Cell culturing method for accelerating synthesis of glossy ganoderma secondary metabolite

A technology for secondary metabolites and cell culture, applied in the field of microbial cell culture, can solve the problems of low yield of Ganoderma lucidum polysaccharide and Ganoderma lucidum acid.

Inactive Publication Date: 2008-07-09
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the defects of low production of Ganoderma lucidum polysaccharide and Ganoderma acid in the existing Ganoderma cell culture method, and

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033]One, the preparation of the fermentation mycelia of Penicillium citrinum (purchased from China Center for Type Culture Collection, its number is AF93032):

[0034] Medium (g / L): 30 sucrose, 3 sodium nitrate, 0.5 magnesium sulfate heptahydrate, 0.5 potassium chloride, 0.01 ferrous sulfate tetrahydrate, 1 dipotassium hydrogen phosphate, 15 agar (agar is not added for liquid culture);

[0035] Culture conditions: 250 ml shake flask, liquid volume is 50 ml; culture temperature 25°C; 120 rpm;

[0036] Two, prepare 3 kinds of fungal elicitors by Penicillium citrinum mycelia:

[0037] (1) The preparation method of the all-component elicitor: the Penicillium citrinum mycelium was collected by filtration and washed twice with deionized water, then soaked in a 0.1 mol / liter acetate buffer solution with a pH value of 5.6 and homogenized, and the homogenate was Centrifuge at 5,500 rpm at high speed, take the supernatant, and sterilize it under high pressure;

[0038] (2) The prepa...

Embodiment 2

[0053] The added elicitors are three kinds of elicitors prepared by Chinese truffle (Tuber sinense);

[0054] The preparation method of Chinese truffle (Tuber sinense) mycelium is as follows:

[0055] (1) Incline cultivation: PDA, 25°C, culture for 6 days;

[0056] (2) First-level liquid seed culture: culture medium (g / L): glucose 35, peptone 5, yeast powder 2.5, magnesium sulfate heptahydrate 0.5, potassium dihydrogen phosphate 1, vitamin B 1 0.05, pH 5.5; culture conditions: 50ml medium / 250ml shake flask, 25°C, 120 rpm.

[0057] (3) Secondary liquid seed culture: culture medium (g / L): glucose 35, peptone 5, yeast powder 2.5, magnesium sulfate heptahydrate 0.5, potassium dihydrogen phosphate 1, vitamin B 1 0.05, pH5.5; culture conditions: 200ml medium / 500ml shake flask, 25°C, 120 rpm;

[0058] (4) Liquid submerged fermentation: fermentation medium (g / L): glucose 35, peptone 5, yeast powder 5, magnesium sulfate heptahydrate 0.5, potassium dihydrogen phosphate 1, vitamin B ...

Embodiment 3

[0064] Compared with Example 2, the elicitor of Example 3 was prepared from the mycelium of Tuber melanosporum, and other conditions were the same as in Example 2. The experimental results are shown in Table 3.

[0065] Table 3 Contents and yields of secondary metabolites of Ganoderma lucidum induced by three kinds of truffle elicitors

[0066] control group

[0067] It can be seen from Table 3 that, compared with the control group (without adding the elicitor), after adding the polysaccharide protein elicitor prepared by the black spore truffle (Tubermelanosporum) mycelium, the content of ganoderma acid was greatly improved, which was higher than that of the control group. 89.9%, which indicates that the addition of this elicitor during the submerged fermentation of Ganoderma lucidum can effectively promote the biosynthesis of Ganoderma acid.

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PUM

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Abstract

The invention discloses a cell culture method used for accelerating biological synthesis of ganoderam lucidum karst secondary metabolite, which comprises the following procedures: inoculating ganoderam lucidum karst bacterial to the oblique plane culture medium in order to culture; proceeding with one-stage liquid breed culture, two-stage liquid breed culture and liquid submerged cultuse sequentially, wherein definite quantity induction zygotic which is prepared by funginite mycelium is added in order to induce the biological synthesis of ganoderam lucidum karst polysaccharide and ganoderam lucidum karst acid during the liquid submerged cultuse process. The ganoderam lucidum karst polysaccharide content and ganoderam lucidum karst acid content of cell culture are determined separately with concentrated sulfuric acid-phenol method and ultraviolet spectrophotometry method, the result shows that the method can accelerate he biological synthesis of ganoderam lucidum karst polysaccharide and ganoderam lucidum karst acid in the culture and provides the basis for industry production and commercial production of ganoderam lucidum karst polysaccharide and ganoderam lucidum karst acid.

Description

technical field [0001] The invention relates to a microbial cell culture method, in particular to a ganoderma cell culture method for promoting the biosynthesis of ganoderma polysaccharides and ganoderma acid by adding elicitors in the ganoderma liquid submerged fermentation process, and belongs to the field of microbial fermentation. Background technique [0002] Ganoderma lucidum (Leyss ex Fr.) Krast, belonging to Basidiomycetes, Polyporaceae, and Ganoderma genus, is a precious medicinal fungus with a wide range of biological activities, such as anti-tumor, anti-oxidation, etc., and has been widely used in China. It has a history of more than two thousand years. Since ancient times, it has been regarded as a treasure to prolong life. It is a precious traditional Chinese medicine that nourishes and strengthens, strengthens the body, and helps maintain the balance of the body. Ganoderma lucidum has a long history of medicinal use in Southeast Asia, and the people of our cou...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/04C12P7/40C12R1/645
Inventor 汤亚杰朱丽雯李冬生
Owner HUBEI UNIV OF TECH
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