KR101594490B1 - Method for Manufacturing Fermented Sea Cucumber, and Fermented Sea Cucumber Produced Thereby - Google Patents

Abstract

The present invention relates to a method for producing a fermented sea cucumber and a fermented sea cucumber prepared by the method, in which a component contained in sea cucumber is reduced in molecular weight, decomposed and solubilized so that an oil component of sea cucumber can be more absorbed into the body.
The fermented sea cucumber according to the present invention has a two-step hydrolysis process optimized for the characteristics of the proteolytic enzyme, whereby the protein of the polymer is decomposed into peptides of low molecular weight, thereby improving the absorption of the proteins into the body. It is possible to improve the absorption rate of calcium by inhibiting insolubilization in the human body by binding with phosphorus. Furthermore, hydrolysis of sea cucumber into proteolytic enzymes and fermentation with lactic acid bacteria having high beta-glucosidase activity result in a low digestion and absorption rate The saponin glycoside can be converted into a saponin non-glycoside having a high digestion and absorption rate, so that the human body absorption rate of sea cucumber-containing saponin can be improved.

Description

TECHNICAL FIELD The present invention relates to a fermented sea cucumber and a fermented sea cucumber produced from the fermented sea cucumber,

The present invention relates to a method for producing a fermented sea cucumber and a fermented sea cucumber prepared by the method, in which a component contained in sea cucumber is reduced in molecular weight, decomposed and solubilized so that an oil component of sea cucumber can be more absorbed into the body.

Sea cucumber (sea 蔘) is an alkaline food with an efficacy enough to be called ginseng from the sea. It is divided into three kinds of red ginseng, blue ginseng and black ginseng according to color.

Sea cucumbers are rich in proteins such as arginine and cystine, and contain polysaccharides and minerals such as calcium, iron, phosphorus and iodine. They contain vitamin B1, B2, amino acids and fatty acids such as palmitic acid, stearic acid, oleic acid and linolenic acid .

In particular, the sea cucumbers have a high content of calcium and iron, which helps the hematopoiesis of the young and pregnant women in the growing season and is known as an excellent food source as a calcium source since the amount of phosphorus is in an ideal ratio to help absorption of calcium, And skeletal formation, normal muscle contraction, blood coagulation and many other physiological functions are involved.

In addition, sea cucumber contains chondroitin sulfate, which is a main component of viscous polysaccharide which stimulates the follicle of the colon epithelium and increases mucous secretion amount, and saponin which is involved in the control of immune and inflammatory reaction. And wound healing effect, antibacterial, anti-cancer, anti-inflammatory effect.

However, when the ingredients of sea cucumber are specifically examined, it is difficult to absorb digestion due to protein 21.5%, sugar 1.4% and fat 0.3%, and the rest is mostly composed of collagen. When the glycoprotein extracted from sea cucumber is taken with various protein sources, Studies have also shown that digestibility decreases by ~ 11%.

A protein is a complex molecule of complex structure made up of relatively simple molecules called amino acids. The amino acids that make up the protein are connected to each other through chemical bonds to form a polypeptide.

In academia, it is reported that peptides are more absorbed in the intestinal tract than amino acids, and protein deficiency is solved when proteins are supplemented by protein in the case of protein deficiency patients that do not absorb amino acids. Also, peptides have protein Antimicrobial effect, anticancer effect, and cholesterol lowering effect.

Therefore, the ingestion of protein-rich sea cucumber is advantageous for hydrolyzing the protein and converting the peptide into a peptide having a small number of amino acid residues, thereby improving digestion and absorption rate. Proteinase is generally used for this purpose.

For example, Korean Patent Publication No. 10-0311903 discloses a method for producing sea cucumber peptides. The dried sea cucumber powder is extracted with hot water and ethanol, and then proteinaceous proteins of papain, bromelain, pancreatin, To digestion and absorption rate of sea cucumber.

However, the sea cucumber contains a useful saponin component in addition to protein, and the saponin exists in the form of a glycoside linked to a sugar contained in sea cucumber. The glycosides bonded with sugar and saponin have a low digestion and absorption rate, Saponin is not absorbed by the human body when there is no microorganism to decompose.

From this viewpoint, the above-mentioned invention can decompose the protein into peptides to improve the digestion and absorption rate of proteins, but does not solve the problem of low digestion and absorption rate of saponin because it does not decompose glycosides bonded with sugar and saponin.

Korean Patent Laid-Open Publication No. 10-2014-0016461 discloses a method for converting a macromolecule compound of sea cucumber into a low molecular compound by using Bacillus subtilis, lactic acid bacterium, and yeast. These microorganisms decompose carbohydrates, sugars and proteins Or decomposing enzymes thereof, thereby converting the polymeric substance of sea cucumber into a low molecular substance, whereby a substance useful for skin can be obtained.

The microorganisms used in the above-mentioned invention include Bacillus subtilis, Lac tobacillus sakei), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus flange tareom (Lactobacillus plantarum , Lactococcus lactis), Bacillus core Tangerine Lawrence (Bacillus coagulance , and Saccharomyces cerevisiae . These microorganisms can degrade carbohydrates, sugars and proteins and convert them into skin-friendly substances. However, these microorganisms are not selective for all proteins exposed to the cell membrane The fermented sea cucumber is degraded and the nutritional value of fermented sea cucumbers is lowered. The saccharin - bound sugars are not decomposed well enough to improve digestion and absorption rate of saponin component of sea cucumber.

The present invention has been made to solve the problems caused by the limitations and disadvantages of the related art as described above. It is an object of the present invention to provide a fermented sea cucumber which can improve the water uptake of useful components contained in sea cucumbers by hydrolyzing and then fermenting sea cucumbers And fermented sea cucumber produced by this method.

In order to solve the above-mentioned problems, the present invention provides a method for preparing a sea cucumber, comprising grinding sea cucumbers, boiling water for 3 to 7 times and boiling for 20 to 40 minutes; Cooling the boiled sea cucumber, adding 0.01 to 2.00% by weight of an alkaline enzyme, and performing primary hydrolysis for 0.5 to 3.0 hours; Adding 0.01 to 2.00% by weight of a flavosome enzyme to the primary hydrolyzed sea cucumber and performing secondary hydrolysis for 1 to 24 hours; Heating the secondary hydrolyzed sea cucumber at 110 to 130 ° C for 10 to 20 minutes and cooling; 0.05 to 3.00% by weight of Pediococcus pentosaceus which is a lactic acid bacterium extracted from Kimchi in the cooled sea cucumber; And fermenting the sea cucumber inoculated with the lactic acid bacteria at 30 to 50 ° C for 15 to 30 hours.

At this time, the first and second hydrolysis steps are preferably performed at a pH of 6 to 8 and a temperature of 45 to 65 ° C.

The method of extracting lactic acid bacteria from the kimchi comprises the steps of filtering the kimchi soup of kimchi to remove solid matter; Culturing the Kimchi soup in which the solid content has been removed, in an MMS liquid culture medium; Continuously diluting the cultured medium with saline and then culturing the BEA culture medium in a plate medium to obtain a single culture; Obtaining a single cell population around the bacterial colonies by referring to the colonies again on a BEA medium; And isolating the bacterial populations to obtain a strain.

It is preferable that the kimchi is aged at 3-7 캜 for 5 ~ 6 months after immersion.

In addition, after the fermentation step, it is preferable that the fermented sea cucumber is heated at 110 to 130 ° C. for 10 to 20 minutes, and then the excipient is mixed and dried, followed by preparing powder, granule, capsule and tablet form .

The present invention also provides a fermented sea cucumber produced by the above method.

The fermented sea cucumber according to the present invention has a two-step hydrolysis process optimized for the characteristics of the proteolytic enzyme, whereby the protein of the polymer is decomposed into peptides of low molecular weight, thereby improving the absorption of the proteins into the body. It is possible to inhibit insolubilization by phosphorus bonding in the human body, thereby improving the human body absorption rate of calcium.

In addition, by hydrolyzing sea cucumber with proteolytic enzymes and fermenting them with lactic acid bacteria having high beta-glucosidase activity, saponin glycosides with low digestibility and absorption rate are converted into saponin non-glycosides with high digestion and absorption rate, .

The present invention relates to a method for converting a protein of a sea cucumber into a protein hydrolyzing enzyme to convert a protein having a low digestibility and absorption rate into a low molecular weight peptide having a high digestibility and absorption rate and then fermenting it with a lactic acid bacterium extracted from kimchi to digest saponin glycosides To saponin non-glycosides with high water absorption.

First, crush raw sea cucumber, add 3 ~ 7 times of water and simmer for 20 ~ 40 minutes.

The sea cucumber raw material can be used by grinding or drying the collected sea cucumber as it is or by pulverizing the dried sea cucumber which is circulated, and the collagen component of the sea cucumber turns into gelatin during boiling, and it is dissolved into water and becomes paste state. , The sea cucumber can not be sufficiently dissolved so that the subsequent process of decomposing and fermenting the sea cucumber can not proceed smoothly. If it exceeds 40 minutes, various physiologically active substances and functional ingredients of sea cucumber may be destroyed.

Next, the above-mentioned sea cucumber paste is cooled to room temperature, and proteolytic enzyme is used to decompose the sea cucumber protein into a small molecule peptide.

The above-mentioned sea cucumber paste is added with an alcalase enzyme in an amount of 0.01 to 2.00 wt% based on the total weight of the mixture, subjected to primary hydrolysis for 0.5 to 3.0 hours, and the flavorzyme enzyme is then added in an amount of 0.01 to 2.00 By weight, and then subjected to secondary hydrolysis for 1 to 24 hours.

As described above, the hydrolysis of the sea cucumber paste by the enzymes of alkalase is shortened before the hydrolysis by the flavosome enzyme, so that the activity of the flavosome enzyme is maximized, and the protein including the collagen contained in the sea cucumber is converted into a small molecule peptide And the flavosome enzyme also plays a role in preventing the bitter taste, coagulation and the like caused by the enzyme.

It is preferable to increase the activity of the proteolytic enzyme by adjusting the pH to 6 to 8 and the temperature to 45 to 65 ° C during the primary and secondary hydrolysis. When the hydrolysis is completed, the hydrolyzate is heated at 110 to 130 ° C for 10 to 20 The enzyme is inactivated by heating for a minute and then cooled.

The temperature, pH and time during the first and second hydrolysis are in a range for optimizing the activity of the enzyme, and the protein of the polymer having a low digestibility and absorption rate in the human body is decomposed into low molecular protein having a high digestion and absorption rate by the hydrolysis It is possible to improve the absorption rate of useful components contained in sea cucumbers.

In addition, the sea cucumber contains a lot of minerals such as calcium, iron, phosphorus and iodine. Calcium is the most abundant minerals in the human body. It constitutes the skeletal structure and takes charge of physiological regulation. Most of them are calcium phosphate, which is present in bones and teeth, and the rest constitutes all tissues as phospholipids and nucleic acids, and also plays an important role in metabolism in living organisms.

However, in order for these minerals to be absorbed by the human body, calcium and phosphorus must be in a water-soluble state in the intestinal tract. However, calcium and phosphorus bind to each other in the body to form an insoluble salt or calcium carbonate form and can not be absorbed by the human body. The present invention provides an effect of improving the absorption rate of the human body by inhibiting the binding of calcium and phosphorus and keeping them in a water-soluble state.

Next, the cooled hydrolyzate is fermented with lactic acid bacteria extracted from kimchi to convert saponin glycoside of sea cucumber having low digestion rate into saponin non-glycoside with high digestion and absorption rate.

Saponin indicates that aglycone, which is a non-sugar part of glycosides widely present in plants, is composed of various ring compounds, and triterpene saponin is contained in sea cucumber.

Saponin has a chemical structure similar to cholesterol, which inhibits cholesterol absorption and helps release, inhibits cell destruction by free radicals and inhibits mutations, thus reducing the chance of developing colon cancer.

However, saponin contained in sea cucumbers is a glycoside in the form of a sugar-linked glycoside. Since glycosides must be hydrolyzed in the body to be absorbed into the body, there is a disadvantage in that digestion absorption rate is low. In the present invention, the saponin glycoside of sea cucumber is decomposed, Thereby converting it into an easily non-glycoside.

For this purpose, microorganisms producing β-glucosidase, which converts saponin glycosides of sea cucumber to non-glycosides, are separated from kimchi and used for the conversion of saponin glycosides to non-glycosides. There is no side effect to the human body by using the microorganism extracted from Kimchi as a food which is usually consumed frequently.

Kimchi, which is a fermented food, is a source of various minerals and vitamins. It has a function of succulent action (lactic acid action) by lactic acid bacteria, inhibits the action of other harmful bacteria in the intestines, inhibits abnormal fermentation, And cancer.

First, the kimchi is immersed and aged at 3-7 ° C. for 5 to 6 months. Then, the kimchi of the kimchi is filtered to remove the solid content. This aging process enriches the production of lactic acid bacteria in kimchi.

In the temperature decreasing method (temperature drop method) in which the genus Leuconostoc and the genus Pediococcus are produced energetically at the aging temperature and the aging temperature is lowered to about 3 to 7 ° C at about 10 ° C, Lactobacillus spp . ( Pediococcus pentosaceus ) is mainly composed of Pseudomonas aeruginosa , Lactobacillus sp., Lactobacillus sp., And Pediococcus sp .

Next, microorganisms producing beta-glucosidase are isolated and identified from the Kimchi soil from which the solid content has been removed.

The microbial separation is carried out by a conventional method of separating a colony by using a culture medium. In a solid medium containing esculin, a microorganism-produced beta-glucosidase is added to the culture colonies It is preferable to use a Bile esculin agar (BEA) method.

The identification of the microorganisms is performed by measuring the beta-glucosidase enzyme activity of the isolated microorganisms and selecting the strain with the highest activity.

For example, p-nitrophenyl [beta] -D-glucopyranoside (p-NPGlu) can be used as a method for measuring the enzyme activity, Can be measured.

When a strain having high beta-glucosidase activity is prepared, it is inoculated into the cooled hydrolyzate at a concentration of 0.05 to 3.00 wt% based on the total weight, and then fermented at 30 to 50 DEG C for 15 to 30 hours.

During the fermentation, the beta-glucosidase enzyme of the strain extracted from kimchi transforms saponin glycosides contained in sea cucumber into non-glycosides that are easily absorbed into the body, thereby increasing the digestion and absorption rate of sap ginseng saponin. The fermentation temperature and fermentation Time is the range in which saponin glycosides contained in sea cucumbers are converted to non-glycosides at the highest rate.

When the fermentation is completed, the preparation of the fermented sea cucumber according to the present invention is completed, and then the enzyme is preferably inactivated by heating at 110 to 130 ° C for 10 to 20 minutes.

Since the fermented sea cucumber is in the form of a liquid, it is easily spoiled and difficult to be stored and handled. Therefore, the fermented sea cucumber is preferably dried and pulverized. The drying can be carried out by a known method such as vacuum drying, freeze drying, vacuum freeze drying Spray drying and the like.

In addition, the fermented sea cucumber according to the present invention can be ingested in the form of liquid or powder as described above. However, the fermented sea cucumber according to the present invention can be manufactured into various formulations for convenience of ingestion, for example, granules, tablets, capsules, The excipient such as dextrin may be mixed. The excipient may be mixed before the fermentation sea cucumber is dried to dry a mixture of the fermented sea cucumber and the excipient.

Hereinafter, the present invention will be described in more detail with reference to the following examples and test examples.

It is to be understood, however, that the invention is not to be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Will be apparent to those skilled in the art to which the present invention pertains.

Example 1 Isolation and Identification of Microorganisms Producing Beta-Glucosidase in Kimchi

Chinese cabbage was pickled and seasoned in a conventional manner, and the kimchi was prepared by aging for 6 months at 5 ° C in an Onggi jar.

Kimchi was homogenized after 60 days, 100 days, 140 days, 180 days, and 220 days in the aging period of the kimchi and juice was obtained to obtain Kimchi juice. Then, the Kimchi juice was filtered to remove 5 solid Kimchi juice samples .

1 ml each of the above samples was taken and placed in a 100 ml Man-Rogosa-Sharpe (MRS) liquid medium (Difco, USA) and cultured at 37 ° C for 24 hours under anaerobic conditions.

The cultured samples were serially diluted in 0.85% saline, spread on a BEA medium, and incubated for 36 hours at 37 ° C in an anaerobic condition to confirm the presence of a single black colonies.

The colonies were again streaked on the BEA medium to isolate only a single black cell population around the colonies.

The isolated microbial community was identified using API 50 CHL kit (Biomerieux, France). As a result, Leuconostoc mesenteroides (60 days and 100 days), a kind of lactic acid bacteria, and Pediococcus pentosus Souse (140 days, 180 days, 220 days) was identified as the main species.

<Test Example 1> Measurement of beta-glucosidase enzyme activity of the isolated microorganism

1) Extraction of enzyme

Each isolated single cell population of Example 1 above was stationary cultured under anaerobic conditions for 24 hours at a temperature of 37 DEG C in MRS medium.

The culture supernatant was divided into a culture supernatant and a pellet by centrifugation (10000 x g, 10 min, 4 캜), and the supernatant was stored. The pellet was resuspended in a buffer (5 mM Na ₂ HPO ₄ -citric acid, pH 5.8 , McIlvaine buffer). After washing, the cells were subjected to cell disruption with a 500-mg glass bead for 10 minutes for 1 minute.

After centrifugation (10000 × g, 10 min, 4 ° C), the supernatant was taken and mixed with the supernatant. Four times as much ethyl alcohol was added thereto, followed by stirring at 4 ° C for 30 minutes.

After stirring, the supernatant was discarded by centrifugation (10000 x g, 10 minutes, 4 캜), and the pellet was dissolved in the buffer (McIlvaine buffer) to prepare an enzyme for measuring beta-glucosidase activity.

2) Measurement of beta-glucosidase activity

45 μl of the extracted enzyme and 15 μl of 10 mM p-NPGlu were mixed, reacted at 37 ° C for 15 minutes, and then 900 μl of 0.25 M sodium carbonate (Na ₂ CO ₃ ) was added to terminate the reaction.

In order to observe the change of absorbance at 400 nm, 135 μl of buffer (McIlvaine buffer) and 45 μl of 10 mM p-NPGlu were added as a control group, and the reaction was performed at 37 ° C for 15 minutes. The relative activity was measured on the 60th day The results are shown in Table 1 below.

Results of beta-glucosidase activity measurement Aging period 60 days 100 days 140 days 180 days 220 days Relative activity (%) 100 121 137 143 148

As can be seen from the above Table 1, the activity of beta-glucosidase was increased as the aging period was increased, and the decrease was observed after 180 days.

Example 2 Preparation of fermented sea cucumber

The dried sea cucumbers distributed in the market were cleaned, crushed, boiled for 5 minutes, and cooled naturally.

Alkaline enzyme was added to the cooled sea cucumber to be contained in an amount of 1 wt%, and the mixture was heated at a pH of 7 and a temperature of 55 ° C for 2 hours to obtain a primary hydrolyzate.

A flavosome enzyme was added to the primary hydrolyzate in an amount of 1 wt%, and the mixture was heated at a pH of 7 and a temperature of 55 ° C for 12 hours to obtain a secondary hydrolyzate.

Next, the secondary hydrolyzate was heated at 121 DEG C for 15 minutes to inactivate the enzymes, followed by natural cooling.

The cooled secondary hydrolyzate was inoculated with 1.5% by weight of the lactic acid bacteria extracted from the 180-day sample of Example 1 having high beta-glucosidase activity, and then fermented at 40 ° C for 24 hours to prepare a fermented sea cucumber.

Thereafter, the fermented sea cucumber was heated at 121 ° C for 15 minutes to inactivate the lactic acid bacteria, followed by natural cooling.

&Lt; Test Example 2 > Measurement of degree of hydrolysis

In Example 2, boiled sea cucumber, primary hydrolyzate, secondary hydrolyzate and fermented sea cucumber were taken, and then 20% trichloroacetic acid (TCA) was added in the same amount, followed by centrifugation, and a predetermined amount of the supernatant was taken After measuring the amount of 10% TCA soluble protein according to the Lowry method (1951), the degree of hydrolysis of the protein was calculated using the following equation.

Hydrolysis (%) = 10% TCA soluble protein amount / total protein amount x 100

Hydrolysis rate (%) Boiled sea cucumber Primary hydrolyzate Secondary hydrolyzate Fermented sea cucumber Hydrolysis degree 11.7 43.6 88.2 91.5

As shown in Table 2, 43.6% of total proteins were hydrolyzed by primary hydrolysis but 88.2% of proteolysis was achieved due to secondary hydrolysis, and proteolysis was slightly increased by fermentation .

Thus, the protein of the polymer can be converted into the hydrolyzed low-molecular protein through two-step hydrolysis with the enzyme alkalase and the flavozymes.

&Lt; Test Example 3 > Measurement of protein digestibility

The protein digestibility of the test sample in Test Example 2 was measured. The assay was performed by the Association of Official Analytical Chemists (AOAC) method modified by Satterlee et al. (1979). The control protein was ANRC (Animal Nutrition Research Council casein was used.

Protein digestibility (%) Boiled sea cucumber Primary hydrolyzate Secondary hydrolyzate Fermented sea cucumber digestibility
(in vitro) 23.1 67.8 85.3 88.4

The results of Table 3 are similar to those of Table 2. It can be seen that the protein digestibility is increased by the sequential hydrolysis of the enzymes alkalase and flavozymes, It can be judged that most of the protein of the polymer having a low digestion rate is converted into a peptide having a low digestibility and a high digestibility.

<Test Example 4> Analysis of the content of saponin

In Example 2, saponin content was analyzed for boiled sea cucumber, primary hydrolyzate, secondary hydrolyzate and fermented sea cucumber. For comparison, the boiled seaweed was not hydrolyzed with the enzymes of alkaline and flavonoid enzymes Fermented sea cucumber fermented with lactic acid bacteria was analyzed as a control group.

5 g of each sample was placed in a round flask, 100 ml of 100% methanol was added, and the mixture was extracted for 2 hours in a water bath at 80 ° C equipped with a reflux condenser. The extraction procedure was repeated two more times to prepare an extract.

The extracts were combined, filtered through filter paper, concentrated under reduced pressure at 60 ° C, dissolved in 10 ml of water, purified with an OASIS HLB cartridge, and filtered through a 0.2 μm filter to obtain an analytical sample.

The OASIS HLB cartridge purification conditions are as follows.

Conditioning: 4 ml of methanol, 4 ml of ultrapure water

Sample loading: 10 ml

Washing: 4 ml of 5% methanol

Elution: 4 ml of methanol

The analytical sample prepared above was measured for the content of ginsenoside components using a high performance liquid chromatography / mass spectrometer (HPLC / MS, Waters Quattro micro VE). The analytical conditions were as shown in Table 4 below, Table 5 shows the results.

Chromatography / Mass Spectrometry Analysis Conditions Separation column ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 2.1 mu m particle size Flow rate 0.4 ml / min Column temperature 30 ℃ Sample injection solution 2 μl Detection wavelength 203 nm Mobile Conditions Time (minutes) Water (%) ACN (%) Curve 0 85 15 One 0.5 85 15 6 18 70 30 6 24 65 35 6 38 45 55 6 39 10 90 6 39.5 10 90 One 40 85 15 One

Analysis of saponin content (mg / g) Boiled sea cucumber Primary hydrolyzate Secondary hydrolyzate Fermented sea cucumber Control group Rg1 0.014 0.014 0.013 0.010 0.012 Re 0.007 0.006 0.006 0.004 0.005 Rb1 0.004 0.004 0.004 0.002 0.002 Rc 0.020 0.017 0.015 0.007 0.010 Rd 0.005 0.004 0.003 0.001 0.003 Rh1 + Rg2 0.011 0.015 0.019 0.035 0.027 Rg3 0.0004 0.0012 0.0013 0.0025 0.0020

Ginsenoside, ginsenoside, is a derivative of triterpene saponin contained in sea cucumber. In general, saponins such as Rg1, Re, Rb1, Rc and Rd are not easily decomposed and absorbed in the body and Rh1, Rg2, Rg3 Saponin has been reported in the academia as being easy to absorb and have a variety of unique behaviors.

Rg1, Re, Rb1, Rc, and Rd saponins having low digestion and absorption rates are gradually lowered as the hydrolysis and fermentation proceeds, and the contents of Rh1, Rg2, and Rg3 saponin having high digestion and absorption rates gradually increase, When the fermentation was carried out with lactic acid bacteria having high β - glucosidase activity extracted from kimchi, the digestion and absorption rate of sea cucumber saponin was improved.

The fermented sea cucumbers of the control group fermented without hydrolysis had a relatively low increase in the amounts of Rh 1, Rg 2 and Rg 3 saponins compared to the fermented sea cucumber of Example 2, so that decomposition of the polymer proteins of the sea cucumber into low molecular weight peptides, It is preferable to improve digestion and absorption rate.

&Lt; Test Example 5 > Calcium absorption enhancement effect measurement

In general, calcium in the body is solubilized by gastric acid and absorbed in the intestinal tract. Soluble calcium binds with phosphate ions present in the intestinal tract and forms precipitates that are not absorbed into the body.

Therefore, in order for the calcium to be absorbed in the intestine, it is necessary to have a large amount of calcium solubilized. To measure the solubilized calcium concentration, the solubilized calcium concentration of the test sample of Test Example 4 is measured and shown in Table 6 below.

For the measurement, 6 ml of 20 mM sodium phosphate (Na ₂ HPO _{4 )} was added to 1 ml of the sample, and then 5.5 ml of 20 mM calcium chloride (CaCl ₂ ) was added and the mixture was reacted at 37 ° C for 2 hours. × g, 5 minutes), and the calcium concentration in the supernatant was measured by an atomic absorption spectrometer.

Solubilized calcium concentration (ppm) Boiled sea cucumber Primary hydrolyzate Secondary hydrolyzate Fermented sea cucumber Control group Calcium concentration 46.5 68.2 82.7 85.1 57.9

As shown in Table 6, the solubilized calcium concentration was significantly increased by the hydrolysis in the second step and slightly increased in the fermentation process. In the control group without hydrolysis, the hydrolysis showed lower values than the first hydrolyzate.

From the above results, it is considered that the low molecular peptide formed by the hydrolysis inhibits the binding of calcium to phosphorus, inhibiting the insolubility of calcium and increasing the solubilizing effect of calcium by fermentation. The higher the solubilized calcium concentration, As a result, the calcium absorption rate of the fermented sea cucumber according to the present invention was improved.

Claims (7)

Crushing the sea cucumber, adding 3 to 7 times of water and boiling for 20 to 40 minutes;
Cooling the boiled sea cucumber, adding 0.01 to 2.00% by weight of an alkaline enzyme, and performing primary hydrolysis for 0.5 to 3.0 hours;
Adding 0.01 to 2.00% by weight of a flavosome enzyme to the primary hydrolyzed sea cucumber and performing secondary hydrolysis for 1 to 24 hours;
Heating the secondary hydrolyzed sea cucumber at 110 to 130 ° C for 10 to 20 minutes and cooling;
0.05 to 3.00% by weight of Pediococcus pentosaceus which is a lactic acid bacterium extracted from Kimchi in the cooled sea cucumber; And
And fermenting the sea cucumber inoculated with the lactic acid bacteria at 30 to 50 DEG C for 15 to 30 hours. The method according to claim 1,
Wherein the primary and secondary hydrolysis steps are performed at a pH of 6 to 8 and a temperature of 45 to 65 ° C. The method according to claim 1,
The method of extracting lactic acid bacteria from the above-
Filtering the kimchi of the kimchi to remove solid matter;
Culturing the Kimchi soup in which the solid content has been removed, in an MMS liquid culture medium;
Continuously diluting the cultured medium with saline and then culturing the BEA culture medium in a plate medium to obtain a single culture;
Obtaining a single cell population around the bacterial colonies by referring to the colonies again on a BEA medium; And
And isolating the bacterial community to obtain a strain. The method of claim 3,
Wherein the kimchi is aged at 3-7 ° C. for 5 to 6 months after immersed in the fermented sea cucumber. delete The method according to claim 1,
Characterized in that the fermented sea cucumber is heated at 110 to 130 ° C for 10 to 20 minutes and then mixed with an excipient and dried to prepare a powder, granule, capsule and tablet form. Method of manufacturing sea cucumber. A fermented sea cucumber produced by the method of any one of claims 1 to 4 or claim 6.