Key Technology Of Mycobacterium Degrading Soybean Oil By-product Phytosterol To Produce Androstenedione

As an indispensable intermediate,androstenedione is widely used in drug manufacturing,especially steroidal drugs.However,the chemical manufacturing process of androstenedione is generally complicated and difficult,and it will cause serious environmental pollution in the production process.The biological method for the production of androstenedione has a very promising development prospect,because it is more economical and environmentally friendly than chemical methods.As one of the important components of China’s food industry,products from soybean processing will produce a large amount of soybean meal and distillate in the production process,and phytosterol is the main component of soybean processing distillate,which can be It is deeply utilized by the production of androstenedione as a substrate for biotechnology.In order to better produce androstenedione on a large scale and to solve the imbalance between supply and demand,androstenedione was produced by using a two-aqueous phase system to increase the substrate concentration and using mycobacterium to transform soybean oil by-product phytosterol.By comparing a variety of detection methods,it was found that thin layer chromatography as a qualitative detection and high performance liquid chromatography as a quantitative detection of androstenedione are more efficient and accurate.Orthogonal experiments determined that the optimal conditions for the production of androstenedione by microorganisms are: water / sunflower oil in a two-aqueous phase system,temperature 30 ° C,initial pH 6.5,substrate concentration 0.4 g / L.Under such conditions,the yield of androstenedione can be more than 84%.Based on the orthogonal experiment,other factors affecting the yield of androstenedione were explored.Through the design and analysis of the response surface experiment,the best process conditions obtained were speed 250 rpm,inoculation 14.83%,organic comparative example 20.65 %,The liquid volume is 30%.Under this optimal condition,the yield of androstenedione can exceed 85%.Comparing the extraction effects of various organic solvents on androstenedione in the fermentation system,ethyl acetate was selected as the extractant,compared with 1,and the extraction level was 4.After the extraction is completed,74.58% of ethyl acetate can be recovered by distillation under reduced pressure.Isoacetone extracts the oil in the remaining solution,which can realize the oil recovery and recovery rate of up to 82.45%.The preliminary production of androstenedione by microbial method has found suitable process conditions and provided data and theoretical support for its large-scale production.Realized the recycling of resources and the saving of production costs.