Research Technology And Testing Methods For The Preparation Of High Purity RNase A

RNase A(EC 3.1.27.5)as an endonuclease with a molecular weight of 13.7kD,is derived from bovine pancreas.An U-C 3',5'-pHospHodiester bond can be digested by RNA enzyme,forming a 2',3'-cyclic pHospHate derivative oligonucleotides.RNaseA is useful for the treatment of trauma and joint pain,playing a role in inhibiting the growth of influenza and herpes virus as well.Nowadays,the demand for RNaseA is strong in China.While with a poor product quality in China,the RNaseA we used is usually produced by foreign firms.In order to optimize the production technology of RNaseA,and propose a reasonable detecting method of RNaseA,improving the market competitiveness of domestic firms,we performed two methods for RNaseA separation and purification.The influencing factors were analysed as well.As a result,the purification rate of RNaseA we prepared can reach to more than 95%by SDS-PAGE.Adding to this,the average enzyme activity of RNaseA we prepared was as height as 58.69 kU/mg,while the enayme activity of RNaseA produced by other manufactures was less than 100 U.Furthermore,according to the definition and detection methods of RNaseA enzyme activity proposed by some major foreign biotech companies,we established a new one for RNaseA enzyme activity,which would meet the needs of large-scale production of RNaseA.To sum up,this work would make some contributions to forming the industrialization of RNaseA,and improving the market competitiveness of domestic firms.