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Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans
Author(s) -
Noda Mamoru,
Shibata Kenichiro,
Sawa Yoshihiko,
Shimokoube Hirokata,
Watanabe Tsuguo
Publication year - 1994
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1994.tb01750.x
Subject(s) - biology , biochemistry , gel electrophoresis , phosphate , enzyme , polyacrylamide gel electrophoresis , threonine , affinity chromatography , chromatography , ion chromatography , sodium dodecyl sulfate , sepharose , molecular mass , phosphatase , serine , chemistry
An acid phosphatase associated with the cell membranes of Mycoplasma fermentans was released from the membranes with Triton X‐100, then purified by ion‐exchange chromatography on DEAE‐Sephacel and CM‐Sepharose, followed by affinity chromatography on Con A‐Sepharose. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified enzyme revealed a single band with a molecular mass of 31.2 kilodaltons. The enzyme activity toward p ‐nitrophenyl phosphate was enhanced remarkably by Cu 2+ , Co 2+ and Mg 2+ , but the activity was not inhibited by EDTA. The enzyme dephosphorylated O ‐phospho‐ l ‐tyrosine as well as p ‐nitrophenyl phosphate, but not O ‐phospho‐ l ‐threonine, O ‐phospho‐ l ‐serine, glucose‐1‐phosphate, phosphoryl choline and adenosine triphosphate. The level of the O ‐phospho‐ l ‐tyrosine phosphatase activity was the highest in Mycoplasma faucium and the second highest in Mycoplasma fermentans of all tested human mycoplasmas.