NDLI: Purification and characterization of alkaline phosphatase from the gut of sea cucumber Stichopus japonicus

Content Provider	Springer Nature Link
Author	Wu, Hai Tao Li, Dong Mei Zhu, Bei Wei Cheng, Jing Heng Sun, Jin Jian Wang, Feng Lin Yang, Yang Song, Yu Kun Yu, Chen Xu
Copyright Year	2013
Abstract	An alkaline phosphatase was purified from the gut of sea cucumber Stichopus japonicus by n-butyl alcohol extract, ammonium sulfate precipitation, ion exchange chromatography with diethylaminoethyl cellulose, gel filtration chromatography with Sephacryl S-200 and preparative electrophoresis with polyacrylamide gel electrophoresis. The native enzyme was estimated to be 166 ± 9 kDa and produced a single predominant band corresponding to active enzyme on nondenaturing electrophoresis, but showed 2 bands of 97 and 35 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that the native enzyme is composed of two dissimilar subunits. The enzyme displayed maximum activity at pH 11 and 40 °C, showing narrow pH stability (pH 10–12) and thermal instability at temperature higher than 30 °C. The activity of the purified alkaline phosphatase was enhanced by Mg$^{2+}$, whereas inhibited by Zn$^{2+}$, Ca$^{2+}$ and EDTA at 1 and 10 mM, suggesting its activity is in a magnesium ion-dependent manner. The product-analog WO$_{4}$ $^{2−}$ and product HPO$_{4}$ $^{2−}$ showed strong inhibitory effects on the enzyme activity. Using p-nitrophenyl phosphate as substrate, the V $_{max}$ and K $_{m}$ values were 24.45 μmol/L min and 5.76 mM, respectively.
Starting Page	477
Ending Page	485
Page Count	9
File Format	PDF
ISSN	09199268
Journal	Fisheries Science
Volume Number	79
Issue Number	3
e-ISSN	14442906
Language	English
Publisher	Springer Japan
Publisher Date	2013-04-12
Publisher Place	Japan
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Alkaline phosphatase Sea cucumber Stichopus japonicus Gut Purification Characterization Fish & Wildlife Biology & Management Freshwater & Marine Ecology Food Science
Content Type	Text
Resource Type	Article
Subject	Aquatic Science

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