Purification and characterization of Ulva pertusa Kjellm alkaline phosphatase.
The activity of alkaline phosphatase (ALP, EC 3.1.3.1.) was found in seaweeds, including five kinds of green alga, eighteen kinds of red alga, and six kinds of brown alga, collected from the seaside of Dalian in China. The enzyme was purified 1230-fold from Ulva pertusa Kjellm. It had a specific activity of 48.6 U/mg protein and was proven to be homogeneous by SDS-PAGE with a subunit molecular mass of 19.5 kDa. The activity of ALP peaked at pH9.8, and was completely inhibited by DTT and partly by NBS. The Michaelis-Menten constant Km and the maximum reaction velocity Vmax, at pH 9.8 and 37 degrees C were 0.950 mM and 5.00 microM/min, respectively.
Yang
Dong
D
Department of Bioengineering, Dalian University of Technology, Dalian, P. R. China. dyangdong@sohu.com
Wang
Jingyun
J
Bao
Yongming
Y
An
Lijia
L
D000469:Alkaline Phosphatase / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*; D000460:Chlorophyta / Q000201:enzymology; D002846:Chromatography, Affinity / Q000379:methods; D003035:Cobalt / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D000459:Phaeophyceae / Q000201:enzymology; D000461:Rhodophyta / Q000201:enzymology; D012624:Seaweed / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013997:Time Factors; D044445:Ulva / Q000201:enzymology*
D003035:Cobalt; D003300:Copper; D009532:Nickel; D004492:Edetic Acid; D000469:Alkaline Phosphatase; D008274:Magnesium; D004229:Dithiothreitol