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Method for preparing swainsonine by solid fermentation

A solid fermentation method, swainsonine technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of high cost, long time, low purity, etc., to save extraction costs, easy to obtain , The effect of shortening the separation and extraction time

Active Publication Date: 2019-04-12
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In order to overcome the defects of long time, high cost and low purity of the existing swainsonine preparation method, the present invention provides a method for preparing swainsonine by solid fermentation

Method used

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  • Method for preparing swainsonine by solid fermentation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] 1) Source of bacteria

[0076] The fungus used is derived from the endophytic fungus Alternariasect.Undifilumoxytropis of the locoweed plant, provided by the China Industrial Microbiology Culture Collection and Management Center.

[0077] 2) Strain cultivation and fermentation

[0078] PDA solid medium: 200g of potatoes, 20g of glucose and 15-20g of agar, after boiling, add distilled water to 1000mL.

[0079] Rice medium: 60g of rice, 80mL of distilled water, autoclaved at 121°C for 30 minutes and then set aside.

[0080] First, inoculate the purchased strains on PDA solid medium for activation, and store some well-grown strains in refrigerators at 4°C and -20°C respectively for subsequent needs, and the rest grow well on PDA solid medium The strains are reserved for further solid fermentation.

[0081] Secondly, the fungus growing on the PDA medium and in the logarithmic phase was cut into small pieces, inoculated on the rice medium in the ultra-clean workbench, and...

Embodiment 2

[0098] The contrast of embodiment 2 eluents

[0099] 1. TLC sample point selection: take the commercially available swainsonine standard product (purity above 99%), and the alkaline n-butanol part extract prepared according to steps 1) to 3) of Example 1 as the extract containing Crude product of swainsonine;

[0100] 2. Dissolve the swainsonine standard substance with chromatographic methanol, and be prepared with a concentration of 5mg / ml swainsonine standard solution (standard product 1); Each is prepared into a concentration of swainsonine crude product solution of 10mg / ml, which is recorded as crude product 1 and crude product 2;

[0101] 3. Choose the GF254 type of silica gel thin-layer plate, cut it into a suitable size with a glass knife, and wait for the sample to be loaded. Use clean 0.3mm capillary pipettes to absorb standard product 1, crude product 1, and crude product 2 respectively onto the prepared thin-layer plates, and prepare two thin-layer plates that hav...

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Abstract

The invention provides a method for preparing swainsonine by solid fermentation, and relates to the technical field of separation and purification of secondary metabolites of fungi. The method includes performing solid culture on plant endophytes producing swainsonine, performing extraction on the obtained fermentation product with a water-containing organic solvent, followed by extraction with petroleum ether, ethyl acetate and an alkaline water saturated n-butanol solution to obtain an alkaline n-butanol layer; and subjected the alkaline n-butanol layer to silica gel column chromatography and preparative liquid chromatography to obtain the pure product of swainsonine. Compared with the existing swainsonine extraction method, the method provided by the invention shortens the extraction time and saves the extraction cost, and the purity of the prepared swainsonine can reach 99%. The method provided by the invention saves the problems of high cost, long time and great difficulty in obtaining the pure product of swainsonine by the prior art.

Description

technical field [0001] The invention relates to the technical field of separation and purification of fungal secondary metabolites, in particular to a method for preparing swainsonine by a solid fermentation method. Background technique [0002] Fungal secondary metabolites are an important source of active lead compounds in the development of new drugs. Since the 19th century, livestock in the vast grassland areas such as the western frontier of the United States and Northwest my country have suffered from a chronic poisoning disease of nervous system dysfunction. Poisoned livestock show salivation, paralysis, infertility, abortion, weight loss, and serious death. , causing huge economic losses. However, the culprit behind the scenes is that the domestic animals have eaten the notorious "crazy weed", which is the collective name for poisonous plants of the genus Astragalus and Oxytropis genus of the leguminous family. It is the main poisonous weed that endangers the develop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C07D471/04C12R1/645
CPCC07D471/04C12P17/182
Inventor 丁刚谭祥梅张乐
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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