KR101696835B1 - The fermenting method of Codonopsis lanceolata for enhancing recognition ability improvement activity of Codonopsis lanceolata - Google Patents

Abstract

The present invention relates to a fermentation process of dodok for fermentation of dodok, and more particularly to a fermentation process for fermenting dodok which comprises (a) a first step of washing and drying the doduck; (b) a second step of activating the bacteria by inoculating the fermenting microorganism into a culture medium containing the roots extract at 20 to 30 DEG C at pH 6.0-6.8; And (c) inoculating the fermented microorganism activated through the second step in a state where the water-washed and dried dill are not cut, followed by solid fermentation with constant stirring with a stirrer, followed by aging; And a fermented product thereof for promoting cognitive function improving activity. According to the present invention, by enhancing the dissolution of the active substance for improving cognitive function of Doduk, the value of the dodo-fermented product as a food and medicine material is greatly improved since it has safety and low side effects in comparison with a chemical substance used for improving cognitive function , It is possible to minimize the possibility of bacterial contamination and generation of carcinogenic substances, which is a problem caused by liquid fermentation in the past, by virtue of uniformly fermenting the whole dough while keeping it in its original form without cutting it through solid fermentation.

Description

[0002] The fermenting method of Codonopsis lanceolata for enhancing recognition ability of Codonopsis lanceolata

 More particularly, the present invention relates to a fermentation method of roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roasted roast roast.

The ability to maintain and reproduce memorable experiences is called cognitive ability. Decrease in cognitive ability including memory is a serious problem in society because it causes learning deterioration, social activity inactivity, and deterioration of quality of life. Dementia is the most representative disease of memory impairment. It is the most common symptom of memory impairment, which is caused by various causes such as memory, thinking ability, comprehension, calculation ability, learning ability, language and judgment This is the whole of the syndrome of mental brain syndrome in which deterioration occurs. Most geriatric dementias, commonly referred to as Alzheimer's disease, have not been elucidated in detail, but based on the results of many animal studies and clinical trials, it has been shown that deprivation of acetylcholine in the brain, secretase, which is the main cause of toxicity by amyloid protein. Accordingly, the drugs for treating memory disorders include the expression of toxic proteins by inhibiting muscarinic acetylcholine receptor agonists, acetylcholine production promoters, acetylcholinesterase inhibitors or -secretase inhibitors Are being developed. However, such artificial synthetic drugs have the potential to cause side effects such as weight loss, insomnia, nightmares, and muscle spasms, so it is imperative to develop a suitable therapeutic agent for a living body with less side effects.

Codonopsis lanceolata is a perennial herbaceous plant belonging to the genus Chironomidaceae. It is known as white ginseng, red ginseng, rosemary, gaduk, and flea. It is native to Korea, China (Manchuria) and Japan. Is being cultivated.

In particular, it contains triterpenoid saponins, carbohydrates and fibers, and contains amino acids, minerals, sterols, triterpenoids, cycloartenols, N-formylamines (N -formylharman, 1-carbomethoxy-β-carboline, perlolyrine, norharman and volatile flavor components.

In addition to the unique flavor, carbohydrate and protein, it also contains minerals such as calcium, phosphorus and iron, vitamin B1, vitamins B2 is abundant and has been widely used for food.

Studies have shown that the reduction of triglyceride and cholesterol through serum lipid reduction and antioxidant effect, antiviral and antiallergic effects are known. In addition, this effect has been shown to be due to the iridoid glycosides, lignin, isofraxidin and the like contained in the roots. In addition, there are other pharmacological actions such as thymocyte proliferation and increased activity of peritoneal macrophages, and recently, cognitive function improving activity has also been reported.

However, despite the fact that the profitability of cultivated farmers is worse and the growth area of cultivated farms is difficult to increase in the next year, the processing technology of the middle and lower paddy stays almost at the initial level, Pickles, pickles, candies, and some cosmetic materials by simple extraction. In addition, the variation of the preference for the freshness is very severe, and compared with the ginseng, it is perceived as a sub-agricultural product. In addition, there is a problem in utilization of dodok, which is a conventional root crop and has a densely formed structure, which has not exhibited a corresponding activity compared to a high useful ingredient content.

Thus, much effort has been made to promote the active ingredient of the roots through fermentation to date.

Previously, fermentation has been used for food processing by improving the taste, flavor and texture of food by using the microbial metabolism, enhancing useful ingredients, enhancing food storage and nutrition. Korean Patent No. 10-1150179 discloses a fermentation method using a culture medium of lactobacillus lactobacillus bulgaricus for the purpose of enhancing the antimicrobial activity of the fermented soybean curd, -2013-0049586 discloses a method in which the roots of the roots are dried at 55 to 65 ° C for 30 to 40 hours or the roasted sample is subjected to a hot water extraction process at 55 to 65 ° C for 20 to 30 hours, And a method for promoting physiological activity using a fermentation process.

Fermentation of these foods is largely liquid fermentation and solid fermentation. Conventional food processing uses a liquid fermentation method in many cases. When this liquid fermentation method is used, fermentation proceeds in a liquid phase, so there is a high possibility of bacterial contamination and hygiene problems arise. In some fermented products, the possibility of carcinogen release during liquid fermentation has been suggested. In the case of the conventional solid fermentation, since the subject material is continued without grinding processing, there is a problem that it is difficult to uniformly ferment the whole fermented product.

Therefore, it is urgent to establish the fermentation methods and conditions that can minimize the possibility of bacterial contamination and carcinogenic substances caused by liquid fermentation while enhancing the elution of active substances that enhance the cognitive function of Doduck.

Accordingly, the present inventors have been studying to elucidate the activity of the active substance for improving the cognitive function of dodder and to minimize the possibility of bacterial contamination and the generation of carcinogens, which are problems caused by conventional liquid fermentation, and to exhibit uniform fermentation as a whole, Bifidobacterium longum and Leuconestoc mesenteroides can be used to enhance the activity of improving the cognitive function of dodec by using the solid fermentation method, thus completing the present invention.

Korean Patent No. 10-1150179

Accordingly, it is an object of the present invention to provide a fermentation method of dodok for the promotion of the cognitive function improving active substance of doduck.

It is another object of the present invention to provide a fermentation product of the fermentation broth produced by the fermentation method.

In order to achieve the above object,

(a) a first step of washing and drying the roots;

(b) a second step of activating the bacteria by inoculating the fermenting microorganism into a culture medium containing the roots extract at 20 to 30 DEG C at pH 6.0-6.8; And

(c) inoculating the activated fermenting microorganism through the second step in a state of not being washed and dried, and then fermenting the fermented microorganism while continuously stirring the fermented microorganism with a stirrer, and aging the fermented microorganism The present invention also provides a fermentation method of doduk for the promotion of active substances.

In addition, the present invention provides a fermented product of dodo having an enhanced cognitive function improving activity obtained by the fermentation method.

According to the present invention, by enhancing the dissolution of the active substance for improving cognitive function of Doduk, the value of the dodo-fermented product as a food and medicine material is greatly improved since it has safety and low side effects in comparison with a chemical substance used for improving cognitive function , It is possible to minimize the possibility of bacterial contamination and generation of carcinogenic substances, which is a problem caused by liquid fermentation in the past, by virtue of uniformly fermenting the whole dough while keeping it in its original form without cutting it through solid fermentation.

FIG. 1 is a schematic view showing a fermentation method of dodder according to an embodiment of the present invention.
FIG. 2 is a graph showing the average number of protrusions of neurite cells to which a fermented product prepared according to an embodiment of the present invention is added.
FIG. 3 is a graph showing an average length of neurite outgrowth of neurite cells to which a fermented matter prepared according to an embodiment of the present invention is added.
FIG. 4 is a graph showing the number of viable cells of neurite cells to which a rocuronium fermented product prepared according to an embodiment of the present invention is added.
FIG. 5 is a graph showing passive avoidance test results of a mouse administered with a roasted ferret produced according to an embodiment of the present invention. FIG.

In this specification, 'fermented dodec' means a fermented product obtained by fermenting doduck, and 'fermented product of doduck', 'fermented product of doduck' and 'fermented dod' can be used interchangeably.

Hereinafter, the present invention will be described in detail.

According to the present invention,

(a) a first step of washing and drying the roots;

(b) a second step of activating the bacteria by inoculating the fermenting microorganism into a culture medium containing the roots extract at 20 to 30 DEG C at pH 6.0-6.8; And

(c) inoculating the activated fermenting microorganism through the second step in a state of not being washed and dried, and then fermenting the fermented microorganism while continuously stirring the fermented microorganism with a stirrer, and aging the fermented microorganism The present invention also provides a fermentation method of doduk for the promotion of active substances.

The fermentation process of dodok for the enhancement of the cognitive function improving active substance according to the present invention will be described in more detail with reference to FIG.

[Step 1: Washing / Drying Process of Doduk]

Wash the dodok and dry it. In general, the fermentation may be carried out in the living body, but in terms of the fermentation efficiency, it is preferable to dry the roasted roots through a conventional drying method such as hot air or far-infrared irradiation, remove water and then ferment.

In one embodiment of the present invention, the washed dodec was dried in a warm air dryer at 20 to 30 ° C for 2 to 3 days, but it is not limited thereto.

[Step 2: Culture of fermenting microorganisms]

When microorganisms are inoculated into the broodstock, fermenting microorganisms can be cultured in a medium with optimal conditions for more effective and rapid reaction.

At this time, the MRS broth medium can be used as the medium, and it is preferable to add Mulberry extract to the medium for activation of the microorganism. The roe extract may be obtained by a method commonly used in the art, for example, hot water extraction, ultrasonic extraction, but is not limited thereto.

Specifically, it can be cultured by inoculating a fermenting microorganism to a MRS broth medium having a pH of 6.0-6.8 and adding 0.1-0.5% of the total weight of Mulberry root extract at 20-30 ° C.

The fermenting microorganism is Leuconostoc mesenteroides, Bifidobacterium longum, Lactobacillus rhamnosus, Saccharomyces exiguus, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Saccharomyces S. cerevisiae, Rhodopseudomonas palustris, Saccharomyces fragilis, Aspergilus niger, Bacillus stearothermophilus, and Acetobacter xylinum, and preferably Bifidobacterium longum and Leuconostoc mesenteroides can be used.

[Step 3: Fermentation and Aging Process]

The present invention utilizes solid fermentation to minimize the generation of bacterial contamination and carcinogenic substances that may be caused by the liquid fermentation process and to solve the disadvantage of liquid fermentation that does not effectively adapt to microorganisms when finely ground And the microorganism is inoculated with the microorganism in a circular state, thereby making more effective adaptation.

For the solid fermentation, the washed fermented microorganism is inoculated through the second step, without being washed and dried. At this time, the amount of the fermenting microorganism to be inoculated is preferably 1 to 10% based on the total weight. If the amount is outside the above range, the fermentation may not be performed smoothly.

When two or more kinds of fermenting microorganisms are inoculated, sequential inoculation at 20-25 hour intervals rather than simultaneous inoculation is preferable for effective fermentation and synergistic action of the fermentation microorganism.

In one embodiment of the present invention, Bifidobacterium longum was first inoculated and then Leuconestoc mesenteroides was inoculated 24 hours later, but the present invention is not limited thereto.

The fermented microorganisms are fermented for 3 ~ 5 days under 20 ~ 30 ℃ at 30 ~ 70% humidity and aged for 10 ~ 15 days. At this time, it is preferable to continuously stir the fermentation broth for fermentation throughout the fermentation period using a stirrer for uniform fermentation.

In addition, the present invention provides a fermented product of dodecae in which the cognitive enhancing activity substance produced by the fermentation process is promoted.

The fermented product of Dodae according to the present invention is produced by using the fermentation method described above. The common description of the two inventions is omitted in order to avoid the excessive complexity of the specification according to the repetitive description.

The fermented fermented product according to the present invention is a natural product having safety and low side effects as a natural product, increasing the nerve cell protrusion (see FIG. 2), increasing the protrusion length (see FIG. 3 ), Increase the number of living cells in the nerve cells (see Fig. 4), and exhibit a cognitive performance improving effect through passive avoidance experiments in white mice (see Fig. 5) And can be useful for treatment.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

< Example  1> Cognitive ability  Improved Active Substance Fermentation product  Produce

Bifidobacterium longum, cultured at 25 ° C, was inoculated at 5% of the total weight in an MRS broth medium containing 0.2% of Dodec hot-water extract and pH 6.5, and incubated at 25 ° C , The fermented product was fermented for 5 days at 30% humidity and then aged at 20 ° C for 10 days to obtain a fermented product.

< Example  2>

A fermented product of Doduck was obtained by the same method as in Example 1, except that Leuconostoc mesenteroides was inoculated as a fermenting microorganism instead of Bifidobacterium longum.

< Example  3>

Bifidobacterium longum as a fermenting microorganism was inoculated at 10% based on the total weight, and maintained at a temperature of 25 ° C and a humidity of 50%, fermented for 5 days, and then aged for 10 days at 20 ° C. A fermented product of Doduck was obtained.

< Example  4>

A fermented product of Doduck was obtained in the same manner as in Example 3, except that Leuconostoc mesenteroides was inoculated as a fermenting microorganism instead of Bifidobacterium longum.

< Example  5>

Fermented product was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides were mixed as a fermenting microorganism at a ratio of 1: 1.

< Example  6>

A fermented product of Doduck was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides as fermenting microorganisms were inoculated sequentially at a ratio of 1: 1 at intervals of 24 hours.

< Example  7>

A fermented product of Doduck was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides were mixed as a fermenting microorganism at a ratio of 1: 2.

< Example  8>

Fermented product was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides were inoculated as fermenting microorganisms at a ratio of 1: 2 sequentially at intervals of 24 hours.

< Example  9>

Fermented product was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides as fermenting microorganisms were mixed at a ratio of 2: 1 and inoculated.

< Example  10>

A fermented product of Doduck was obtained in the same manner as in Example 3, except that Bifidobacterium longum and Leuconostoc mesenteroides were inoculated as fermenting microorganisms at a ratio of 2: 1 sequentially at intervals of 24 hours.

< Experimental Example  1> In vitro in PC12  Measurement of cell growth promoting activity

To confirm the improvement of cognitive ability of fermented duck meat, the fermented duck meat (fermented duck meat) prepared in Example 6 was pulverized into powders and the following cognitive performance tests were carried out.

PC12 neurons, which were isolated from pheochromocytoma of rats, were used for the measurement of neuronal differentiation activity of each sample including Rhodiola and Fermented Rhizome. PC12 cells were inoculated at 500 占 퐇 in a 24 well plate at a concentration of 5.5 占⁰² cells / well. Twenty-four hours after the inoculation, 100 μl of the sample was added at a concentration of 60 μg / ml, and the cells were cultured in a 5% CO ₂ incubator at 36.5 ° C. The photographs were taken under an inverted microscope (× 200) every day, and the average number of protrusions produced per each cell was measured. The results are shown in FIG. 2. The longest neurite among the cells was extracted The mean length of neurite was measured and the results are shown in FIG. In addition, the number of living cells of the neurite was measured using Trypan Blue Dye Exclusion method (living cells were not stained with this reagent) during the 5-day culture period, and the results are shown in FIG. 4 Respectively.

As shown in FIG. 2, in the case of Staphylococcus aureus, the activity of differentiation of neurons was hardly observed over time. However, in the case of fermented Staphylococcus aureus prepared according to the present invention, differentiation of neurons is actively occurred over time, And the number of protrusions was increased. As the amount of added fermentation increased, the number of protrusions per cell increased further, and the differentiation of neurons was further increased.

Further, as shown in FIG. 3, the average length of the neurites in the case of fermented duck meat prepared according to the present invention was also significantly increased as compared with the case of adding fresh duck meat. As shown in FIG. 4, The number of dendritic cells was also found to be about twice that in the case of fermented duck meat prepared according to the present invention.

Therefore, it can be seen that the fermentation method of Dodok according to the present invention increases the activity of improving the cognitive function of Doduck, thereby increasing the number of neurites of the PC 12, increasing the number of protrusions per cell and the length of neurites.

< Experimental Example  2> Passive avoidance experiment of white rat

To confirm the improvement of cognitive performance of fermented duck meat, the duck meat fermented product (fermented duck meat) prepared in Example 6 was pulverized into powder and the following cognitive performance-related passive avoidance experiment was performed.

 The rats used in this experiment were 4 weeks old SPF male ICR mice. After the acquisition, the adaptation period in the breeding environment was completed for one week. The animals were weighed during the adaptation period and the selected animals were uniformly distributed to the average weight of each group. The animals were given a temperature of 22 ± 2 ℃, a relative humidity of 50 ± 5%, a ventilation frequency of 10 to 15 times / hr, an illumination time of 12 hours (lighting at 7.5 o'clock in the morning from 7.5 o'clock in the afternoon), and an illumination condition of 150 to 300 Lux.

The test group consisted of 167 mg / kg administered group (G3), 333 mg / kg administered group (G4), 1 mg / kg administered group (G2), scopolamine treated group ) And fermented red ginseng (667 mg / kg) (G5), scopolamine, or the like, to the mice which caused the decline of cognitive function, Respectively.

First, I put a rat in a box and turned it on brightly. At the same time, opening the compartment of the other dark chamber instinctively moves the rats to the dark chamber. At this time, in order to memorize electric shock, the door of the dark chamber was closed and 20 mA of electricity was supplied for 3 seconds on the floor, and the electric impact was applied to imprint it. This allowed the rats to learn enough to prevent them from entering the dark chamber even if they were illuminated. Twenty-four hours later, the samples and scopolamine were administered to each test group, and the same procedure was performed as the previous day to measure the retention time taken to the dark chamber. The excipient control group, scopolamine alone group, And the results are shown in Fig.

As shown in FIG. 5, in the case of the rat (G2) in which the cognitive function declined due to the administration of scopolamine, the dark chamber fell into the dark chamber within 50 seconds despite the electric shock. However, (G3-G5) treated with min was increased in the bright chamber than in the administration of scopolamine alone. Particularly, in the case of the rat (G5) administered with 667 mg / kg of the fermented dodec prepared according to the present invention, the rats were not administered with scopolamine, and the rats with normal cognitive function (excipient control group, G1) Time spent was almost the same.

Therefore, the fermented product prepared by the fermentation method of the fungus according to the present invention increases the number of protrusions and cells of nerve cells, increases the length of the fungus, And the like.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (8)

(a) a first step of washing and drying the roots;
(b) a second step of activating the microorganism by inoculating activated fermenting microorganisms, Bifidobacterium longum or Leuconostoc mesenteroide, into a culture medium containing a dodok extract added at a pH of 6.0 to 6.8 and containing 0.1 to 0.5% ; And
(c) Bifidobacterium longum, activated by the second step, is inoculated 1 to 10% of the total weight of the fermented microorganism, Leuconostoc mesenteroide, 1 to 10%, followed by solid fermentation for 3 to 5 days while maintaining the humidity at 30 to 70% at 20 to 30 ° C while continuously stirring with a stirrer, and aging for 10 to 15 days ; Wherein the method comprises the steps of: delete delete delete delete delete delete A fermented product of dodec, prepared according to claim 1, in which the cognitive enhancing active substance is promoted.

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