KR20160007717A - Method for producing Phellinus linteus mycelium pressurized extract with increased antioxidant and whitening activity cultured in Mori ramulus medium - Google Patents

Abstract

The present invention relates to a method for preparing a Phellinus linteus mycelium extract with improved antioxidant and whitening activity, including steps of inoculating and culturing Phellinus linteus mycelium in a Mori Ramulus medium, drying and grinding the culture to obtain Phellinus linteus mycelium culture powder, and adding water to the powder and extracting the mixture under pressure; a Phellinus linteus mycelium extract with improved antioxidant and whitening activity prepared by the method; and a composition, a processed food and a cosmetic product containing the Phellinus linteus mycelium extract.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for producing an extract of pressed mushroom mycelium having increased antioxidative and whitening activity, which is cultured in an upper limb medium,

The invention upper limb (Mori lt ; RTI ID = 0.0 > Phellinus & lt ; / RTI > The present invention relates to a method for preparing an extract of Mycelium of mushroom mycelium enhanced in antioxidant and whitening activity, which comprises culturing the mushroom mycelium by inoculating and culturing the mushroom mycelium, linteus mycelium, A processed mushroom mycelium extract, and a composition containing the mushroom mycelial extract, processed food, and cosmetics, wherein the antioxidant and whitening activity produced by the above method is enhanced.

Excessive development of reactive oxygen species (ROS) or reactive nitrogen species (RNS), which cause aging and disease in the human respiration process, causes a serious imbalance between antioxidant defense systems Which causes oxidant stress and adversely affects the development and progression of various diseases. These free radicals are unstable because they are molecules with unpaired electrons, and they react easily with cellular constituents and cause lipid peroxidation, protein denaturation or inactivation, DNA synthesis inhibition, mutagenesis, cancer induction and enzyme inactivation And so on. The body maintains its homeostasis by producing various antioxidants that can prevent oxidation in the body. However, since the antioxidant system in the body can not defend all the oxides in large amount, addition of antioxidants to prevent oxidation in the body prevents aging and disease prevention very important.

Mushrooms accumulate secondary metabolites such as phenolic compounds, polypeptides, terpenes and steroids, and especially phenolic compounds obtained from mushrooms have excellent antioxidative effects, The antioxidant activity increases depending on the content in a concentration-dependent manner. Interest in mushrooms has been heightened by the report that mushrooms prevent and improve chronic degenerative diseases such as cancer, stroke, heart disease and diabetes. Mushroom mycelium contains more than four times more nutrients than fruiting body. In particular, the mycelium contains 50 ~ 60 times more medicinal components such as anti-cancer component and immune function enhancing component than fruity body. have. Antimicrobial, Antioxidant and Anticancer Activity of Mixed Culture Extract of Mushroom Mycelium by the Development of Mushroom Mycelial Extracts, Mycelial Function of Mushroom Using Mycelia of Mycelial Culture of Ginseng Cultured Rice, Germinated Brown Rice Immersed in Cultured Mycelium of Mycelia, .

Phelinus linteus ) is a white rot fungus belonging to the genus Hymenochaetaceae and Phellinus taxonomically , and a similar type of dried mud mushroom ( Phellinus gilvus ), Phellinus isniarius , Phellinus robustus ), black mud mushroom ( Phellinus nigricans ), larch mushroom ( Phellinus pini ). As a result, many studies have been carried out in the field of mushroom's hot-water extract of fruiting body, known to have an inhibitory effect on cancer of digestive system, and various kinds of physiological activity of mushroom such as anticancer activity and inhibitory effect of intestinal bacterial harmful enzymes such as colon cancer and bladder cancer, .

The upper limb (桑枝, Mori ramulus ) is a deciduous tree belonging to Moraceae ( Morus alba L.), which was cut in late spring and early summer. The constellation is also prepared in a long columnar shape. Its length is not constant but its diameter is 0.51 cm. The surface is yellowish brown or yellowish brown with many yellowish brown papules and thin vertical wrinkles, with traces of slightly semicircular leaves of grayish white color and yellowish brown masses. The vagina is hard, the folded surface is fibrous, and it is distributed throughout Korea and China.

Korean Patent Laid-Open Publication No. 2000-0072608 discloses an antioxidant composition having a antioxidant activity, and Korean Patent No. 1150226 discloses an antioxidant composition containing a carboxymethylhydrolyzate extract. However, And the whitening activity of the mushroom mycelial extract was increased.

The present invention has been made in view of the above needs, and it is an object of the present invention to provide a process for extracting mycelia of mushroom mycelium, comprising the steps of: The present invention aims at establishing a method for preparing a mushroom mycelium extract having improved physiological activity such as antioxidant activity, tyrosinase inhibiting activity and reducing power in the extract.

In order to solve the above problems, the present invention is the upper limbs (Mori lt ; RTI ID = 0.0 > Phellinus & lt ; / RTI > linteus mycelium), culturing the cells, and then adding water to the cultured powder of the dried and ground condition mycelium, followed by pressurized extraction. The method for producing the extract of Mycelium of mushroom mycelia enhanced the antioxidant and whitening activity, to provide.

In addition, the present invention provides a mycelium extract of Mushroom mycelium prepared by the above method and having enhanced antioxidant and whitening activity.

In addition, the present invention provides a composition for promoting antioxidative and whitening activity, which contains the mycelium of mycelia of mushrooms as an active ingredient.

Further, the present invention provides a processed food containing the mycelia of mycelia of mushrooms.

In addition, the present invention provides a cosmetic product containing the mycelium of mycelia of the situation.

The mycelial mycelium extract prepared by the method of the present invention is extracted using a mushroom mycelium cultured using a top medium having a superior antioxidative activity as a solid medium to thereby enhance antioxidative activity and extract it using a pressurized extraction method , It is advantageous in that it is safer, eco-friendly and has a simple extraction process as compared with the conventional organic solvent extraction method. In addition, it is effective to extract antioxidative activity, tyrosinase inhibitory activity, and other functionalities effectively from the mycelia of mycelium of mushroom and effective for anti-aging and whitening. Therefore, the mushroom mycelium extract can be effectively used in food, medicine, .

1 is a graph comparing DPPH, ABTS and superoxide radical scavenging activity of the upper limb extract.
HE: hot water extract, EE: 50% ethanol extract, UE: ultrasonic extract, PE: pressure extract
2 is a graph comparing FRAP and reducing power of the upper limb extract.
HE: hot water extract, EE: 50% ethanol extract, UE: ultrasonic extract, PE: pressure extract
FIG. 3 is a graph comparing DPPH and ABTS radical scavenging activities of mycelia extracts of the mushroom cultured by an extraction process in the upper limb.
HE: hot water extract, UE: ultrasonic extract, PE: pressure extract
FIG. 4 is a graph comparing superoxide radical scavenging activities of mycelium extracts of the mushroom mycelium according to the extraction process cultivated in the upper limbs.
HE: hot water extract, UE: ultrasonic extract, PE: pressure extract
FIG. 5 is a graph comparing the FRAP and the reducing power of the mycelium mycelium extract according to the extraction process cultured in the upper limb.
HE: hot water extract, UE: ultrasonic extract, PE: pressure extract
FIG. 6 is a graph comparing tyrosinase inhibitory activities of mycelium extracts of the mushroom mycelium according to the extraction process cultured in the upper limbs.
HE: hot water extract, UE: ultrasonic extract, PE: pressure extract

According to an aspect of the invention there is provided the upper limbs (Mori lt ; RTI ID = 0.0 > Phellinus & lt ; / RTI > linteus mycelium), culturing the cells, and then adding water to the cultured powder of the dried and ground condition mycelium, followed by pressurized extraction. The method for producing the extract of Mycelium of mushroom mycelia enhanced the antioxidant and whitening activity, to provide.

In the method for producing the mycelium extract of the present invention, the cultivation of the mushroom is generally performed by cutting the log, inoculating the mushroom seeds on the cut surface, culturing the mushroom for about 13 months, removing the seeds, Fruit bodies are formed for 15 to 20 days for artificial cultivation. However, in the present invention, a top culture medium having excellent antioxidative activity was used in order to utilize mycelium more effective than fruiting body. Preferably, the upper medium is prepared by sterilizing the dried upper ground powder at 110-130 ° C and 1.1-1.3 atm for 50-70 minutes, more preferably drying the top powder at 121 ° C And then sterilized at a temperature of 1.2 atm for 60 minutes. Culturing the mycelia of the mushroom using the above-prepared upper culture medium was able to produce the mushroom mycelium extract having enhanced functionality such as antioxidant activity.

In addition, in the method of the present invention for extracting the mycelium of mushroom mycobacteria, the pressurized extraction is preferably carried out at a temperature of 110 to 130 ° C. and a pressure of 1.1 to 1.3 atm for 2 to 4 hours, more preferably at a temperature of 121 ° C. It can be extracted for 3 hours at 1.2 atm. Extracting the mycelium extract of mushroom mycelium under the above conditions not only enhances the extraction yield, total phenol and total flavonoid content but also enhances the physiological activity effects such as antioxidant and whitening activity as compared with extraction by other methods and conditions Mycelium extract could be prepared.

The method for producing the mycelia of mycelia of mushrooms of the present invention, more specifically,

(a) preparing a top culture medium by sterilizing the dried upper ground powder at a temperature of 110 to 130 DEG C and a pressure of 1.1 to 1.3 atm for 50 to 70 minutes;

(b) culturing the mycelia of mushroom on the upper medium prepared in step (a), culturing the mushroom mycelium at 20 ~ 30 ° C for 10 ~ 20 days, drying and pulverizing the mushroom mycelia to prepare a culture conditional mycelium; And

(c) extracting the cultured conditional mycelia powder of step (b) at a temperature of 110 to 130 ° C. and 1.1 to 1.3 atm for 2 to 4 hours,

More specifically,

(a) sterilizing the dried upper ground powder at 121 DEG C and 1.2 atm for 60 minutes to prepare a top culture medium;

(b) cultivating the mycelia of mycelium of mushroom in the upper medium prepared in the step (a), culturing the mushroom mycelium at 25 ° C for 15 days, drying and pulverizing the mushroom mycelia to prepare a culture conditional mycelium; And

(c) extracting the cultured conditional mycelium powder obtained in step (b) at 121 ° C and 1.2 atm for 3 hours.

The present invention also provides a mycelium extract of Mushroom mycelium prepared by the above method and having enhanced antioxidant and whitening activity.

The present invention also provides a composition for promoting antioxidative and whitening activity, which comprises the above mycelium extract of Mycelia mushroom as an active ingredient.

The present invention also provides a processed food containing the mycelium of mycelia of the situation mushroom. There is no particular limitation on the kind of the processed food. Examples of the food to which the mycelium of mycelia of the present invention can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , A drink, an alcoholic beverage, and a vitamin complex, all of which are processed foods in a conventional sense.

The present invention also provides cosmetics containing the above mycelium of mycelia of mushrooms. The mushroom mycelial extract of the present invention, which is an active ingredient of the cosmetic of the present invention, has a tyrosinase-inhibiting activity and thus can be used for whitening purposes and has antioxidant activity, so that it can be used for antioxidant use. The cosmetics of the present invention can be used in cosmetics such as soft longevity, astringent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, ≪ / RTI > body essence, and the like.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

1. Experimental material

The upper limbs used in this experiment (Kuji, Mori ramulus ) were harvested from Korea, and mycelial mycelia ( Phellinus linteus mycelium) was supplied from Ryuchunghyang medicinal mushroom in Andong, Gyeongbuk Province. Mycelial cultivation was performed by cutting and crushing the upper limbs with 30 ~ 40% moisture by natural drying on the shade, sterilization at 121 캜 and 1.2 atm for 1 hour, and used as a solid medium. Liquid seeds were prepared by inoculating 5% (v / w) of liquid seeds in a solid medium and incubating at 25 ° C for 15 days. The cells were hot-air dried at 50 ° C for 24 hours. The dried sample was pulverized with a pulverizer (FM-909W, Hanil, Co., Korea) and stored in a dark room at -20 ° C or lower, and used as a sample for extraction.

2. Preparation of upper limb extract

30 g of distilled water was added to the upper part of the powder at a ratio of 20 times as much as the solids. The mixture was subjected to hot water extraction (HE), 50% ethanol extraction (EE), ultrasonic extraction (UE) Extracts were prepared by PE (pressurized extraction) method.

Extraction of hot water and extraction of 50% ethanol were carried out by adding distilled water or 50% ethanol to the ground sample and heating at 80 ° C for 3 hours using a reflux condenser (CA-1112, Eyela Co., Japan). Ultrasonic extraction was performed for 3 hours in a 40 kHz ultrasonic bath (JAC-4020, KODO Co., Korea) filled with water by adding distilled water to the ground sample. At this time, the bottom surface of the Erlenmeyer flask was not contacted with the ultrasonic bath, and the extract and the solvent in the Erlenmeyer flask were completely immersed in water. Pressurized extraction was carried out at 121 ℃ and 1.2 atm for 3 hours by adding distilled water to the ground sample. Each extract was filtered using a filter paper (No. 1, Whatman International Ltd., England) to remove impurities. The filtered solution was concentrated under reduced pressure using a vacuum evaporator (Model N-1N, Eyela Co., Japan), and lyophilized (FreeZone 2.5, Labconco Co., USA) And used as a sample.

3. Production of Mycelia Extract of Situ Mushrooms Cultured in Upper Limb

30 g of mushroom powder was added 20 times as much as the solid content of distilled water and the extract was prepared by hot water extraction (HE), ultrasonic extraction (UE), and pressured extraction (PE) Respectively.

The hot water extraction was performed by adding distilled water to the ground sample and extracting it at 80 ° C for 3 hours using a reflux condenser (CA-1112, Eyela Co., Japan). Ultrasonic extraction was performed for 3 hours in a 40 kHz ultrasonic bath (JAC-4020, KODO Co., Korea) filled with water by adding distilled water to the ground sample. At this time, the bottom surface of the Erlenmeyer flask was not contacted with the ultrasonic bath, and the extract and the solvent in the Erlenmeyer flask were completely immersed in water. Pressurized extraction was carried out at 121 ℃ and 1.2 atm for 3 hours by adding distilled water to the ground sample. Each extract was filtered using a filter paper (No. 1, Whatman International Ltd., England) to remove impurities. The filtered solution was concentrated under reduced pressure using a vacuum evaporator (Model N-1N, Eyela Co., Japan), and lyophilized (FreeZone 2.5, Labconco Co., USA) And used as a sample.

4. Experimental Method

(1) extraction yield and total phenol content

The extraction yield was determined by lyophilizing each extract to determine the weight of the building and then expressed as a percentage of the amount of raw material used in the preparation of the extract.

The total phenol content was determined by mixing 1 mL of 1 N Folin ciocalteu reagent with 1 mL of the sample according to the Folin-Denis method, adding 1 mL of 20% Na ₂ CO _3, reacting for 30 minutes at room temperature, Absorbance was measured at 725 nm using Ultraspec 2100pro, Amersham Co., Sweden). The total phenolic content was calculated from the standard curve prepared by quantifying tannic acid (Sigma Co., USA).

(2) Total flavonoid content

The total flavonoid content was measured by mixing 150 μl of 5% NaNO ₂ in 1 ml of the sample, reacting at room temperature for 6 minutes, adding 300 μl of 10% AlCl _3, reacting at room temperature for 5 minutes, mixing with 1 ml of 1 N NaOH Absorbance was measured at 510 nm using a spectrophotometer (Ultraspec 2100pro, Amersham Co.). The total flavonoid content was calculated from the standard curve prepared by quantifying the routine (Sigma Co., USA).

(3) Measurement of ORAC (Oxygen radical absorbance capacity)

In this experiment, Neutral Phosphate Buffer (61.6: 38.9 v / v, 0.75 MK ₂ HPO and 0.75 M NaH ₂ PO ₄ ) was used for the dilution of the test and standard solutions and the preparation of experimental samples. To prepare a calibration curve, Trolox (Water soluble analogue of vitamin E, 6-hydroxy-2,5,7,8-tetramethlychroman-2-carboxylic acid, Sigma Chem, Inc., St. Louis, MO , USA) was dissolved in 50 mL of phosphate buffer, and the mixture was excited at 485 nm and emitted at 538 nm.

(4) Measurement of DPPH radical scavenging activity

The DPPH radical scavenging activity was measured by using the reducing power of DPPH (1,1-diphenyl-2-picrylhydrazyl). That is, 5 mL of 4 × 10 ^-4 M DPPH solution (dissolved in 99.9% ethyl alcohol) was added to 0.5 mL of the sample, and the mixture was reacted at room temperature for 15 minutes. Then, the reaction was carried out at 517 nm using a spectrophotometer (Ultraspec 2100pro, Amersham Co.) Absorbance was measured. The DPPH radical scavenging activity was expressed as a percentage before and after the addition of the extract as shown below.

DPPH radical scavenging activity (%) = (1 - sample absorbance / control absorbance) × 100

(5) Measurement of ABTS radical scavenging activity

The ABTS radical scavenging activity was measured by mixing 7.4 mM 2,2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS, Sigma Co., USA) and 2.45 mM potassium persulfate After incubation in room temperature for 24 hours, ABTS + · was formed and diluted with PBS (phosphate buffer saline, pH 7.4) to absorbance value of 0.70 ± 0.02 at 734 nm. 20 μl of the sample was mixed with 180 μl of the diluted solution, reacted for exactly 1 minute, and the absorbance was measured at 732 nm. The ABTS radical scavenging activity was expressed as a percentage before and after the addition of the extract.

ABTS radical scavenging activity (%) = (1 - sample absorbance / control absorbance) × 100

(6) Measurement of superoxide radical scavenging activity

To measure the superoxide radical scavenging activity, 500 μl of a sample was mixed with 100 μl of 0.1 M Tris-HCl buffer (pH 8.5) and 200 μl of 100 μM PMS, followed by 200 μl of 500 μM NBT and 400 μl of 500 μM NADH After incubation at room temperature for 10 minutes, the absorbance was measured at 560 nm using a spectrophotometer (Ultraspec 2100pro, Amersham Co.). The superoxide radical scavenging activity was expressed as a percentage before and after addition of the extract as shown below.

Superoxide radical scavenging ability (%) = (1 - sample absorbance / control absorbance) x 100

(7) Measurement of ferric reducing antioxidant power (FRAP)

The FRAP reagent was prepared by heating 25 mL of acetate buffer (300 mM, pH 3.6) at 37 ° C and adding 10 mM 2,4,6-tris (2-pyridyl) -s-triazine (TPTZ, Sigma Co , USA) and 2.5 mL of 20 mM ferric chloride (FeCl ₃ ). 900 μl of the prepared FRAP reagent and 90 μl of distilled water were added to 30 μl of the sample, reacted at 37 ° C. for 10 minutes, and the absorbance was measured at 510 nm using a spectrophotometer (Ultraspec 2100pro, Amersham Co.). FRAP was calculated from the standard curve prepared by quantifying FeSO _{4 ㎍} · 7H ₂ O (Sigma Co., USA).

(8) Measurement of reducing power

To measure the reducing power, 2.5 mL of 0.2 M phosphate buffer (pH 6.6) and 2.5 mL of 1% potassium ferricyanide solution were added to 1 mL of the sample, followed by reaction at 50 ° C for 30 minutes. After the reaction, 2.5 mL of 10% trichloroacetic acid solution was added, and 2.5 mL of the supernatant was added to 2.5 mL of distilled water and 0.1% FeCl ₃ (Ultraspec 2100pro, Amersham Co.), and the absorbance at 700 nm was measured using a spectrophotometer (Ultraspec 2100pro, Amersham Co.).

(9) Measurement of tyrosinase inhibitory activity

Tyrosinase inhibitory activity was determined by adding 100 μl of a sample, 40 μl of 0.175 M phosphate buffer (pH 6.8), and 40 μl of 5 mM L-DOPA to a 96-well plate in the order of mushroom tyrosinase (2,000 U / ml, Sigma Co., USA), and incubated at 37 ° C for 10 minutes. The resulting DOPA chromium was measured for absorbance at 490 nm using a microplate reader (UVM-340, ASYS Co., Austria). The inhibitory activity of tyrosinase was expressed as a percentage before and after the addition of the extract as shown below.

(%) = (1 - sample absorbance / control absorbance) x 100

(10) Statistical processing

The ANOVA was performed using SPSS (version 19.0, SPSS Inc., Chicago, IL, USA) and the significance of the mean values (p <0.05) was tested by Duncans multiple range test .

Example  1: Extraction yield, total phenol content, total flavonoid content, ORAC

Table 1 shows the extraction yield, total phenol, total flavonoid content and ORAC of the upper limb extract according to the extraction process. The extraction yields were significantly different in hot water extraction (HE), 50% ethanol extraction (EE), ultrasonic extraction (UE), and pressured extraction (PE) , PE (3.07%), UE (1.43%), HE (1.18%) and EE (1.07%) were the highest yields. Total phenol and flavonoid contents were found to be 33.47 g and 3.56 g per 100 g of EE, respectively, and it was confirmed that the extraction efficiency of the phenolic substance was the highest when the upper part was extracted with 50% ethanol. 50% ethanol is considered to be due to the easy dissolution of the phenolic substance contained in the upper part since water and the organic solvent are mixed. The ORAC of the upper limb according to the extraction method was measured at a concentration of 50 ㎍ / mL. As a result of the ORAC measurement, the highest value was 3483.37 uM per g in EE, HE 2687.52 uM, UE 2300.45 uM and PE 2117.62 uM Respectively. As shown in Table 1, the ORAC value was highest in the upper 50% ethanol extract (33.47 g / 100 g) having a relatively high phenolic compound content.

Extraction yield, total phenol, total flavonoid content and ORAC Extraction method ^{1 )} yield
(dry basis,%) Total phenol
(Tannic acid g / 100 g) Total flavonoid
(Routine g / 100 g) ORAC
(Trolox uM / g FW) HE 1.18 ± 0.04 ^{c2 )} 24.71 + - 0.23 ^b 3.53 + - 0.12 ^a 2687.52 ± 221.44 ^b EE 1.07 ± 0.02 ^d 33.47 ± 0.06 ^a 3.56 ± 0.07 ^a 3483.37 ± 189.91 ^a UE 1.43 ± 0.05 ^b 20.66 ± 0.15 ^d 3.45 ± 0.32 ^a 2300.45 ± 176.36 ^c PE 3.07 ± 0.09 ^a 21.08 + - 0.13 ^c 3.36 + 0.24 ^a 2117.62 ± 178.35 ^c

^One) HE: hot water extraction, EE: 50% ethanol extraction, UE: ultrasonic extraction, PE: pressure extraction

²⁾ The same superscript in each column means no significant difference ( p <0.05)

Example  2: Topical extract DPPH , ABTS  And Superoxide Radical  Scavenging activity

In the extraction process The upper DPPH, ABTS and superoxide radical scavenging activities of the upper limb are shown in Fig. The DPPH radical scavenging activity of the upper part of the upper part of the extract was 17.34 ~ 65.84% in the 50% ethanol extract (EE). Especially, the concentration of ascorbic acid (77.49%) as a positive control at 1,000 ㎍ / 0.85 times the activity. The ABTS radical scavenging activity increased with increasing concentrations in all samples. Especially, it showed the highest activity in EE (42.35 ~ 97.52%) and showed similar tendency to DPPH radical scavenging activity. However, it was confirmed that ABTS radical scavenging activity was higher than DPPH radical scavenging activity. In contrast to DPPH radical scavenging activity by receiving hydrogen or electron, in the case of ABTS radical scavenging activity, hydrogen donating antioxidant and chain breaking antioxidant It is generally considered that the DPPH radical scavenging activity is higher than that of DPPH radical scavenging activity. The superoxide radical scavenging activity showed more than 80% scavenging activity from 250 ㎍ / mL concentration, and the scavenging activity was 62.56 ~ 98.26% in all samples. The radical scavenging activity of the upper limb according to the extraction method was found to be different according to the radicals used in each experiment. It is also considered that there is a difference in the elution of substances having inhibitory activity.

Example  3: Topical extract FRAP  And reducing power

The ferric reducing antioxidant power (FRAP) and reducing power of the upper limb according to the extraction process are shown in FIG. The content of FRAP was the highest at 189.00 ~ 974.80 uM in 50% ethanol extract (EE). Especially, at 1,000 ㎍ / mL concentration, FRAP showed about 0.8 times effect of ascorbic acid (1,239.88 uM) used as a positive control. Reducing power was increased in all the samples in a dose - dependent manner. Especially, EE at 0.8 ㎍ / mL showed similar effect to ascorbic acid (0.783) used as a positive control.

As a result of six different antioxidant activity analyzes of Examples 1 to 3, it is considered that all of the extraction methods applied in the present invention are sufficient for extracting antioxidant components. In this study, we propose an effective extraction method for extracting the mushroom mycelium cultured in the upper limb by inoculating the liquid seeds using the upper part of the upper surface showing excellent antioxidant activity as a solid medium, .

Example  4: Extraction yield, total phenol content, total flavonoid content, ORAC

Table 2 shows the extraction yield, total phenol, total flavonoid content and ORAC of the mushroom mycelium cultured in the upper limb according to the extraction process. The extraction yields were 5.44%, 6.03% and 7.73% in hot water extraction (HE), ultrasonic extraction (UE) and pressurized extraction (PE) Pressure extraction, ultrasonic extraction, and hot water extraction. Total phenol and flavonoid contents were highest at 9.87 g and 1.90 g per 100 g of PE, respectively, and the lowest contents were 3.18 g and 1.03 g per 100 g of UE, respectively. Since the phenolic compound has a phenolic hydroxyl group, it has a property of binding to proteins and other macromolecules, and is known to have a physiological activity function such as antioxidant activity. Therefore, when the content of the phenolic compound contained in the extract is high It is considered that the antioxidant activity of PE is high. The ORAC of mushroom mycelium extract was determined at the concentration of 100 ㎍ / mL according to the extraction process cultivated in upper limb. PE was 769.63 uM per g, and HE was 622.96 uM and UE was 249.06 uM. Especially, the content of total phenolics and total flavonoids were high in the extracts, regardless of the high extraction yield.

Extraction yield, total phenol, total flavonoid content and ORAC Extraction method ^{1 )} yield
(dry basis,%) Total phenol
(Tannic acid g / 100 g) Total flavonoid
(Routine g / 100 g) ORAC
(Trolox uM / g FW) HE 5.44 + 0.07 ^{c2 )} 8.30 ± 0.14 ^b 1.71 ± 0.09 ^a 622.96 ± 37.41 ^b UE 6.03 ± 0.14 ^b 3.18 ± 0.05 ^c 1.03 + - 0.20 ^b 249.06 + - 41.59 ^c PE 7.73 ± 0.11 ^a 9.87 ± 0.16 ^a 1.90 + 0.08 ^a 769.63 ± 44.77 ^a

^One) HE: hot water extraction, UE: ultrasonic extraction, PE: pressure extraction

²⁾ The same superscript in each column means no significant difference ( p <0.05)

Example  5: Mycelium extract of mycelia DPPH  And ABTS Radical  Scavenging activity

The DPPH and ABTS radical scavenging activities of the mycelium extracts of the mushroom by the extraction process cultured in the upper limb are shown in FIG. The DPPH radical scavenging activity of the extracts of the mushroom mycelium cultured on the upper limb according to the extraction process showed the highest activity from 13.04 to 35.85% in the pressurized extract (PE) and the lowest activity from 6.34 to 12.91% in the ultrasonic extract (UE) Respectively. The ABTS radical scavenging activity assay is characterized by its ability to measure both the antioxidant activity of polar and nonpolar samples and thus has a broader application range than DPPH radical scavenging activity. The ABTS radical scavenging activity of mushroom mycelial extracts grown on the upper limbs by the extraction process increased with increasing concentrations in all samples. Especially, the concentration of ascorbic acid (92.55%), which was used as a positive control at 1,000 ㎍ / Which is about 0.55 times greater than that of the control. ABTS radical scavenging activity was higher than DPPH radical scavenging activity in all samples because radical scavenging mechanism was different and substrate binding degree was different.

Example  6: Extract of Mycelium of Situ mushroom Superoxide Radical  Scavenging activity

The superoxide radical scavenging activity of the mycelium mycelium extract according to the extraction process cultivated in the upper limb is shown in Fig. Superoxide radicals, which are generated in various electron transport systems in vivo, are known to cause damage and death of cells and tissues as a major causative factor of reactive oxygen species generation. The superoxide radical scavenging activity of mushroom mycelial extracts cultured in the upper limb following the extraction process was found to be 29.16 ~ 96.03% higher in all samples. Especially, at the concentration of 500 ㎍ / mL of all samples, the activity of ascorbic acid (87.02%) was higher than that of ascorbic acid (87.02%) which was used as a positive control, and it was confirmed that superoxide radical scavenging activity of the mycelia extract was superior.

Example  7: Mycelium extract of mycelia FRAP  And reducing power

The ferric reducing antioxidant power (FRAP) and the reducing power of the mycelia extract of the mushroom according to the extraction process cultivated in the upper limb are shown in FIG. The content of FRAP of the mycelium extract of the mushroom cultured in the upper limb was the highest at 180.60 ~ 607.93 uM in pressurized extract (PE), 126.39 ~ 453.44 uM in hydrothermal extract (HE) and 70.40 ~ 140.13 uM , Which was increased in a concentration-dependent manner in all samples. The reducing power of culture supernatant of the mushroom mycelium by extraction was 0.51, 0.44 and 0.27 in PE, HE and UE at 1,000 ㎍ / mL, respectively. As the sample concentration increased, the reducing power was significantly increased and showed similar tendency to FRAP.

It is considered that the excellent antioxidant activity of the pressurized extract is due to the content of phenol and flavonoid according to the extraction process. Considering that it is an extract which is not a single component, it is possible to obtain an antioxidant .

Example  8: Tyrosinase elimination activity of mycelium extract

The tyrosinase inhibitory activity of the mycelium mycelium extract according to the extraction process cultivated in the upper limb is shown in Fig. Melanin production of skin causing spots or erythema is a series of oxidative polymerization reactions starting from tyrosine, a relatively stable material. In this process, tyrosinase plays an important role and is oxidized to melanin pigment through DOPA chromium, which is an intermediate produced by the action of this enzyme. Therefore, it is considered that the activity of tyrosinase enzyme is inhibited or the oxidation reaction of its intermediates is inhibited, thereby reducing the melanin pigment. The tyrosinase inhibitory activity of mushroom mycelial extracts cultured on the upper legs according to the extraction process was 24.05%, 19.43% and 13.52% at 500 ㎍ / mL in the pressurized extract (PE), hot water extract (HE) and ultrasonic extract % Of total extracts. As the sample concentration increased, the tyrosinase inhibitory activity was significantly increased, which was similar to the antioxidant activity. Ascorbic acid, known as a natural substance with high tyrosinase inhibitory activity, showed a high activity of 78% or more at 250 ㎍ / mL. Therefore, it was confirmed that the hot - water extract of Mycelia mushroom cultivated in the upper limb showed high antioxidative and tyrosinase inhibitory activity during the pressure extraction process.

Taken together, these results indicate that the pressurized extraction process of the mycelia of mushroom cultured on the upper limb, which exhibits high antioxidant activity and tyrosinase inhibitory activity, can be used to develop antioxidant-functional food materials and cosmetic materials capable of inhibiting melanin biosynthesis .

Claims (7)

Mori lt ; RTI ID = 0.0 &gt; Phellinus & lt ; / RTI &gt; linteus mycelium), culturing the cells, and then adding water to the culture powder obtained by drying and pulverizing the mycelium, followed by pressure extraction. [Claim 2] The method according to claim 1, wherein the pressure extraction is performed at 110-130 &lt; [deg.] &Gt; C and 1.1-1.3 atm for 2-4 hours. 3. The method of claim 2,
(a) preparing a top culture medium by sterilizing the dried upper ground powder at a temperature of 110 to 130 DEG C and a pressure of 1.1 to 1.3 atm for 50 to 70 minutes;
(b) culturing the mycelia of mushroom on the upper medium prepared in step (a), culturing the mushroom mycelium at 20 ~ 30 ° C for 10 ~ 20 days, drying and pulverizing the mushroom mycelia to prepare a culture conditional mycelium; And
(c) adding water to the cultured conditional mycelial powder prepared in step (b), and extracting the mixture at 110 to 130 ° C. and 1.1 to 1.3 atm for 2 to 4 hours. A method for producing an extract of Mycelia mushroom having enhanced whitening activity. 4. An extract of Mycelium of mushroom having enhanced antioxidant and whitening activity, prepared by the method of any one of claims 1 to 3. A composition for promoting antioxidative and whitening activity, which comprises the extract of Mycelia mushroom of claim 4 as an active ingredient. Processed food containing the mycelium of mycelia extract of claim 4. A cosmetic product containing the mycelium of mycelia of the situation of claim 4.

Images