A method for preparing 7α, 15α-dihydroxydhydroepiandrosterone by using Trichospora flaxensis

Abstract

The invention discloses a method for preparing 7alpha,15alpha-dyhydroxyldehydroepiandrosterone by utilizing flax colletotrichum mould. The method comprises the following steps: (1) performing spore culture, namely taking flax colletotrichum mould ST-1 as a production strain, performing slant culture, thereby obtaining mature spores; (2) performing mycelium culture, namely performing mycelium culture in a mycelium culture medium in which the spores obtained in the step (1) are inoculated, thereby obtaining the mycelium culture liquid; (3) performing biotransformation, namely adding carboxymethyl oleoyl-chitosan and DHEA into the mycelium culture liquid in the step (1), wherein the final concentration of the DHEA is 10.5-12g / L, the weight ratio of the carboxymethyl oleoyl-chitosan to DHEA is (2-3):1, transforming under the stirring and rotating speed of 200-220 revolutions per minute for 36-60 hours, thereby obtaining the transformation liquid; and (4) extracting the product, extracting and concentrating the transformation liquid by using ethyl acetate, thereby obtaining the 7alpha,15alpha-dyhydroxyldehydroepiandrosterone.

Description

technical field [0001] The steroidal compound biotransformation method of the present invention specifically designs a hydroxylation method. Background technique [0002] Drospirenone is a new type of progestin, because it has a certain mineralocorticoid effect, when it is used to prepare oral short-acting contraceptives, it can overcome the side effect of weight gain caused by general progesterone. 7α, 15α-dihydroxy dehydroepiandrosterone (ie 7α, 15α-dihydroxy dehydroepiandrosterone, referred to as 7α, 15α-diOH-DHEA, CAS: 2963-69-1) is an important Intermediates are generally produced by biotransformation or dehydroepiandrosterone (DHEA) due to the difficulty of synthesis. [0003] Chinese patent CN102703558B discloses a method of converting DHEA into 7α-hydroxyl-dehydroepiandrosterone and 7α, 15α-dihydroxydehydroepiandrosterone by using Colletotrichum lini ST-1 (preservation number is CGMCC No.6051). Hydroepiandrosterone and the conversion method adopt the method of pre-...

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Problems solved by technology

[0003] Chinese patent CN102703558B discloses a method of converting DHEA into 7α-hydroxyl-dehydroepiandrosterone and 7α, 15α-dihydroxydehydroepiandrosterone by using Colletotrichum lini ST-1 (preservation number is CGMCC No.6051). Hydroepiandrosterone and the conversion method adopt the method of pre-inclusion of the substrate and the methyl-β-cyclodextrin of equimolar ratio before feeding, and the formation can improve the solubility and conversion efficiency of the substrate. After our Experimental method, this method has high selectivity for strains, that is, when different strains deal with the substrate pre-included with methyl-β-cyclodextrin, the results are not consistent. It does not work well with methyl-β-cyclodextrin pre-inclusion substrates, and the product obtained by this method is a mixture of 7α-hydroxydehydroepiandrosterone and 7α, 15α-dihydroxydhydroepiandrosterone , according to the published data of the patent, only 60% of the conversion products are 7α, 15α-dihydroxydhydroepiandrosterone, and the separation of these two products is relatively difficult

[0004] Chinese patent CN 102757997A discloses a method for preparing 7α, 15α-dihydroxyandrostenone (i.e. 7α, 15α-dihydroxydhydroepiandrosterone) by a substrate-inducing method, adding substrates in stages to make the substrate The final addition concentration of the method reaches 8-10g / L, and can increase the conversion rate to about 90%. However, through our experiments, we found that this method needs to prolong the total fermentation time, thereby affecting production efficiency, and the inoculum size used in this method 8-10%, larger inoculation volume will actually restrict the expansion of production scale

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