Novel salt-resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius

It was shown that the alkaline phosphatase (AP) isolated from eggs of the sea urchin Strongylocentrotus intermedius was a homodimer with a molecular mass of 150 kDa and that it exhibited maximal activity at a рН of 8.1–8.5 and a temperature of 45°С. Mg 2+ , Ca 2+ , and Mn 2+ ions, as well as DTT, ar...

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Bibliographic Details
Published in:Applied biochemistry and microbiology 2017, Vol.53 (1), p.11-19
Main Authors: Seitkalieva, A. V., Menzorova, N. I., Vakorina, T. I., Dmitrenok, P. S., Rasskazov, V. A.
Format: Article
Language:English
Subjects:
Aquatic life
Biochemistry
Biological activity
Biomedical and Life Sciences
Chemical analysis
Eggs
Enzymatic activity
Ions
Life Sciences
Mass spectrometry
Medical Microbiology
Microbiology
Peptides
Phosphatase
Salt
Seawater
Sodium chloride
Water analysis
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Summary:It was shown that the alkaline phosphatase (AP) isolated from eggs of the sea urchin Strongylocentrotus intermedius was a homodimer with a molecular mass of 150 kDa and that it exhibited maximal activity at a рН of 8.1–8.5 and a temperature of 45°С. Mg 2+ , Ca 2+ , and Mn 2+ ions, as well as DTT, are activators of AP activity, while Zn 2+ , Cu 2+ , Cd 2+ , Pb 2+ , and Ni 2+ ions, as well as EDTA, N-EMI, and p -CMB, inhibit its activity. The enzyme demonstrates unique salt resistance and can hydrolyze the substrate in seawater. It was shown by fluorescence methods and circular dichroism that an increase of the NaCl concentration above 1.0М caused noticeable changes in the secondary and tertiary structures of the protein and a decrease in enzyme activity. Analysis of the molecular masses of tryptic peptides conducted by mass spectrometry with the MASCOT program (which is based on the NCBI and SWISS-PROT databases) did not allow identification of the investigated protein. The uniqueness of the source and of the properties of the investigated AP, which were not characteristic for nonspecific AP, as well as the difficulties in primary structure identification with the fingerprinting technique, suggested that this enzyme was a nontypical AP with a novel structure.
ISSN:0003-6838
1608-3024
DOI:10.1134/S0003683817010173