Screeing And Transformation Conditions Of 11α-hydroxylation Of 17α-hydroxy Progesterone

11α,17α-diOH-progesterone is an important intermediate product in steroids hormone biotransformation,and it can be used as the raw material to produce many other kinds of corticosteroids,especially in the steroids biotransformation production from phytosterol.Now,there are little reports are about the biotransformation from 17α-diOH-progesterone to 11α,17α-diOH-progesterone,and there still remains many problems in this biotranformation reaction,such as few strains of bioconversion,low conversion of substrate and too much side-product etc.Thus,this study aimed at selecting out the high quality strains to perform the biotransformation of 17α-diOH-progesterone,then optimizing the conversion process condition to increase the yield of 11α,17α-diOH-progesterone.Aspergillus ochraceus CICC41473 and Cunninghamella elegans CICC40250 were singled out from five common 11α hydroxylation strains,which including Cunninghamella elegans CICC40250,Beauveria bassiana ACCC30728,Aspergillus ochraceus CICC41473,Rhizopus nigricans CGMCC33900 and Metarrhizium anisopliae ACCC30194.A.ochraceus CICC41473 and C.elegans CICC40250 canperform the hydroxylation at C-11α position of 17α-hydroxy progesterone efficiently.When the substrate concentration was 10g/L,the conversion rate of 11α,17α-diOH-progesterone formation reached 70.4%and 68.3%,respectively.The conversion process conditions of A.ochraceus CICC 41473 were optimized,and the result were:spore concentration 106/mL,temperature 28℃,rotation speed of shake flask 200 r/min,the volume of liquid 50 mL/250 mL,induction concentration of substrate 0.5 g/L,transformation of reaction media ethanol,concentration of substrate 25 g/L,ratio of dilution 0.5:1.The yield of the product was 86.1%,which increased15.7%,and the substrate concentration increased by 2.5 times of the control.The conversion process conditions of C.elegans CICC40250 were also optimized,and the result were:spore concentration 106/mL,temperature 28℃,rotation speed of shake flask 180 r/min,the volume of liquid 50 mL/250 mL,transformation of reaction media Tween80,fermentaion periods 24h,coenzyme precursor glucose,concentration of substrate 30 g/L.The conversion rate of 11α,17α-diOH-progesterone formation reached 84.2%,which increased 15.9%,and the substrate concentration increased by 3 times of the control.This experiment results provide the data basis for industrial production of 11α,17α-dihydroxy progesterone.