KR101121570B1 - Fermentation method of starfish extract using microorganisms and hangover curing agent comprising starfish extract fermented by the same - Google Patents

Abstract

PURPOSE: A method for fermenting starfish extract using effective microorganisms is provided to enhance value as a food and to enhance alcohol metabolic enzyme activity. CONSTITUTION: A method for fermenting starfish extract comprises: a step of sterilizing a medium containing 1-5 wt% of starfish extract at 121°C and 1.5 psi for 20 minutes and cooling; a step of inoculating Saccharomyces cerevisiae or Bacillus subtilis; and a step of culturing at pH 4.0-6.0 and 30-37°C for 24-48 hours. The extract is a hot water extract of starfish. An agent for relieving hangover contains the fermentation as an active ingredient.

Description

Fermentation method of starfish extract using microorganisms, and hangover releasing agent containing starfish fermentation product produced thereby

The present invention relates to a method for producing a functional raw material for improving fermentation using starfish microorganisms and alcohol metabolizing enzyme activity using the same.

Starfish is a marine benthic creature belonging to the echinoderm family, and about 1,700 species have been reported all over the world, and about 200 species inhabit the offshore of Korea.

In particular, Asterina amurensis ( Asterina amurensis ) has abundant shellfish such as abalone, clams, shellfish, and scallops as a prey, and is causing great damage to the shellfish farming industry. .

In order to solve this problem, fishermen are capturing enormous amounts of starfish, but most of them are discarded and some small amounts are used as fertilizers, but they are not used industrially.

There are some examples of currently used starfish as a raw material for calcium or some compost, but since it is not properly utilized on land, it is mostly treated as waste, causing odor and environmental pollution with secondary decay, and This is a time consuming situation.

Alcohol metabolism in the body is first degraded to acetaldehyde by alcohol dehydrogenase (ADH) present in liver cells and 80 to 90% of alcohol is metabolized by aldehyde dehydrogenase (ALDH) enzyme After forming the acetic acid, it is hydrolyzed with carbon dioxide and water to undergo a complete decomposition process.

Most of the hangover-relieving products developed so far have been focused on promoting ADH enzyme activity, but since the hangover symptoms actually felt after drinking are known to be caused by acetaldehyde, it should be focused on promoting ALDH enzyme activity. Therefore, the most desirable hangover resolution in the human body is to promote not only ADH enzyme activity but also ALDH enzyme activity.

Accordingly, the present inventor has developed a fermentation method that effectively destroys the marine ecosystem through fermentation using useful microorganisms and usefully converts starfish, which is difficult to discard due to odor or the like, even after capture. And to develop a hangover reliever raw material that increases the activity of ALDH.

As described above, the captured starfish is an example that is used as a raw material for calcium or some compost, but since it is not properly utilized on land, it is mostly treated as waste, causing odor and environmental pollution with secondary decay, and waste treatment The situation is consuming the cost and time.

Therefore, it is thought that various methods for efficiently processing and using the captured starfish should be sought, and to improve the functionality of the starfish through microbial fermentation to develop a useful resource-recycling method. Particularly, it is involved in alcohol metabolism in the human body when alcohol is consumed. As it shows the effect of increasing the activity of the enzyme, a functional food raw material using the same is developed.

The method for fermenting starfish using microorganisms of the present invention is a method in which the growth rate and medium pH of each bacterium are grown in a liquid medium to which starfish is added by using useful microorganisms Bacillus subtilis and Saccharomyces cerevisiae , which ensure safety for use in food. By measuring the degree of fermentation through measurement, the optimum starfish fermentation conditions of each microorganism are determined. The determination of the optimum fermentation conditions determines the concentration of starfish addition, determines the optimum pH, determines the optimum fermentation temperature, and determines the optimum fermentation conditions by measuring the degree of fermentation according to the inoculation amount of microorganisms during fermentation.

Using the starfish fermented product fermented under the optimum conditions determined as described above, the activity of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) extracted from yeast is compared to the control without the sample added to measure the increase in enzyme activity. .

The invention configured as described above can develop a method to increase the usability of starfish and the value as a food processing raw material through granting the functionality of the starfish captured with the fishery during the fishing process, and collect the collected starfish voluntarily through purchase. It is possible to protect fishery resources (increase in shellfish production) and create sustainable economic benefits (new income sources). In addition, by using the effect of increasing the alcohol metabolizing enzyme activity of the starfish fermentation, it can be used as a raw material to relieve hangover and develop a functional hangover reliever to help alleviate hangover symptoms caused by excessive alcohol consumption by modern people.

1 is a view measuring the aldehyde dehydrogenase (aldehyde dehydrogenase, ALDH) activity of the organic solvent fraction of the starfish fermentation according to the present invention [SFE (5%): Starfish extract 5%, SFE (5%) + BS: Starfish Extract 5% + archaea, SFE (5%) + SC: starfish extract 5% + yeast].
Figure 3 is a view of measuring the activity of alcohol dehydrogenase (alcohol dehydrogenase, ADH) and aldehyde dehydrogenase (ALDH) of starfish extract and starfish fermentation according to the present invention [SFE (1%): starfish extract 1%, , SFE(5%): Starfish extract 5%, SFE(1%)+BS: Starfish extract 1% + Archaea, SFE(5%)+BS: Starfish extract 5% + Archaea, SFE(1%)+SC: Starfish extract 1% + yeast fungus, SFE (5%) + SC: starfish extract 5% + yeast fungus].

In the present invention, the Amur Bulgari taken from the coast of the East Sea was used in the invention. The collected starfish was stored frozen (-20°C) to prevent spoilage, and used in the invention.

Add 1 kg of starfish to the extraction bottle, pour distilled water and extract it 3 times for 2 hours at a temperature of 80 to 121°C, or add 1 kg of starfish to 95 to 99% concentration of ethanol and methanol in a volume of 1 to 5 times and cool the condenser. Extracted by heating at 70~80℃ for 5 hours in the attached extractor. The extract was filtered under reduced pressure with paper filter paper only, and the remaining liquid was concentrated under reduced pressure in a concentrator equipped with a cooling condenser and dried with hot air (60~80℃). Hot air dried starfish hot water extract was used for fermentation experiments.
The degree of fermentation of starfish was determined by measuring the growth degree of two microorganisms (Optical density, OD-an index that indirectly measures the growth rate of microorganisms) according to each fermentation condition.

To determine the optimum fermentation conditions for starfish, the concentration of the starfish hot water extract was adjusted by adjusting the concentration by 1 to 5%. The medium adjusted to each concentration was inoculated with 0.2% of Bacillus subtilis , or yeast ( Saccharomyces cerevisiae ) after 121°C, 1.5 psi, and sterilized for 20 minutes, and cooled for 24 to 48 hours at a temperature of 37°C and 30°C. After incubation, UV-vis spectrophotometer (Optizen, Mecasys Co.) was used to measure the growth rate of archaea or yeast and pH after fermentation using a pH meter (Suntex Co.).

Growth rate and pH after fermentation of archaea or yeast according to the added concentration of starfish extract Concentration of starfish extract (%) Bacillus subtilis Saccharomyces cerevisiae OD _660nm PH after fermentation OD _660nm PH after fermentation One 2.17 8.34 0.27 7.27 2 2.4 8.41 0.6 7.31 3 2.47 8.23 1.01 7.47 4 2.22 8.08 1.02 7.39 5 3 8.01 1.44 7.49
As shown in Table 1, as the concentration of the starfish extract increases during fermentation of the starfish extract, both the Bacillus ( B. subtilis ) and yeast ( S. cerevisiae ) grow at a turbidity (Optical density, OD-microorganism) at 660 nm. It can be seen that the growth of microorganisms increases as the value increases).

The optimum pH of the starfish fermentation was determined using a medium to which 1% of the previously determined starfish hot water extract was added. The pH range was determined within the range of 4.0 to 10.0, and was adjusted using 1N HCl and 1N NaOH. Each medium whose pH is adjusted is 121℃, 1.5 psi, sterilized for 20 minutes, inoculated with 0.2% of Bacillus or Homobacteria after cooling, and cultured at 37°C and 30°C for 24 to 48 hours, and the degree of growth of Bacillus or yeast. The optimum fermentation pH condition during fermentation of the starfish extract was determined by measuring the pH after fermentation with.

Growth rate and pH after fermentation of archaea or yeast according to the initial pH of fermentation of starfish extract Initial pH Bacillus subtilis Saccharomyces cerevisiae OD _660nm PH after fermentation OD _660nm PH after fermentation 4 0.13 4.17 0.53 4.22 5 0.68 6.97 0.41 5.59 6 2.08 8.2 0.31 6.5 7 1.75 8.28 0.32 6.94 8 1.37 8.32 0.13 7.48 9 0.73 7.94 0.05 8.3 10 0.03 9.11 0.05 9.09
As shown in Table 2, when fermentation of starfish extract, B. subtilis was used, the growth of microorganisms was most excellent at pH 6.0, and when yeast ( S. cerevisiae ) was used, microorganisms at pH 4.0 Showed the best growth rate.

After adjusting the medium to which the starfish hot water extract 1% was added to the optimum fermentation pH of 6.0 and the yeast fermentation optimal fermentation pH of 4.0 for archaea, the optimum fermentation temperature during fermentation of the starfish extract is determined by using the pH-adjusted medium. Did. The optimum temperature was determined at 17, 24, 30, and 37°C, and each medium was 121°C, 1.5 psi, sterilized for 20 minutes, and 0.2% inoculated with Bacillus or Homobacteria after cooling for 24 to 48 hours at each temperature. After cultivation, the degree of growth of archaea or yeast bacteria and pH after fermentation were measured to determine the degree of fermentation.

Growth rate and pH after fermentation of archaea or yeast according to fermentation temperature of starfish extract Fermentation temperature Bacillus subtilis Saccharomyces cerevisiae OD _660nm PH after fermentation OD _660nm PH after fermentation 17℃ 0.14 6.15 0.03 4.15 24℃ 1.36 7.77 0.33 4.21 30℃ 1.6 8.17 0.78 4.57 37℃ 1.94 8.57 0.65 4.46
As shown in Table 3, during the fermentation of the starfish extract of Bacillus subtilis ( B. subtilis ) In the case of 37 ℃, the growth rate of the microorganism was the best, and the yeast ( S. cerevisiae ) In the case of 30 ℃, the growth rate of the microorganism was the best.

Determination of the optimum inoculation amount of fermented bacteria for optimum fermentation conditions when fermenting the starfish extract is 0.2, 0.4, 0.8, 1.6, 3.2, 5.0 based on the total weight of the medium with the inoculation amount of archaea or yeast on the medium to which 1% starfish hot water extract is added. %. The medium is 121°C, 1.5 psi, sterilized for 20 minutes, cooled, and inoculated with the Bacillus or Homobacteria according to the above concentrations, cultured at each temperature for 24 to 48 hours, and the growth degree of the Bacillus or Yeast bacteria and the pH after fermentation are measured. The optimal inoculum amount of archaea or yeast was determined.

Growth rate and pH after fermentation depending on the inoculation amount of archaea or yeast bacteria during fermentation of starfish extract Inoculum or yeast inoculation amount Bacillus subtilis Saccharomyces cerevisiae OD _660nm PH after fermentation OD _660nm PH after fermentation 0.2% 1.36 7.02 0.74 4.41 0.4% 1.68 7.2 1.13 4.45 0.8% 2.01 7.8 1.43 4.8 1.6% 1.77 7.5 1.49 4.59 3.2% 1.76 7.4 1.57 4.65 5.0% 2.02 8.0 1.67 4.54
As shown in Table 4, B. subtilis used during fermentation of starfish extract and Of yeast ( S. cerevisiae ) In the case, it was confirmed that the growth rate of microorganisms was excellent at the inoculation amount of 0.8% or more.

The effect of starfish extract and fermented starfish fermented under optimal conditions on the activity of alcohol metabolizing enzyme alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) was measured. First, distilled water is added to the starfish fermentation product prepared according to the fermentation conditions of Examples 1 to 4, and then sequentially fractionated with an equal amount of hexane and chloroform, and then concentrated by using a reduced pressure concentrator (Eleyea Co.). Did. Each concentrated fraction was placed in a dry heat sterilizer (Vision Co.) and the organic solvent capable of remaining at 80° C. for 5 hours was completely volatilized to obtain a hexane fraction, a chloroform fraction, and a water fraction of the starfish fermentation.
Measurement of ADH and ALDH activity of the starfish hot water extract (SFE), starfish fermentation product (SFE+BS or SFE+SC), and the hexane fraction, chloroform fraction, and water fraction of the starfish fermentation was performed according to the Worthington manual method. The absorbance was measured at 340 nm, and the amount of NADH was measured and compared with the relative activity to the control group. That is, the composition of the reaction solution is 500 μl of 1.0 M Trizma buffer, 250 μl of 25 mM β-NAD, 2.0 M substrate (ADH: ethanol, ALDH: acetaldehyde), 50 μl enzyme (ADH or ALDH), and 50 μl sample is placed in a cuvette. , UV-Vis spectrophotometer (Optizen, Mecasys Co.) was used to measure the change in absorbance at 340 nm for 2 minutes. At this time, the relative activity (%) was shown as a control without adding a sample.

Relative enzyme activity (%) = [1-(sample absorbance change)/(control absorbance change)]×100

Aldehyde dehydrogenase (ALDH) activity of the organic solvent fraction of the starfish fermentation product according to the present invention is shown in FIG. 1, alcohol dehydrogenase (alcohol dehydrogenase, ADH) and aldehyde dehydrogenation of the starfish extract and the starfish fermentation product according to the present invention The enzyme (aldehyde dehydrogenase, ALDH) activity is shown in FIG. 3.
As shown in FIG. 3, the aldehyde dehydrogenase (ALDH) activity of the starfish fermentation product (SFE+BS or SFE+SC) according to the present invention is 13.1% in the case of [SFE(1%) + archaea], [SFE(5 %) + Archaea], which was found to be higher than the starfish extract (1%: 14.4%, 5%: 22.0%). However, in the case of alcohol dehydrogenase (ADH), the starfish extract concentration and fermentation were not significantly affected.
In addition, as shown in FIG. 1, the aldehyde dehydrogenase (ALDH) activity of the organic solvent fraction of the starfish fermentation product (SFE+BS or SFE+SC) with high aldehyde dehydrogenase (ALDH) activity also increased compared to the starfish extract. Confirmed. In particular, when the starfish extract was fermented using yeast, it was confirmed that the aldehyde dehydrogenase (ALDH) activity of the water fraction of the starfish fermentation was highest (112%).

delete

delete

Claims (5)

The medium in which the starfish extract was added at a concentration of 1 to 5% by weight relative to the total weight of the medium was sterilized and cooled at 121°C and 1.5 psi for 20 minutes, and then cooled by Bacillus subtilis or Bacillus subtilis, which is yeast. Fermentation method of starfish extract, characterized by inoculating Cerebicae (Saccharomyces cerevisiae) at 0.8 to 5.0% by weight relative to the total weight of the medium and culturing for 24 to 48 hours under conditions of pH 4.0 to 6.0 and temperature of 30 to 37°C. .
According to claim 1, The starfish extract is characterized in that the starfish hot water extract, fermentation method of the starfish extract.
delete A hangover releasing agent containing a fermented starfish fermented by the method of claim 1 as an active ingredient.
The hangover release agent according to claim 4, wherein the starfish fermentation product has an aldehyde dehydrogenase (ALDH) activity of 1.3 to 3.5 times higher than that of a starfish extract.

Images