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Method for increasing yield of phellinus linteus total triterpene compound

A technology for total triterpenes and compounds, applied in the field of biological fermentation engineering, can solve problems such as the lack of a clear verification conclusion, and achieve the effects of promoting and improving effects, simple preparation methods, and simple processes and extraction methods.

Inactive Publication Date: 2012-08-08
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 verification conclusion and mechanism elucidation at home and abroad.
Therefore, there are very few research reports on the various applications of expansin, and the application of expansin in the liquid fermentation production of Phellinus chinensis to improve the output of secondary metabolites such as Phellinus triterpenoids has not yet been seen at home and abroad. to report

Method used

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  • Method for increasing yield of phellinus linteus total triterpene compound

Examples

Experimental program
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Effect test

preparation example Construction

[0034] The preparation steps of the expansin solution described in the examples are as follows:

[0035] Soybean (Glycine max L.Merr.CV.M40; purchased from Jinan Weili Seed Industry Co., Ltd.) or cucumber (Cucumissativus 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

[0044] A liquid fermentation method for improving the output of total triterpenoids of Phellinus Phellinus by using expansin, comprising the following steps:

[0045] (1) Phellinus bacterium is inoculated on the PDA plate culture medium, carries out activation culture, culture temperature 25 ℃, culture time 5 days, then get the bacterial block 6 pieces of diameter 0.3cm, inoculate in containing PD liquid fermentation medium Carry out fermentation culture in the Erlenmeyer flask (every 500mL Erlenmeyer flask contains 150mL of PD liquid fermentation medium), the liquid fermentation culture conditions are: temperature 25°C, shaker culture for 2 days, shaker speed 150r / min, to obtain the primary fermentation liquid;

[0046] (2) Add the expansin solution to the initial fermentation broth prepared in step (1), so that the final concentration of the expansin is 0.2 mg / mL, and then continue culturing at a temperature of 25°C and a shaker speed of 150r / min. 5 days, centrifuged under t...

Embodiment 2

[0066] The liquid fermentation method as described in Example 1, the difference is that the concentration of expansin in step (2) is 0.4mg / mL;

[0067] After testing and calculation, every milliliter of fermented liquid contains 952.12 μg of total extracellular triterpenoids of Phellinus radix, that is, 952.12 mg of total extracellular triterpenoids of Phellinus radix per liter of fermented liquid; every gram of mycelia (dry weight) Contains 17.26mg of total triterpenoids in Phellinus Phellinus cells, which corresponds to 333.47mg of total triterpenoids in Phellinus Phellinus cells per liter of fermentation broth. The result is as figure 1 shown.

[0068] Therefore, the output of total triterpenoids of Phellinus per liter of fermentation broth is: 952.12mg+333.47mg=1285.59mg.

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Abstract

The invention relates to a liquid fermentation method for increasing the yield of a phellinus linteus total triterpene compound. The method comprises the following steps of: (1) inoculating a phellinus linteus strain onto a plating medium, activating, culturing, and inoculating a strain block into a liquid fermentation culture medium for fermenting and culturing to obtain a primary zymotic fluid; (2) adding an expansion protein solution into the primary zymotic fluid, and continually culturing for 3-5 days to obtain a phellinus linteus mycelium and a zymotic fluid; and (3) extracting a phellinus linteus extracellular total triterpene compound from the zymotic fluid, extracting a phellinus linteus intracellular total triterpene compound from the phellinus linteus mycelium, and mixing the phellinus linteus extracellular total triterpene compound and the phellinus linteus intracellular total triterpene compound to obtain the phellinus linteus total triterpene compound. In the method, an expansion protein is applied to liquid fermentation of the phellinus linteus total triterpene compound, so that the yields of the phellinus linteus extracellular total triterpene compound and the phellinus linteus intracellular total triterpene compound are increased by 2.52 times and 2.19 times in comparison to the prior art, and the phellinus linteus total triterpene compound has good industrial application prospect.

Description

technical field [0001] The invention relates to a liquid fermentation method for increasing the yield of total triterpenoids of Phellinus radix, belonging to the technical field of biological fermentation engineering. Background technique [0002] Phellinus igniarius belongs to Basidiomycotina Basidiomycotina, Hymenomycetes Hymenomycetes, Polyporales Polyporales, Hymenochaetacae Prussiaceae, Phellinus fungus, and is a precious large medicinal fungus. Traditional Chinese medicine believes that Phellinus is sweet in nature and bitter in taste, and it is used to treat metrorrhagia, leukorrhea, amenorrhea, spleen deficiency and diarrhea. Modern pharmacological studies have shown that Phellinus has anti-tumor, immune-enhancing, anti-hepatic fibrosis and other effects. Phellinus is reported to be the medicinal fungus with the highest anti-tumor ability at present. As early as 1968, Japanese scholar Tetsuro Ikekawa et al. conducted cell experiments with the water extract of Phell...

Claims

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Application Information

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IPC IPC(8): C12P1/02C12R1/645
CPCC12P33/00C12P1/02C07J63/00C12P5/007C12R1/645C12P15/00A61P35/00A61P37/02C07J63/008C12N1/145C12R2001/645
Inventor 姚强宫志远单洪涛韩建东王琦高能孙涛万鲁长任鹏飞李瑾曲玲
Owner INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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