CN112094753A - Nutrient solution for culturing ganoderma lucidum and method for culturing ganoderma lucidum by fermentation process - Google Patents

Abstract

The invention discloses a mythic fungus culture nutrient solution and a method for culturing mythic fungus by using a fermentation process, wherein the nutrient solution comprises the following raw materials: astragalus, chromium-rich yeast, cordyceps sobolifera powder, red dates, medlar, fructose and mineral water. The method breaks through the bottleneck of slow growth of ganoderma lucidum, realizes the rapid culture of ganoderma lucidum, has stable production period, can obtain the ganoderma lucidum product with high effective active substance content and high nutritional and medicinal values, and makes the industrialized production possible; the nutrient solution can shorten the culture period of the pacific, improve the growth state of the pacific, improve the content of various beneficial active substances in the pacific obtained by culture, and further improve the nutritional and health-care effects of the pacific.

Description

Nutrient solution for culturing ganoderma lucidum and method for culturing ganoderma lucidum by fermentation process

Technical Field

The invention relates to the technical field of ganoderma lucidum cultivation, in particular to a ganoderma lucidum cultivation nutrient solution and a method for cultivating ganoderma lucidum by using a fermentation process.

Background

Taisui, which is called meat ganoderma lucidum in the traditional Chinese medicine theory of China. Is read as a bright wonderful flower in a vast Chinese medicine treasury and is regarded as the symbol of natural, luckiness, riches and longevity since ancient times. Modern scientists believe that myxomycete complex is a large-scale myxomycete complex, but why its cellular structure forms and aggregates into such a regular morphology, and its medical value remains a mystery. The compendium of materia Medica records: meat sesame like meat. Attached to the large stone, with the head and tail, is also a living thing. Red such as coral, white such as fat, black such as sun euphorbia, green such as jade feather, yellow such as purple gold, all of which are clear and have strong holes, such as strong ice. Dongjin Daojia Kudzuvine flood is recorded in the 'republic of Pupuzi', and the smashing of various glossy ganoderma into powder or the water dissolving of the product is helpful for people to lose weight and grow old. "is a long-life and impatient immortal drug considered by ancient people.

In the huge transition of the sea-vicarious field in billions of years, many species are annihilated and cultivated, and the species can propagate to the present by the age of the old, so that the strong vitality of the species is unparalleled. The biological fertilizer grows in an anaerobic environment with the ground bottom of 20-100 meters, lives in soil and survives by water, so that the biological fertilizer cannot rot or deteriorate when placed in water. Meanwhile, the slime mold is mainly propagated by spores and hyphae, has strong activity and can be regenerated by random cutting. But never the so-called endless growth, and naturally not rare if it can grow rapidly.

Through the detection of state authority, the aged people are rich in 46 active substances and nutrient substances required by human bodies, such as immune PQQ (pyrroloquinoline quinone), nucleotide, chitin, flavone, polysaccharide, glutathione, saponin, chitin, unsaturated fatty acid, SOD, trace elements, amino acid and the like. Has special effects of resisting tumor, resisting aging, enhancing immunity, etc.

The research shows that the myxobacteria is a polymer mainly composed of three types of lactobacillus, myxobacteria and fungi. In natural environment, due to the limitation of growth conditions, the formation of the Taisui complex is difficult to realize, the difference of active ingredients of natural Taisui in different producing areas is large, and due to the extremely high medicinal and health-preserving values, the price of the naturally formed Taisui is extremely high, and generally about 5 ten thousand yuan per 500 g. The wild Taisui is expensive, the wild Taisui is rare in quantity and is short in supply and demand, and artificial breeding becomes an effective method for improving the supply of the Taisui due to the limited quantity of the wild Taisui. However, the ganoderma lucidum obtained by the existing artificial culture mainly has the defects of slow growth of thalli, long culture period, low nutritional value, low content of effective components and the like. A reliable solution is now required.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a nutrient solution for culturing ganoderma lucidum and a method for culturing ganoderma lucidum by using a fermentation process, aiming at the defects in the prior art.

In order to solve the technical problems, the invention adopts the technical scheme that: a nutrient solution for culturing the pacific years, which comprises the following raw materials: astragalus, chromium-rich yeast, cordyceps sobolifera powder, red dates, medlar, fructose and mineral water.

Preferably, the addition amount of the other raw materials per 1L of mineral water is: 10g of astragalus membranaceus, 2g of chromium-enriched yeast, 5g of cordyceps sobolifera and cordyceps sinensis powder, 30g of red dates, 20g of medlar and 100g of fructose.

Preferably, the preparation method of the nutrient solution comprises the following steps:

1) weighing radix astragali, chromium-rich yeast, Cordyceps sobolifera powder, fructus Jujubae, fructus Lycii, fructose and mineral water in proportion;

2) washing radix astragali, fructus Jujubae and fructus Lycii with tap water, cleaning with purified water for 2 times, soaking in 10g/L NaCl solution for 30min, draining, pulverizing, adding into a container together with chromium-rich yeast and Cordyceps cicada powder, adding mineral water, stirring, filtering, and adding fructose into the filtrate to obtain nutritional liquid.

The invention also provides a method for culturing the myxomycete by using a fermentation process, which comprises adding the nutrient solution as defined in any one of claims 1 to 3 into a myxomycete culture solution.

Preferably, the volume ratio of the nutrient solution to the culture solution for the third year is 7: 3.

Preferably, the method comprises the steps of:

1) cleaning wild pacific years old with purified water, putting the cleaned wild pacific years old into a first container, adding mineral water and fructose, culturing for 10-15 days, keeping the temperature at 26-28 ℃ and the pH value at 5-7.5;

2) when the meat of the third party grows to be 8-12 mm thick on the surface of the culture solution of the first container and the pH value of the culture solution reaches 4.0-4.5, taking out the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the purification culture is finished, transferring the meat and the culture solution to a fermentation tank, adding a nutrient solution into the fermentation tank for amplification culture, and culturing for 10-15 days at the temperature of 26-28 ℃ and the pH value of 5-7.5;

4) when the pH value of the solution in the fermentation tank reaches 4.0-4.5, completing a culture period, and taking out all meat of the myxomycete; part of the culture solution was taken out and supplemented with the same volume of nutrient solution for repeated culture.

Preferably, the method comprises the steps of:

1) cleaning 1000g of wild pacific years old with purified water, putting the cleaned wild pacific years old into a first container with the volume of 5L, adding 4L of mineral water and 150g of fructose, culturing for 10-15 days, and keeping the temperature at 26 ℃ and the pH value at 5-7.5;

2) when the meat of the third generation with the thickness of 10mm grows on the surface of the culture solution in the first container and the pH value of the culture solution reaches 4.5, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution for 10-15 days, keeping the temperature at 26 ℃ and the pH value at 5-7.5;

4) when the pH value of the solution in the fermentation tank reaches 4.0-4.5, completing a culture period, and taking out all meat of the myxomycete; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

Preferably, the myxomycete cultured by the method contains at least the following active substances: PQQ, nucleotide, fungal polysaccharide, vitamin C, vitamin E, free amino acid, chitin, flavone, glutathione and saponin.

The invention has the beneficial effects that: the method breaks through the bottleneck of slow growth of ganoderma lucidum, realizes the rapid culture of ganoderma lucidum, has stable production period, can obtain the ganoderma lucidum product with high effective active substance content and high nutritional and medicinal values, and makes the industrialized production possible;

the nutrient solution can shorten the culture period of the pacific, improve the growth state of the pacific, improve the content of various beneficial active substances in the pacific obtained by culture, and further improve the nutritional and health-care effects of the pacific.

Detailed Description

The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

The invention firstly provides a nutrient solution for culturing the myxomycete, which comprises the following raw materials: astragalus, chromium-rich yeast, cordyceps sobolifera powder, red dates, medlar, fructose and mineral water.

In a preferred embodiment, the addition amount of the other raw materials per 1L of mineral water is as follows: 10g of astragalus membranaceus, 2g of chromium-enriched yeast, 5g of cordyceps sobolifera and cordyceps sinensis powder, 30g of red dates, 20g of medlar and 100g of fructose.

The invention also provides a preparation method of the nutrient solution, which comprises the following steps:

1) weighing fresh and high-quality astragalus membranaceus, chromium-rich yeast, cordyceps sobolifera powder, red dates, medlar, fructose and mineral water in proportion;

2) washing radix astragali, fructus Jujubae and fructus Lycii with tap water, cleaning with purified water for 2 times, soaking in 10g/L NaCl solution for 30min, draining, pulverizing, adding into a container together with chromium-rich yeast and Cordyceps cicada powder, adding spring water, stirring, filtering, and adding fructose into the filtrate to obtain nutritional liquid.

The invention also provides a method for rapidly culturing the myxomycete by using the modern biological fermentation process, which adds the nutrient solution into the myxomycete culture solution. And in a preferred embodiment, the volume ratio of the nutrient solution to the culture solution for the third year is 7: 3.

In a preferred embodiment, the method for culturing myxomycete using a fermentation process comprises the steps of:

1) cleaning wild pacific years old with purified water, putting the cleaned wild pacific years old into a first container, adding mineral water and fructose, culturing for 10-15 days, keeping the temperature at 26-28 ℃ and the pH value at 5-7.5;

2) when the meat of the third party grows to be 8-12 mm thick on the surface of the culture solution of the first container and the pH value of the culture solution reaches 4.0-4.5, taking out the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the purification culture is finished, transferring the meat and the culture solution to a fermentation tank, adding a nutrient solution into the fermentation tank for amplification culture, and culturing for 10-15 days at the temperature of 26-28 ℃ and the pH value of 5-7.5;

4) when the pH value of the solution in the fermentation tank reaches 4.0-4.5, completing a culture period, and taking out all meat of the myxomycete; a part of the culture solution was taken out and supplemented with the same volume of nutrient solution, thereby enabling repeated culture.

In an alternative embodiment, in the purification culture, 3 types of flora (including 2 types of myxobacteria, 10 types of lactobacillus and 5 types of yeast) required for growth of the myxomycete are separated, each type of flora is subjected to purification culture, and the strain which grows most rapidly and can metabolize the most effective components is selected as the production bacteria. Determining the preferable proportion (mass ratio) of each fungus as myxobacteria: lactobacilli: yeast =2:6: 2.

The detection shows that the myxomycete cultured by the method at least contains the following active substances: PQQ, nucleotide, fungal polysaccharide, vitamin C, vitamin E, free amino acid, chitin, flavone, glutathione and saponin.

Further experimental detection shows that in the stock solution prepared by the method, the content of PQQ is 65-75 mu g/mL, the content of nucleic acid is 370-390 mu g/mL, the content of fungal polysaccharide is 2600-2700 mu g/mL, the content of vitamin C is 90-95 mu g/mL, the content of vitamin E is 8.5-9.5 mu g/mL, the content of free amino acid is 40-45 mu g/mL, the content of chitin is 200-220 mu g/mL, and the content of chromium is 550-720 mu g/mL.

The nutrient solution shortens the culture period of the pacific, improves the growth state of the pacific, improves the content of various beneficial active substances in the pacific obtained by culture, and further improves the nutritional and health-care value of the pacific.

The method of the invention realizes the rapid culture of the myxomycete, has stable production period, and can obtain the myxomycete product with high effective active substance content and high nutritional and medicinal value.

The foregoing is a general idea of the invention, and specific examples are provided below to further illustrate the invention.

Example 1

Firstly, preparing a culture nutrient solution for the myxomycete by the following method in advance:

1) weighing 200g of fresh and high-quality astragalus root, 40g of chromium-enriched yeast, 100g of cordyceps cicadae flower, 600g of red date, 400g of medlar, 2000g of fructose and 20L of mineral water according to a proportion;

2) washing radix astragali, fructus Jujubae and fructus Lycii with tap water, cleaning with purified water for 2 times, soaking in 10g/L NaCl solution for 30min, draining, pulverizing, adding into a container together with chromium-rich yeast and Cordyceps cicada powder, adding mineral water, stirring, filtering, and adding fructose into the filtrate to obtain nutritional liquid (about 20L for use).

Secondly, the myxomycete complex is quickly cultured by applying a modern biological fermentation process:

1) cleaning 1000g of wild pacific years old by using purified water, putting the cleaned wild pacific years old into a glass culture bottle with the volume of 5L, adding 4L of mineral water and 150g of fructose, keeping the temperature at 26 ℃ and the pH value at 6-6.5;

2) after 12 days of culture, growing meat of the third year with the thickness of 10mm on the surface of the culture solution of the first container, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution, and keeping the temperature at 26 ℃ and the pH value at 6-6.5;

4) after 13 days of culture, the pH value of the solution in the fermentation tank reaches 4.3, and all meat of the myxomycete is taken out to complete one culture period; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

Example 2

A method for rapidly culturing the myxomycete by using a modern biological fermentation process comprises the following steps:

1) cleaning 1000g of wild pacific years old by using purified water, putting the cleaned wild pacific years old into a glass culture bottle with the volume of 5L, adding 4L of mineral water and 120g of fructose, keeping the temperature at 26 ℃ and the pH value at 6-6.5;

2) after 12 days of culture, growing meat of the third year with the thickness of 10mm on the surface of the culture solution of the first container, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution, and keeping the temperature at 26 ℃ and the pH value at 6-6.5;

4) after 13 days of culture, the pH value of the solution in the fermentation tank reaches 4.2, and all meat of the myxomycete is taken out to complete one culture period; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

The culture medium for the myxomycete was the same as in example 1.

Example 3

A method for rapidly culturing the myxomycete by using a modern biological fermentation process comprises the following steps:

1) cleaning 1000g of wild pacific years old by using purified water, putting the cleaned wild pacific years old into a glass culture bottle with the volume of 5L, adding 4L of mineral water and 180g of fructose, and keeping the temperature at 26 ℃ and the pH value at 6-6.5;

2) after 12 days of culture, growing meat of the third year with the thickness of 10mm on the surface of the culture solution of the first container, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution, and keeping the temperature at 26 ℃ and the pH value at 6-6.5;

4) after 13 days of culture, the pH value of the solution in the fermentation tank reaches 4.5, and all meat of the myxomycete is taken out to complete one culture period; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

The culture medium for the myxomycete was the same as in example 1.

Experimental example a, the growth state and culture period of the pacifier in the pacifier culture method of the present invention were previously analyzed in connection with comparative example 1 and comparative example 2 below.

Comparative example 1

A method for rapidly culturing the myxomycete by using a modern biological fermentation process comprises the following steps:

1) cleaning 1000g of wild pacific years old by using purified water, putting the cleaned wild pacific years old into a glass culture bottle with the volume of 5L, adding 4L of mineral water and 150g of fructose, keeping the temperature at 26 ℃ and the pH value at 6-6.5;

2) after 12 days of culture, growing meat of the third year with the thickness of 10mm on the surface of the culture solution of the first container, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the mineral water, keeping the temperature at 26 ℃, and keeping the pH value at 6-6.5;

4) after 13 days of culture, the pH value of the solution in the fermentation tank reaches 4.3, and all meat of the myxomycete is taken out to complete one culture period; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

The difference from example 1 is mainly that the culture medium was replaced with mineral water.

Comparative example 2

Firstly, preparing a culture nutrient solution for the myxomycete by the following method in advance:

1) weighing 200g of fresh and high-quality astragalus root, 600g of red date, 400g of medlar, 2140g of fructose and 20L of mineral water according to a proportion;

2) washing radix astragali, fructus Jujubae and fructus Lycii with tap water, cleaning with purified water for 2 times, soaking in 10g/L NaCl solution for 30min, draining, pulverizing, adding into container, adding mineral water, stirring, filtering, and adding fructose into the filtrate to obtain nutritional liquid (about 20L for use).

Secondly, rapidly culturing the myxomycete by applying a modern biological fermentation process:

1) cleaning 1000g of wild pacific years old by using purified water, putting the cleaned wild pacific years old into a glass culture bottle with the volume of 5L, adding 4L of mineral water and 150g of fructose, keeping the temperature at 26 ℃ and the pH value at 6-6.5;

2) after 12 days of culture, growing meat of the third year with the thickness of 10mm on the surface of the culture solution of the first container, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution, and keeping the temperature at 26 ℃ and the pH value at 6-6.5;

4) after 13 days of culture, the pH value of the solution in the fermentation tank reaches 4.3, and all meat of the myxomycete is taken out to complete one culture period; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

The difference from example 1 is mainly in the composition of the culture broth.

After one culture period (25 days) was reached, the growth of the meat of Taisui obtained in examples 1-3 and comparative examples 1-2 was observed, and the thickness of the meat of Taisui was measured, and the statistical results are shown in Table 1 below.

TABLE 1 growth of the pacifier

Index (I)	Example 1	Example 2	Example 3	Comparative example 1	Comparative example 2
						Growth of hypha	White and dense	White and dense	White and dense	White and not dense	Whiter and denser
Thickness of meat of Taisui Degree/mm	23	22	22	17	19
						Texture of fruiting body	The skin and the shell are good in layer and compact in texture, specific gravity of heavy	The skin and the shell are good in layer and compact in texture, specific gravity of heavy	Leather caseThe layer is good, the texture is tight, specific gravity of heavy	Loose texture and no appearance Cooked and light hand feeling	Loose texture and no appearance Has light cooked and hand feeling

As can be shown by the results in Table 1, the growth cycle of the pacific is shortened and the growth state of the pacific and the quality of the finished product are improved by adding the culture solution with a unique formula in the invention.

Experimental example B the main active ingredients in meat of pacific years obtained in examples 1-3 were tested and compared with commercially available meat of pacific years, and the specific results are shown in table 2 below.

TABLE 2 analysis of the ingredients

Detecting items	Example 1	Example 2	Example 3	Commercial Taisui
					PQQ content μ g/mL	75	73	70	34
Nucleic acid content μ g/mL	388	375	380	280
					The content of fungus polysaccharide is mug/mL	2660	2630	2670	2330
Vitamin C content mu g/mL	92	90	95	85
					Vitamin E content mu g/mL	9.4	9.2	9.2	6.3
The content of free amino acid is mu g/mL	45	41	43	26
					Chitin content μ g/mL	218	215	220	190
Chromium content μ g/mL	650	672	645	45

The detection method comprises the following steps: the pacifies obtained in examples 1-3 and commercially available pacifies were prepared as pacifies stock solutions and the contents of each component in the table above were measured using conventional measurement methods. Specifically, for example:

detecting the PQQ content by adopting an NBT-Gly method; detecting the content of nucleic acid by adopting a phosphorus determination method; detecting the content of fungal polysaccharide by adopting a phenol-sulfuric acid method; detecting the content of vitamin C and vitamin E by adopting a display reaction method; detecting the content of free amino acid by a colorimetric method; detecting the chromium content by an ammonium ferrous sulfate titration method; for the detection of chitin content, reference is made to the method described in chinese patent 200910143787.6.

The detection results in the table 2 show that the contents of PQQ, nucleic acid, fungal polysaccharide, free amino acid and chromium in the cultured myxomycete are all obviously higher than those in the commercial myxomycete, which indicates that the contents of various effective active substances in the myxomycete cultured by the method provided by the invention are improved, and the nutritional and medicinal values of the substances are further improved.

Test example C study of hypoglycemic Effect of the third decade obtained by the method of the present invention

Through a large number of experimental researches, the myxomycete obtained by the method has obvious effect of reducing blood sugar, and the blood sugar reducing effect is superior to that of the myxomycete sold in the market. Specific test methods and data are provided below.

1. The test method comprises the following steps:

animal grouping and administration: the SD model rats were randomly grouped, i.e. blank group, model group, pacifier group (a-d), metformin control group, 10 per group. Wherein the product obtained in examples 1-3 was dried to a powder form for the third year. Gavage (ig) was administered at the following doses: the dosage of the medicine is 0.60 g/kg/day; metformin is 0.050 g/kg/day. The blank group and the model group are perfused with 5 per mill CMC-Na solution and continuously administrated for 60 days.

Experimental drugs:

pacifier group a: the product of example 1;

group b: the product of example 2;

c, Taisui group: the product of example 3;

the third year of age group d: commercial products of the pacific year;

a metformin control group;

in the group of Taisui, the raw materials are decocted with water for oral administration.

2. Observation index

(1) Determination of fasting blood glucose: after six weeks of administration, the experimental animals are fasted for 8 hours, tail veins are used for blood sampling, fasting blood glucose values of rats in each group are measured by using a Roche glucometer and corresponding test paper and recorded, and the measurement principle is that enzyme on the test paper is recombined and expressed by escherichia coli E, glucose in blood is converted into gluconolactone, and then harmless direct current is generated, so that the blood glucose values are read.

(2) Oral glucose tolerance: after each group is administrated for six weeks, experimental animals are fasted for 8 hours, the animals are weighed to be body mass, after fasting blood glucose is measured, each group is administrated with 40% glucose solution by stomach irrigation according to the dose of 2g/kg, blood glucose values are measured by 30min, 1h and 2 tail vein blood taking after stomach irrigation, and the change of the area under the blood glucose curve of each time point of each group is observed and calculated:

area under the blood glucose curve is 1/2 × (0 hour blood glucose value +0.5 hour blood glucose value) × 0.5+1/2 × (2 hour blood glucose value +0.5 hour blood glucose value) × 1.5

3. Results of the experiment

3.1 Effect of the age of Taisui of the invention on fasting glucose

Referring to table 3, as a result of the influence of the pacific age on fasting glucose, according to the SPSS18.0 analysis of variance (one way ANOVA), it was shown that the pacific age group and the metformin group significantly decreased fasting blood glucose values in the model rats compared to the model group, and the results were significantly different (p < 0.05).

TABLE 3 results of the effect of pacific year on fasting plasma glucose (n ═ 10)

Group of	Fasting blood glucose level (mmol/L)
		Blank group	5.1±0.6
Model set	30.8±4.9
		Metformin control group	24.5±4.2*
Taisui group a	15.3±5.1*
		Taisui group b	17.5±5.4*
Taisui group c	16.9±3.6*
		Taisui group d	23.1±5.5*

Note: *. compared with the model control group, there is a significant difference (p <0.05)

3.2 Effect of the age of the Taisui of the invention on oral glucose tolerance

See table 4 for the effect of the age of pacific on oral glucose tolerance.

Table 4 effect of myxomycete on oral glucose tolerance results (n ═ 10)

Group of	Area under the blood glucose curve
		Blank group	16.2±2.8
Model set	82.5±7.3
		Metformin control group	47.1±5.8*
Taisui group a	31.5±2.9*
		Taisui group b	34.7±4.4*
Taisui group c	32.2±5.1*
		Taisui group d	45.6±6.2*

Note: *. compared with the model control group, there is a significant difference (p <0.05)

After each group is administrated for six weeks, experimental animals are fasted for 8 hours, the animals are weighed to be body mass, after fasting blood glucose is measured, each group is administrated with 40% glucose solution by stomach irrigation according to the dose of 2g/kg, blood glucose values are measured by 30min, 1h and 2 tail vein blood taking after stomach irrigation, and the change of the area under the blood glucose curve of each time point of each group is observed and calculated:

the area under the blood glucose curve is 1/2 x (0 hour blood glucose value +0.5 hour blood glucose value) x 0.5+1/2 x (2 hour blood glucose value +0.5 hour blood glucose value) x 1.5, and the sps 18.0 analysis of variance shows that the formula groups and the metformin control group have significant difference (p is less than 0.05) compared with the model group, which indicates that the taisui group has the effect of obviously improving the oral glucose tolerance of the model rat. The results are shown in Table 4.

The test C mentioned above, the taisui obtained by the method of the present invention was found to have a significant hypoglycemic effect, which is superior to that of the commercially available taisui. Test analysis shows that the nutrient solution added in the culture process of the invention has important influence on improving the blood sugar reducing effect of the pacific, and especially the cordyceps sobolifera powder and the chromium-rich yeast added in the nutrient solution carry out analysis test on the effects of the two components.

Test example D Effect of the addition of Cordyceps sobolifera powder on the hypoglycemic Effect of the invention

1. The test method comprises the following steps:

in the same manner as in test example C, the following groups were added to this test example, except for the blank group, the model group, the pacifier group (a-d), and the metformin control group: comparative groups 1-3. That is, the data of comparative groups 1 to 3 were also added to those of test example C.

Wherein, the comparison group 1 is a comparison myxomycete 1, which specifically comprises the following components: the nutrient solution added in the myxomycete culture is different from the myxomycete group a only in that the myxomycete powder is not included, and the myxomycete product obtained in this case is dried and made into powder; the dosage is 0.60 g/kg/day;

comparative group 2 was: the dosage of the cordyceps sobolifera powder is 0.60 g/kg/day;

comparative group 3 was: cordyceps cicadae powder + comparative myxomycete 1, wherein the dosage of the Cordyceps cicadae powder is N g/kg/day, wherein N is equal to the amount of the Cordyceps cicadae powder added in the culture process per 0.6g of myxomycete in myxomycete group a; ensuring that the dosage of the cordyceps sobolifera powder is basically the same as the equivalent of the cordyceps sobolifera powder contained in the myxomycete; the dosage for comparison of the pacifier 1 was (0.6-N) g/kg/day.

The results of the effects of comparative groups 1 to 3 on fasting plasma glucose in test example D were obtained in the same manner as in test example C, and are shown in tables 5 to 6.

Table 5 results on fasting glucose (n ═ 10)

Group of	Fasting blood glucose level (mmol/L)
		Blank group	5.1±0.6
Model set	30.8±4.9
		Metformin control group	24.5±4.2*
Taisui group a	15.3±5.1*
		Taisui group b	17.5±5.4*
Taisui group c	16.9±3.6*
		Taisui group d	23.1±5.5*
Comparative group 1	23.2±4.6*
		Comparative group 2	22.5±3.7*
Comparative group 3	21.7±2.8*

Note: *. compared with the model control group, there is a significant difference (p <0.05)

See table 5 for the results on fasting glucose. Cordyceps cicadae powder has been proved to have hypoglycemic effect, and the results of comparative group 2 can also be proved. The comparison results of the groups 1, 2 and 3, the myxocypris group a and the like also show that the hypoglycemic effect obtained by the scheme of singly adopting the cordyceps sobolifera powder or adopting the combination of the cordyceps sobolifera powder and the common myxocypris is obviously inferior to that obtained by the scheme of the invention, the main reason is that the myxocypris obtained by adding the cordyceps sobolifera powder serving as the raw material of the culture medium into the nutrient solution for myxocypris culture, and the hypoglycemic effective components in the cordyceps sobolifera powder are organized in the myxocypris culture process, so that the hypoglycemic effective components can be fully absorbed and utilized by organisms, and the better effect is exerted. The cordyceps sobolifera powder can play a certain role in reducing blood sugar when being directly taken, but the blood sugar efficacy is worse than that of the cordyceps sobolifera powder due to insufficient absorption and the like.

Test example E Effect of chromium-enriched Yeast addition on hypoglycemic Effect of the invention

1. The test method comprises the following steps:

in the same manner as in test example C, the following groups were added to this test example, except for the blank group, the model group, the pacifier group (a-d), and the metformin control group: comparative groups 4-6. That is, the data of comparative groups 4 to 6 were also added to those of test example C.

Wherein, the comparison group 4 is the comparison myxomycete 4, which specifically comprises the following components: the nutrient solution added in the culture of the pacifier is different from that of the pacifier group a only in that chromium-rich yeast is not included therein, in which case the pacifier product obtained in this case is dried and powdered; the dosage is 0.60 g/kg/day;

comparative group 5 was: the chromium-rich yeast powder has the dosage ensuring that the dosage of chromium is as follows: 2 mug/kg/day;

comparative group 6 was: chromium-rich yeast powder + comparative pacific 1, wherein the amount of chromium-rich yeast powder administered is M g/kg/day, wherein M is equal to the amount of chromium-rich yeast powder added during the culturing process per 0.6g of pacific in pacific group a; ensuring that the dosage of the cordyceps sobolifera powder is basically the same as the equivalent of the chromium-rich yeast powder contained in the myxomycete; the dose for comparison of the pacifier 1 was (0.6-M) g/kg/day.

The results of the effects of comparative groups 1 to 3 on fasting plasma glucose in test example D were obtained in the same manner as in test example C, and are shown in tables 5 to 6.

Table 6 results on fasting glucose (n ═ 10)

Group of	Fasting blood glucose level (mmol/L)
		Blank group	5.1±0.6
Model set	30.8±4.9
		Metformin control group	24.5±4.2*
Taisui group a	15.3±5.1*
		Taisui group b	17.5±5.4*
Taisui group c	16.9±3.6*
		Taisui group d	23.1±5.5*
Comparative group 1	20.9±3.9*
		Comparative group 2	26.1±4.2*
Comparative group 3	19.3±3.4*

Note: *. compared with the model control group, there is a significant difference (p <0.05)

See table 6 for the results of the effect on fasting glucose. Trivalent chromium is widely present in human bones, muscles, hair, skin, subcutaneous tissues, major organs (except lungs) and body fluids, and achieves hypoglycemic effects mainly through the synergistic and insulin-potentiating effects of GTF. But the absorption and utilization rate of the human body to inorganic chromium is extremely low and is less than 1 percent; the utilization rate of the human body to the organic chromium can reach 10-25%. The chromium-rich yeast is edible functional yeast with chromium element function, is a natural blood sugar regulating agent, plays a special role in the sugar metabolism and the lipid metabolism of a body, and is a trace element-chromium which is necessary for the human body activates the insulin activity, regulates the blood sugar, inhibits the conversion of the sugar into the fat, and has higher absorption and utilization rate.

The important effect of the addition of the chromium-rich yeast on the blood sugar reducing effect of the obtained pacific product can be embodied by the comparison group 5, and the results of the comparison groups 6 and 7, the pacific group and the like can also show that the effect of the direct administration of the chromium-rich yeast or the direct administration of the chromium-rich yeast and the common pacific is obviously inferior to that of the pacific obtained by the scheme of the invention.

In this example, the absorption rate of chromium in rats in chromium-rich yeast, commercial pacifier, a combination of chromium-rich yeast powder and commercial pacifier (mass ratio of chromium-rich yeast powder to commercial pacifier is 1: 100) and pacifier group a was also measured, and the measurement results are shown in table 7 below.

Group of	Chromium-rich yeast	Commercial Taisui	Chromium-rich yeast powder and commercial myxomycete	Taisui of Taisui group a
					Absorption rate of chromium%	22	29	38	26

Table 7 chromium absorption results

Test of degree of organization of chromium in chromium-enriched Yeast, commercial "Taisui" and "Taisui group a

10g of the dry powder of each sample is dissolved in 30ml of deionized water, and the mixture is respectively leached for 5 hours, 10 hours and 20 hours, then centrifuged, dried and precipitated, and the chromium content of the precipitate is measured to express the organization degree of chromium. The results show that the yeast, the commercial pacifier and the pacifier group a all have the biotransformation capability to the chromium, the added chromium is not mechanically physically adsorbed on the surface of the thallus, but is organically biotransformed to form the firmly combined complex organic chromium, the chromium-rich yeast has the chromium organization degree of 84.8 percent, the commercial pacifier has the chromium organization degree of 87.4 percent, and the pacifier group a has the chromium organization degree of 90.2 percent.

The experimental example E fully shows that the addition of the chromium-rich yeast can obviously enhance the hypoglycemic effect of the invention obtained by the invention, and further, the chromium-rich yeast is added into the nutrient solution as the raw material of the culture medium for carrying out the culture of the invention, and the chromium is organized by the invention in the culture process of the invention, so that the absorption and utilization rate of the chromium by the human body can be improved, and the hypoglycemic effect of the chromium is enhanced.

While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.

Claims (8)

1. The culture nutrient solution for the pacific years is characterized by comprising the following raw materials: astragalus, chromium-rich yeast, cordyceps sobolifera powder, red dates, medlar, fructose and mineral water.

2. The culture nutrient solution for the pacifier of claim 1, wherein the amount of the other raw materials added per 1L of mineral water is: 10g of astragalus membranaceus, 2g of chromium-enriched yeast, 5g of cordyceps sobolifera and cordyceps sinensis powder, 30g of red dates, 20g of medlar and 100g of fructose.

3. The culture nutrient solution for the pacifier of claim 2, wherein the preparation method of the nutrient solution comprises the following steps:

1) weighing radix astragali, chromium-rich yeast, Cordyceps sobolifera powder, fructus Jujubae, fructus Lycii, fructose and mineral water in proportion;

2) washing radix astragali, fructus Jujubae and fructus Lycii with tap water, cleaning with purified water for 2 times, soaking in 10g/L NaCl solution for 30min, draining, pulverizing, adding into a container together with chromium-rich yeast and Cordyceps cicada powder, adding mineral water, stirring, filtering, and adding fructose into the filtrate to obtain nutritional liquid.

4. A method for culturing the myxomycete by using a fermentation process, wherein the nutrient solution as claimed in any one of claims 1 to 3 is added to the myxomycete culture solution.

5. The method for cultivating the pacifier according to the process of claim 4, wherein the volume ratio of the nutrient solution to the pacifier culture solution is 7: 3.

6. The method of claim 5, comprising the steps of:

1) cleaning wild pacific years old with purified water, putting the cleaned wild pacific years old into a first container, adding mineral water and fructose, culturing for 10-15 days, keeping the temperature at 26-28 ℃ and the pH value at 5-7.5;

2) when the meat of the third party grows to be 8-12 mm thick on the surface of the culture solution of the first container and the pH value of the culture solution reaches 4.0-4.5, taking out the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the purification culture is finished, transferring the meat and the culture solution to a fermentation tank, adding a nutrient solution into the fermentation tank for amplification culture, and culturing for 10-15 days at the temperature of 26-28 ℃ and the pH value of 5-7.5;

4) when the pH value of the solution in the fermentation tank reaches 4.0-4.5, completing a culture period, and taking out all meat of the myxomycete; part of the culture solution was taken out and supplemented with the same volume of nutrient solution for repeated culture.

7. The method of claim 6, comprising the steps of:

1) cleaning 1000g of wild pacific years old with purified water, putting the cleaned wild pacific years old into a first container with the volume of 5L, adding 4L of mineral water and 150g of fructose, culturing for 10-15 days, and keeping the temperature at 26 ℃ and the pH value at 5-7.5;

2) when the meat of the third generation with the thickness of 10mm grows on the surface of the culture solution in the first container and the pH value of the culture solution reaches 4.5, taking out the culture solution, transferring the culture solution to a second container for purification culture, and separating out harmful microorganism populations;

3) after the completion of the purification culture, the meat of the third year was transferred to a fermenter together with the culture broth, and a mixture of the meat of the third year and the culture broth was added to the fermenter in a volume ratio of 7:3, carrying out amplification culture on the nutrient solution for 10-15 days, keeping the temperature at 26 ℃ and the pH value at 5-7.5;

4) when the pH value of the solution in the fermentation tank reaches 4.0-4.5, completing a culture period, and taking out all meat of the myxomycete; the culture solution with the volume fraction of 70% is taken out and supplemented with the nutrient solution with the same volume for repeated culture.

8. The method of claim 7, wherein the cultured product contains at least the following active ingredients: PQQ, nucleotide, fungal polysaccharide, vitamin C, vitamin E, free amino acid, chitin, flavone, glutathione and saponin.