Characteristic Analysis And Purification Of Ulva Pertusa Kjellm Aminocacylase And Alkaline Phosphatase

The distributions of aminoacylase and alkaline phosphatase had been studied in the common seaweeds in Dalian area, and the U/va pertusa Kjel/m had been chosen as the target for isolation and purification. These two enzymes had been purified for a single band in the SDS-PAGE and their basic characteristics had been studied, too. In addition, the inhibition of EDTA against the alkaline phosphatase had been testified to be the complexing irreversible inhibition.1.The distribution of aminoacylase had been studied in thirty-one seaweeds including seven green seaweeds, seven brown seaweeds and seventeen red seaweeds, and the specific activity of arninoacylase changed with the changeable factors including the place, the season and the increment stage.2.The purified U/va pertusa KjeIlm aminoacylase, which became a single band in the SDS-PAGE, had been obtained by (NH4)2S04 fractionation and two steps of gel chromatography. Its specific activity was IO4U/mg.3.The U/va pertusa KjeI/m aminoacylase, whose molecular weight is l56kD, was comprised by two same subunits. The ideal temperature was 370C. The optimal pH was 7.0. The specific activity of N-Ac-DL-Ala was the highest in the six substrates. Both Cc> and Zn2 can increase the enzyme activity. PMSF didn抰 affect the enzyme activity. Th& salts of K~ and NO~ didn抰 affect the enzyme activity within 1M, while the salts of Na and S04 inhibited the enzyme activity when their concentrations were low.4.The distribution of alkaline phosphatase had been studied in twenty-nine seaweeds including five green seaweeds, six brown seaweeds and eighteen red seaweeds, and the specific activity of aminoacylase changed with the changeable factors including the place, the season and the increment stage.5.The purified U/va pertusa KjeI/m alkaline phosphatase, which became a single band in the SDS-PAGE, had been obtained by (NH4)2S04 fractionation and affinity chromatography. Its specific activity was 48.6U/mg.6.The U/va pertusa Kje/lm alkaline phosphatase, whose molecular weight was76kD, was comprised by four same subunits. The ideal temperature was 370C. The optimal pH was 9.9. Km and Vmax were 0.95mM and 5.OOpM/min, respectively. 2 didn抰 affect the enzyme activity, while Co>, Cu2 and Ni> inhibited the enzyme activity, obviously. The enzyme activity can inhibit completely when the concentration of DTT was 2.5mM, which illustrated S-S bond in the molecule was key. The NBS can also inhibit the enzyme activity, which demonstrated Trp was a component in the activity center.II7.The EDTA inhibition dynamics of U/va pertusa KjeUm alkaline phosphatase and calf intestine alkaline phosphatasc were the complexing irreversible inhibition, but these two were different in the inhibition process.