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Method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis)

An endophytic fungus and extraction method technology, applied in the field of chemical treatment of natural medicines, can solve the problems of difficulty in purification, low efficiency, restriction of swainsonine, etc., and achieve the effects of easy industrial production, simple process conditions, and expansion of raw material resources.

Pending Publication Date: 2022-05-13
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, due to the long extraction period, low efficiency, complex artificial synthesis process, and difficult purification of pure swainsonine, the price of pure swainsonine is extremely expensive, as high as 1400 yuan / mg, which seriously restricts swainsonine. In-depth research as a drug

Method used

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  • Method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis)
  • Method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis)
  • Method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Extraction method of swainsonine in locoweed endophytic fungus (U.oxytropis)

[0035] The locoweed endophyte (U. oxytropis) was inoculated in the PDA medium, and cultured in a constant temperature incubator at 25°C for 30 days until recovery. Use the inoculation loop to pick the mycelia into a sterile tissue grinder, add sterile water to grind to make a bacterial suspension for later use; transfer 500 μL of the bacterial suspension to a 300 mL Erlenmeyer flask filled with 150 mL of sterilized potato dextrose water, Co-cultivate 32 bottles, wrap the mouth of the bottle with air-permeable sealing film, put it in a constant temperature shaker at 27°C, 130r / min, and culture continuously for 24 days. After the cultivation, use filter paper to filter the fermented mycelium balls, rinse them with ultrapure water for 3 times, then transfer the filtered mycelium balls into a glass plate, and dry them in an oven at 45°C until they reach the quality. No change. The dr...

Embodiment 2

[0038] Embodiment 2: single factor experiment

[0039] Taking the yield of swainsonine as the evaluation index, the effects of four parameters including extraction time, extraction temperature, concentration of formic acid in the solvent, and solid-liquid ratio on the yield of swainsonin were investigated respectively, and six levels were set for each factor to conduct a single factor experiment. The experimental design is as follows:

[0040] ① Comparison of extraction time: the concentration of formic acid in the solvent methanol is 0, the ratio of solid to liquid is 1:30g / mL, the extraction temperature is 60°C, and the extraction time is set to 20, 40, 60, 80, 100 and 120 min respectively;

[0041]②Comparison of extraction temperature: the concentration of formic acid in solvent methanol is 0, the extraction time is 40min, the ratio of solid to liquid is 1:30g / mL, and the extraction temperature is set to 30, 40, 50, 60, 70 and 80°C respectively;

[0042] ③Comparison of th...

Embodiment 3

[0045] Example 3: Model building

[0046] 1. Response surface design

[0047] According to the results of the single factor experiment, the four factors of extraction time (A), extraction temperature (B), formic acid concentration (C), and solid-liquid ratio (D) were selected as variables. Using Design-Expert 8.0.6 software, -1, 0 , 1 represents the variable level, and the four-factor three-level response surface experiment of the response surface design is carried out. The true values ​​and coding levels of the independent variables are shown in Table 1.

[0048] Table 1 Test factors and levels

[0049]

[0050] Accurately weigh 5 parts of each 0.2g sample, extract swainsonine according to the modified extraction process conditions, process the test solution of the extract according to the experimental steps in Example 1, and measure the swainsonine content according to the method in Example 2 , Calculate the extraction rate of swainsonine.

[0051] 2. Box-Behnken test ...

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Abstract

The invention relates to the field of chemical treatment of natural medicines, in particular to a method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis), which comprises the following steps: (1) preparing the barbadosweed endophytic fungi into a bacterial suspension, and culturing; (2) after culture of the bacterial suspension in the step (1) is finished, performing suction filtration, rinsing, drying, grinding and wall breaking, and collecting dry powder; (3) weighing the dry powder collected in the step (2), transferring the dry powder into a centrifuge tube, adding methanol, extracting, centrifuging, collecting supernate, evaporating to dryness, redissolving, filtering and diluting; according to the extraction method, the yield of swainsonine reaches 220 mu g / g and is remarkably improved compared with the prior art, the method is simple in process condition, safe, low in cost and easy for industrial production, a basis is provided for development and utilization of swainsonine produced through microbial fermentation, raw material resources of swainsonine products are expanded, and the method is suitable for industrial production. Wide application prospects are realized.

Description

technical field [0001] The invention relates to the field of chemical treatment of natural medicines, in particular to a method for extracting swainsonine from locoweed endophytic fungi (U. oxytropis). Background technique [0002] Locoweed is a collective term for poisonous plants of the genus Oxytropis and Astragalus that contain swainsonine (SW) and can cause neurological symptoms and pathological changes typical of locoweed poisoning in animals. Animals can cause neurological diseases "Localism" or "Peastruck" after eating. As early as 1984, Tulsiani DR et al proved for the first time that the toxic substance of locoweed is mainly swainsonine. Toxicological studies have shown that swainsonine can inhibit the activity of lysosome α-mannosidase and Golgi mannosidase II in mammalian cells, causing intracellular oligosaccharide metabolism and glycoprotein synthesis disorders, resulting in cell vacuoles Degeneration and tissue organ dysfunction. When people were worrying a...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/645
CPCC12P17/182
Inventor 刘宇郝宝成贺炯杰崔东安杨珍张红娟王玲尚若锋莫亚男王学红王胜义
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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