CN111607525B - Ganoderma lucidum mycelium culture medium and culture method thereof - Google Patents
Ganoderma lucidum mycelium culture medium and culture method thereof Download PDFInfo
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Abstract
The invention discloses a ganoderma lucidum mycelium culture medium which comprises a seed culture medium and a fermentation culture medium, wherein the culture medium can provide nutrient substances required by the growth of ganoderma lucidum mycelium, promote the growth of ganoderma lucidum mycelium, improve the triterpenoids and ganoderma lucidum polysaccharide of the ganoderma lucidum mycelium, and also improve the tolerance of ganoderma lucidum to metal. The method is simple and effective, is easy to operate, can improve the production efficiency, and is favorable for the industrial production of the ganoderma lucidum mycelia with high yield of triterpenoids and polysaccharides.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to a ganoderma lucidum mycelium culture medium and a culture method thereof.
Background
Ganoderma (Ganoderma lucidum) belonging to Basidiomycetes, Polyporaceae, Ganoderma genus has various effective components, such as: polysaccharides, triterpenes, proteins, amino acid polypeptides, nucleosides, trace components and the like, wherein the ganoderma lucidum triterpenes and ganoderma lucidum polysaccharides are important components in ganoderma lucidum and have various effects of resisting virus, resisting tumor, resisting oxidation, neurasthenia, enhancing immunity, reducing hypertension, delaying aging and the like.
The traditional cultivation of ganoderma lucidum is easily influenced by external factors such as environment, artificial planting mode and the like, and has long growth period, thus being difficult to realize large-scale industrial production. The liquid fermentation ganoderma lucidum mycelium is a culture mode which can ensure the increase of the quality, the quantity and the polysaccharide content of the mycelium and quickly generate a large amount of secondary metabolites by introducing sterile air into a shake flask or a fermentation tank under the control of a proper culture medium and culture parameters, and has the advantages of high yield, short period, low cost, suitability for industrial production and the like.
The ganoderma triterpene and the ganoderma polysaccharide are used as main biological active components of the ganoderma, the further research on the ganoderma triterpene and the ganoderma polysaccharide is beneficial to searching the effective components in the ganoderma and further exploring new activity and clarifying the pharmacological activity mechanism of the ganoderma triterpene and the ganoderma polysaccharide, and the technical problem which needs to be solved at present is how to increase the yield of the ganoderma triterpene and the ganoderma polysaccharide by adjusting a fermentation culture medium and culture conditions.
Disclosure of Invention
The invention aims to provide a ganoderma lucidum mycelium culture medium which can provide nutrient substances required by the growth of ganoderma lucidum mycelium, promote the growth of ganoderma lucidum mycelium, improve triterpenoids and ganoderma lucidum polysaccharide of ganoderma lucidum mycelium and improve the tolerance of ganoderma lucidum to metal.
The above object of the present invention is achieved by the following technical solutions:
a ganoderma lucidum mycelium culture medium for high-yield triterpene comprises a seed culture medium and a fermentation culture medium, wherein 1L of the seed culture medium comprises the following components: 1-5g of glucose, 2-5g of peptone, 0.5-8g of yeast powder, 0.1-1.0g of monopotassium phosphate, 0.01-0.25g of magnesium sulfate heptahydrate, 0.05-0.05 g of vitamin B10.001, 1-5g of sodium deoxycholate, 0.01-0.05g of heme peptide and the balance of water;
the fermentation medium is 1L of culture medium which comprises 10-25g of galacto-oligosaccharide, 5-20g of aureobasidium polysaccharide, 5-10g of peptone, 0.5-8g of yeast powder, 0.1-1.0g of monopotassium phosphate, 0.01-0.5g of magnesium sulfate heptahydrate, 10.05-0.1g of vitamin B, 0.01-0.05g of heme peptide and the balance of water.
Experiments show that the yield of ganoderma triterpenes in ganoderma lucidum mycelia can be increased by taking galactooligosaccharide and aureobasidium pullulans as carbon sources and taking peptone and heme peptide as nitrogen sources; in addition, the addition of rosiglitazone into the fermentation medium can generate combined action with heme peptide, so that the yield of ganoderma triterpene can be further promoted, and the addition of aureobasidium polysaccharide into the fermentation medium can obviously increase the conversion rate of ganoderma polysaccharide.
Preferably, the seed culture medium is 1L culture medium composed of the following components: 2g of glucose, 3g of peptone, 1.5g of yeast powder, 0.3g of monopotassium phosphate, 0.15g of magnesium sulfate heptahydrate, 10.03g of vitamin B, 2.5g of sodium deoxycholate, 0.025g of heme peptide and the balance of water;
the fermentation medium is 1L of culture medium which comprises 25g of galacto-oligosaccharide, 18g of aureobasidium polysaccharide, 8g of peptone, 5g of yeast powder, 0.5g of potassium dihydrogen phosphate, 0.25g of magnesium sulfate heptahydrate and vitamin B10.03g, 0.035g heme peptide and the balance of water.
Another object of the present invention is to provide a method for culturing Ganoderma lucidum mycelia, comprising the steps of:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution;
and S2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.005-0.05g of rosiglitazone into the fermentation culture medium during seed solution inoculation, and filtering after fermentation to obtain the ganoderma lucidum mycelia.
Preferably, in step S1, the ganoderma lucidum mycelia are inoculated into the seed culture medium in an inoculation amount of 0.5-2%.
Preferably, the inoculation amount of inoculating the seed solution into the fermentation medium in the step S2 for fermentation culture is 2-15%.
Preferably, the conditions for inoculating the ganoderma lucidum mycelia into the seed culture medium in the step S1 are as follows: the temperature is 22-25 ℃, the rotating speed is 100-180r/min, the pressure is 0.03-0.05MPa, the air flow is 1:0.3-0.4v/v.min, and the dark culture is carried out for 2-5 days.
Preferably, the conditions for inoculating the ganoderma lucidum mycelia into the seed culture medium in the step S1 are as follows: the temperature is 25-30 ℃, the rotating speed is 80-160r/min, the pressure is 0.03-0.05MPa, the air flow is 1:0.5-1.0v/v.min, and dark culture is carried out for 4-6 days.
The addition of proper rosiglitazone into the culture medium can greatly improve the dissolved oxygen concentration and the permeability of cell membranes in the culture medium, does not influence the production capacity of strains, and changes the process of exchanging substances between cells and the environment, thereby promoting the synthesis of ganoderma triterpenoids from ganoderma mycelia; the addition of heme peptide and Aureobasidium pullulans in the fermentation medium can promote the formation of extracellular polymer of Ganoderma and increase Fe content of Ganoderma2+And Se2+The tolerance of the ganoderma lucidum is improved, and the growth of ganoderma lucidum hyphae is promoted; the addition of sodium deoxycholate in the culture medium can selectively inhibit the growth of bacteria, further promote the growth of ganoderma lucidum hyphae, and the aureobasidium pullulans polysaccharide and galacto-oligosaccharide can synergistically promote the conversion of ganoderma lucidum polysaccharide.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the invention, galactooligosaccharide and aureobasidium pullulans are used as carbon sources, peptone and heme peptide are used as nitrogen sources and used as fermentation culture media to culture ganoderma lucidum mycelia, so that the yield of ganoderma lucidum polysaccharides can be remarkably increased, and a proper amount of rosiglitazone is added in the fermentation process, so that the yield of ganoderma lucidum triterpenoids in ganoderma lucidum mycelia can be greatly increased.
(2) The invention adopts the culture medium to add the heme peptide and the aureobasidium polysaccharide to culture the ganoderma lucidum mycelia, can greatly improve the tolerance of the ganoderma lucidum mycelia to metal ions and promote the growth of the ganoderma lucidum mycelia.
(3) The method is simple and effective, is easy to operate, can improve the production efficiency, and is favorable for the industrial production of high-yield triterpenoid ganoderma lucidum mycelia.
Detailed Description
The present invention will be described in further detail below.
The ganoderma lucidum strain is purchased from China general microbiological culture collection center with CCGMC No. 5.616.
Example 1
Seed culture medium: 1g of glucose, 2g of peptone, 3g of yeast powder, 0.1g of monopotassium phosphate, 0.015g of magnesium sulfate heptahydrate, 10.007g of vitamin B, 2g of sodium deoxycholate, 0.03g of heme peptide and the balance of water.
Fermentation medium: 10g of galacto-oligosaccharide, 5g of aureobasidium pullulans, 5g of peptone, 5g of yeast powder, 0.1g of monopotassium phosphate, 0.014g of magnesium sulfate heptahydrate, 10.007g of vitamin B, 0.03g of heme peptide and the balance of water.
The method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 0.5%; culturing at 22 deg.C, rotation speed of 100r/min, pressure of 0.03MPa, airflow rate of 1:0.3v/v.min in dark for 5 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.005g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 5%, the temperature is 25 ℃, the rotating speed is 80r/min, the pressure is 0.03MPa, the airflow is 1:0.5v/v.min, and dark culture is carried out for 5 days.
Example 2
Seed culture medium: 5g of glucose, 3.5g of peptone, 4g of yeast powder, 0.5g of monopotassium phosphate, 0.25g of magnesium sulfate heptahydrate, 10.05g of vitamin B, 5g of sodium deoxycholate, 0.01g of heme peptide and the balance of water.
Fermentation medium: 25g of galacto-oligosaccharide, 5g of aureobasidium pullulans, 10g of peptone, 4g of yeast powder, 0.5g of monopotassium phosphate, 0.25g of magnesium sulfate heptahydrate, 10.05g of vitamin B, 0.05g of heme peptide and the balance of water.
The method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 2%; the temperature is 23 ℃, the rotating speed is 180r/min, the pressure is 0.04MPa, the airflow is 1:0.35v/v.min, and dark culture is carried out for 4 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.01g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 2%, the temperature is 26 ℃, the rotating speed is 90r/min, the pressure is 0.04MPa, the airflow is 1:0.6v/v.min, and dark culture is carried out for 5 days
Example 3
Seed culture medium: 2.5g of glucose, 2g of peptone, 5g of yeast powder, 0.3g of monopotassium phosphate, 0.01g of magnesium sulfate heptahydrate, 10.001g of vitamin B, 1g of sodium deoxycholate, 0.05g of heme peptide and the balance of water.
Fermentation medium: 15g of galacto-oligosaccharide, 13g of aureobasidium pullulans, 6g of peptone, 6g of yeast powder, 0.6g of monopotassium phosphate, 0.03g of magnesium sulfate heptahydrate, 10.06g of vitamin B, 0.03g of heme peptide and the balance of water;
the method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 1%; culturing at 24 deg.C, rotation speed of 140r/min, pressure of 0.04MPa, airflow rate of 1:0.3v/v.min in dark for 4 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.0075g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 15%, the temperature is 27 ℃, the rotating speed is 160r/min, the pressure is 0.05MPa, the airflow is 1:0.7v/v.min, and dark culture is carried out for 4 days.
Example 4
Seed culture medium: 3g of glucose, 2.5g of peptone, 6g of yeast powder, 0.55g of monopotassium phosphate, 0.15g of magnesium sulfate heptahydrate, 10.025g of vitamin B, 3g of sodium deoxycholate, 0.35g of heme peptide and the balance of water.
Fermentation medium: 20g of galacto-oligosaccharide, 10g of aureobasidium pullulans, 7g of peptone, 8g of yeast powder, 0.7g of monopotassium phosphate, 0.4g of magnesium sulfate heptahydrate, 10.08g of vitamin B, 0.035g of heme peptide and the balance of water;
the method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 2%; culturing at 25 deg.C, rotation speed of 160r/min, pressure of 0.03MPa, airflow rate of 1:0.3-0.4v/v.min in dark for 2-5 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.02g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 7%, the temperature is 28 ℃, the rotating speed is 120r/min, the pressure is 0.03MPa, the airflow is 1:0.6v/v.min, and dark culture is carried out for 5 days.
Example 5
Seed culture medium: 2g of glucose, 3g of peptone, 1.5g of yeast powder, 0.3g of monopotassium phosphate, 0.15g of magnesium sulfate heptahydrate, 10.03g of vitamin B, 2.5g of sodium deoxycholate, 0.025g of heme peptide and the balance of water.
Fermentation medium: 25g of galactooligosaccharide, 18g of aureobasidium pullulans polysaccharide, 8g of peptone, 5g of yeast powder, 0.5g of monopotassium phosphate, 0.25g of magnesium sulfate heptahydrate, 0.035g of vitamin B, 0.05g of heme peptide and the balance of water.
The method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 1.5%; culturing at 25 deg.C, rotation speed of 140r/min, pressure of 0.04MPa, airflow rate of 1:0.4v/v.min in dark for 3 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.025g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 10%, the temperature is 29 ℃, the rotating speed is 135r/min, the pressure is 0.04MPa, the airflow is 1:0.7v/v.min, and dark culture is carried out for 5 days.
Example 6
Seed culture medium: 4g of glucose, 3g of peptone, 8g of yeast powder, 1.0g of monopotassium phosphate, 0.25g of magnesium sulfate heptahydrate, 10.05g of vitamin B, 1g of sodium deoxycholate, 0.01g of heme peptide and the balance of water.
Fermentation medium: 17g of galacto-oligosaccharide, 20g of aureobasidium pullulans polysaccharide, 3g of peptone, 8g of yeast powder, 1.0g of monopotassium phosphate, 0.25g of magnesium sulfate heptahydrate, 10.05g of vitamin B, 0.025g of heme peptide and the balance of water.
The method for culturing the ganoderma lucidum mycelia with high triterpene yield comprises the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution, wherein the inoculation amount is 1.3%; culturing at 25 deg.C, rotation speed of 150r/min, pressure of 0.04MPa, airflow rate of 1:0.4v/v.min in dark for 4 days;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.05g of rosiglitazone into the fermentation culture medium during seed solution inoculation, filtering after fermentation to obtain ganoderma lucidum mycelia, wherein the inoculation amount is 15%, the temperature is 30 ℃, the rotating speed is 160r/min, the pressure is 0.05MPa, the airflow is 1:1.0v/v.min, and dark culture is carried out for 5 days.
Comparative example 1
The difference from example 5 is that the seed medium and the fermentation medium were not supplemented with the heme peptide and the deletion portion was replaced with a corn oligopeptide.
Comparative example 2
The difference from example 5 is that no sodium deoxycholate was added to the seed medium, and the mass of the missing part was supplemented with glucose.
Comparative example 3
The difference from example 5 is that the galactooligosaccharide of the fermentation medium is replaced by lactose.
Comparative example 4
The difference from example 5 is that the galactooligosaccharides of the fermentation medium are replaced by glucose.
Comparative example 5
The difference from example 5 is that no Aureobasidium pullulans was added to the fermentation medium and the mass of the missing part was made up with galacto-oligosaccharides.
Comparative example 6
The difference from example 5 is that the fermentation medium is not supplemented with galacto-oligosaccharides and the missing mass is filled with pullulan.
Comparative example 7
The difference from example 5 is that in the culture method, rosiglitazone was not added at the time of inoculation of the seed solution.
Test example 1 measurement of hyphal Biomass, intracellular triterpene content and ganoderan
Measurement of hyphal biomass: the fermented mycelia of examples 1 to 6 and comparative examples 1 to 7 were mixed with water, filtered, the mycelia and the fermentation broth were separated, the obtained mycelia were dried at 60 ℃ to constant weight, and the biomass of each strain was calculated (in g of dry weight of mycelia per 100mL of fermentation broth).
Determination of the yield of intracellular triterpenes: the fermented ganoderma lucidum mycelia of examples 1-6 and comparative examples 1-7 were washed with 95% (v/v) ethanol for 2 times, centrifuged, and mycelia were collected, and an appropriate amount of mycelia was taken and added to 95% (v/v) ethanol at 1g/50mL and disrupted with a cell disrupter (1 min. times.4), extracted at 60 ℃ for 1h at 400W, and repeated once, and the supernatants were combined and fixed to volume to obtain solutions to be tested of examples 1-6 and comparative examples 1-7. The content of the triterpene is determined by adopting a vanillin perchloric acid method.
Vanillin-perchloric acid assay method: respectively putting 0.1mL of the solution to be detected of the examples 1-6 and the comparative examples 1-7 in a test tube, heating at 70 ℃ to volatilize the solvent, adding 0.2mL of 5% (m/v) vanillin glacial acetic acid solution and 0.5mL of perchloric acid, uniformly mixing, putting in a water bath at 60 ℃, preserving heat for 20min, taking out, putting in cold water for 10min, finally adding 5mL of glacial acetic acid, and determining the light absorption value at 550 nm.
And (3) measuring the content of the ganoderma lucidum polysaccharide: the measurement is carried out by a phenol-sulfuric acid method, 10mg of the ganoderma lucidum mycelia of examples 1-6 and comparative examples 1-7 are weighed in a 100ml volumetric flask, about 80ml of water is added, the mixture is heated to assist dissolution and then cooled to room temperature, and the volume is determined in the 100ml volumetric flask. Precisely sucking a sample of 1.0m1, placing in a 15ml test tube, respectively adding phenol and sulfuric acid reagents, fully reacting, measuring the absorbance value at 490nm, and calculating the sugar content of the sample by taking glucan as a standard substance.
The measurement results of the hyphal biomass and the intracellular triterpene content are shown in Table 1.
TABLE 1 measurement of hyphal biomass and intracellular triterpene content
| Hypha biomass g/100mL | Yield of intracellular triterpene mg/g | The yield of ganoderma lucidum polysaccharide is mg/g | |
| Example 1 | 6.05 | 11.11 | 14.77 |
| Example 2 | 5.14 | 13.41 | 15.25 |
| Example 3 | 6.69 | 10.37 | 16.96 |
| Example 4 | 6.36 | 11.69 | 15.14 |
| Example 5 | 8.56 | 15.78 | 18.12 |
| Example 6 | 7.08 | 12.99 | 17.84 |
| Comparative example 1 | 5.01 | 9.61 | 11.11 |
| Comparative example 2 | 4.11 | 8.62 | 12.01 |
| Comparative example 3 | 5.89 | 7.32 | 14.11 |
| Comparative example 4 | 5.09 | 6.13 | 13.78 |
| Comparative example 5 | 4.39 | 5.51 | 8.74 |
| Comparative example 6 | 4.86 | 6.44 | 9.01 |
| Comparative example 7 | 4.05 | 5.99 | 8.45 |
As can be seen from Table 1, the present example can significantly improve the biomass of Ganoderma lucidum mycelia, the yield of Ganoderma lucidum intracellular triterpenes and the yield of Ganoderma lucidum polysaccharides, wherein the biomass of Ganoderma lucidum mycelia reaches 5.14-8.56g/100mL, the yield of Ganoderma lucidum intracellular triterpenes reaches 10.37-15.78mg/g, and the yield of Ganoderma lucidum polysaccharides reaches 14.77-18.12mg/g, wherein example 5 is the best example.
Test example 2 measurement of Ganoderma lucidum hypha Biomass under Metal stress
Slowly shaking Ganoderma mycelia obtained from fermentation culture of examples 1-6 and comparative examples 1-7, without filtering, subpackaging into 500mL triangular flasks with 200mL each, and respectively inoculating 800mg/L sterilized Fe2+Solution and Se at a final concentration of 100mg/L2+After the solution is continuously cultured for three days, accurately measuring 100ml of mycelium fermentation liquor in a conical flask, adding water into the mycelium, uniformly mixing, carrying out suction filtration, separating the mycelium and the fermentation liquor, taking the amount of wet mycelium as the biomass, and obtaining the measurement result shown in table 2.
TABLE 2 determination of Ganoderma mycelium biomass under metal stress
As shown in Table 2, this example can significantly improve the Fe content in the Ganoderma mycelium2+And Se2+Tolerance of (3), Fe2+Under the stress, the biomass of ganoderma lucidum hypha reaches 7.23-9.94g/100mL, and Se is2+Under the stress, the biomass of ganoderma lucidum mycelia reaches 7.15-9.67g/100mL, wherein example 4 is the best example.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. All cooked
Those skilled in the art can modify and change the above-described embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (6)
1. A ganoderma lucidum mycelium culture medium comprises a seed culture medium and a fermentation culture medium, and is characterized in that the seed culture medium is characterized in that 1L of the culture medium consists of the following components: 1-5g of glucose, 2-5g of peptone, 0.5-8g of yeast powder, 0.1-1.0g of monopotassium phosphate, 0.01-0.25g of magnesium sulfate heptahydrate, 0.05-0.05 g of vitamin B10.001, 1-5g of sodium deoxycholate, 0.01-0.05g of heme peptide and the balance of water;
the fermentation culture medium is 1L culture medium composed of galacto-oligosaccharide 10-25g, aureobasidium polysaccharide 5-20g, peptone 5-10g, yeast powder 0.5-8g, potassium dihydrogen phosphate 0.1-1.0g, magnesium sulfate heptahydrate 0.01-0.5g, vitamin B10.05-0.1g, 0.01-0.05g of heme peptide and the balance of water.
2. A method for culturing ganoderma lucidum mycelia is characterized by comprising the following steps:
s1, inoculating the ganoderma lucidum mycelia into a seed culture medium for culture to obtain a seed solution;
s2, inoculating the seed solution into a fermentation tank filled with a fermentation culture medium for fermentation tank culture, adding 0.005-0.05g of rosiglitazone into the fermentation culture medium during seed solution inoculation, and filtering after fermentation to obtain ganoderma lucidum mycelia;
the seed culture medium is characterized in that 1L of the culture medium consists of the following components: 1-5g of glucose, 2-5g of peptone, 0.5-8g of yeast powder, 0.1-1.0g of monopotassium phosphate, 0.01-0.25g of magnesium sulfate heptahydrate, 0.05-0.05 g of vitamin B10.001, 1-5g of sodium deoxycholate, 0.01-0.05g of heme peptide and the balance of water;
the fermentation culture medium is 1L culture medium composed of galacto-oligosaccharide 10-25g, aureobasidium polysaccharide 5-20g, peptone 5-10g, yeast powder 0.5-8g, potassium dihydrogen phosphate 0.1-1.0g, magnesium sulfate heptahydrate 0.01-0.5g, vitamin B10.05-0.1g, 0.01-0.05g of heme peptide and the balance of water.
3. The method according to claim 2, wherein the Ganoderma lucidum mycelia is inoculated into the seed medium in the step S1 in an amount of 0.5-2%.
4. The culture method according to claim 2, wherein the inoculation amount of the seed solution into the fermentation medium for fermentation culture in step S2 is 2-15%.
5. The culture method according to claim 2, wherein the conditions for inoculating Ganoderma lucidum mycelia into the seed culture medium in step S1 are as follows: the temperature is 22-25 ℃, the rotating speed is 100-180r/min, the pressure is 0.03-0.05MPa, the air flow is 1:0.3-0.4v/v.min, and the dark culture is carried out for 2-5 days.
6. The culture method according to claim 2, wherein the conditions for inoculating Ganoderma lucidum mycelia into the seed culture medium in step S1 are as follows: the temperature is 25-30 ℃, the rotating speed is 80-160r/min, the pressure is 0.03-0.05MPa, the air flow is 1:0.5-1.0v/v.min, and the dark culture is carried out for 3-5 days.
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| CN110172492A (en) * | 2019-06-13 | 2019-08-27 | 上海市农业科学院 | A kind of method of liquid submerged fermentation production ganodenic acid |
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