Glossy ganoderma cell culturing method for improving ganoderic acid content in cells

A cell culture, ganoderma lucidum acid technology, applied in the fields of botanical equipment and methods, horticulture, agriculture, etc., can solve the problems of restricting the research and wide application of ganoderma acid activity and action mechanism, difficult separation and purification, and low ganoderma acid content.

Active Publication Date: 2010-04-14
TAIXING DONGSHENG FOOD TECH
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  • Description
  • Claims
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Problems solved by technology

However, Ganoderma acid has a wide range of pharmacological activities. Further research on Ganoderma lucidum will facilitate the search for active ingredients in Ganoderma lucidum, further explore new activities and clarify its pharmacological activity mechanism. The content of Ganoderma acid in Ganoderma lucidum cells is generally very low , separation and purification are also difficult, which limits the research and wide application of the activity and mechanism of ganoderma acid

Method used

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  • Glossy ganoderma cell culturing method for improving ganoderic acid content in cells
  • Glossy ganoderma cell culturing method for improving ganoderic acid content in cells
  • Glossy ganoderma cell culturing method for improving ganoderic acid content in cells

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Embodiment 1

[0023] step one,

[0024] Prepare seed medium, the seed medium is specifically, 1L medium is composed of the following components, glucose 35g, peptone 5g, yeast powder 2.5g, potassium dihydrogen phosphate 1.0g, MgSO4 7H2O 0.5g, vitamin B1 0.05g , the balance is water;

[0025] Preparation of fermentation medium, the fermentation medium is specifically, 1L medium is composed of the following components, lactose 35g, peptone 5g, yeast powder 5g, potassium dihydrogen phosphate 1.0g, MgSO4 7H2O 0.5g, vitamin B1 0.05g, The remainder is water;

[0026] Preparation of 0.2M phenobarbital solution: Weigh 0.0464 g of phenobarbital and dissolve it in 1 ml of DMSO.

[0027] Step 2, in a 250mL shake flask, insert 10mL of Ganoderma lucidum CGMCC NO.5616 mycelia into 40mL of seed culture medium, and cultivate at 30°C and 120rpm for 6 days to obtain a primary seed culture solution;

[0028] Step 3: In a 250mL shake flask, add 5mL of the primary seed culture solution to 45mL of the seed me...

Embodiment 2

[0038] The steps of this embodiment are the same as those in Embodiment 1, the difference is that:

[0039] In step one, prepare the phenobarbital solution:

[0040] 2M phenobarbital solution: Weigh 0.4645 g of phenobarbital and dissolve it in 1 ml of DMSO.

[0041] 0.2M phenobarbital solution: take 100 μl of the above 1.0 mM phenobarbital solution, and add 900 μl DMSO.

[0042] 0.02M phenobarbital solution: take 100 μl of the above 0.1 mM phenobarbital solution, and add 900 μl DMSO.

[0043] Step 5, prepare four parts of fermented cells (50ml each) obtained in step 4, place them in petri dishes with a diameter of 10cm respectively, and carry out static culture; the first part of fermented cells does not add phenobarbital, and the remaining three parts of fermented cells are in On the fifth day, 2M, 0.2M, and 0.02M phenobarbital solutions were added, so that the final concentrations of phenobarbital in the fermented cells were: 1.0mM, 0.1mM, and 0.01mM; the four fermentation...

Embodiment 3

[0048] The steps of this embodiment are the same as those in Embodiment 1, the difference is that:

[0049] In step one, prepare miconazole solution:

[0050] 0.2M miconazole solution: Weigh 0.096g of miconazole and dissolve it in 1ml of DMSO.

[0051] 0.02M miconazole solution: Take 100 μl of the above 0.2M phenobarbital solution and add 900 μl DMSO.

[0052] Step two to step four are all the same as embodiment 1,

[0053] Step five, prepare four parts of fermented cells obtained in step four (50ml each), place them in culture dishes with a diameter of 10cm respectively, and carry out static culture; the first fermented cells are not added to miconazole during static culture; in the second part Add 0.02M miconazole on the second day of static culture of fermented cells, so that the final concentration of miconazole is 0.1mM; The final concentration of azole was 0.1mM; 0.02M miconazole was added on the seventh day of static culture of the fourth fermented cells, so that the...

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Abstract

A glossy ganoderma cell culturing method for improving ganoderic acid content in cells in the biological technical field comprises the following steps: preparing seed culture medium and fermentation culture medium, switching ganoderma lucidum bodies into the seed culture medium, culturing, obtaining first-grade seed culture fluid, taking the first-grade seed culture fluid, switching into the seed culture fluid, culturing, obtaining second-grade seed culture fluid, taking the second-grade seed culture fluid, switching into the formation culture medium, culturing, obtaining fermentation cells, placing the fermentation cells into a static culture vessel, statically culturing, adding aphenyletten or miconazole in the cell growth logarithmic phase, continuing to culture statically until the harvest period, and collecting super cells. The method of the invention enables total ganoderic acid contents in the ganoderma lucidum to achieve 41.4mg / g stem cells, and simultaneously the contents of monomer ganoderic acid contents are also improved by 15% to 60%.

Description

technical field [0001] The invention relates to a method in the field of biotechnology, in particular to a ganoderma cell culture method for increasing the content of ganoderma acid in cells. Background technique [0002] Ganoderma lucidum is a higher fungus belonging to Basidiomycetes, Polyporaceae, and Ganoderma lucidum. It has been used as a traditional Chinese medicine for thousands of years. Pharmacological studies have shown that Ganoderma lucidum has a wide range of pharmacological effects, such as anti-HIV virus, immune regulation, anti-myocardial ischemia, blood lipid regulation, hypoglycemia, sedation, liver protection, anti-radiation and anti-chemotherapy, anti-hypoxia and anti-aging effects Wait. In modern times, people began to study and obtain a large amount of Ganoderma lucidum mycelium and its active ingredients through artificial cultivation and submerged fermentation technology, which provided sufficient raw materials for the pharmacological research and p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G1/04C05G1/00
Inventor 钟建江梁翠霞李英波王家乐
Owner TAIXING DONGSHENG FOOD TECH
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