Separation method of swainsonine-producing endophytic fungi in glabrous crazyweed

Abstract

A separation method of swainsonine-producing endophytic fungi in glabrous crazyweed comprises the following steps of: acquiring mycelium of the strain and a broth through a bio-fermentation method, carrying out ultrasonic extraction, quantitatively and qualitatively detecting swainsonine by thin-layer chromatography and gas chromatography technologies after solvent extraction, and identifying swainsonine-producing endophytic fungi through morphological and molecular biology methods. The result shows that one out of six separated strains of endophytic fungi can generate swainsonine and the output is 16.7338 mg / L. After identification, the strain is fusarium proliferatum. In comparison with present strains for the production of swainsonine, swainsonine provided by the invention has high output. The bio-fermentation method for the production of swainsonine doesn't influence the meadow ecological environment. The separation method provided by the invention solves the problem of swainsonine sources and provides strain supports, thus establishing a theoretical foundation for future industrial production of swainsonine. The invention has a wide development prospect.

Description

technical field [0001] This patent belongs to the field of biopharmaceuticals, and specifically relates to a method for isolating endophytic fungi producing swainsonine from Oxytropis florida and the obtained swainsonine-producing strains. Background technique [0002] Swainsonine is a polyhydroxyindolizidine alkaloid, which is a very strong competitive inhibitor of α-mannosidase, which leads to abnormal synthesis of glycoproteins and accumulation of partially synthesized oligosaccharides in lysosomes. Cause cell vacuolar degeneration, cause organ tissue damage and dysfunction and poisoning. However, swainsonine has pharmacological effects such as enhancing the body's immune function, anti-virus, anti-cancer, anti-tumor, and antibacterial. It is worth mentioning that the anti-tumor activity of swainsonine has become a hot spot in the screening of tumor backup drugs in recent years. However, the main problem that restricts the research and development process of domestic sw...

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Problems solved by technology

Regarding the biosynthesis of swainsonine, there are 6 patents that have been applied in China, namely "Technology for Purifying Swainsonin by Biological Fermentation" (02114591.1), "Technology for Purifying Swainsonin by Metarhizium Anisopliae Fermentation" (200610043120.5) ,"Beauveria bassiana fermentation process for the production of swainsonin" (200710199249.X), "A kind of Ehrigiospora spinosa for the synthesis of swainsonin and its preparation method and application" (200910021082.7), "A "Ehrigia spinosa FEL5-AS1 for the synthesis of swainsonin and its application" (200910021858.5), "A type of Ehrigia spinosa FEL6-AS2 for the synthesis of swainsonin and its application" (200910021861.7), Its deficiencies and defects are:

[0010]④ "A kind of Ehrigia spinosa for synthesizing swainsonine and its preparation method and application", "A kind of Echinacea for synthesizing swainsonin The common deficiency of the three patents "E. faeci FEL5-AS1 and its application" and "A kind of E. spinosa FEL6-AS2 for the synthesis of swainsonin and its application" is that there is no type of strain disclosed, and The disclosed numbered strain has a low yield of swainsonine

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