微生物學(xué)通報 AUG 20,2010,37(8):1153～1158
香灰菌凝集素的純化及其部分性質(zhì)
佀國涵1 周瑋婧2 馬愛民2*
(1. 湖北省農(nóng)業(yè)科學(xué)院植保土肥所 湖北 武漢 430064)
(2. 華中農(nóng)業(yè)大學(xué)食品科技學(xué)院 湖北 武漢 430070)
摘 要: 香灰菌菌絲體經(jīng)磷酸緩沖液抽提,、20% 70%飽和濃度的硫酸銨沉淀、DEAE-Cellulose和Sephadex G-100柱層析純化得到香灰菌凝集素(Hypoxylon sp. lectin, 簡稱HSL),。HSL經(jīng)PAGE檢測為單一蛋白條帶, SDS-PAGE測得其亞基分子量為15.9 kD,。過碘酸-Schiff染色法表明HSL為一種糖蛋白, 糖基的含量為15.5%, β-消去反應(yīng)測得其糖和蛋白質(zhì)的連接鍵為O-型糖肽鍵,。HSL能凝集多種動物紅細(xì)胞和人的紅細(xì)胞, 在所測試的紅細(xì)胞中, 對兔紅細(xì)胞的凝集作用最強,。HSL對熱較敏感, 經(jīng)50°C處理10 min, 其凝集活性明顯降低, 其在堿性環(huán)境中較穩(wěn)定, 而在酸性環(huán)境中較不穩(wěn)定,。HSL的凝集活性受Al3+,、Fe3+,、Ca2+和Zn2+等陽離子的影響,。對鼠紅細(xì)胞的凝集作用可被半乳糖和乳糖所抑制,。
關(guān)鍵詞: 香灰菌, 凝集素, 純化, 性質(zhì)
Purification and Partial Characterization of the Lectin from Hypoxylon sp.
SI Guo-Han1 ZHOU Wei-Jing2 MA Ai-Min2*
(1. Plant Protection and Soil Fertilizer Institute, Hubei Academy of Agricultural Sciences, Wuhan, Hubei 430064, China)
(2. College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China)
Abstract: The novel lectin was isolated from the mycelium of Hypoxylon sp. by phosphoric acid buffer, precipitation of 20% 70% (NH4)2SO4, DEAE-Cellulose and Sephadex G-100 chromatography. It turned out to be a single band in PAGE. SDS-PAGE showed the subunit of Hypoxylon sp. lectin (HSL) was 15.9 kD. HSL was glycoprotein through dyed by Periodic acid-Schiff reaction, and the carbohydrate content was 15.5%. β-Elimination revealed the bond between the polysaccharide and the protein of HSL was O-type glucopeptide one. The HSL could agglutinate erythocytes regardless of blood type or animal species. HSL was less thermostable, and the hemagglutinating activity declined obviously after being heated 10 min under 50°C. The HSL was alkali-stable but not acid-stable. Its activity was affected by Al3+, Fe3+, Ca2+ and Zn2+. The hemagglutination of the lectin on mouse erythrocytes was inhibited by galactose and lactose among the sugars tested.
Keywords: Hypoxylon sp., Lectin, Purification, Characterization
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