KR20150094952A - Fermented extract of gastrodia elata with excellant antioxident activity and taste, and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a fermented extract of Gastrodia elata tubers and a preparing method thereof. The present invention provides a fermented Gastrodia elata tuber extract which is obtained by inoculating lactic acid bacteria to a Gastrodia elata tuber extract and fermenting the Gastrodia elata tuber extract and has an antioxidant activity and palatability. The method provided by the present invention includes: a mixture preparing step of mixing 400-800ml of distilled water with each 100g of Gastrodia elata tubers and grinding the mixture to prepare the mixture; an extract obtaining step of extracting the mixture with hot water to obtain the Gastrodia elata tuber extract; a cooling and filtering step of cooling and filtering the Gastrodia elata tuber extract; a sterilizing step of sterilizing the filtered Gastrodia elata tuber extract; an inoculating step of inoculating the lactic acid bacteria in a fermentation medium added with the sterilized Gastrodia elata tuber extract; a fermenting step of fermenting the fermentation medium at 20-30°C for 10-15 hours; and a fermented extract obtaining step of centrifuging the fermented extract centrifugal separator to obtain the supernatant as the fermented Gastrodia elata tuber extract. The present invention has high content of the functional ingredients unique to the Gastrodia elata tubers such as gastrodin and vanillin, has high absorption of the active ingredients and excellent antioxidant activity. The present invention is beneficial to health, has excellent taste, flavor, and palatability to be easily consumed.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a fermented Chunma extract having excellent antioxidant activity and preference, and a method for producing the fermented Chunma extract.

More particularly, the present invention relates to a fermented Chumma extract and fermented fermented milk bacterium having excellent antioxidative activity and favorable taste, and a process for producing the same.

In general, Gastrodia elata is a perennial parasitic herbaceous plant belonging to the orchidaceous plant. It is a rhizomorphic form of the root-related plant. It temporarily has leaves and roots, but is degenerated and only roots are developed. It is a symbiotic bulb with mushroom mycelium.

This kind of gum has been shown to be effective in "Dongbibo" due to paralysis, paralysis, seizures, arthritis, back pain, epilepsy and dizziness due to paralysis, and eliminating headaches and dizziness in " And has been used as a medicinal herb traditionally used in Korea and China since ancient times, for example, called "New medicine to treat wind" to be called "Jungpungcho".

Recent studies have shown that Gastrodin (ρ-hydroxymethyl phenyl- β-D-glucopyranoside), 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde, , Vanillin, vanillyl alcohol, and the like, all of which are phenolic compounds and are well known as active ingredients involved in antioxidative activity functions.

In addition, gastrodine is a representative physiologically active substance and exhibits properties such as improvement of circulation action, treatment of headache and dizziness, and anticonvulsant effect, and 4-hydroxybenzyl alcohol, 4-hydroxybenzylaldehyde and vanillin cause hippocampal CA1 cell death Have been reported to be active against GABAergic neuromodulation and active against anti-ischemic dementia.

However, in Korea, there have been restrictions on the development of food products that are prohibited from being used as food ingredients by the Korea Food and Drug Administration. However, as of September 1, 2000, have.

Korean Patent Laid-Open Publication No. 10-2013-0034540 (published on April 5, 2013) discloses a method for fermenting chunmean using a lactobacillus fermentum strain JS for long time storage A method for producing fermented chondrocyte granules having beneficial lactic acid bacterium alive and having an effect of digestion and absorption of chunmean is disclosed.

However, the above-mentioned conventional technology has a problem that the concentration of the active ingredient contained in the food is low and the low absorption rate of the active ingredient makes it difficult to exhibit useful functionalities in the actually applied food, resulting in poor economical efficiency and the unique taste and flavor There is a problem that the degree of preference is lowered.

DISCLOSURE OF THE INVENTION The present invention has been conceived to solve the above-mentioned problems, and it is an object of the present invention to provide a cosmetic composition containing high functional ingredients such as gastrodine and vanillin, high absorption rate of active ingredient and excellent antioxidative activity, The present invention also provides a fermented Chunmease extract having excellent taste and taste and easy to ingest.

In order to solve the above problems, the present invention provides a fermented Chumma extract having excellent antioxidative activity and favorable taste, which is obtained by fermenting lactic acid bacteria in a chymase extract solution.

At this time, the chymase extract is characterized in that it is obtained by mixing 400 to 800 ml of distilled water per 100 g of horse chestnut and hydrolyzing the ground mixture.

Furthermore, the above mixture is also characterized in that 1 to 5 g of Angelica gigas per cent of 100 g of horse chestnut and 1 to 5 g of Angelica gigas are further added.

In addition, the lactic acid bacteria may be any one of Lactobacillus plantarum (Accession No .: KCCM 11542), Lactobacillus plantarum (Accession No .: KCCM 12116), Bifidobacterium adolesentis (Accession No .: KCCM 11206), Bifidobacterium breve (Accession No .: KCCM 11208) There are also features.

In addition, 3% by weight of oligosaccharide is added to the whole.

The present invention also relates to a method for preparing a mixture, which comprises mixing 400 to 800 ml of distilled water per 100 g of horse meat and subjecting the mixture to milling to prepare a mixture; Obtaining an extract to obtain a chymase extract by hot water extraction of the mixture; A cooling and filtration step of cooling and filtering the chymase extract; A sterilization step of sterilizing the filtered cholama extract; An inoculation step of inoculating lactic acid bacteria into a fermentation medium to which sterilized chymase extract is added; A fermentation step of fermenting the mixture at a temperature of 20 to 30 DEG C for 10 to 15 hours in a fermenter; And a step of obtaining a fermentation broth obtained by centrifuging the fermentation broth by centrifugation in a centrifugal separator to obtain a supernatant fermentation broth.

Here, the preparation of the mixture is characterized by addition of 1 to 5 g of citron gum and 1 to 5 g of citron gum per 100 g of gum.

In addition, the step of obtaining the extract is characterized by being performed at 100 to 120 ° C for 3 to 5 hours.

In addition, the sterilization step is also characterized in that the sterilization is performed at 110 to 130 ° C for 10 to 20 minutes.

Further, the lactic acid bacteria inoculated in the inoculation step may be selected from the group consisting of Lactobacillus plantarum (Accession No .: KCCM11542), Lactobacillus plantarum (Accession No .: KCCM 12116), Bifidobacterium adolesentis (Accession No .: KCCM 11206), Bifidobacterium breve Is also a feature of the present invention.

The fermentation step is also characterized in that it is carried out at 25 ° C for 12 hours.

Also, the step of obtaining the fermented extract is characterized by centrifuging at 14,400 x g for 5 to 15 minutes.

In addition, the method further comprises the step of adding 3% by weight of the oligosaccharide to the whole of the fermented extract to obtain sterilized oligosaccharide.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a cosmetic composition which is highly effective in absorbing active ingredients and has excellent antioxidative activity, such as gastrodine and vanillin, .

FIG. 1 is a flowchart showing a method for producing a fermented Chunmease extract according to the present invention
FIG. 2A is a graph showing changes in pH of an extract of fermented Chamaian milk according to fermentation time in Example 1. FIG.
FIG. 2B is a graph showing the change in sugar content of fermented Chunmung extract according to fermentation time in Example 1. FIG.
FIG. 2C is a graph showing changes in DPPH radical scavenging activity of the fermented Chunmung extract according to fermentation time in Example 1. FIG.
FIG. 2 (d) is a graph showing changes in iron ion-reducing power of the fermented Chunmung extract according to fermentation time in Example 1. FIG.
FIG. 2E is a graph showing the change of the reducing power of the fermented Chromosome extract according to the fermentation time in Example 1. FIG.
FIG. 2f is a graph showing the change in total phenol content with fermentation time in Example 1. FIG.
Fig. 2G is a comparative graph showing the results of sensory evaluation of the aged layer in Example 1. Fig.
FIG. 2h is a graph showing the change in the content of gastrin in the fermented Chunmass extract according to fermentation time in Example 1. FIG.
FIG. 2I is a graph showing changes in vanillin content of the fermented Chunmung extract according to fermentation time in Example 1. FIG.
FIG. 3A is a graph showing the change of DPPH radical scavenging activity of the fermented Chumma extract according to fermentation time in Example 2. FIG.
FIG. 3B is a graph showing changes in iron ion-reducing power of the fermented Chunmung extract according to fermentation time in Example 2. FIG.
FIG. 3c is a graph showing changes in the reducing power of the fermented Chunmung extract according to fermentation time in Example 2. FIG.
FIG. 3D is a graph showing changes in the SOD-like activity of the fermented Chunmung extract according to fermentation time in Example 2. FIG.
FIG. 3E is a graph showing the change in total phenol content with fermentation time in Example 2. FIG.
FIG. 3f is a graph showing changes in total flavonoid content with fermentation time in Example 2. FIG.
FIG. 3G is a graph showing changes in the content of gastrin in the fermented Chunmung extract according to fermentation time in Example 2. FIG.
FIG. 3h is a graph showing changes in the content of vanillin in the fermented Chunmung extract according to fermentation time in Example 2. FIG.
4A is a bar graph showing the DPPH radical scavenging activity of each sample in Example 4. Fig.
FIG. 4B is a bar graph showing the iron on reducing power of each sample in Example 4. FIG.
4C is a bar graph showing the reducing power of each sample in Example 4. Fig.
4D is a bar graph showing the SOD-like activity of each sample in Example 4. Fig.
4E is a bar graph showing the total phenol content of each sample in Example 4. Fig.
FIG. 4f is a bar graph showing the total flavonoid content per sample in Example 4. FIG.
Fig. 4g is a bar graph showing the content of gastrin in each sample in Example 4. Fig.
4H is a bar graph showing the vanillin content of each sample in Example 4. Fig.

Hereinafter, the present invention will be described in detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, but includes modifications of equivalent technical ideas.

1. Obtaining fermented milk extract

After adding 600ml of distilled water to 95.24g of horse chestnut, the mixture was prepared by adding 2.38g of Angelica gigas and 2.38g of Angelica gigas, which are well suited to the rhizome, to control the unique taste and aroma of the horse. The mixture was subjected to hot water extraction at 110 DEG C for 4 hours to obtain a chymase extract. After cooling, the mixture was filtered and diluted to 700 ml. Then, the mixture was sterilized at 121 DEG C for 15 minutes, and lactic acid bacteria were inoculated (1%, v / v). Lactobacillus plantarum (Accession No .: KCCM11542), Lactobacillus plantarum (Accession No .: KCCM 12116), Bifidobacterium adolesentis (Accession No .: KCCM 11206) and Bifidobacterium breve (Accession No .: KCCM 11208) were used as the lactic acid bacteria. Then, samples were taken at 6 hours intervals while being fermented at 37 ° C for 24 hours, followed by centrifugation (14,400 × g, 10 minutes) to obtain a supernatant liquid of the fermented Chromosome extract, and analyzed under the following conditions.

2. Analysis method

The pH was measured directly with a pH meter (Model: Orion Star Series, USA) and the sugar content was directly measured with a sugar content meter (model: PAL-3, Japan). The chromaticity was measured with a colorimeter (Model: SP-80, Tokyo Denshoku, Japan ) Were expressed as L (lightness), a (redness) and b (yellowness) according to the Hunter scale.

Antioxidant activity samples were prepared by dilution with 0.02% BHT (butylated hydroxytoluene).

The DPPH radical assay was carried out by Williams et al. (1995), and 1.0 ml of a 2-fold dilution of ethanol was added to 3 ml of 60 mM DPPH , And the reaction was carried out in the dark for 15 minutes. The absorbance at 517 nm was measured. The DPPH radical scavenging activity was calculated by the following equation and expressed as a percentage.

The ferric ion reducing antioxidant power (FRAP) assay was performed using the method of Heo et al. (2006), 300 mM acetate buffer (pH 3.6): 10 mM TPTZ solution: 20 mM FeCl ₃ .6H ₂ O solution at a ratio of 10: 1: 1 was added to 1.9 ml of the reaction mixture, and the reaction solution was reacted for 20 minutes. The absorbance of the solution was measured at 590 nm. The FRAP was FeSO ₄ · 7H ₂ O , And converted to mg% in terms of a calibration curve.

The reducing power was determined by the method of Oyaizu et al. (1986). 2.5 ml of 0.2 M phosphate buffer (pH 6.6) and 2.5 ml of 1% K ₃ Fe (CN) ₆ And 2.5 ml of 10% trichloroacetic acid was added thereto. After centrifugation, 2.5 ml of supernatant was obtained. 2.5 ml of distilled water and 0.5 ml of 0.1% FeCl ₃ were added to the supernatant. The absorbance at 700 nm was measured, and the difference in absorbance between the sample added and the non-added sample was expressed as a percentage.

The total phenolic compound (TPC) assay was performed using Dewanto et al. (2002) method. 5.0 ml of distilled water and 0.5 ml of folin-ciocalteu's phenol reagent were added to 0.1 ml of sample The reaction was carried out for 1 minute and then 1.5 ml of 10% Na ₂ CO ₃ was added. The reaction was carried out for 1 hour in a dark room and the absorbance was measured at 765 nm. The total phenol content (TPC) ㎍ / ml), and expressed as mg%.

Sensory evaluation was performed on the taste, smell, color, preference, etc. The young people of 23 college students and graduate students were asked about their taste sensations when they ingested Chunma extract and lactic acid fermented juice obtained from lactic acid fermentation. (1 point: very weak or very poor, 9 points: very strong or very good), and the sensory test was conducted on 9 points scale (dissatisfactory, arine, ) Were used. Also, two samples with the most favorable taste were selected and the reason why they were selected was described.

For the elderly, 10 sensory evaluations were carried out on four samples selected from the sensory evaluation of the young people. The sensory evaluation was performed on the basis of color, taste (sweetness, sour taste), smell , 5 points scale method (1 point: extremely weak or extremely disliked, 5 points: extremely strong or extremely good) were used for overall likelihood.

In addition, the functional materials were analyzed for the four samples selected by the sensory evaluation of the young adults. The same amount of 70% methanol was added to the samples, and the mixture was centrifuged (14,400 × g, 10 minutes) filer, and analyzed by HPLC under the conditions shown in Table 1 below.

[Table 1]

3. Results

As shown in FIG. 2A, the pH of the fermented milk extract gradually decreased from 4.49 to 3.15 to 3.57 after 24 hours of fermentation.

As shown in FIG. 2B, the sugar content of the fermented milk fermented extract showed a constant level in the range of 2.4 to 2.7 Brix during the fermentation time of 24 hours.

As shown in the following Table 2, the chromaticity of the fermented Chunmung extract was in the range of 23.88 ~ 26.17, the a value (redness) in the range of 0.79 ~ 1.87, the b value The degree of yellowness was in the range of 11.41 ~ 16.99.

[Table 2]

As shown in FIG. 2C, the DPPH radical scavenging activity of the 2-fold diluted fermented Chumma extract was slightly reduced compared with that at the initial stage of fermentation for 24 hours.

As shown in FIG. 2 (d), the iron reducing power of the fermented Chunmung extract was found to be 17.75 mg% at the 12th hour of fermentation at the fermentation time of Lactobacillus plantarum (accession number: KCCM 12116), and Bifidobacterium adolesentis 11206), the maximum value of 18.31 mg% was obtained at 18 hours after fermentation, and Bifidobacterium breve (KCCM 11208) was the highest at 17 hours after fermentation at 12 hours. On the other hand, Lactobacillus plantarum (accession number: KCCM 11542) was found to be slightly decreased compared to the initial fermentation time.

As shown in FIG. 2E, the reducing power of the fermentation broth of Lactobacillus plantarum (accession number: KCCM 12116) showed a maximum value of 50.50% at 6 hours of fermentation at fermentation, and that of Bifidobacterum adolesentis (accession number: KCCM 11206) As a result, the maximum value was 54.27% at 18 hours after fermentation and maintained until 24 hours of fermentation. In addition, Bifidobacterium breve (Accession No .: KCCM 11208) showed a maximum of 51.13% at 12 hours after fermentation. On the other hand, Lactobacillus plantarum (accession number: KCCM 11542) was found to be slightly decreased compared to the initial stage of fermentation.

As shown in FIG. 2F, the total phenol content of the fermented Chunmung extract was in the range of 48.05 ~ 53.99 mg% during the fermentation time, and that of Lactobacillus plantarum (accession number: KCCM 12116) was 53.76 mg Bifidobacterum adolescentis (Accession No .: KCCM 11206) showed the maximum value of 53.99 mg% at 24 hours after fermentation, and Bifidobacterium breve (accession number: KCCM 11208) The highest value was 54.26 mg%. On the other hand, Lactobacillus plantarum (accession number: KCCM 11542) was found to be slightly decreased compared to the initial stage of fermentation.

As a result of sensory evaluation of 23 college students and graduate students, no statistically significant difference was observed except for fresh taste as shown in Table 3 below, but the taste tended to be improved as a whole. The disgusting taste slightly increased with fermentation, but the arine taste, the bad taste and the bitter taste were weakened. As shown in Table 4 below, the sample with the highest degree of preference was K, which was fermented with Lactobacillus plantarum (accession number: KCCM 12116) for 18 hours. The next highest preference was fermented for 12 hours with fermented Chunma extract (A), Lactobacillus plantarum (accession number: KCCM 11542), Factic acid (F) and Lactobacillus plantarum (accession number: KCCM 12116) (G). As a result, the strain Lactobacillus plantarum (accession number: KCCM 11542) and the strain Lactobacillus plantarum (accession number: KCCM 12116) were judged to be suitable strains for fermentation of chum, and the fermentation time was judged to be suitable from 12 to 18 hours. Therefore, G and K fermented with F and J, and Lactobacillus plantarum (accession number: KCCM 12116) fermented for 12 hours and 18 hours with Lactobacillus plantarum (accession number: KCCM 11542) The sensory evaluation of the elderly was carried out.

[Table 3]

[Table 4]

As a result of sensory evaluation of the four samples selected from the young people's sensory evaluation of 10 elderly people, as shown in the following Table 5 and Fig. 2g, when comparing A12 and A18, fermentation was added in A12, The smell became stronger, but the overall preference increased due to the increase of sweetness and sour taste. In addition, when B12 and B18 were compared, the addition of fermentation at B12 resulted in poor necking, strong smell, and almost similar sweetness and sourness, indicating a decrease in overall acceptability.

[Table 5]

As shown in FIG. 2h, the gastronine content of the fermented milk extract of Lactobacillus plantarum (accession number: KCCM 11542) was 6.72 mg% at the 18th hour of fermentation at fermentation. Likewise, Lactobacillus plantarum KCCM 12116) showed a maximum value of 7.96 mg% at 18 hours after fermentation.

As shown in FIG. 2 (i), the vanillin content of the fermented milk extract of Lactobacillus plantarum (accession number: KCCM 11542) was slightly lowered as the fermentation progressed during fermentation, but fermentation into Lactobacillus plantarum (accession number: KCCM 12116) The maximum value of 63.07 mg% was obtained at 6 hours after fermentation.

Finally, based on the results of the sensory evaluation and the functional component analysis of Example 1, it can be seen that Lactobacillus plantarum KCCM 12116 is the most suitable strain for the fermented milk fermentation extract.

1. Obtaining fermented milk extract

600ml of distilled water was added to 95.24g of horse chestnut, and the mixture was ground by adding 2.38g of Angelica gigantei and 2.38g of Angelica giganteus. The mixture was subjected to hot water extraction at 110 DEG C for 4 hours to obtain a chymase extract. After cooling, the mixture was filtered and diluted to 700 ml. Then, the mixture was sterilized at 121 DEG C for 15 minutes, and Lactobacillus plantarum (accession number: KCCM 12116) was inoculated (1%, v / v) as lactic acid bacteria. Then, samples were collected every 6 hours while being fermented at 25 占 폚 and 37 占 폚 for 24 hours, followed by centrifugation (14,400 占 g, 10 minutes) to obtain supernatant liquids for the fermented Chumma extract.

2. Analysis method

Antioxidant activity samples were prepared by dilution with 0.02% BHT (butylated hydroxytoluene).

The DPPH radical assay was performed by Williams et al. (1995), and 1.0 ml of a 10-fold dilution of ethanol was added to 3 ml of 60 mM DPPH , And the reaction was carried out in the dark for 15 minutes. The absorbance at 517 nm was measured. The DPPH radical scavenging activity was calculated by the following equation and expressed as a percentage.

The ferric ion reducing antioxidant power (FRAP) assay was performed using the method of Heo et al. (2006), 300 mM acetate buffer (pH 3.6): 10 mM TPTZ solution: 20 mM FeCl ₃ .6H ₂ O solution at a ratio of 10: 1: 1 was added to 1.9 ml of the reaction mixture, and the reaction solution was reacted for 20 minutes. The absorbance of the solution was measured at 590 nm. The FRAP was FeSO ₄ · 7H ₂ O , And converted to mg% in terms of a calibration curve.

The reducing power was determined by the method of Oyaizu et al. (1986). 2.5 ml of 0.2 M phosphate buffer (pH 6.6) and 2.5 ml of 1% K ₃ Fe (CN) ₆ And 2.5 ml of 10% trichloroacetic acid was added thereto. After centrifugation, 2.5 ml of supernatant was obtained. 2.5 ml of distilled water and 0.5 ml of 0.1% FeCl ₃ were added to the supernatant. The absorbance at 700 nm was measured, and the difference in absorbance between the sample added and the non-added sample was expressed as a percentage.

SOD-like activity was measured by Marklund and Marklund (1974). To 0.2 ml of the sample, 3.0 ml of tri-HCl buffer and 0.2 ml of 7.2 mM pyrogallol were added and the mixture was incubated for 10 minutes After the reaction, 1.0 ml of 1N HCl was added to stop the reaction, and the absorbance at 420 nm was measured. The SOD-like activity was calculated by the following formula and expressed as a percentage.

The total phenolic compound (TPC) assay was performed using Dewanto et al. (2002) method. 5.0 ml of distilled water and 0.5 ml of folin-ciocalteu's phenol reagent were added to 0.1 ml of sample The reaction was carried out for 1 minute and then 1.5 ml of 10% Na ₂ CO ₃ was added. The reaction was carried out for 1 hour in a dark room and the absorbance was measured at 765 nm. The total phenol content (TPC) ㎍ / ml), and expressed as mg%.

Total flavonoid content was determined by adding 0.075 ml of 5% NaNO _{2 to} 0.5 ml of the sample, reacting the mixture at room temperature for 5 minutes, adding 0.05 ml of 10% AlCl ₃ for 5 minutes, adding 0.5 ml of ₁ M NaOH, And the absorbance at 510 nm was measured. The calibration curve was prepared using epicatechin as a reference material and expressed as mg%.

The sensory evaluation was evaluated on the taste, smell, color, preference, etc. The young people of 9 college students and graduate students were asked about their sensory qualities about the taste of Chunmae extract obtained from Chunma extract and lactic acid fermented by lactic acid fermentation (1 point: Extremely weak or extremely disgusting, 5 points: Extremely strong), and 5 point scale (1 point: Extremely weak or extremely disgusting, 5 points: Extremely strong Or extremely good).

The contents of Gastrodin and Vanillin were determined by mixing the same amount of 70% methanol in the sampled samples and centrifuging (14,400 × g, 10 minutes). The supernatant was filtered with a 0.22 μm syringe filter, 1 and HLPC, respectively.

3. Results

As shown in FIG. 3 A, the DPPH radical scavenging activity of the fermented extract of Chumma lean diluted 10-fold was maintained at a constant level without noticeable change with fermentation time of 24 hours, and no difference was observed according to the fermentation temperature.

As shown in FIG. 3B, the iron reducing power of the fermented Chunmung extract increased gradually with fermentation at 25 ° C., and reached 17.65 mg% at 18 hours after fermentation and gradually increased at 37 ° C. The highest value was 15.71 mg% at 24 hours after fermentation and higher than that at fermentation at 25 ℃.

As shown in FIG. 3C, the reducing power of the fermented Chunmung extract increased until 12 hours after fermentation at 25 ° C, showing a maximum activity of 44.15% and then slightly decreased. On the other hand, fermentation progressed at 37 ° C And the maximum value was 44.35% at the 18th hour after fermentation and then maintained at the 24th hour after fermentation.

As shown in FIG. 3D, the SOD-like activity of the fermented Chunmung extract tended to increase with fermentation, and the fermentation at 37 ° C was slightly higher than that at 25 ° C.

As shown in FIG. 3E, the total phenol content of the fermented Chunmung extract was maintained at a constant level without any significant increase or decrease with fermentation, and there was little difference in fermentation temperature.

As shown in FIG. 3F, the total flavonoid content of the fermented Chunmung extract was slightly increased when fermented at 25 ° C, and was highest at 5.50mg% at 6 hours of fermentation, whereas when fermented at 37 ° C The content was slightly decreased.

The sample of the sensory evaluation was excluded from the 18 hour fermentation sample and the 24 hour fermentation sample which were too acidic through the preliminary sensory evaluation. As shown in Table 6 below, statistically significant differences were observed in color, smell, and sweetness among 9 university students and graduate students. However, the difference in sour taste was slightly different, and it was judged that the extract having no lactic acid fermentation had the weakest sour taste and the sour taste was stronger with increasing fermentation time. Overall acceptability showed the highest score at 6 ℃ fermentation at 25 ℃, but no statistically significant difference was observed.

[Table 6]

As shown in FIG. 3g, the content of gastronine in the fermented extract of Chunmaj was decreased to 18.86 mg% at 12 hours after fermentation at 25 ° C and then decreased gradually at 37 ° C And showed a maximum value of 19.39 mg% at 24 hours after fermentation.

As shown in FIG. 3h, the content of vanillin in the fermented Chunmung extract increased until 12 hours after fermentation at 25 ° C. and showed a maximum content of 21.58 mg%. After fermentation at 37 ° C., Hour, and showed the highest content of 21.51 mg% and then decreased again.

In order to determine the optimal fermentation temperature and fermentation time, the antioxidant activity of the Lactobacillus plantarum (accession number: KCCM 12116) fermented with the fermentation broth was measured for 24 hours at 25 ° C and 37 ° C for 6 hours, As a result of functional analysis and sensory evaluation, the antioxidant activity was higher in fermentation at 25 ℃ than in 37 ℃ fermentation. When the fermentation time was less than 12 hours, the content of gastrodine was the highest at 25 ℃ for 12 hours. Therefore, optimum fermentation temperature and fermentation time at 25 ℃ were 12 hours.

As shown in Table 7, the antioxidative activity of the fermented Chunma fermented extract before fermentation and after fermentation showed that the DPPH radical scavenging activity and total phenol content were decreased by 2.18% and 1.14%, respectively, but the decrease was not significant , The reducing power was increased by 16.81%, the reducing power was increased by 12.57%, the SOD-like activity was increased by 207.62%, the total plono-void content was increased by 10.82%, and the lactic acid fermentation significantly increased the antioxidant activity there was.

As a result of the change of the functional components before and after the fermentation of the fermented Chunmung extract, the content of gastronin was increased by 16.20% and the content of vanillin was increased by 3.40% .

[Table 7]

1. Obtaining fermented milk extract

600ml of distilled water was added to 95.24g of horse chestnut, and the mixture was ground by adding 2.38g of Angelica gigantei and 2.38g of Angelica giganteus. The mixture was subjected to hot water extraction at 110 DEG C for 4 hours to obtain a chymase extract. After cooling, the mixture was filtered and diluted to 700 ml. Then, the mixture was sterilized at 121 DEG C for 15 minutes, and Lactobacillus plantarum (accession number: KCCM 12116) was inoculated (1%, v / v) as lactic acid bacteria. After the fermentation at 25 ° C. for 12 hours, a sample was collected and centrifuged (14,400 × g, 10 minutes) to obtain a supernatant of the fermented Chumma extract. The oligosaccharide was added thereto under the conditions as shown in Table 8 below As a result of preliminary experiments to improve the taste, it was judged that the added amount of oligosaccharide was less than 5%, and the sensory evaluation was performed by adding oligosaccharide to the fermented yeast extract at 0%, 3%, and 5%, respectively.

[Table 8]

2. Analysis method

Ten students of college and graduate students were examined for their taste sensations when they were ingested the extract of Chunmae, the lactic acid fermented extract obtained from lactic acid fermentation, and the Chungma fermented extract added with oligosaccharide.

The sensory test is a 5 point scale method (1 point: extremely weak or extremely disliked, 5 points: extremely strong or extremely good) for color, taste (sweetness, sour taste), smell Were used for the sensory evaluation.

3. Results

As shown in Table 9, no significant difference was observed in color and smell, and there was a difference in the addition of oligosaccharide in sweet and sour taste, and 3% of oligosaccharide was added The overall acceptability of the city was highest.

[Table 9]

1. Obtaining fermented milk extract

600ml of distilled water was added to 95.24g of horse chestnut, and the mixture was ground by adding 2.38g of Angelica gigantei and 2.38g of Angelica giganteus. The mixture was subjected to hot water extraction at 110 DEG C for 4 hours to obtain a chymase extract. After cooling, the mixture was filtered and diluted to 700 ml. Then, the mixture was sterilized at 121 DEG C for 15 minutes, and Lactobacillus plantarum (accession number: KCCM 12116) was inoculated (1%, v / v) as lactic acid bacteria. After the fermentation at 25 ° C for 12 hours, a sample was collected and centrifuged (14,400 × g, 10 minutes) to obtain a supernatant liquid of the fermented Chromosome extract. The oligosaccharide was added thereto in an amount of 3% ≪ / RTI > for 10 minutes.

2. Analysis method

The free sugar content was determined by mixing HLPC with 70% methanol in the same amount as the sample, centrifuging (14,400 × g, 10 minutes), filtering the supernatant with 0.22 μm syringe filter, Respectively.

[Table 10]

The supernatant obtained by centrifugation (14,400 × g, 10 minutes) was filtered with a 0.22 μm syringe filter and analyzed by HLPC under the conditions shown in Table 11 below. Respectively.

[Table 11]

Antioxidant activity samples were prepared by dilution with 0.02% BHT (butylated hydroxytoluene).

The DPPH radical assay was performed by Williams et al. (1995), and 1.0 ml of a 10-fold dilution of ethanol was added to 3 ml of 60 mM DPPH , And the reaction was carried out in the dark for 15 minutes. The absorbance at 517 nm was measured. The DPPH radical scavenging activity was calculated by the following equation and expressed as a percentage.

The ferric ion reducing antioxidant power (FRAP) assay was performed using the method of Heo et al. (2006), 300 mM acetate buffer (pH 3.6): 10 mM TPTZ solution: 20 mM FeCl ₃ .6H ₂ O solution at a ratio of 10: 1: 1 was added to 1.9 ml of the reaction mixture, and the reaction solution was reacted for 20 minutes. The absorbance of the solution was measured at 590 nm. The FRAP was FeSO ₄ · 7H ₂ O , And converted to mg% in terms of a calibration curve.

The reducing power was determined by the method of Oyaizu et al. (1986). 2.5 ml of 0.2 M phosphate buffer (pH 6.6) and 2.5 ml of 1% K ₃ Fe (CN) ₆ And 2.5 ml of 10% trichloroacetic acid was added thereto. After centrifugation, 2.5 ml of supernatant was obtained. 2.5 ml of distilled water and 0.5 ml of 0.1% FeCl ₃ were added to the supernatant. The absorbance at 700 nm was measured, and the difference in absorbance between the sample added and the non-added sample was expressed as a percentage.

SOD-like activity was measured by Marklund and Marklund (1974). To 0.2 ml of the sample, 3.0 ml of tri-HCl buffer and 0.2 ml of 7.2 mM pyrogallol were added and the mixture was incubated for 10 minutes After the reaction, 1.0 ml of 1N HCl was added to stop the reaction, and the absorbance at 420 nm was measured. The SOD-like activity was calculated by the following formula and expressed as a percentage.

The total phenolic compound (TPC) assay was performed using Dewanto et al. (2002) method. 5.0 ml of distilled water and 0.5 ml of folin-ciocalteu's phenol reagent were added to 0.1 ml of sample The reaction was carried out for 1 minute and then 1.5 ml of 10% Na ₂ CO ₃ was added. The reaction was carried out for 1 hour in a dark room and the absorbance was measured at 765 nm. The total phenol content (TPC) ㎍ / ml), and expressed as mg%.

Total flavonoid content was determined by adding 0.075 ml of 5% NaNO _{2 to} 0.5 ml of the sample, reacting the mixture at room temperature for 5 minutes, adding 0.05 ml of 10% AlCl ₃ for 5 minutes, adding 0.5 ml of ₁ M NaOH, And the absorbance at 510 nm was measured. The calibration curve was prepared using epicatechin as a reference material and expressed as mg%.

The contents of Gastrodin and Vanillin were determined by mixing the same amount of 70% methanol in the sampled samples and centrifuging (14,400 × g, 10 minutes). The supernatant was filtered with a 0.22 μm syringe filter, 1 and HLPC, respectively.

3. Results

As shown in Table 12 below, the pH of the chymus extract was 5.05, which decreased to 4.38 upon fermentation, and pH was not affected when oligosaccharide was added thereto. The sugar content was 2.3Brix in Chunma extract and decreased to 2.2Brix during fermentation and increased to 4.5Brix at 3% addition of oligosaccharide. The L value of Chumama extract was 16.94, the a value was -1.51 and the b value was -0.19. There was no significant difference in fermentation and addition of oligosaccharide after fermentation.

[Table 12]

As shown in Table 13 below, fructose, glucose and sucrose were detected in the free sugars of Chunma extract, and the free sugar content was decreased during fermentation. When 3% oligosaccharide was added thereto, the fructose content was 242.06 mg% 222.75mg%, sucross content was 182.17mg% and total free sugar content was 646.98mg%.

[Table 13]

Citric acid, malic acid, and succinic acid were detected as organic acids in the extract of Chumma extract as shown in Table 14, and the total organic acid content was increased and lactic acid was newly produced during fermentation. When 3% oligosaccharide was added, citric acid was 40.42mg%, malic acid was 141.15mg%, lactic acid was 56.13mg%, succinic acid was 16.54mg% and total organic acid content was 258.24mg%.

[Table 14]

As shown in FIG. 4A, the DPPH radical scavenging activity of the 10-fold diluted fermentation broth of Chumma showed no significant difference according to the addition of 3% oligosaccharide.

As shown in FIG. 4B, the iron reducing power of the fermented Chunmung extract decreased slightly from 17.02 mg% to 16.50 mg% when 3% oligosaccharide was added.

As shown in FIG. 4C, the reducing power of the fermented Angelica keiskei extract was 47.17% when 3% oligosaccharide was added, and the addition of 3% oligosaccharide did not show any significant effect.

As shown in FIG. 4D, the SOD-like activity of the fermented milk extract was 3.45% when 3% oligosaccharide was added, and the effect of adding 3% oligosaccharide was not significant.

As shown in FIG. 4E, the total phenol content of the fermented milk extract was 37.36 mg% when 3% of the oligosaccharide was added, and the addition of 3% of the oligosaccharide did not show any significant effect.

As shown in FIG. 4F, the total flavonoid content of the fermented milk extract was 4.83 mg% when 3% oligosaccharide was added and 0.51 mg% when 3% oligosaccharide was added.

As shown in FIG. 4g, the gastroin content of the fermented milk extract was 18.32 mg% when 3% oligosaccharide was added, and 0.72 mg% when 3% oligosaccharide was added.

As shown in FIG. 4h, the vanillin content of the fermented milk extract was 20.49 mg% when 3% oligosaccharide was added, and 0.93 mg% when 3% oligosaccharide was added.

As shown in Table 15 below, the antioxidant activity of Chunma beverage containing 3% of oligosaccharide added to the fermented Chunmung extract showed a slight decrease in the total phenol content compared with the chunma extract, but the DPPH radical scavenging activity, iron ion reductivity, , SOD - like activity, total flourboronide content were higher in.

Gastroin content in the functional ingredients of the beverage was 14.14% higher than that of the chymase extract and vanillin content was 1.69% higher.

[Table 15]

Finally, the fermentation of Chunma extract with lactic acid bacteria Lactobacillus plantarum (accession number: KCCM 12116) enhanced the content of antioxidant activity and functional ingredient, and also showed the effect of improving intestinal intolerance of Chunma.

As can be seen from the foregoing examples, the fermented milk extract and its preparation method having excellent antioxidative activity and favorable taste according to the present invention have high intrinsic functional ingredients such as gastronin and vanillin, It has high antioxidant activity and is beneficial to health. It is easy to ingest because it has excellent taste and flavor.

The present invention is not limited to the above-described embodiments. Anything having substantially the same constitution as the technical idea described in the claims of the present invention and achieving the same operational effect is included in the technical scope of the present invention.

Claims (13)

Which is obtained by inoculating lactic acid bacteria in an extract of Chunmei and fermenting the extract.
The method according to claim 1,
Wherein the chymase extract is obtained by hydrolyzing a mixture obtained by mixing 400 to 800 ml of distilled water per 100 g of horse chestnut.
3. The method of claim 2,
Wherein the mixture comprises 1 to 5 g of Angelica gigas per 100 g of horse chestnut and 1 to 5 g of Angelica gigas per se.
The method according to claim 1,
Wherein the lactic acid bacteria are any one of Lactobacillus plantarum (Accession No .: KCCM 11542), Lactobacillus plantarum (Accession No .: KCCM 12116), Bifidobacterium adolesentis (Accession No .: KCCM 11206), Bifidobacterium breve (Accession No .: KCCM 11208) Fermented Chungma Extract with Excellent Antioxidant Activity and Preferences.
The method according to claim 1,
And 3% by weight of an oligosaccharide as a whole.
400 to 800 ml of distilled water per 100 g of horse meat is mixed and ground to prepare a mixture;
Obtaining an extract to obtain a chymase extract by hot water extraction of the mixture;
A cooling and filtration step of cooling and filtering the chymase extract;
A sterilization step of sterilizing the filtered cholama extract;
An inoculation step of inoculating lactic acid bacteria into a fermentation medium to which sterilized chymase extract is added;
A fermentation step of fermenting the mixture at a temperature of 20 to 30 DEG C for 10 to 15 hours in a fermenter;
And obtaining a fermentation broth obtained by centrifuging the fermentation broth by centrifugation to obtain a supernatant fermentation broth.
The method according to claim 6,
Wherein the preparation of the mixture comprises adding 1 to 5 g of Angelica keiskei per 100 g of horse chestnut and 1 to 5 g of Angelica keiskei kansei.
The method according to claim 6,
Wherein the step of obtaining the extract is carried out at 100 to 120 ° C for 3 to 5 hours, wherein the extract has an antioxidant activity and a favorable taste.
The method according to claim 6,
Wherein the sterilization step is performed at 110 to 130 ° C for 10 to 20 minutes.
The method according to claim 6,
The lactic acid bacteria inoculated in the inoculation step are any of Lactobacillus plantarum (Accession No .: KCCM 11542), Lactobacillus plantarum (Accession No .: KCCM 12116), Bifidobacterium adolesentis (Accession No .: KCCM 11206), Bifidobacterium breve (Accession No .: KCCM 11208) Wherein the fermented milk is fermented in a fermentation broth.
The method according to claim 6,
Wherein the fermentation step is carried out at 25 DEG C for 12 hours, wherein the fermentation step has excellent antioxidative activity and favorable taste.
The method according to claim 6,
Wherein the step of obtaining the fermented extract is centrifuged at 14,400 x g for 5 to 15 minutes.
The method according to claim 6,
Wherein the fermentation broth is obtained by adding 3% by weight of oligosaccharide to the whole of the fermented extract, and then sterilizing the fermented broth.

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