ISSN:
1618-2545
Keywords:
acid phosphatase
;
N-terminal amino acid sequence
;
Pholiota nameko
;
phosphate-deficiency
;
purification
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Activity of acid phosphatase secreted by mycelia ofPholiota nameko on cultivation for 30d in Pi-depleted medium was 88-fold higher than the corresponding activity in the Pi-supplied medium. One isozyme of the secreted acid phosphatases was purified from the culture filtrate of Pi-depleted medium by ammonium sulfate fractionation and cation exchange chromatography. The purified enzyme was homogeneous on electrophoresis. Gel filtration analysis showed change chromatography. The purified enzyme was homogeneous on electrophoresis. Gel filtration analysis showed that the native molecule had a molecular weight of 117,000. The molecular weight on gel electrophoresis with SDS was 52,000, indicating that the native form of the enzyme was a homodimer. The optimum pH and temperature of the enzyme were, 5.5 and 45°C, respectively, and the isoelectric point of the enzyme was pH 6.9. Adsorption on Con A-Sepharose and periodic-Schiff stain suggested that the enzyme is a glycoprotein. The enzyme hydrolyzed a wide variety of phosphate esters, nucleoside phosphates, sugar phosphates, and phosphorylated amino acids. Cu2+, Fe2+, Hg2+, iodoacetate, molybdate, tartaric acid, and SDS inhibited the enzyme activity. Fe3+ (1 mM), Triton X-100, methanol, and ethanol activated it. Fifteen residues of the N-terminal amino acid sequence were determined.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02461459
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