Alkaline phosphatase(AKP)was isolated and purified from the skin of Tylototriton taliangensis Liu and its kinetic property was examined.The partially purified alkaline phosphatase was purified by salting\|out,CM\|cellulose ion\|exchange column and gel filtration with Sephadex G\|150.The purified enzyme from skin moved as a single electrophoretic band in PAGE.The specific activity was 90.26 units/mg protein.Its subunit weight was 42.0 kD as determined with SDS\|PAGE.The optimum pH value for the enzyme was pH9.0.The optimum temperature was about 34℃.The Michealis\|Menton constant( K \-m)was 0.83 mmol/L on the disodium phenyl phosphate.Activity differences of the enzymes were determined when metal ions effected on the AKP.The results showed that K + was found to have no inhibition of AKP activity.Mg 2+ ,Ca 2+ ,Cu 2+ could activate the AKP and the higher the metal ions concentration were,the more the activity of AKP increased.When 3.0 mmol/L Cu 2+ was used,the activity of AKP could rise to 187%.Ni 2+ ,Zn 2+ and Ag + could inhibit the AKP and the higher the metal ions concentration were,the more the activity of AKP decreased.When 3.0 mmol/L Ag + was used,the activity of AKP retained 23% only.
Atomic absorption and plasma arc spectroscopy were used to determine the contents of Ca,Mn,Cu,Ni,Co,Pb,Zn,Fe,Mg,Hg,Mo,Cr,totally 12 macro and trace elements in yellow,white and black pearls.The results showed that all the three kinds of pearls contained Ca of more than 3×104 mg/100g.The contents of Mn,Cu,Fe,Mg were above 1mg/100g.Ni,Co,Pb,Mo,Cr,Zn had less contents in the pearls,and Hg had the least.It was also showed that Mn,Cu,Pb,Zn,Mg,Co and Cr contents in different colorpearls varied greatly.
