Technique for purifying spherosinin by fermenting green muscardine fungus

Abstract

The invention discloses a process of purifying spherosin by fermentation of metarrhizzium anisopliae. The metarrhizium anisopliae fungus is purchased from the microorganism conservation center of the Chinese academy of agricultural sciences, and through fermenting and culturing to obtain mycelium which contains rich spherosin and fermented liquid, through ultrasonic and solution extracting, and treating by thin-film chromatography, D101 resin, silica column chromatography and gradient sublimation etc. technology to separate pure spherosin from the mycelium and fermented liquid. The spherosin produced by the process is white needle crystal with molecular weight of 173, fusion point 144-145 DEG C, and purity being not less than 98%. The process does not damage present plant resources, and has no problem of lacking resources. It has short production period, low cost, big productivity, high purity. Therefore, it is suitable to be industrialized, and it can promote the industrialization of R&D that adopts the spherosin as anti-tumour medicine and immunity intensifier.

Description

technical field [0001] The invention belongs to the field of bioengineering, and specifically relates to microbiology, biological fermentation, natural product extraction, separation and identification methods, in particular to a process for purifying swainsonine by fermentation of Metarhizium anisopliae. Background technique [0002] There are three sources of swainsonine: (1) extracting and separating swainsonine from plants: the plants known to contain swainsonin include ① plants of the leguminous genus swainsonine, such as swainsonine, gray bittern Swainsonacanescens, the plant of this genus is distributed in Australia and my country, and is the first plant to isolate swainsonin; ② Astragalas and Oxytropis plants of the legume family, this Plant-like plants are the most studied plants at home and abroad. They are distributed all over the world, especially in North America and my country, and are very rich in resources. According to incomplete statistics, there are 44 spec...

AI Technical Summary

Problems solved by technology

However, the key to restricting the research and development of swainsonine as an anti-tumor drug, the industrialization of the product, and the final clinical application is that the source of pure swainsonine is very scarce, which is far from meeting the needs of people's research.

The content of swainsonine in plants is very low, only about 0.02%, and plant resources are greatly affected by natural factors, making the production of swainsonine very scarce

Due to the limitation of synthetic route and separation technology, the sources of swainsonine are also very limited in chemical synthesis, which seriously restricts the research of swainsonine's anti-tumor activity and its product development

Extracting and separating swainsonine from fungal mycelium and its culture solution, Chinese Patent Publication No. CN1396263, published on February 12, 2003, the name of the invention is a process for purifying swainsonine by biological fermentation, which adopts The microbial strain is the Rhizoctonia leguminosa 7-3 strain (code name) that is isolated and screened from natural legumes and can produce swainsonine, but does not disclose the specific acquisition of the Rhizoctonia leguminum 7-3 strain The method and approach have not disclosed the specific characteristics, standards, and quality identification methods of the Rhizoctonia leguminosa 7-3 strain, and those of ordinary skill in the field cannot repeat the realization according to the contents of the instructions; in addition, adopt biological fermentation to extract bitter horse Swainsonine not only contains swainsonine in the mycelium, but also contains a large amount of swainsonine in the culture medium, and the process of this invention only involves the extraction of swainsonine in the mycelia, and the culture medium is indeed lost The extraction of swainsonine in medium reduces the efficiency of extracting swainsonine

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