KR101290795B1 - Manufacturing methods of fermentation bellflower increased aneffective ingredient - Google Patents

Abstract

The present invention relates to a method for producing fermented bellflower with an increased active ingredient, and more particularly, to a method for producing fermented bellflower with an increased content of Platicodin D, including culturing the bellflower with yeast or fermenting it with yeast. .
According to the present invention, by using the fermented bellflower production method of the present invention to which the Takju production process is applied, it is possible to produce a fermented bellflower with an increased content of Platicodin D. In the present invention, when the cultivated pulverized bellflower is cultured with yeast isolated from yeast, fermented with yeast or fermented with yeast after cultivating with yeast, the content of Platicodin D is significantly increased as compared to general bellflower extract. When the bellflower fermented product was included in the cycle with dextrin or dextrin, the bitter taste peculiar to the bellflower could be removed. Therefore, the fermented bellflower composition with increased content of Platicodin D prepared through the preparation method of the present invention may be usefully used in functional food compositions or pharmaceutical compositions.

Description

Manufacturing method of fermented bellflower with increased active ingredient {Manufacturing methods of fermentation bellflower increased an effective ingredient}

The present invention relates to a method for producing fermented bellflower with an increased active ingredient, and more particularly, to a method for producing fermented bellflower with an increased content of Platicodin D, including culturing the bellflower with yeast or fermenting it with yeast. .

The bellflower is called Gilgyeong (桔梗) in Oriental medicine, and it is a perennial herb belonging to the lantern family. It grows in Korea, China, Japan, etc. The number of oriental prescriptions has a variety of pharmacological effects, including 273 in consent and one in 49. The major pharmacological component of bellflower is triterpenoid-based saponin (saponin). In animal experiments, it is anti-inflammatory for antitussive, expectorant, central nervous system (sedative, analgesic, antipyretic) and acute inflammation. Action, anti-ulcer and gastric secretion suppression, blood vessels dilate blood pressure lowering anticholinergic, hypoglycemic action, cholesterol metabolism improvement, etc. have been found. In addition, the hot water and ethanol extract of bellflower inhibit the apratoxin of the fungus, and the inulin fraction has potent antitumor effect against phagocytosis and solid and ascites cancers, and 40% bellflower extract has been shown to inhibit alcohol absorption. .

Bellflower is used in the form of grilling raw bellflower or boiling dried bellflower for coughing and relief. In addition, a recent study reported that the bellflower has pharmacological and antimicrobial effects, and to verify this, the antimicrobial activity against fermented bellflower and fermented bellflower was investigated. Bronchial diseases are mainly caused by colds, air pollution, smoking, and drying, and when mucosal cells in the bronchus weaken, secondary infections by pathogenic bacteria progress, causing inflammation, and asthma and pneumonia persist. It is known to lead to pulmonary tuberculosis. In particular, Klebsiella pneumonia accounts for 0.5 to 5.0% of all pneumonia and 6 to 8.6% of socially-infectious pneumonia, and occurs in men in their 40s with diabetes or chronic lung disease, especially chronic alcohol abuse of Klebsiella pneumonia. It is widely known as a risk factor group. The causative agent is Klebsiella As a pneumoniae, it is an aerobic Gram-negative bacillus and is difficult to treat because it is surrounded by a thick membrane capsule that can protect itself from macrophages. Most of the clinical symptoms are very fast-paced acute respiratory symptoms with high fever, cough, hemoptysis, and sputum discharge of Curreant-Jelly type. K diagnosis in sputum and blood cultures. It is possible to identify pneumoniae bacteria. Klebsiella Pneumonia is rare, but in the case of very fast and widespread pulmonary necrosis, medical treatment alone can have very fatal consequences such as acute respiratory failure, sepsis and multiple organ failure. Parallel is an essential method for patient survival.

According to the herbal medicine (Han Dae-seok, 1998), the main component of the extract of Platycodin D, which has several effects, is Platycodin D. I see it as a relationship. As such, complete or partial deglycosyldation of saponin glycosides can be used as an approach to the search for new biomes with alleviation of side effects.

Deglycosyldation of saponins in humans or animals must be mediated by gastric acid or intestinal bacteria deglycosidase to be released into fully hydrolyzed free prosapogenin. Unlike chemical hydrolysis, partial hydrolysis of glycoside chains by enzymes helps to separate glycosidic bonds and produce new outputs. It can be a good means for structural modification and functional improvement of such natural saponins. Plytycodin, a saponin of Gilt, has a distinctive arine and bitter taste despite its known pharmacological effects.

On the other hand, Nuruk supplies various hydrolytic enzymes including amylase to decompose starch raw materials such as rice in the brewing of traditional liquor, Takju and Yakju, and plays an important role as inoculator of fermentation-related microorganisms including yeast.

In general, Takju was prepared by saponifying starch by fermenting starch using artificially propagated molds on the increased starch raw material and producing yeast fermented by administering yeast. Takju undiluted solution is prepared by immersing one stage by administering water and immigration to the lower liquor, then immersing with two stages by administering water and steamed rice.

Therefore, the present inventors have made diligent research efforts to overcome the problems of the prior art, when fermenting the bellflower by using the process of fermenting the koji in goosebap (steamed rice), which is a conventional Takju manufacturing method, the active ingredient of bellflower It was confirmed that it can effectively increase the content of phosphorus Platicodin D, and completed the present invention.

Therefore, the main object of the present invention is to provide a method for producing fermented bellflower with increased content of Platicodin D.

It is another object of the present invention to provide a fermented bellflower composition in which the content of Platicodin D using the preparation method is increased and the bellflower characteristic is reduced.

According to one aspect of the present invention, the present invention provides a method for producing a fermented bellflower with an increased content of Platicodin D, comprising the following steps:

a) grinding the bellflower;

b) sterilizing the ground bellflower;

c) fermenting the sterilized bellflower with yeast;

d) filtering and concentrating the fermented bellflower; And

e) lyophilization to powder.

In the preparation method of the present invention, 5 to 30% of yeast, preferably 50 to 100 g of yeast per 1 kg of the sterilized bellflower in step c), and incubated for 2 to 4 days at 25 to 35 ° C It features. Here, Nuruk means that the fungus having an alcohol-making fungus is propagated in cereals, and in the embodiment of the present invention, the Nuruk used for brewing Takju and Yakju, which are traditional liquors of Uri, is used by grinding.

In the production method of the present invention, further comprising the step of culturing the sterilized bellflower before yeast step c).

Yeast used for the culture is isolated from yeast, Saccharomyces cerevisiae , Saccharomyces ellipsoideus ( Saccharomyces) ellipsoideus ), Saccharomyces coreanus ), Saccharomyces carlsbergensis ), Zygosaccharomyces major ), Saccharomyces lactics ), Saccharomyces rouxii ), and Hansenula anomala .

In the specific example of the present invention, when the fermented bellflower was prepared by applying the above-described manufacturing method, that is, the preparation process of Takju, it was confirmed that the content of Platicodin D was increased. More specifically, when the pulverized bellflower is fermented with Nuruk, or when the pulverized Bellflower is incubated with yeast and fermented with Nuruk, the content of Platicodin D is two or more times and four times or more, respectively, compared to the control (flower bellflower). Increased (see Example 2 and FIG. 2).

In the production method of the present invention, the concentration of step d), characterized in that it further comprises the step of inclusion.

The inclusion is characterized in that the inclusion of the dextrin selected from the group consisting of alpha, beta, gamma cyclodextrin and dextrin, more specifically the inclusion is added 1 to 40 w / w% of dextrin relative to the fermented bellflower solids, 0.5 It is characterized by performing by stirring at 50 to 500 rpm for 5 hours.

After cultivating the bellflower pulverized by yeast culture in a specific embodiment of the present invention, the fermented bellflower fermented with yeast fermented with various dextrins, and then subjected to high-evaluation of the inclusion sample, when the inclusion of alpha or gamma cyclodextrin high removal rate It was confirmed that this is measured at 80% or more (see Example 3 and Figure 3).

According to another aspect of the present invention, the present invention provides a method for preparing fermented bellflower with an increased content of Platicodin D, comprising the following steps:

a) grinding the bellflower;

b) sterilizing the ground bellflower;

c) incubating the sterile bellflower with yeast;

d) filtering and concentrating the fermented bellflower; And

e) lyophilization to powder.

In the production method of the present invention, inoculated at a concentration of 3 to 7% of the yeast seed solution per kg of the bellflower sterilized in step c), preferably inoculated at 150 to 300 mL, 2 to 4 at 25 to 35 ℃ It is characterized by culturing daily.

In the production method of the present invention, the concentration of step d), characterized in that it further comprises the step of inclusion. The inclusion is characterized in that the inclusion of the dextrin selected from the group consisting of alpha, beta, gamma cyclodextrin and dextrin, more specifically the inclusion is added 1 to 40 w / w% of dextrin relative to the fermented bellflower solids, 0.5 It is characterized by performing by stirring at 50 to 500 rpm for 5 hours.

Yeast used for the culture is isolated from yeast, Saccharomyces ( Saccharomyces cerevisiae), as a Saccharomyces ellipsis Soi Deus (Saccharomyces ellipsoideus), Saccharomyces Korea Augustine (Saccharomyces coreanus), saccharose in MRS car's Bergen sheath (Saccharomyces carlsbergensis), as is Saccharomyces Mazu Le (Zygosaccharomyces major), in a MRS lactic (Saccharomyces lactics), Saccharomyces Saccharomyces roksiyi (Saccharomyces rouxii), and a century nyulra cyano dry ( Hansenula anomala ) is characterized in that it is selected from the group consisting of.

As confirmed in the embodiment of the present invention, when culturing the ground bellflower using the yeast, it was confirmed that the content of Platicodin D effectively increased (see Example 2 and Figure 2).

According to an aspect of the present invention, the present invention is fermentation with an increase in the content of Platicodin D, characterized in that it is prepared according to any one of the manufacturing method of the fermentation dodo method with increased content of the Platicodin D It provides a bellflower composition.

Platicodine D is a saponin with major pharmacological components of bellflower, and is known to be excellent in anti-apoptosis, anti-obesity and anti-inflammation. It is known as a valuable single component. Therefore, the fermented bellflower composition with increased content of Platicodin D through the preparation method of the present invention may be usefully used in functional food compositions or pharmaceutical compositions.

In addition, the composition of the present invention is characterized in that it is possible to reduce the bitterness of the bellflower by enclosing the bellflower intrinsic bitter component molecule.

In the present invention, the inclusion is characterized in that the fermented bellflower extract prepared in Examples 1-4 was included by stirring with dextrin selected from the group consisting of alpha, beta, gamma cyclodextrin and dextrin.

Specifically, when the addition of 1% to 40 w / w% of alpha, beta, gamma cyclodextrin or dextrin relative to the solid content of the fermented bellflower extract and stirred in a condition of 50 to 500 rpm for 0.5 to 5 hours, It can enclose the bellflower's unique bitter taste molecule, thereby removing only the bellflower's bitter taste while maintaining the bellflower's inherent flavor, and at the same time, increasing the solubility and stability of the bellflower extract and improving transparency during liquid formulation. It can improve the overall quality of the product.

In the present invention, the composition is characterized in that it has an antimicrobial effect against pneumococcal bacteria.

Klebsiella pneumoniae is the causative agent of Klebsiella pneumonia It is characterized by being pneumoniae .

In a specific embodiment of the present invention, K. As a result of measuring the antimicrobial effect of the fermented bellflower according to the present invention against pneumoniae , it was confirmed that there is an antimicrobial activity against the bacterium, and in particular, it was confirmed that the antibacterial effect of the fermented bellflower was superior to the fermented bellflower extract (Example 4 and FIG. 4).

As described above, according to the present invention, by using the fermented bellflower production method of the present invention to which the Takju production process is applied, fermented bellflower with increased content of Platicodin D can be prepared.

According to an embodiment of the present invention, when the cultivated crushed bellflower in yeast isolated from yeast, fermented with yeast or fermented with yeast after incubating with yeast, the content of Platicodin D is higher than that of general bellflower extract. It was confirmed that the increase significantly, and when the fermented bellflower is enclosed in the cyclodextrin or dextrin can be removed the bitter taste peculiar to the bellflower. Therefore, the fermented bellflower composition with increased content of Platicodin D prepared through the preparation method of the present invention may be usefully used in functional food compositions or pharmaceutical compositions.

Figure 1 shows a schematic diagram of the production of fermented bellflower.
Figure 2 is the result of measuring the Platicodin D content of the fermented bellflower prepared according to each manufacturing process.
Figure 3 is the result of measuring the change in the taste of the fermented bellflower (sample of Example 1-4) by inclusion of the cyclodextrin or dextrin.
4 is a result of measuring the antibacterial effect of fermented bellflower (sample of Example 1-4).

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example  1. Fermented Bellflower Production Using Takju Production Process

1-1. Bellflower extract manufacturer

The bellflower was pulverized, 1.5 kg of pulverized bellflower and 15 liters of 10-fold amount of ionized water were placed in a fermenter and extracted under the condition of 95 ° C. for 3 hours. The extract was then filtered, concentrated and lyophilized.

1-2. Yeast Fermented Bellflower Production

The bellflower was pulverized as described above, and 1.5 kg of pulverized bellflower and 15 L of ionized water 15 L were put in a fermenter and sterilized at 121 ° C. for 30 minutes, and then the temperature was lowered to 30 ° C. Thereafter, the bellflower samples sterilized in the fermenter were put into an incubator with 100 g of pre-crushed Nuruk and incubated for 3 days while stirring at 30 rpm to 150 rpm. After the culture was completed, the solution was filtered and concentrated to lyophilize.

1-3. Production of Fermented Bellflower Cultured in Yeast

The bellflower was pulverized as described above, and 1.5 kg of pulverized bellflower and 15 L of ionized water 15 L were put in a fermenter and sterilized at 121 ° C. for 30 minutes, and then the temperature was lowered to 30 ° C. The yeast seed solution incubated for 1 day at 30 ℃ in YM medium (Yeast Extract 3.0 g / L, Malt Extract 3.0 g / L, Peptone 5.0 g / L and Dectrose 10.0 g / L) in the bellflower sterilized in the fermenter ( Saccharomyces Isolated from Yeast cerevisiae strain) was inoculated with 300 mL, and then put into the incubator and incubated for 3 days with stirring at 150 rpm at 30 ℃. After the culture was completed, the solution was filtered and concentrated to lyophilize.

1-4. Fermented Bellflower Production after Yeast Culture and Nuruk Continuous Culture

The bellflower was pulverized as described above, and 1.5 kg of pulverized bellflower and 15 L of ionized water 15 L were put in a fermenter and sterilized at 121 ° C. for 30 minutes, and then the temperature was lowered to 30 ° C. After inoculating 300 mL of yeast seed solution incubated in YM medium at 30 ° C. for 1 day in a bellflower sample sterilized in the fermenter, it was added to the incubator and incubated for 3 days with stirring at 150 rpm at 30 ° C. After completion of the culture, crushed Nuruk was added to the culture sample, and then aged at 30 ° C for 3 days. After the culture was completed, the solution was filtered and concentrated to lyophilize.

Example  2. Bellflower according to the manufacturing process Fermented Platicodine  D ( Plyticodin  D) Content change measurement

In order to measure the Platicodin D content contained in the fermented bellflower powder prepared according to the respective preparation processes in Example 1, liquid chromatography (High Liquid Chromatography, HPLC, Waters) was used. 1 g of each fermented bellflower powder was quantitatively placed in a 50 mL mass flask, dissolved in ionized water, filtered using a 0.45 mm membrane filter, and used as an HPLC sample. In the case of HPLC measurement, as standard, Platicodin D was dissolved in methanol at a concentration of 1 mg / mL, respectively, to prepare a stock solution of 1.0, 0.1, and 0.01 mg / mL as a stock solution. A calibration curve was prepared. The analysis conditions and mobile phase conditions are shown in Table 1 below.

Table 1. HPLC Analysis Conditions

As a result of HPLC measurement, as shown in FIG. 2, the content of Platicodin D contained in the bellflower extract (control) not cultured with yeast or yeast was measured as 2.56 mg / g, and the omission was inoculated only for 3 days. Fermented bellflower, which had been fermented), increased slightly to 4.33 mg / g by 1.5 times than the control group which was not cultured with the strain. In addition, the content of Platicodin D in the fermented bellflower fermented with yeast isolated from Takju for 3 days was 8.05 mg / g, which was about three times higher than the fermented product (control). After 2 days of culturing yeast bacteria isolated from Takju, it was confirmed that the content of placodin D was increased 4.3 times compared to the control in the fermented bellflower fermented with fermentation (fermentation) for 3 days.

Yeast are used in Aspergillus oryzae and Asp. niger mut. Kawachii and Asp. usami mut. Some other, including Shirousamii Fungi and Saccharomyces of the Genus Rhizopus , Mucor , Monascus Takju yeast of the cereviase type, lactobacillus, lactobacillus , Bacillus It is a complex microorganism containing subtilis , including anaerobic Bacillus spp. and other cocci.

Saccharomyces, yeast pure from yeast When the fermentation process was performed with the cerevisiae strain and the fermentation process with the yeast, the content of placodin D was increased, indicating that the single strain of Saccharomyces serevisiae was more fermented as the fermentation proceeds than the complex strain in the yeast. It can be seen that it has a relatively more influence on the production rate of din D, which is Saccharomyces serevisiae It is considered that the strain is inhibited by the inhibiting strain present in the complex strain in the growth.

The fermentation of Platycodon D production by fermenting the bellflower, Saccharomyces serevisiae primarily, increased the production of Platicodin D secondly with the increase of the complex strain present in the yeast. The production capacity of din D is thought to be better with the help of various enzymes produced by the complex strain.

Example  3. Fermented Bellflower Enclosure  after High evaluation

Thereafter, in order to remove the bitter taste of the bellflower, it was enclosed with dextrin or the like during fermented bellflower preparation. The fermented bellflower sample used in the inclusion experiment was a yeast fermentation sample (Example 1-4) after yeast culture in which the production rate of Platicodin D was most increased.

After pulverizing and sterilizing bellflower in the same manner as in Example 1-4, after the culture of yeast culture and yeast fermentation was filtered, the concentration of alpha, beta, gamma cyclodextrin or dextrin was about 10% based on the solid content. It was added and stirred at 150 rpm for 30 minutes to make a clathrate reaction and dried. Thereafter, sensory evaluation was performed to determine how effective the inclusion of dextrin was in removing the bitter taste while maintaining the inherent flavor of the fermented bellflower. Fermented bellflower clath powder prepared at a concentration of 5 to 30%, respectively, was measured based on the fermented bellflower clath powder and a control (fermented bellflower concentrate without adding anything) to five trained sensory inspectors. At this time, the control group was set to a value of 10, and was evaluated relative to the 9-step scoring method as an average value or an average value ± standard deviation, and repeated analysis three times or more, and verified the significance by Duncan's multi-range verification method.

As a result of sensory evaluation, as shown in Figure 3, among the fermented bellflower clad with the various dextrins, it was observed that the delicacy of fermented bellflower was significantly reduced when the cladding with alpha or gamma cycle dextrin.

Example  4. No fermentation  Antimicrobial Activity of Bronchial Bacteria from Fermented Bellflower Powder

In order to separate the useful material from the bellflower, using a H ₂ O as an extraction solvent extracted at 90 ℃, 3 hours and lyophilized to prepare a dry fermented bellflower powder. As the fermented bellflower powder, a sample prepared in the same manner as the method for preparing bellflower fermented product yeast was cultured after the yeast culture (Example 1-4) was used. Then, to determine the antimicrobial effect of bellflower on bacteria causing bronchial diseases, K. Nutrient agar was prepared as a culture medium for pneumoniae , and fermented bellflower or fermented bellflower powder was mixed with dilution water to prepare a concentration of 0.05, 0.5, 1, 3, and 5%, respectively. K to the culture medium to which each fermented and fermented bellflower powder prepared for each concentration was added. After inoculation with pneumoniae was incubated in the culture conditions of 37 ℃, 24 hours, and after the completion of the culture to determine the antimicrobial effect according to the treatment of bellflower powder.

Bronchial disease-causing bacterium K. As a result of measuring the antimicrobial effect of the non-fermented and fermented bellflower extract powder against pneumoniae , the fermented bellflower extract powder showed better antibacterial activity than the non-fermented bellflower extract powder as shown in FIG. In particular, when the fermented bellflower powder was added at a concentration of 0.05%, it showed an inhibitory effect of 80% or more, and when it was added 5% or more, the growth of the bacteria did not occur, indicating an inhibitory effect of 100%. In addition, the fermented Dorado also exhibited a 60% inhibitory effect when added at a concentration of 0.5%, and more than 80% when inhibited. Through the above results, bronchial disease-inducing bacteria K. The antimicrobial activity of the extracts of p . paniculata against pneumoniae may be due to the fact that the fermented bellflower and fermented bellflower powder samples contain a large number of pharmacologically useful substances that inhibit the growth of bacteria. It was found to have an excellent antibacterial effect.

Claims (14)

Fermented bellflower manufacturing method having an increased content of Platicodin D comprising the following steps:
a) grinding the bellflower;
b) sterilizing the ground bellflower;
c) fermenting the sterilized bellflower with yeast;
d) filtering and concentrating the fermented bellflower; And
e) lyophilization to powder.
According to claim 1, wherein 50 to 100 g of yeast per kg of the sterilized bellflower in step c) is added, and the content of Platicodin D is increased, which is incubated for 2 to 4 days at 25 to 35 ℃ Fermented Bellflower Production Method.
The method of claim 1, further comprising the step of incubating the sterilized bellflower with yeast before step c).
According to claim 3, wherein the yeast is isolated from yeast Saccharomyces ( Saccharomyces) cerevisiae), as a Saccharomyces ellipsis Soi Deus (Saccharomyces ellipsoideus), Saccharomyces Korea Augustine (Saccharomyces coreanus ), Saccharomyces carlsbergensis ), Zygosaccharomyces major ), Saccharomyces lactics ), Saccharomyces rouxii ), and Hansenula anomala ( Hensenula anomala ) is a method for producing a fermented bellflower with an increased content of Platicodin D, characterized in that selected from the group consisting of.
The method of claim 1, wherein the concentration of the step d), the method of producing a fermented bellflower with an increased content of Platicodin D, characterized in that it further comprises the step of inclusion.
[Claim 6] The method of claim 5, wherein the inclusion is alpha, beta, gamma cyclodextrin and dextrin selected from the group consisting of dextrin selected from the group consisting of fermented bellflower production method with an increased content of Platicodin D.
According to claim 5, The clathrate is 1 to 40 w / w% of dextrin to the solid content of fermented bellflower, and the content of Platicodin D is increased, characterized in that the stirring at 50 to 500 rpm for 0.5 to 5 hours Fermented bellflower manufacturing method.
Fermented bellflower manufacturing method having an increased content of Platicodin D comprising the following steps:
a) grinding the bellflower;
b) sterilizing the ground bellflower;
c) incubating the sterile bellflower with yeast;
d) filtering and concentrating the fermented bellflower; And
e) lyophilization to powder.
10. The method according to claim 8, inoculating 150 to 300 mL of the yeast seed solution per kg of the bellflower sterilized in step c), the content of the Platicodin D is characterized in that for 2 to 4 days incubation at 25 to 35 ℃ Increased fermented bellflower manufacturing method.
According to claim 8, wherein the yeast of step c) is isolated from yeast Saccharomyces cerevisiae ), Saccharomyces ellipsoideus ), Saccharomyces coreanus ), Saccharomyces carlsbergensis , Zygosaccharomyces major ), Saccharomyces lactics ), Saccharomyces rouxii ), and Hansenula anomala anomala ) fermented bellflower manufacturing method with increased content of Platicodin D, characterized in that selected from the group consisting of.
A fermented bellflower composition with increased content of Platicodin D, which is prepared according to any one of claims 1 to 8.
12. The fermented bellflower composition according to claim 11, wherein the composition is reduced by the inclusion of bellflower bitter component molecules.
The fermentation bellflower composition according to claim 12, wherein the inclusion of the fermented bellflower extract prepared according to the method of claim 1 is carried out by stirring with dextrin selected from the group consisting of alpha, beta, gamma cyclodextrin, and dextrin.
The fermented bellflower composition according to claim 11, wherein the composition has an antimicrobial effect against pneumococcal bacteria.

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