Enzyme Hydrolysis Of Saikosaponin Sugar-moiety

In order to obtain the microorganisms which can produce saikosaponinase, enzymes areproduced by sp.42（fungi）; sp.GS514-saiko（bacterium afforded by kyunghee university）,andRhodanobacter sp.GS3054（afforded by KAIST）.I compared the enzymes’ hydrolyzing capabilities of saikosaponin, ginsengsaponin andbarrenwortsaponin. Fungi（sp.42） induced by Flo sophorae, fermented at28℃,for5days.Theproduct-saikosaponinase was purified by DEAE-cellulose column, After being detected bySDS-PAGE, there was a single submit albumen, and the molecular weight of it is about59KDa.The purified enzyme （Tube NO.16） can hydrolyze the saiko sugar-moiety and theenzyme, about57KDa, can hydrolyze PPD, the optimum react condition is under40℃, pH5.The mental ion (Ca²⁺, Mg²⁺, and Zn²⁺)has no infection on the reaction with PPD. But,Ca²⁺&Mg²⁺can accelerate the reaction on saikosaponin, except the Zn²⁺.The rough enzyme produced by bacterium sp.GS514, can convert ginsenoside Rb1to Rd,Rg3, Rh2, and can also hydrolyze saikosaponin. Separated by DEAE-cellulose column, theenzyme can only convert Rb1to Rd, and only can hydrolyze tenth0.2%consistence ofsaikosaponin. After being tested by SDS-PAGE, there are two strips, the molecular weight ofthem are67KDa and59KDa separately.It shows that the activities of the enzymes may be reduced during the conservation orpurification, and it needs further research.The leavening of Rhodanobacter sp.GS3054, has a low act on saikosaponin, but thecapability of hydrolyzing can be increased after being concentrated for5times, The enzymeinduced by0.4%Rutin has a high saikosaponinase capability.Also the sp.GS3054can produce the barrenwortsaponinase under the condition of25℃,6days.Being compared the three kinds of microorganism, the fungi-sp.42has a betterproduction of saikosaponinase, but the enzyme produced by bacterium sp.GS514issignificant to convert ginsenoside PPD to Rg3and Rh2; The rhodanobacter sp.GS3054canhydrate not only saikosaponin, but also barrenwortsaponin, which needs to be discussed.