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正红菇菌丝体经磷酸缓冲液浸提、硫酸铵分级沉淀、DEAE-Sepharose FF离子交换层析和Sephadex G-100分子筛层析纯化得到红菇凝集素(Russula vinosa Lectin,RVL)。经SDS-PAGE检测为单一蛋白带,其亚基相对分子质量为55kDa,Sephadex G-100凝胶过滤测得相对分子质量为55.25kDa,提示RVL分子只有一个亚基。RVL中性糖含量为3.87%,经酸水解测定含15种氨基酸。温度在20–60℃、pH在5–9的范围内,凝集活性保持相对的稳定。RVL的凝血活性受Mn2+、Zn2+、Ca2+的影响。糖抑制实验表明,在供试的11种糖中,D-甘露糖强烈抑制RVL的凝血活性。抑菌实验显示,RVL对供试的细菌没有抑制作用,对稻瘟病菌、绿色木霉、红色链包霉、黑曲霉菌丝生长有显著的抑制作用。
The mycelial growth of Russula was extracted by phosphate buffer, ammonium sulfate fractionation, DEAE-Sepharose FF ion exchange chromatography and Sephadex G-100 molecular sieve chromatography to obtain Russula vinosa Lectin (RVL). The single protein band was detected by SDS-PAGE. The relative molecular mass of the subunit was 55 kDa. The relative molecular mass of Sephadex G-100 gel filtration was 55.25 kDa, suggesting that there is only one subunit of RVL molecule. RVL neutral sugar content of 3.87%, determined by acid hydrolysis containing 15 kinds of amino acids. Agglutination activity remained relatively stable at a temperature in the range of 20-60 ° C with a pH in the range of 5-9. The coagulation activity of RVL is affected by Mn2 +, Zn2 + and Ca2 +. Sugar inhibition experiments showed that among the 11 kinds of sugars tested, D-mannose strongly inhibited the coagulation activity of RVL. Bacteriostasis experiments showed that RVL had no inhibitory effect on the tested bacteria and inhibited the growth of M. grisea, Trichoderma viride, Cladosporium rubrum and Aspergillus niger.