摘要
Acid phosphatases (ACPs) are marker enzymes for the detection of lysosomes in cell fractions. However, ACPs in sea creatures are less studied than those on land. An acid phosphatase was partially purified from pearl oyster Pinctada fucata by chromatography on Sephadex G 150 and Con A Sepharose 4B. The specific activity was 1719 U·mg -1 and with optimum pH (5.0) and temperature (60℃). The enzyme was strongly inhibited competitively by product analog WO 3- 4 and MoO 3- 4, but less inhibited by product analog AsO 3- 4. The enzyme could also be strongly inhibited by heavy metal ions, such as Ag + and Cu 2+ , but was not affected by Pb 2+ . High concentrations of ethanol (64%) and NaF (10 -3 mol·L -1 ) could inhibit the enzyme while low concentration of NaF (<10 -4 mol·L -1 ) could slightly activate the enzyme. Other haloids (Cl -, Br -, I -) and EDTA did not have any effect on this enzyme, while tartrate and some chemical modification reagents (bromoacetic acid, formaldehyde and dithiothreitol) could inhibit the enzyme. It is concluded that the properties of the enzyme are different from many fresh water mollusks.
Acid phosphatases (ACPs) are marker enzymes for the detection of lysosomes in cell fractions. However, ACPs in sea creatures are less studied than those on land. An acid phosphatase was partially purified from pearl oyster Pinctada fucata by chromatography on Sephadex G 150 and Con A Sepharose 4B. The specific activity was 1719 U·mg -1 and with optimum pH (5.0) and temperature (60℃). The enzyme was strongly inhibited competitively by product analog WO 3- 4 and MoO 3- 4, but less inhibited by product analog AsO 3- 4. The enzyme could also be strongly inhibited by heavy metal ions, such as Ag + and Cu 2+ , but was not affected by Pb 2+ . High concentrations of ethanol (64%) and NaF (10 -3 mol·L -1 ) could inhibit the enzyme while low concentration of NaF (<10 -4 mol·L -1 ) could slightly activate the enzyme. Other haloids (Cl -, Br -, I -) and EDTA did not have any effect on this enzyme, while tartrate and some chemical modification reagents (bromoacetic acid, formaldehyde and dithiothreitol) could inhibit the enzyme. It is concluded that the properties of the enzyme are different from many fresh water mollusks.
基金
Supported by the National Natural Science Foundation of China(No. 30 170 72 3)
