Production Of 11?-Hydroxyandrosta-1,4-diene- 3,17-dione From Phytosterol By Mixed Cultivation Of Mycobacterium And Aspergillus Ochraceus

In the pharmaceutical industry, steroidal drugs are the second largest category of drugs, second only to the production of antibiotics, there is a very wide range of applications in the medical field. In the production of steroids, multiple groups of female steroid nucleus need be modified. Phytosterols side-chain degradation preparation of Androst-4-ene-3,17-dione (AD) or Androsta-1,4-diene-3,17-dione (ADD) are the important intermediates steroid hormone drugs, almost all drugs steroid can be prepared of ADD or AD as starting material. In the biotransformation process, one of the most important of which is the reaction of 11?-hydroxylation, the reaction can be introduced into a hydroxyl on the ADD nucleus C11? position, can significantly improve the anti-inflammatory activity of steroid drugs.This article first to preparation ADD, mycobacteriumMF006 degradation of phytosterols was studied, and the best conditions were determined as follows: fermentation medium initial pH 7.0, fermentation temperature was 30?, inoculation size of mycobacterium was 10%, biotransformation time was 144 h. In order to increase the solubility of the substrate, the experiments with several kinds of organic solvents, where DMF can make phytosterols conversion efficiency reached 50.34%; When the surfactant sulfobutyl ether-?-cyclodextrin (SBE-?-CD) and phytosterols molar ratio of 2:1, phytosterols conversion efficiency reached 88.79%; Fourier infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) studies have shown that inclusion complex was formed between P-sitosterol and sulfobutyl ether-?-cyclodextrin. Second, Aspergillus ochraceusMF007 hydroxylate ADD preparation of lla-OH-ADD was studied, the results showed that the hydroxylation best glucose concentration was 20.0 g/L, corn starch concentration was 9.0 g/L; Biotransformation condition were:inoculation size of Aspergillus ochraceus was 10%, fermentation temperature was 28?. On the basis of those researchs, mycobacteriumMF006 and Aspergillus ochraceus MF007 mixed cultivation of phytosterols preparation of 11?-OH-ADD was studied, and optimized the transformation conditions of mixed cultivation, the temperature was 30?, initial pH 7.0, inoculation size of Aspergillus ochraceus was 20%, and Aspergillus ochraceus vaccination time was 48 h. Finally, P-sitosterol as one kind of phytosterols with organic silica hollow spheres adsorption inclusion complex has carried on the mixed cultivation research, the experimental results showed that added organic silica hollow spheres could improved the conversion efficiency of ?-sitosterol, when ?-sitosterol concentration was 1.0 g/L, the concentration of 11?-OH-ADD was 0.631 g/L, conversion efficiency reached 87.11%, compared with no organic silica hollow spheres, ?-sitosterol conversion efficiency increased by 28.44%. Scanning electron microscope (SEM) analysis showed that closely contact with the cell surface of the organic silica hollow spheres, Fourier transform infrared spectroscopy and thermogravimetric analysis (TGA) showed that adsorption inclusion complex was formed between ?-sitosterol and organic silica hollow spheres, the adsorption ability of organic silica hollow spheres was 0.22 g/g.