Screening Of Endophytes Producing Saponin Hydrolase From Dioscrea Zingiberensis C.H. Wright And Preliminary Separating Enzyme

Diosgenin is an important precursor compound for the synthesis of steroid hormone and steroid contraceptive,which itself also has significant biological activity.Diosgenin exists in plants in the form of steroidal saponins,and the traditional production method is acidic hydrolysis which would generate a lot of waste water caused great environment pollution and difficult to handle.Enzymatic hydrolysis mild,environmental protection,is a new approach to production of diosgenin.In this paper,the Dioscorea zingiberensis C.H as research object,on the production of Dioscorea zingiberensis diosgenin enzyme endophytic bacteria screening,production of saponin enzyme bacteria screening,high efficient bacteria growth and enzyme production relations,saponin enzyme enzymolysis efficiency were studied,and the extraction and separation of saponins enzyme were studied.The main results are as follows:(1)Five endophytic bacterial strains and two strains of Endophytic fungi were isolated from Dioscorea zingiberensis plants by the coating method and coating method.Were used to culture medium with saponins of substrate on the endophytic bacteria were cultured in solid media and liquid shake flask fermentation strains,obtained from the product by extraction method;and by thin layer chromatography and high performance liquid chromatography detection to extract diosgenin,found that 2 strains of endophytic fungi(SYfx1 13.1、SYfx2 3.2)and 2 strains endophytic bacteria(SYbx313.3、SYbx413.4)in the product are diosgenin.(2)The production of saponin were analyzed by high performance liquid chromatography,SYfx213.2 bacteria transformed the saponin content was the highest,determine the strain SYfx213.2 is efficient bacteria producing saponin enzyme.Morphology,structure characteristics of high efficient bacteria by using optical microscope and scanning electron microscope,identified as aspergillus.(3)Studyed the relationship between SYfx213.2 bacteria growth and yield between saponin enzyme,cell growth rate reached a maximum at sixth days for 2.6g/L,saponin hydrolysis saponins saponin from generation eighth days had the highest rate of 65.71%;broken by ultrasonic method,grinding method,ammonium sulfate fractionation method of separation the process of extraction,saponin enzyme,ammonium sulfate saturation good separation effect for 60%.