Purification of Schizophyllum commune Protein of 30 kDa with Antitumor and Immunostimulative Activities.
Date Issued
2008
Date
2008
Author(s)
Chuang, Po-Yen
Abstract
From the fermentation liquid of the mushroom Schizophyllum commune, a monomer protein, BCⅡ, with immunostimulative and antitumor activities had been isolated. The purification procedure was involved (NH4)2SO4 precipitation, hydrophobic interaction chromatography on Phenyl sepharose, ion exchange chromatography on Mono-Q and gel filtration on Superdex 75 by fast protein liquid chromatography (FPLC). The molecular mass of BCⅡ was 30 kDa as estimated via SDS-PAGE and gel filtration. Further staining using the periodic acid / Schiff reagents showed that BCⅡ was not a glycoprotein. BCⅡshowed antiproliferative activity toward mouse Lewis lung carcinoma (LLC) at BCⅡ concentration of 50, 25 μg/ml, respectively. And showed no cytotoxicity to mouse spleen cells by MTT assay. The immunostumulative activities of BCⅡ was also demonstrated in this study, which activate directly cytotoxicity of nature killer (NK) cell and macrophage RAW 264.7 to produce nitric oxide and TNF-α. BCⅡ stimulated macrophage RAW 264.7 to produce nitric oxide in a dose-dependent effect. The highest level of TNF-α was exhibited at BCⅡ concentration of 25μg/ml. Based on the results obtained in the present study, BCⅡ is a potent immune stimulant and could develop potentially using in medicine field.
Subjects
mushroom protein
purification
atitumor
immunostimulative
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