Song K S, Cho S M, Lee J H, Kim H M, Han S B, Ko K S, Yoo I D
Korea Research Institute of Bioscience & Biotechnology. Taejon, Korea.
Chem Pharm Bull (Tokyo). 1995 Dec;43(12):2105-8. doi: 10.1248/cpb.43.2105.
Hot water extract prepared from the mycelial culture of mushroom Phellinus linteus stimulated polyclonal antibody production in an in vitro culture system. The active fraction PLP was purified from the extract ca. 1030-fold by ethanol precipitation followed by DEAE-cellulose and gel permeation chromatography. PLP contained 13.2% (w/w) peptide and 82.5% (w/w) carbohydrate. About 6.8% (w/w) of the total carbohydrate was uronic acid. The molecular weight distribution of PLP was found to be nearly homogeneous (153 kDa) in gel permeation HPLC analysis. Neutral sugar composition analysis revealed Ara (7.5%), Xyl (3.7%), Glc (21.1%), Gal (24.1%) and Man (44.2%). Uronic acid was identified as a glucuronic acid by gas chromatography. Ten amino acids were detected and Asp and Glu were the major components. In our assay system, the half-maximal concentration of PLP for B-lymphocyte stimulation was ca. 3 micrograms/ml. Partial acid hydrolysis as well as sodium periodate treatment of PLP decreased the activity significantly, suggesting that both the full molecular size and the sugar moiety were essential. However, proteinase K treatment for up to 48 h did not affect the activity.
从桑黄菌丝体培养物中制备的热水提取物在体外培养系统中刺激多克隆抗体的产生。活性组分PLP通过乙醇沉淀，然后经DEAE - 纤维素和凝胶渗透色谱从提取物中纯化约1030倍。PLP含有13.2%（w/w）的肽和82.5%（w/w）的碳水化合物。总碳水化合物的约6.8%（w/w）是糖醛酸。在凝胶渗透HPLC分析中发现PLP的分子量分布几乎均匀（153 kDa）。中性糖组成分析显示含有阿拉伯糖（7.5%）、木糖（3.7%）、葡萄糖（21.1%）、半乳糖（24.1%）和甘露糖（44.2%）。通过气相色谱法鉴定糖醛酸为葡萄糖醛酸。检测到十种氨基酸，其中天冬氨酸和谷氨酸是主要成分。在我们的测定系统中，PLP刺激B淋巴细胞的半数最大浓度约为3微克/毫升。PLP的部分酸水解以及高碘酸钠处理显著降低了活性，这表明完整的分子大小和糖部分都是必不可少的。然而，蛋白酶K处理长达48小时并不影响活性。
