Method for producing androstane dienedione through fermentation of arthrobacter simplex

Abstract

The invention belongs to the technical field of preparation of steroid hormone drugs, and particularly relates to a method for producing androstane dienedione by fermentation of arthrobacter simplex, which mainly comprises the following steps: optimizing fermentation culture medium components, feeding points and substrates for whole-cell transformation of arthrobacter simplex to obtain an ADD crude product with high conversion rate, and extracting and refining by mixing acetone and water to obtain the androstane dienedione. And an ADD fine product is obtained. The method effectively solves the problems that in the prior art, the conversion efficiency is not high, byproducts are difficult to separate, the process is tedious and not suitable for industrial mass production, and the like, the fermentation feeding concentration is increased to 30-60 g / L, and the conversion rate is increased to 97% or above.

Description

technical field [0001] The invention belongs to the technical field of preparation of steroid hormone drugs, in particular to a high-efficiency method for Arthrobacter simplex CPCC 140451 to ferment and produce androstadienedione (ADD), especially the medium components used, the optimization of feeding points and Optimization of substrate pretreatment. Background technique [0002] Steroidal drugs have strong anti-infection, anti-allergic, anti-viral and anti-shock pharmacological effects, and are an important class of drugs in clinical practice. Most of the raw materials for the early synthesis of steroidal drugs are directly extracted from animal tissue fluid, the recovery rate is low, the cost is high, and it cannot meet the production needs; the later synthesis method uses dioscin as the starting material and undergoes chemical synthesis methods, which is the industrialization of steroidal drugs Production provides abundant raw materials, but with the increasing depleti...

AI Technical Summary

Problems solved by technology

Most of the raw materials for the early synthesis of steroidal drugs are directly extracted from animal tissue fluid, the recovery rate is low, the cost is high, and it cannot meet the production needs; the later synthesis method uses dioscin as the starting material and undergoes chemical synthesis methods, which is the industrialization of steroidal drugs Production provides abundant raw materials, but with the increasing depletion of dioscin, more abundant resources and methods are urgently needed to replace them; and the emergence of phytosterols has brought new changes to the field of steroids. It is obtained from various crops, and the important steroid drug intermediates androstenedione (AD) and androstenedione (ADD) are obtained by using biological selective side chain degradation technology, and then androstenedione and androstenedione Steadienedione is used as the starting material for further preparation to obtain steroid drugs, wherein ADD is an important starting material for the synthesis of estrogens

[0003] The main method of preparing ADD at present is: one, adopt mycobacteria to degrade phytosterols to obtain AD and ADD, such as U.S. Patent USP4345029, Chinese Patent CN102477404A, but these technologies all have long fermentation period (150h), and the shortcoming that transformation efficiency is not high, transformation The highest rate is only 70%, and a large amount of by-products such as AD, phytosterol, and testosterone are produced at the same time, which is not conducive to subsequent separation and purification

2. Use Arthrobacter simplex, Mycobacterium, Bacillus subtilis, and Nocardia simplex to use AD to carry out oxidative dehydrogenation to produce ADD, such as Chinese patent documents CN103266161A, CN106191190A, CN104531746A, CN108456666A Chinese invention document CN106636160A, these process advantages are obviously higher than The first method, but because most of the bacterial strains are transgenic recombinant bacterial strains, there is uncontrollability in the enlarged production, and it is not suitable for industrial production; Solvent soybean oil not only increases the cost of auxiliary materials, but also has a cumbersome post-processing process, which is not suitable for large-scale industrial production

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