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Liquid fermentation method for improving yield of cordyceps sinensis polysaccharide by utilizing expansin

A technology of liquid fermentation and protein expansion, which is applied in the direction of using fungi, methods based on microorganisms, fermentation, etc., can solve the problems of few reports on protein expansion, achieve good industrial application prospects, promote and improve fermentation production, and increase production Effect

Inactive Publication Date: 2012-08-01
INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism of action of expansin is still mostly speculation and inference, and there is no clear research conclusion and mechanism elucidation at home and abroad, so there are few reports on the practical application of expansin
The application of expansin in the liquid fermentation production of Cordyceps militaris to increase the production of secondary metabolites such as Cordyceps polysaccharides has not been reported at home and abroad.

Method used

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  • Liquid fermentation method for improving yield of cordyceps sinensis polysaccharide by utilizing expansin

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0037] The preparation steps of the expansin solution described in the examples are as follows:

[0038] Soybean (Glycine max L.Merr.CV.M40; purchased from Jinan Weili Seed Industry Co., Ltd.) or cucumber (Cucumis sativus L.CV. Jinnian No.6; purchased from Jinan Weili Seed Industry Co., Ltd.) seeds were subjected to 0.1 wt% HgCl 2 Disinfect for 5 minutes, rinse with running water for 6 hours, then plant in wet vermiculite, and culture in dark at 27°C for 5 days. Cut off the top 3-4cm of the hypocotyl of the seedlings, that is, about 100g of the growth area, pre-cool in a -20°C refrigerator for 0.5h, add a homogenization buffer pre-cooled to 4°C, and after high-speed homogenization, filter with a 70μm nylon mesh. Wash the filter residue with homogenization buffer, then add the filter residue to the homogenization buffer, and stand at room temperature for 2 hours to obtain a standing solution; add the extract to the standing solution, extract at 4°C for 48 hours, filter, and th...

Embodiment 1

[0051] A liquid fermentation method for improving the yield of Cordyceps polysaccharide by utilizing expansin, comprising the following steps:

[0052] (1) Inoculate the strain of Cordyceps militaris (CGMCC No.5.699) into PD liquid fermentation medium, and carry out activation culture. Cultivate for 96 hours, then transfer the inoculum to the seed fermentation medium according to the volume percentage of 15%, and carry out seed cultivation. The seed cultivation conditions are: temperature 25°C, shaker cultivation for 3 days, to obtain seed liquid;

[0053] (2) Inoculate the seed solution prepared in step (1) in a liquid fermentation medium at a volume ratio of 10%, and carry out liquid fermentation culture for 1 day at a temperature of 25° C., and then add the expansin solution to increase the concentration of the expansin 0.15mg / mL, then continue to cultivate for 9 days, and centrifuge for 10min under the condition of 15000r / min to obtain Cordyceps militaris mycelium and ferm...

Embodiment 2

[0071] The liquid fermentation method as described in Example 1, the difference is that in step (2), under the condition of a temperature of 25°C, the liquid fermentation culture was carried out for 3 days, and then the expansin solution was added to make the concentration of the expansin 0.5 mg / mL, and then continue to culture for 7 days,

[0072] After detection and calculation, each milliliter of fermentation broth contains 7.83 mg of exopolysaccharide of Cordyceps militaris, that is, 7.83 g of exopolysaccharide per liter of fermentation broth; 260.15 mg of intracellular polysaccharide of Cordyceps militaris is contained in each gram of mycelium (dry weight). That is, the mycelia produced by each liter of fermentation broth contains 1.73g of intracellular polysaccharide of Cordyceps militaris. The result is as figure 1 shown.

[0073] Therefore, the total yield of Cordyceps polysaccharide is 7.83+1.73=9.56g per liter of fermentation broth.

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Abstract

The invention relates to a liquid fermentation method for improving yield of cordyceps sinensis polysaccharide by utilizing expansin. The liquid fermentation method includes inoculating cordyceps militaris strains to a PD liquid fermentation medium for activated culture, inoculating the strains to a strain fermentation medium with 10-15% inoculum amount for strain culture to obtain a strain liquid; inoculating the strain liquid to the liquid fermentation medium for liquid culture, adding an expensin solution for continuous culture, separating the liquid to obtain cordyceps militaris mycelium and a fermentation broth; extracting cordyceps militaris extracellular polysaccharide from the fermentation broth, extracting cordyceps militaris intracellular polysaccharide from the cordyceps militaris mycelium, and mixing the cordyceps militaris extracellular polysaccharide and the cordyceps militaris intracellular polysaccharide to obtain the cordyceps sinensis polysaccharide. According to the liquid fermentation method for improving the yield of the cordyceps sinensis polysaccharide by utilizing the expansin, the expansin is applied to liquid fermentation production of the cordyceps militaris polysaccharide, compared with the prior art, yields of the intracellular polysaccharide and the extracellular polysaccharide are improved by more than 1.5 times and 6.9 times respectively, and the industrial application prospect is bright.

Description

technical field [0001] The invention relates to a liquid fermentation method for increasing the yield of cordyceps polysaccharide by using expansin, and belongs to the technical field of biological fermentation engineering. Background technique [0002] Cordyceps militaris is the model species of Ascomycota, Hypocreales, Clavicipitaceae, Cordyceps. The scientific name is Cordyceps militaris, also known as Cordyceps militaris, Cordyceps militaris, Cordyceps militaris, etc. It is a complex composed of two parts, the subseapotta (that is, the grass part) and the sclerotia (that is, the dead body part of the insect). It is distributed worldwide but the amount of natural resources is very small. few. Traditional Chinese medicine believes that Cordyceps enters the two meridians of the lung and kidney, which can not only nourish the lung yin, but also nourish the kidney yang. Chinese medicine for yin and yang. A large number of modern pharmacological studies have shown that Cord...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12R1/645
CPCC12P19/04C12R1/645C12N1/14C12N1/38
Inventor 姚强宫志远单洪涛韩建东王琦高能孙涛万鲁长任鹏飞赵军胜曲玲
Owner INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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