day ^−1. The sugar consumption rate and alcohol production rate were slightly higher than the batch fermentation using the device, and it was able to produce semi-continuously for 13 days with 5 kg of soy sauce containing 1.92% (v / v) of alcohol and 0.06% of residual sugar. Could. In addition, in the fermentation apparatus of the present invention in which C. vertilis is fixed, as shown in FIG. 7, the specific alcohol production rate and the specific consumption rate of the soy sauce did not differ from 0.0029 day ^-1 and 0.0081 day ^-1 batch fermentation, respectively. 1.79% (v / v). Z.rouxii showed slightly lower production capacity than immobilized.

Example 3 Continuous Fermentation of Conventional Soy Sauce Using the Inventive Fermentation Device Filled with Immobilized Yeast

In Example 2, the semi-continuous fermentation was converted into a continuous fermentation method, and the fermentation was carried out. In continuous fermentation, the dilution rate per day was 0.25 day ⁻¹ like the semi-continuous fermentation, and the substrate soy was continuously subdivided into a constant peristaltic pump. It recovered continuously, supplying (21). At this time, since the specific gravity of soy sauce was measured as 1.154, 180 mL and 90 mL of soy sauce were continuously supplied as the hourly dose feed rate, respectively, so that the soy sauce corresponding to 5 kg / day and 2.5 kg / day was continuously fed into the fermenter 10 of the present invention. Continuous alcohol fermentation of the soy sauce while supplying, the foam discharge prevention device 40 and the locking device (23, 43) is kept open continuously to continuously inject the raw soy sauce and recover the fermented soy alcohol concentration of the recovered soy sauce Was measured over time. Alcohol fermented soy sauce produced during the continuous fermentation as described above was to be recovered to the fermentation broth recovery tank 50 through the present invention foam discharge prevention device (40). At this time, the bubbles generated by the gas generated during the fermentation and the injected air bubbles are removed through the foam discharge prevention device 40 and only the fermentation broth is transferred to the recovery tank 50.

As a result, as shown in FIG. 8, a fixed alcohol was produced upon fermentation for 17 days. In other words, Z.rouxii obtained 1.51% (w / v) alcohol and 0.047% (w / v) soy sauce, while C. versatilis had 1.44% (w / v) alcohol and 0.06 Soy sauce containing% of the residue was obtained.

As shown in Table 1, when the fermentation apparatus of the present invention filled with immobilized Z.rouxii and C.versatilis was fermented by conventional, batch, semi-continuous, and continuous fermentation methods , the alcohol production rate and non-sugar consumption rate were greatly increased. Shows no difference. Therefore, in view of the efficiency and convenience of work, it was judged that it is advantageous to use the continuous fermentation method.

Comparison of Alcohol Production According to Fermentation Method in Alcohol Fermentation of Traditional Soy Sauce by Fermentation Device Filled with Immobilized Yeast Batch type Semi-continuous Continuous expression Z.rouxii C.versatilis Z.rouxii C.versatilis Z.rouxii C.versatilis Non-alcoholic production rate (day ^-1 ) 0.0033 0.0031 0.00451 0.00288 Consumption rate (day ^-1 ) -0.00874 -0.00874 -0.01 -0.00811 Alcohol production over time Y ^* = 0.377X + 0.008 Y = 0.362X + 0.0048 Y = 0.421X + 1.097 Y = 0.332X + 1.087 Timely consumption formula Y = -1.0082X + 3.9734 Y = -1.0082X + 3.9734 Y = -1.175X + 1.234 Y = -0.936X + 0.961 Fermented soy sauce average alcohol content (%, v / v) 1.91 (1.51%, w / v) 1.81 (1.43%, w / v) 1.92 (1.52%, w / v) 1.79 (1.42%, w / v) 1.91 (1.51%, w / v) 1.79 (1.42%, w / v) Fermented soy sauce average residue content (%) 0.057 0.033 0.059 0.02 0.047 0.06 Note: Y:% alcohol content, X: fermentation time

Example 4 Continuous Fermentation of Traditional Soy Sauce Using Yeast Seed Culture Solution in the Fermentation Apparatus of the Present Invention

Continuous fermentation was performed by culturing the yeast seedling culture medium without inoculating immobilized enzyme into the fermentation apparatus of the present invention and inoculating it in the liver. Z.rouxii and C.versatilis were inoculated in liquid medium and incubated at 30 ° C. for 5 days to prepare yeast seed culture medium.

The present invention is then filled with 13.0L, 26L of soy sauce raw material in the fermenter 10 by using fermentation apparatus of 15L and 30L respectively, and aseptically inoculated 0.5L and 1.0L of yeast seed culture medium prepared aseptically, respectively. It was fermented in a fermentation chamber air-conditioned at 28 ± 0.5 ° C while injecting sterile air at a rate of 0.5vvm through the porous plate 14, and 1.8% (v / v) and 1.6% ( v / v) was found to stably produce alcoholic fermented soy sauce containing alcohol, and then immediately converted to continuous fermentation. Substrate soy sauce was recovered from the raw material storage tank 20 using a peristaltic pump 21 at a constant rate so that the daily dilution rate was 0.25 day ⁻¹ during continuous fermentation. During continuous fermentation, the foam release preventing device 40 and the locking devices 23 and 43 of the present invention were kept open, and the same amount of fermented soy sauce was continuously collected while continuously injecting the raw materials.

Each dose per hour feed rate of Z.rouxii and C.versatilis is the case of the supply using a tick Perry Stahl the liver of 281mL and 140.5mL pump (21) Z.rouxii include alcohol content of 1.8% (v / v) Soy sauce was obtained, and C.versatilis reliably produced soy sauce with an alcohol content of 1.60% (v / v).

Experimental Example 2: Investigation of the Components of the Fermented Soy Sauce of the Present Invention

The lactic acid, glucose content, total nitrogen, color, salt, alcohol content and free amino acid content of sterilized conventional soy sauce and alcoholic fermented soy sauce used as raw materials of the present invention were investigated. First, the measurement of lactic acid and glucose content was carried out by diluting the soy sample at a constant rate according to the method of Yoda et al., Filtering with 0.45 μm membrane filter, and then using the YSI 2700 Selective Biochemistry Analyzer using an enzyme membrane. Was analyzed by the mechanism as shown in the following formula (VI) and formula (iii).

The total nitrogen content was measured according to the AOAC method, and the sample was decomposed by a decomposition device and the NH ₄ H ₂ BO ₃ , which was obtained by diluting NH3 distilled using a Keltec device with 5% boric acid water, was converted to 0.1N HCl. The amount was calculated by converting the number of mL consumed by titration into total nitrogen. The chromaticity of the liver was centrifuged at 7,200 g for 30 minutes, and the supernatant was taken to measure the value at 500 nm using a spectrophotometer. The amount of saline and net extract was diluted 100 times with 1 mL of soy sauce, 20 mL of the diluted solution was added to a 100 mL Erlenmeyer flask, and 1 mL of 2% K ₂ CrO ₄ solution was added as an indicator, followed by pale orange color with 0.1 N AgNO ₃ solution. Titration was carried out until the amount of salt was calculated as shown in the following formula (iii).

0.1 N AgNO ₃ 1 mL = 0.005845 g NaCl... (Ⅷ)

Pure solids content was obtained by distillation of purified nasal dish at 105 ± 1 ° C., and then a certain amount of sample was taken to evaporate water to determine the content of extract, and then the amount of salt was subtracted to quantify pure solids. In alcohol analysis, the soy sample was filtered by 0.45μm membrane filter and injected directly into the GC. The column filler was Forapack QS, and the inlet temperature was 210 ° C and the detector (FID) temperature. Was 220 ℃, the carrier gas was maintained at a rate of 20 mL / min using nitrogen and the column temperature was 150 ℃. For free amino acid analysis, the liver samples were diluted 20-fold with lithium citrate buffer for amino acid analysis, and then filtered by 0.45 μm membrane filter.

As a result, as shown in Table 2, there was no change between before and after sterilization of raw soy sauce, but the fermented soy sauce after alcohol fermentation increased 1.1-1.4% of pure solids and 0.8% of OD (optical density). It was. This resulted in the increase of the net solids content and the OD value due to Maillard reaction between sugar and free amino acid during the alcohol fermentation period.

Changes in the Components of Soy Sauce, Sterilized Soy Sauce and Alcoholic Fermented Soy Sauce Ingredients / Soy Sauce Raw soy sauce Sterilized Soy Sauce ^* Z.rouxii fermented soy sauce C.versatilis fermented soy sauce CZ Soy Sauce ^** moisture 74.90 74.99 74.04 73.73 73.83 NaCl 19.58 19.43 19.29 19.29 19.28 Net solid content 5.52 5.58 6.67 6.98 6.89 Total nitrogen content 1.06 1.05 1.11 1.12 1.12 Lactic acid 0.681 0.708 0.691 0.685 0.644 pH 5.09 5.08 5.24 5.24 5.29 OD ^*** 2.29 2.19 3.02 3.00 3.00 [Note] * at 111~113 ℃ 8.48 bungan continuous hot sterilization soy ** CZ liver: the C.versatilis Z.rouxii liver and liver after alcoholic fermentation 1: Soy sauce mixed in a weight ratio of 2 *** OD: at 500nm Absorbance

In addition, as shown in Table 3, the amino acid composition of raw soy sauce, sterilized raw soy sauce and alcoholic fermented soy sauce increased the total amino acid content in alcoholic fermented soy sauce.

Changes in Amino Acid Contents of Raw Soy Sauce, Sterilized Raw Soy and Alcoholic Fermented Soy Sauce Amino acid type / soy sauce type Raw soy sauce Sterilized Soy Sauce ^* Fermented soy sauce Z.rouxii C.veratilis cz soy sauce ^** Aspartic acid 75.85 74.01 85.36 85.86 87.58 Treonine 87.64 82.04 90.62 90.03 86.93 Serin 141.34 136.93 147.27 148.85 140.68 Glutamic acid 607.65 580.64 612.76 615.15 592.36 Proline 121.14 116.29 127.47 123.76 119.51 Glycine 77.62 74.67 79.12 77.84 75.44 Alanine 215.83 207.59 206.72 207.47 196.89 Cystin 22.14 21.56 21.36 21.54 20.15 Val Lin 133.58 132.26 135.32 137.98 126.81 Methionine 33.30 31.73 31.85 32.04 29.86 Isoleucine 130.96 128.21 133.52 136.11 126.53 Luxin 189.90 191.47 199.95 203.51 190.09 Tee 66.64 67.70 68.52 67.79 63.30 Phenylalanine 109.44 107.46 111.66 112.11 107.85 Histidine 9.73 9.10 13.73 11.56 14.74 Lai Sin 143.72 140.69 149.10 148.53 144.87 Arginine 3.73 3.62 4.97 4.75 5.62 Sum 2170.23 2105.98 2219.30 2224.89 2129.21 GA / TA (%) ¹⁾ 28.00 27.57 27.61 27.65 27.82 (Note) 1) GA / TA (%): glutamic acid content / total amino acid content * 8.48 min high temperature sterilization soy sauce at 111-113 ° C. ** cz soy sauce: C.versatilis soy sauce and Z.rouxii soy sauce after alcohol fermentation Soy sauce mixed in a weight ratio of 2:

Experimental Example 3: Aroma Component and Flavor Test of Alcohol-Fermented Soy Sauce of the Present Invention

The alcoholic and fermented soy sauce of the present invention was investigated for its flavor and flavor in preparation for soy sauce made at home. The analysis of the fragrance component silver was carried out using a Nikerson continuous steam distillation apparatus improved according to Schultz et al. 200 mL of soy sauce was mixed with 800 mL of distilled water, and placed in a container and extracted for 2 hours under normal pressure. 100 mL of a mixture of the same amount of n-pentane and ethyl ether was used as an extraction solvent, and anhydrous sodium sulfate was added and left at 5 ° C. for 24 hours to remove moisture. After transferring to a vial for GC, the solution was concentrated to 100 μL with nitrogen gas to prepare an analytical sample of GC / MSD. The fragrance component analysis conditions of the prepared analysis sample are as shown in Table 4.

GC analysis conditions of flavor components of raw soy sauce, sterilized soy sauce and alcoholic fermented soy sauce Gas chromatography Device HP 6890 GC (HewlettPackard co., USA) Carrier gas helium column HP-FFAP (0.32mm × 50m) Temperature 50 ° C (2 minutes) -10 ° C / min-280 ° C (15 minutes) Flux 1.5 psi Mass selectivity detector Device HP6890 Gas-Spectrometer (HP co.USA) Ion mode Electron impact ionization (EI) Temperature 190 ℃ Electronic energy 70 eV library Wiley275

The soy sauce palate survey was conducted by selecting 20-25 test subjects familiar with the taste of soy sauce and scoring the taste of each sample by the 10-point method. The evaluation items were divided into color, fragrance, sweet taste, salty taste, sweetness, sourness, and overall preference. The score was 10 points for good, 8 points for good, 6 points for normal, 4 points for bad, and 2 points for bad. Was taken as the taste test score. SPSS program was used for statistical analysis of the results of the taste test, and variance analysis and Duncan's multiple verification method were used to test the significance.

As a result, as shown in Table 5, Z. rouxii alcohol fermented soy sauce, C. versatilis alcohol fermented soy sauce, Z. rouxii soy sauce, and C. versatilis soy sauce were mixed with conventional soy sauce prepared by the house. There was no significant difference in color and alcohol fermented soy sauce produced by fermentation using the present fermentation device showed a 5% significant difference and was superior to soy sauce in fragrance, sweet taste, salty taste, sour taste, sweet taste and overall preference.

Dietary Characteristics of Alcohol Fermented Soy Sauce Prepared by Strain by the Fermentation Device of the Present Invention Filled with Immobilized Yeast CV soy sauce ZR Soy Sauce CZ Soy Sauce Soy sauce F value color 6.90 ± 1.97 ^a 6.79 ± 1.50 ^a 7.03 ± 1.57 ^a 6.59 ± 1.80 ^a 0.35 Flavor 6.48 ± 1.82 ^a 7.24 ± 1.64 ^a 6.38 ± 1.97 ^a 3.93 ± 1.89 ^b 17.79 ^** Sweet taste 6.55 ± 2.13 ^a 6.41 ± 1.45 ^a 6.31 ± 1.63 ^a 4.48 ± 1.90 ^b 8.56 ^** Salty 6.62 ± 2.01 ^a 6.86 ± 1.77 ^a 6.38 ± 1.74 ^a 5.28 ± 2.74 ^b 3.22 ^* Sour taste 6.24 ± 1.77 ^a 6.45 ± 1.50 ^a 5.69 ± 1.39 ^ab 5.07 ± 2.10 ^b 3.78 ^* sweetness 7.17 ± 1.65 ^a 6.41 ± 1.80 ^ab 6.10 ± 1.86 ^b 3.90 ± 1.70 ^c 18.66 ^** Comprehensive Preference 7.03 ± 1.74 ^a 7.17 ± 1.65 ^a 6.10 ± 1.70 ^ab 3.72 ± 1.75 ^b 25.23 ^** (Note) Mean ± Standard Deviation * P <0.05, ** P <0.01

In addition, the flavor component was found to be much higher than the soy sauce fermented soy sauce as shown in Table 6. This seems to affect the palatability of other taste components because of the fragrance component which is generated by the alcoholic fermentation of conventional soy sauce. Therefore, it was found that the alcoholic fermentation by the fermentation apparatus of the present invention can improve the preference of conventional soy sauce.

Flavor Components of Alcohol-Fermented Soy Sauce of the Present Invention ingredient RT MF area % Raw soy sauce Sterilized Soy Sauce ^* CV soy sauce ZR Soy Sauce CZ Soy Sauce ^** Acetic acid 22.48 C ₂ H ₄ O ₂ 2.61 5.27 0.01 0.28 0.49 Propionic acid 24.67 C ₃ H ₆ O ₂ 0.10 1.17 - - - 2-methyl-propanoic acid 25.32 C ₄ H ₈ O ₂ 1.12 1.78 0.01 0.01 0.01 Butyric acid 26.82 C ₄ H ₈ O ₂ 0.85 1.70 0.01 0.01 0.01 Hexane salt 31.74 C ₆ H ₁₂ O ₂ 0.02 0.10 0.01 0.01 0.01 Octane salt 36.07 C ₈ H ₁₆ O ₂ 0.11 0.26 0.03 0.03 0.01 Decane salt 40.47 C ₁₀ H ₂₀ O ₂ - 0.24 0.01 0.01 0.01 Palmitic acid 47.27 C ₁₆ H ₃₂ O ₂ 2.21 1.25 0.26 0.15 0.08 Stearic acid 49.74 C ₁₈ H ₃₆ O ₂ - - 0.01 0.03 0.01 Alcohol ethanol 10.06 C ₂ H ₆ O 48.11 37.52 90.81 85.51 88.01 1-propanol 11.89 C ₂ H ₈ O 0.22 0.35 0.35 0.99 0.27 2-methyl-1-propanol 13.00 C ₄ H ₁₀ O - 0.60 0.27 0.50 0.04 1-butanol 14.25 C ₄ H ₈ O ₂ 0.51 1.17 0.22 0.05 0.03 3-methylbutanol 15.83 C ₅ H ₁₂ O 1.24 1.10 0.81 1.56 1.42 2-pentanol 29.17 C ₅ H ₁₂ O 0.22 0.37 - 0.03 0.01 Benzene Methanol 33.00 C ₇ H ₈ O 0.17 0.32 0.02 0.03 0.03 Phenyl ethanol 33.85 C ₈ H ₁₀ O 0.93 0.79 1.61 3.71 3.00

Flavor Components of Alcohol-Fermented Soy Sauce of the Present Invention ingredient RT MF area % Raw soy sauce Sterilized Soy Sauce ^* CV soy sauce ZR Soy Sauce CZ Soy Sauce ^** Aldehyde ethanol 8.12 C ₂ H ₄ O ₂ 1.45 1.36 0.86 0.78 0.68 Propanol 8.72 C ₃ H ₆ O ₂ 0.14 0.12 0.12 0.13 0.10 3-methyl-pentenal 16.37 C ₅ H ₈ O 0.28 0.42 0.05 0.06 0.01 Fufural 23.43 C ₅ H ₄ O ₂ 5.10 4.84 0.67 0.72 0.86 Benzaldehyde 25.29 C ₇ H ₆ O - - 0.03 0.04 0.04 5-methyl furfural 26.33 C ₆ H ₆ O ₂ 0.22 0.23 0.03 0.03 0.04 Phenylacetaldehyde 28.18 C ₈ H ₈ O 2.52 1.97 0.02 0.23 0.25 Ester Ethyl Formate 8.98 C ₃ H ₆ O ₂ 0.61 0.51 0.10 0.10 0.09 Ethyl acetate 9.64 C ₄ H ₈ O ₂ 3.86 3.14 0.43 0.37 0.62 1-methoxy-2-methyl-mexiisobutyl ester 18.92 C ₅ H ₁₂ O - 0.15 0.12 0.04 0.05 Benzene Acetic Acid Ethyl Ester 25.29 C ₇ H ₆ O - - 0.01 0.01 0.01 Dimethyl phthalate 42.54 C ₁₀ H ₁₀ O` - 0.13 0.01 0.01 0.01 2-ethylhexyl phthalate 42.88 C ₂₄ H ₃₈ O ₄ - - 0.05 0.01 0.01 Furan2-furanmethanol 27.74 C ₅ H ₆ O ₂ 7.26 10.96 0.45 0.53 0.47 5-ethyldihydro 2-furanone 29.82 C ₆ H ₁₀ O ₂ 0.09 0.12 0.01 0.01 0.01 5-butyldihydro-2-furanone 36.66 C ₈ H ₁₄ O ₂ - - - - 0.02 Dihydro-5-2-octenyl-2 (3H) -furanone 45.46 C ₁₂ H ₂₀ O ₂ - - 0.01 0.01 0.01 Hydrocarbonbenzene 10.61 C ₆ H ₆ 0.19 0.19 - - - toluene 12.47 C ₇ H ₈ 0.30 0.48 0.03 - 0.02 Hydroperoxide 13.48 C ₅ H ₁₂ O ₂ 0.11 0.22 - 0.29 0.01 5-methyl-3 (1-methylethyl) -chlorohexene 27.23 C ₁₀ H ₁₆ 0.02 0.04 0.01 0.01 0.01

Flavor Components of Alcohol-Fermented Soy Sauce of the Present Invention ingredient RT MF area % Raw soy sauce Sterilized Soy Sauce ^* CV soy sauce ZR Soy Sauce CZ Soy Sauce ^** Ketone 1 -cyclopropyl-ethanone 14.46 C ₅ H ₈ O 0.12 0.04 0.10 0.20 0.33 3-hydrocy-2-butanone 18.66 C ₄ H ₈ O ₂ 1.51 1.41 0.11 0.12 0.09 1- (1H-pyrrole-2yl) -ethanone 35.14 C ₆ H ₇ NO 0.13 0.93 0.03 0.04 0.04 Nitrogen-containing compounds Carbosylmethylamide Hemihydrogloide 12.97 C ₂ H ₅ NO ₃ 0.45 - - - - Azabenzene 16.04 C ₅ H ₆ N ₂ 0.46 1.69 - - - Methylpyrazine 18.18 C ₅ H ₆ N ₂ 0.94 1.07 0.13 0.07 0.07 N- (3-methylbutyl) acetamide 32.33 C ₇ H ₁₅ NO - 0.07 0.04 - - N-hexyl-acetamide 32.34 C ₈ H ₁₇ NO - 0.09 - 0.03 0.02 Phenolic 2-methoxyphenol 32.80 C ₇ H ₈ O ₂ 0.14 0.11 0.02 0.02 0.02 4-vinyl-2-methoxy-phenol 39.81 C ₉ H ₁₀ O ₂ 0.18 0.47 0.03 0.04 0.03 Sulfur-containing compound dithiabutane 13.32 C ₂ H ₆ S ₂ 0.19 0.38 - - 0.87 2-methyl-trisulfite 21.47 C ₂ H ₆ S ₃ - - - - 0.08 S-methyl methylthiosulfonate 21.52 C ₂ H ₆ O ₂ S ₂ - - 0.85 - - Transdihydro-2 (4or5) -dimethyl-3 (2H) -tritophenone 31.71 C ₆ H ₁₀ OS 0.07 - - - - 4-Methyl-1,3-dioxolane 10.93 C ₄ H ₈ O ₂ 0.08 0.07 - - - Octadecan 22.93 C ₁₈ H ₃₈ - - 0.01 - 0.01 1,4,7,10,13,16-Hesanoaclooctadecane 34.55 C ₁₂ H ₂₄ O ₆ - - 0.04 0.01 0.03 * 8.48 min high temperature sterilized soy sauce at 111-113 ℃ ** CZ soy sauce: After alcohol fermentation, C.versatilis soy sauce and Z.rouxii soy sauce are mixed in a weight ratio of 1: 2.

As described above, the fermentation apparatus of the soy sauce and liquid food of the present invention and the fermentation method using the apparatus of the present invention disperse the sterilized air into fine bubbles by mounting a porous plate at the lower end of the fermenter to dissolve the oxygen during fermentation. It has the effect of increasing the fermentation is completed and the fermentation broth is discharged to complete the fermentation broth by using the present invention to prevent the mixing of bubbles in the fermentation broth to prevent the mixing of the foam and the soy sauce and liquid using the fermentation apparatus of the present invention It is a very useful invention in the food processing industry because it has the effect of efficiently performing batch fermentation, semi-continuous fermentation and continuous fermentation of foodstuffs.

Claims (4)

Fermentation substrate (10) to accommodate the fermentation substrate soy sauce or liquid food and the raw material storage tank 20 for supplying the substrate and the air supply device (30) for mixing the air supply and fermentation substrate and to prevent the mixing of bubbles during fermentation liquid recovery Soy sauce and liquid fermentation device characterized in that consisting of a foam discharge prevention device 40 and the fermentation broth recovery tank (50). According to claim 1, wherein the fermenter (10) is covered with a fermenter lid (11) for sealing the fermenter, the cooling tube (13) is installed on the upper end to prevent volatilization of alcohol and volatile fragrance components, fermenter The fermentation apparatus of soy sauce and liquid food, characterized in that the lower portion of the (10) is equipped with a porous plate (14) to increase the rate of oxygen dissolution into the fermentation broth by making air into fine bubbles. According to claim 1, wherein the foam discharge preventing device 40 is equipped with a residual fermentation liquid discharge pipe 41, the fruit fermentation pipe 42 is mounted on the top of the residual fermentation liquid discharge pipe and the inlet of the juice pipe is to be immersed in the fermentation liquid and the end The two-strand siphon phenomenon prevention pipe (42) is mounted higher than the fermentation broth and the fish tube, and the locking devices (43, 44) are mounted to prevent the siphon phenomenon and the bubbles generated during the fermentation are fermentation liquid recovery tank ( 50) fermentation device of soy sauce and liquid foods, characterized in that it is formed so as not to flow into. (a) immobilizing yeast on the porous alumina ceracic beads; (b) filling the fermentation tank 10 with 25 to 35% (v / v) of the fermentation tank capacity of the immobilized yeast prepared in step (a); (c) supplying the substrate soy sauce to the fermentation tank (10) filled with the immobilized yeast of step (b) and continuously fermenting alcohol in a fermentation chamber air-conditioned at 25 to 30 ° C. while sterile air of 0.05vvm; (e) continuously subdividing the substrate soy sauce at a constant rate so that the daily dilution rate is 0.20-0.30day- ¹ during the continuous alcoholic fermentation of step (c) Fermentation method of soy sauce and liquid food, characterized in that consisting of.