Clostridium perfringensのヌクレオシドフォスフォトランスフェラーゼ活性の螢光測定法
1976 年 50 巻 12 号 p. 625-627
詳細
1976 年 50 巻 12 号 p. 625-627
A new method for determination of the nucleoside phosphotransferase activity from Cl. perfringens was developed, which was consisted of simple ion exchange chromatography and fluorometric measurement by the modified method of Roberts et al.
After the enzyme reaction, the reaction mixture was charged on a Dowex 1×8 column (5×10mm) and then the nucleotides were eluted with 0.1 N HCl. The nucleotide fraction thus prepared contains the TMP formed by the enzyme reaction and the AMP added as phosphate donor. The TMP content in the nucleotide fraction was determined fluorometrically without any interference from AMP.
Under the optimal conditions studied, the TMP formed by the enzyme reaction could be determined up to about 2 μ moles. The result of this work suggests that this procedure is useful for kinetic research of the nucleoside phosphotransferase.
