Separation method of swainsonine-producing endophytic fungi in glabrous crazyweed

An endophytic fungus, swainsonine technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of low yield of swainsonine and undisclosed strain types, etc.

Inactive Publication Date: 2012-10-17
鄂尔多斯市普众生物技术有限公司
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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" (200910021

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  • Separation method of swainsonine-producing endophytic fungi in glabrous crazyweed
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  • Separation method of swainsonine-producing endophytic fungi in glabrous crazyweed

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Embodiment Construction

[0052] 1) Isolation of endophytic fungi:

[0053] Wash off the dust on the surface of Oxytropis florida (collected from Alxa League, Inner Mongolia) with deionized water, separate the leaves, stems, and flowers of the plant with sterilized tweezers, and then wrap the leaves, stems, and flowers with gauze respectively for surface disinfection , the surface disinfection procedure is as follows: first soak in 75% ethanol for 30s, then soak in sodium hypochlorite solution with 1% available chlorine for 3min, and finally rinse with sterilized deionized water twice, each time for 1min, the deionized Water is inoculated on the surface of the newly prepared PDA agar medium, and observed for 3 days to see whether there is fungal growth on the surface of the medium, so as to test the effect of surface disinfection;

[0054] Use sterilized filter paper to absorb the water on the surface-sterilized plant tissue, then use sterilized scissors to cut leaves, stems, flowers, etc. into 3mm tis...

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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...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/68C12Q1/04G01N30/90G01N30/02C12R1/77
Inventor 赵宝玉刘生武苏东卢围陈基萍胡波刘彤陈燕刘媛
Owner 鄂尔多斯市普众生物技术有限公司
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