Spermidine synthetase

SURVEY OF ENZYMATIC ACTIVITIES ASSAYED BY THE HPLC METHOD 

The enzyme was prepared from mouse brain homogenized in phosphate buffer. An S-100 solution was prepared and used as the source of the synthetase. 

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The enzyme spermidine synthetase catalyzes the conversion of putrescine to spermidine. The separation of the substrate from the product was accomplished on a reversed phase column (Ci8) using a mobile phase of 60% methanol (v/v). The compounds in the eluent were detected by scintillation counting or by UV at 254 nm as shown in Figure 9.101 (labeled Blank). 

Figure 9.56 Mouse brain spermidine synthetase activity as a function of time. Both the absorbance (O) and the radiometric ( ) determination of product formation represent the mean value of duplicate assays. The reproducibility was within 10%. (From Porta et al., 1981.)...

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