KR20050054696A - Basidiomycetes-fermented cereal effective in adjusting blood sugar level - Google Patents

Abstract

The present invention is prepared by culturing Paecilomyces japonica (Pecilomyces japonica) in grains, comprising a mushroom culture grains, a method for producing an electric mushroom culture grains and an electric mushroom culture grains exhibiting a glycemic control effect, blood glucose lowering It relates to a food product having an effect. Mushroom culture grains of the present invention inhibits the production of glucose, and lowers the influx of the generated glucose in the body, it can be widely used in the development of functional foods exhibiting a glycemic control effect.

Description

Mushroom culture grains showing glycemic control effect {Basidiomycetes-fermented Cereal Effective in Adjusting Blood Sugar Level}

The present invention relates to a mushroom culture grain exhibiting a glycemic control effect, more specifically, the present invention is prepared by culturing Paecilomyces japonica in grains, mushroom culture grains exhibiting a glycemic control effect, electric The present invention relates to a method for producing a mushroom culture grain, and a food containing an electric mushroom culture grain as a main agent and exhibiting a hypoglycemic effect.

Modern medicine has not yet been developed to treat diabetes, and it is known that the best treatment is to keep blood sugar at a normal level (see Strowig, S. and Raskin, P., Diabetes Care). , 15: 1126, 1992).

Currently, as a treatment method for diabetes, drug therapy is being attempted along with diet and exercise therapy, and drugs used in clinical practice include insulin preparation, sulfonylurea preparation, biguanide preparation, and troglitazone preparation. The sulfonylurea preparation, which is mainly used as an oral hypoglycemic agent, may cause β-cell exhaustion in the long term, and the risk of hypoglycemia as a side effect, and the biguanide metformin is also an excellent drug, but sometimes it may cause fatal side effects such as lactic acid. Groop, LC, Diebetes Care, 15: 737-754, 1992). In addition, insulin injection therapy is not only inconvenient for patients, but also has a low risk of hypoglycemia and promotes obesity in long-term use, which may cause difficulty in managing long-term blood glucose levels in patients with insulin-independent diabetes (see Koivisto, VA, Diabetes Care, 16:29). -39, 1993).

Recently, there is a tendency to maintain blood sugar in a normal state through folk remedies or natural foods, in addition to drug treatments that are concerned about side effects, and researches to develop blood glucose lowering materials from natural products and food ingredients have been actively conducted. It has been confirmed that the extracts of chicken berry grass, arborella and Hyunsam, which have been widely used in folk medicine, have a hypoglycemic effect in normal or hyperglycemic-induced mice. Effects have been identified (Kim, OK, Kor. J. Pharmacogn., 22: 225-232, 1991; Kim, OK, Kor. J. Pharmacogn., 23: 117-119, 1992; Kim, JY , JK Diabetes Assoc., 18: 377-381, 1994).

More than 8,000 mushroom resources are among the higher fungi, and 20 of them have been used for food and medicinal use since ancient times. Recently, researches on bioactive substances and functional ingredients of mushrooms have been actively conducted worldwide. , Blood pressure lowering, thrombolysis, and blood cholesterol lowering have been scientifically proven, and some medicinal mushrooms have been reported to have hypoglycemic effects, and efforts are being made to study mushrooms for the treatment of diabetes. See: Lucas, EH and Ringler, RL, Antibiotics and Chemotheraphy, 7: 1-4, 1957; Chihara, G. et al., Cancer Res., 30: 2776-2781, 1970; Nagata, J. et al., J. Agric. Food Chem., 49: 4965-4970, 2001; Zhuang, C. et al., J. Jpn. Soc. Food Sci.Technol., 41: 724-732, 1994; Watanabe, T. et al. ., J. Jap. Soc. Food Sci. Technol., 49: 166-173, 2002; Sumi, H. et al., J. Jpn. Soc. Food Sci.Technol., 43: 318-321, 19. 96; Cheung, PCK, Nutr.Res., 16: 1953-1957, 1996; Sugiyama, K. et al., J. Jap.Soc.Nutr.Food Sci., 45: 265-270, 1992; Kurasawa, SI et al., Nutr. Reports International, 26: 167-173, 1982; Ukawa, Y. et al., J. Nutr. Sci. Vit., 48: 73-76, 2002; Yuan, Z. et al., Biosci. Biotechnol. Biochem., 62: 1898-1903, 1998; Kiho, T. et al., Carbohydr.Res., 251: 81-87, 1994).

However, until now, mushrooms have not been directly used for the treatment of diabetes mellitus, which is not easy for common sense of medicinal mushrooms, and there are very small amounts of active ingredients that have a hypoglycemic effect in medicinal mushrooms, It takes time. However, if the mushroom can be effectively used for the treatment of diabetes, it is expected to be able to treat diabetes by diet without administering a separate therapeutic agent, and research on this is being accelerated, but there is no progress yet.

Therefore, there is a constant need to develop a method for treating diabetes using mushrooms.

Accordingly, the present inventors have made intensive studies to develop a method for treating diabetes by using mushrooms. As a result of inoculating mushrooms into grains, mushroom culture grains prepared by using a solid culture method of cultivating significantly lower blood sugar. It was confirmed that the present invention can be completed.

After all, the main object of the present invention is to provide a mushroom culture grain exhibiting a hypoglycemic effect.

Another object of the present invention is to provide a method for producing an electric mushroom culture grain.

Still another object of the present invention is to provide a food containing an electric mushroom culture grain as a main ingredient and exhibiting a glycemic control effect.

Method for producing a mushroom culture grains exhibiting a glycemic control effect of the present invention is a step of immersing and sterilizing grains; And inoculating and incubating Paecilomyces japonica (KCTC 9817) on the electrosterilized grains to obtain mushroom culture grains, wherein the grains are not particularly limited, but brown rice, barley It is preferable to use Yulmu, and the like, and the method of sterilization is not particularly limited thereto, but it is preferable to use methods such as high temperature sterilization and high pressure autoclaving, and conditions for culturing Paesilomyses japonica are particularly limited thereto. However, it is preferable to incubate for 2 to 4 weeks at 20 to 30 ℃ as in the conventional mushroom cultivation method.

First, the inventors of the present invention, in order to search for a mushroom that exhibits a glycemic control effect, known medicinal mushroom Agaricus mushroom (Agaricus arvensis, Agaricus blazei), Cordyceps sinensis (Cordyceps militaris, Cordyceps ophioglossoides, Cordyceps sinensis, Paecilomyces japonica), Ganoderma lucidum mushroom (Ganoderma applanatum, Ganoderma lucidum, Ganoderma valesiacum), Mud Mushrooms (Phellinus baumi, Phellinus chrysoloma, Phellinus igniarius, Phellinus japonica, Phellinus linteus) And Refuge (Wolfiporia cocos, Wolfiporia extensa) And other mushrooms (Coriolus azureus, Stereum gausapatum, Gloeophyllum abietinum, Grifora frondosa, Laetiporus sulphureus, Lentinus edodes, Lenzites betulina, Merulius tremellosus, Mycoacia fuscoatra, Oxyporus latemarginata, Amanita aspera cla, Clacetusadinocabe , Trametes versicolor, Flammulina velutipes, Fomes fomentarius), The mycelia extracts were obtained, and the activity inhibitory effect, α-glucosidase inhibitory effect, glucose permeation inhibitory effect, and glucose load degree of α-glucosidase of the obtained extract were measured. Paesilomyses Japonica (Paecilomyces japonica) Was the best.

In addition, it can be seen that the Paesylomyces japonica culture grains prepared by inoculating the strains previously selected and cultured with brown rice and barley effectively inhibit the activity of α-glucosidase. In order to confirm the above results, mushroom cultures in which Paesilomyces japonica was cultured in brown rice and barley as a result of measuring glucose permeation delay effect in laboratory environment and glucose loading effect in in vivo environment Brown rice and mushroom cultured barley showed an excellent glycemic control effect.

Accordingly, the mushroom culture grains of the present invention can be used to control blood sugar by feeding on itself, and when the food is produced based on the mushroom culture grains of the present invention, the prepared food has a glycemic control effect. Because it is expected to be represented, the mushroom culture grains of the present invention may be widely used in the development of functional foods exhibiting a glycemic control effect.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1 Activity Inhibition Effect of α-Glucosidase of Mushrooms

Various mushrooms were cultivated, mycelia extracts were obtained therefrom, and the degree of activity inhibition of α-glucosidase of the obtained mycelia extracts was measured.

Example 1-1 : Mushroom Mycelia Extract

Known medicinal mushrooms Agaricus arvensis, Agaricus blazei , Cordyceps militaris, Cordyceps ophioglossoides, Cordyceps sinensis, Paecilomyces japonica , Ganoderma applanatum, Ganoderma lucidum, Ganoderma valesiacum Phellinus chrysoloma, Phellinus igniarius, Phellinus japonica, Phellinus linteus ) and Bokyeong ( Wolfiporia cocos, Wolfiporia extensa ) and other mushrooms ( Coriolus azureus, Stereum gausapatum, Gloeophyllum abietinum, Grifora frondosa, Laetiporus Lentinulius benzustensulus phus) fuscoatra, the Oxyporus latemarginata, Amanita aspera, Clavicorona pyxidata, Clitocybe clavipes, Peniophora quercina, Phlebia radiata, Pleurotus ostreatus, Pycnoporus cinnabarinus, Trametes versicolor, Flammulina velutipes, Fomes fomentarius), respectively, a certain amount of time the culture, and 1cm ² on the agar plate medium Obtain slices by size, rotating shake vessel Incubated for 4 weeks under dark conditions at 100 rpm in both phases. After the mycelium is sufficiently grown, the mycelium is cultured under reduced pressure to obtain the mycelium. The mycelium is lyophilized, and the mycelium extract is repeatedly extracted twice at a temperature of 85 ° C. with 75% (v / v) aqueous methanol solution for 3 hours. Was obtained, dissolved in DMSO at a concentration of 10 mg / ml, and used as a sample. At this time, the culture conditions of each mushroom are shown in Table 1.

Culture condition of mushroom Strain KCTC No. badge Culture temperature (℃) Agaricus arvensis Agaricus blazei Cordyceps militaris Cordyceps ophioglossoides Cordyceps sinensis Paecilomyces japonica Ganoderma applanatum Ganoderma lucidum Ganoderma valesiacum Phellinus baumi Phellinus chrysoloma Phellinus igniarius Phellinus japonica Phellinus linteus Wolfiporia cocos Wolfiporia extensa Amanita aspera Clavicorona pyxidata Clitocybe clavipes Coriolus azureus Flammulina velutipes Fomes fomentarius Gloeophyllum abietinum Grifora frondosa Laetiporus sulphureus Lentinus edodes Lenzites betulina Merulius tremellosus Mycoacia fuscoatra Oxyporus latemarginata Peniophora quercina Phlebia radiata Pleurotus ostreatus Pycnoporus cinnabarinus Stereum gausapatum Trametes versicolor 6851 1693264732613798171680062836513168821688416890 6719648066636490160426727160006367636363562615363586733635467626756666167586759 672567096714 PDAPDAYMBPSAMEAMEAMEAMEAMEAMEAMEAMEAPDAPDAPDAPSAMEAPDAMEAPDAMEAPDAPDAYMBMEAYMBPDAYMBMEAMEAYMBMEAMEAPDAPDAPDAPDA 212126232623242424212326262626262621262123232621232626212126212126262626

Example 1-2 : Activity Inhibition Effect of α-Glucosidase

Inhibitory activity of the mycelium extract on α-glucosidase was p-nitrophenyl α-D-glucopyranoside (6 mM), α-glucosidase (1.4 mM / mL) and After mixing mycelial extract (1.0mg / mL) and reacting at 37 ° C for 30 minutes, 0.4M glycine buffer (pH 10.4) was added, and the absorbance at 400 nm was measured for the supernatant obtained by centrifugation. By measuring, the change in absorbance during the reaction time was calculated from the ratio of the change in absorbance of the control group with DMSO (see Table 2). At this time. As a control, acarbose (acarbose) was used as an α-glucosidase inhibitor.

Inhibitory Effect of α-Glucosidase on Mushrooms Strain KCTC No. Deactivation (%) Control Agaricus arvensis Agaricus blazei Cordyceps militaris Cordyceps ophioglossoides Cordyceps sinensis Paecilomyces japonica Ganoderma applanatum Ganoderma lucidum Ganoderma valesiacum Phellinus baumi Phellinus chrysoloma Phellinus igniarius Phellinus japonica Phellinus linteus Wolfiporia cocos Wolfiporia extensa Amanita aspera Clavicorona pyxidata Clitocybe clavipes Coriolus azureus Flammulina velutipes Fomes fomentarius Gloeophyllum abietinum Grifora frondosa Laetiporus sulphureus Lentinus edodes Lenzites betulina Merulius tremellosus Mycoacia fuscoatra Oxyporus latemarginata Peniophora quercina Phlebia radiata Pleurotus ostreatus Pycnoporus cinnabarinus Stereum gausapatum Trametes versicolor 6851 1693264732613798171680062836513168821688416890 6719648066636490160426727160006367636363562615363586733635467626756666167586759 672567096714 94.4 ± 2.5-10.5 ± 2.117.2 ± 3.3-20.5 ± 7.416.3 ± 2.1-15.8 ± 2.5--13.6 ± 2.2--15.9 ± 3.6 ----------- 9.3 ± 5.5--18.8 ± 3.5 --- 10.7 ± 1.4 ---

As shown in Table 2, it was found that the activity of inhibiting the activity of α-glucosidase in several kinds of mushrooms, 10 kinds of mushrooms ( Agaricus blazei) exhibiting good activity inhibitory effect of α-glucosidase , Cordyceps militaris, Cordyceps sinensis, Paecilomyces japonica, Ganoderma lucidum, Phellinus chrysoloma, Phellinus linteus, Lentinus edodes, Mycoacia fuscoatra, Pleurotus ostreatus ).

Example 1-3 Glucose Permeation Delay Effect

The glucose permeation delay effect was measured under the experimental conditions (in vitro), targeting the mushrooms showing a good effect in the results of Example 1-2. For this purpose, after the dialysis membrane is put in aqueous solution of dialysis, the amount of glucose remaining in the dialysis membrane is measured, and the more glucose remaining, the glucose permeation is considered to be delayed. In this case, the absorption of glucose in the body It can be inferred that it is inhibited.

In other words, 300 ppm and 1% (w / v) glucose of mushroom mycelium extract exhibiting good effects in Example 1-2 were dissolved in a dialysis membrane (MWCO 6000-8000, Spectrum Medical Industries, Inc., USA). 10 mL of the mixed solution was added and placed in a 500 mL beaker and shaken at 25 rpm at 100 rpm for 0.1 mL of the dialysis membrane external solution at regular intervals (0, 30 or 120 minutes) to measure glucose concentration. It was converted into the amount of glucose remaining in the dialysis membrane solution (see Table 3). At this time, the concentration of glucose was measured using a glucose analysis kit (Megazyme Pty, Ltd., Ireland), and as a control, pectin (pectin, Sigma Chemical Co., USA) was used at the same concentration.

Delayed Effect of Glucose on Mushrooms Different kinds of mushrooms % Of residual glucose after time passes 0 minutes 30 minutes 120 minutes passed Control Agaricus blazei Cordyceps militaris Cordyceps sinensis Paecilomyces japonica Ganoderma lucidum Phellinus chrysoloma Phellinus linteus Lentinus edodes Mycoacia fuscoatra Pleurotus ostreatus 100 100 100 100 100 100 100 100 100 100 100 40.755.645.454.952.152.155.066.556.850.152.8 8.524.513.126.128.113.423.441.824.015.228.9

As shown in Table 3, the extract of most mushroom mycelium showed the effect of delaying the permeation of glucose compared to the control group, there was a difference in the degree of delay, mud mushroom ( Phellinus linteus ) showed the highest permeation delay effect. Therefore, four kinds of mushrooms ( Cordyceps sinensis, Paecilomyces japonica, Phellinus linteus, Pleurotus ostreatus ) , which exhibited a glucose permeation delay effect at a concentration of 25% or more after 120 minutes , were selected.

This glucose permeation delay effect was predicted because the water-soluble dietary fiber contained in the sample collected glucose by forming a highly viscous gel.

Example 1-4 : Measurement of the degree of sugar load

In order to confirm the glycemic control ability of the mushrooms selected in Example 1-3 in vivo, blood glucose levels after diet were measured and compared.

That is, streptozotocin, a diabetes-causing drug, was administered to the mice in an amount of 40 mg / kg body weight, and after 1 day, only mice having a blood glucose level of 250 mg / 100 mL or more were selected. After fasting the selected mice for 1 day, 2 ml of aqueous maltose solution (0.35 g / ml) was orally administered, and the mycelia extracts of the mushrooms selected in Examples 1-3 were administered in an amount of 300 mg / kg body weight, respectively. The experimental group was set. Immediately after administration of the maltose solution, at 30, 60, 90 or 120 minutes after administration, blood was collected from each experimental group and the blood collected was applied to a blood glucose meter (Precision Plus Electrodes, Medisense Contract Manufacturing Ltd., UL). Blood glucose was measured (see Table 4). In this case, an experimental group in which pectin was administered in the same amount was used as a positive control, and an experimental group in which no mushroom was administered was used as a negative control.

Hypoglycemic Effect by Various Mushroom Extracts Experimental group % Of blood glucose after time 0 min 30 minutes 60 minutes 90 minutes 120 minutes Positive control negative control Cordyceps sinensis Paecilomyces japonica Phellinus linteus Pleurotus ostreatus 100100100100100100 165.3194.1148.9143.2145.7147.8 141.8157.8128.8129.9129.1157.6 118.5150.4131.4118.3110.8125.5 105.1141.3115.595.4104.1128.7

As shown in Table 4, after the administration of maltose solution, the blood sugar rapidly increased and then gradually decreased, but after 120 minutes, only the experimental group to which Paecilomyces japonica was administered was given blood sugar. It turned out that it is lower than before this administration.

The hypoglycemic effect of paesillomyces japonica is analyzed as a result of synergistic effect of the inhibitory effect of α-glucosidase and the trapping effect of glucose in the water-soluble dietary fiber contained in the sample.

Example 2 Preparation and Effect of Mushroom Culture Grains

Mushroom culture grains were prepared using the electroselected Paesylomyces japonica, and the activity inhibitory effect, glucose permeation inhibitory effect, and blood glucose inhibitory effect of α-glucosidase were measured.

Example 2-1 Preparation of Mushroom Culture Grains

The final selected Paesylomyces japonica in Example 1-4 was incubated to obtain mycelium, and then inoculated into grains to prepare mushroom culture grains: 100 g of brown rice, barley, soybean, red beans, or even sweet radish at 15 ° C. Immersed in water for 3 hours, water was removed, sterilized for 1 hour at 121 ℃, 10 atm, inoculated with mycelium of Paesilomyses japonica, respectively, and incubated for 3 weeks in a culture room fixed at 28 ℃ Each mushroom culture grain was obtained. Subsequently, the mushroom culture grains thus obtained were repeatedly extracted two times for three hours with a 10-fold volume of 75% (v / v) ethanol aqueous solution, and concentrated to dryness at a temperature of 40 ° C. to prepare a sample.

Example 2-2 Activity Inhibition Effect of α-Glucosidase of Mushroom Culture Grains

Using the sample prepared in Example 2-1, the activity inhibitory effect of α-glucosidase was measured by the method of Example 1-2 (see Table 5). At this time, the control group was used grains that were not cultured mycelium of Paesilomyses japonica.

Inhibitory Effect of α-Glucosidase on Various Mushroom Cultures Types of grain Deactivation (%) Control Brown Rice Control Barley Control Bean Control Control Red Bean Control Yulmu ----- Mushroom CultureBrown Mushroom CultureBarley Mushroom CultureBean Mushroom CultureRed Mushroom Culture 24.3 ± 5.75.9 ± 1.3 ---

As shown in Table 5, in the case of mushroom cultured brown rice and mushroom cultured barley showed the activity inhibitory effect of α-glucosidase, in the case of mushroom culture beans, mushroom cultured red beans or mushroom culture rate of α-glucosidase It showed little activity inhibitory effect.

Example 2-3 Glucose Permeation Delay Effect of Mushroom Culture Grains

The glucose permeation delay effect was measured by the method of Example 1-3 using 10 mL of the mixed solution of each sample prepared in Example 2-1 and glucose of 1% (w / v) concentration (see Table: 6). At this time, pectin was used at the same concentration as the positive control group, and mycelial extract of Paesillomyces japonica was used as the negative control group.

Delayed Effect of Glucose by Various Mushroom Culture Grains Experimental group % Of residual glucose after time passes 0 minutes 30 minutes 120 minutes passed Mushroom cultureBrown mushroom cultureCultivation 100100100100100100100 40.753.560.725.930.130.152.1 8.543.840.119.923.230.713.4

As shown in Table 6, in the case of mushroom cultured brown rice and mushroom cultured barley, the glucose permeation delay effect was obvious, but in the case of mushroom culture beans, mushroom cultured red beans, or mushroom culture rate, glucose permeation delay effect was hardly observed. Could know.

In addition, since most mushroom culture grains exhibited a better glucose permeation delay effect than the negative control group using the mycelial extract of the previously selected strains, it was found that culturing the strains in grains was essential.

Example 2-4 : Measurement of sugar load degree of mushroom culture grains

Using each sample prepared in Example 2-1, the degree of sugar load was measured by the method of Example 1-4 (see Table 7). At this time, the experimental group administered with the same amount of pectin was used as a positive control group, and the experimental group administered with the same amount of mycelial extract of paesilomyses japonica was used as a negative control group, and the mycelia of paesomyomyces japonica were used. Were compared using uncultivated grains.

Hypoglycemic Effect by Various Mushroom Culture Grains Experimental group % Of blood glucose after time 0 min 30 minutes 60 minutes 90 minutes 120 minutes Positive control Negative control control Brown rice control Barley control Bean control Red bean control Yulmu 100100100100100100100 165.3143.2145.2150.8148.4144.3141.2 141.8129.9157.5164.6159.4161.7160.2 118.5118.3183.2152.4148.6146.9178.2 105.195.4152.0143.8141.4139.2156.4 Mushroom CultureBrown Mushroom CultureBarley Mushroom CultureBean Mushroom CultureRed Mushroom Culture 100100100100100 153.5116.5148.1145.3142.7 173.7131.8157.1160.9162.1 149.2147.9150.9149.1145.5 143.3120.5144.1140.8155.6

As shown in Table 7, the mushroom cultured brown rice and mushroom cultured barley were maintained at a significantly lower blood sugar level than the brown rice and barley of the control, but the mushroom cultured beans, mushroom cultured red beans and mushroom culture rate of the control beans, beans and red beans Since blood glucose levels were maintained at a similar level, mushroom cultured brown rice and mushroom cultured barley showed a glycemic control effect.

In addition, the blood sugar level of the grains and mushroom culture grains of the control group showed a high level compared to the positive control group and the negative control group, which is due to the sugar contained in the grains.

As described and demonstrated in detail in the above, the present invention is produced by culturing Paesilomyces japonica in grains, including mushroom culture grains having a blood sugar control effect, a method for producing an electric mushroom culture grains and electric mushroom culture grains as a main To provide a food exhibiting a hypoglycemic effect. Mushroom culture grains of the present invention inhibits the production of glucose, and lowers the influx of the generated glucose in the body, it can be widely used in the development of functional foods exhibiting a glycemic control effect.

Claims (5)

(Iii) soaking and sterilizing grain; And, (Ii) inoculating and incubating Paecilomyces japonica (KCTC 9817) on the electrosterilized grain, to obtain a mushroom culture grain. The method of claim 1, Grain is characterized in that the brown rice or barley Method for producing mushroom culture grains. The method of claim 1, Paesilomyses japonica is incubated for 2 to 4 weeks at 20 to 30 ℃ Method for producing mushroom culture grains. Mushroom cultures produced by the method of claim 1, which exhibits a hypoglycemic effect. Food containing a mushroom culture grain disclosed in claim 4 as a main, and exhibits a hypoglycemic effect.