Study on the Identification and Potency Determination of L-asparaginases from 2 Kinds of Strain / 中国药房

ABSTRACT

OBJECTIVE: To discuss the identification method of L-asparaginases prepared from E.coliASI.357 and Erwinia carotovora and the optimal enzymatic reaction conditions of potency determination.METHODS: HPLC method and isoelectric focusing electrophoresis were applied for the identification of L-asparaginase from 2 kinds of strain.The effects of category of buffer solution and pH value on enzymatic reaction of potency determination of L-asparaginase were investigated.RESULTS: HPLC chromatogram of L-asparaginases from E.coli ASI.357 was different from that from Erwinia carotovora.The retention time of the peaks were 11.0 min and 11.8 min.The isoelectric point (PI) of L-asparaginase produced from E.coliASI.357 was within 4.65～5.1 and that produced from Erwinia carotovora was within 7.1～8.20.The optimal enzymatic reaction conditions of potency determination of L-asparaginase produced from E.coliASI.35 were Tris-HCl (pH=9.0) as buffer and that produced from Erwinia carotovora was 0.2 mol?L-1 phosphate (pH=8.0) as buffer.CONCLUSION: The isoelectric point (PI) of L-asparaginases produced from 2 kinds of strain is different from each other as well as their optimal enzymatic reaction conditions of potency determination.The L-asparaginases from 2 kinds of strains should be controlled as 2 different categories.