Production Of 11?,17?-hydroxyprogesterone From 17?-hydroxyprogesterone By Aspergillus Ochraceus MF04

Steroid drugs are the second biggest drugs besides antibiotics,which have a very wide range of applications in the pharmaceutical industry in the world.17 hydroxy progesterone acts as an important pharmaceutical intermediates of steroid hormone drugs.11?hydroxylation on the 17a-hydroxy progesterone can significantly improve its anti-inflammatory activity.The use of microbial transformation method is efficient for the 11?hydroxylation of.17?-hydroxy progesterone.This paper prepared 11?,17? hydroxyprogesterone by Aspergillus ochraceus,the following 4 aspects were studied:(1)the optimization of fermentation conditions(2)the effect of organic solvent and surfactant dispersion method on substrate conversion rate(3)the effect of porous coating material(porous starch and attapulgite)on the conversion rate of substrate in media(4)the attempt of 5 L tank fermentation.The best strain of the bacteria isolated from laboratory preservation was MF04,and the optimum composition of fermentation medium was as follows:glucose 20 g/L,corn syrup 20 g/L,peptone 10 g/L,NH4H2PO4 4 g/L,chitin 2 g/L,inoculation volume]0%,rotate speed 200rpm,feeding time 18h.When the smallest average particle size was 5.071 ?m,the highest final molar conversion rate was 80.2%.In order to increase the solubility of the substrate,the organic solvents,surfactants and the mixture of them were utilized in this experiment.The best method of them is adding the mixture of 1%DMSO and 0.2%tween 80 in which condition the conversion rate of,17 a-hydroxy progesterone reached 88.9%.The scanning electron microscope(SEM)analysis showed that Aspergillus ochraceus would break and even form a dense cake when the amount of DMSO was increased by more than 3%.In order to improve the distribution of the substrate,we have carried out a part of the research in the aspect of the embedding material including using the Attapulgite and porous starch.To adsorpt and encapsulate the substrate 17?-hydroxy progesterone.When the add amount of 200 mesh attapulgite was 0.5%,the molar conversion rate was the highest,which reached 87.1%.It was observed that the combination of the attapulgite with the mycelium is helpful to the integration of the substrate into the cell through the SEM.Fourier infrared spectroscopy(FTIR)and the rmogravimetric analysis(TGA)showed that 17 a-hydroxy progesterone were embedded in attapulgite and,the adsorption rate was 0.5040g/g.The molar conversion rate reached 83.7%when the ratio of the porous starch and substrate was 1:1 in the embedded object.Thermogravimetric analysis(TGA)showed that the embedding rate of the porous starch was 0.6476g/g.In the experiment of 5L fermentation tank,Under the optimal culture conditions,17 a-hydroxy progesterone molar conversion rate reached 81.7%.In addition of organic solvent 1%DMSO and 0.2%tween 80,17 a-hydroxy progesterone molar conversion rate reached 87.8%and the time when.the equivalent conversion rate was reached,was shorten 20h than the control.