筛选担子菌中的凝集素及分离出点柄乳菇凝集素。

Screening for lectins from basidiomycetes and isolation of Punctularia atropurpurascens lectin.

机构信息

Cátedra de Microbiología, Departamento de Biociencias, Facultad de Química, UdelaR, Montevideo, Uruguay.

出版信息

J Basic Microbiol. 2014 Feb;54(2):89-96. doi: 10.1002/jobm.201200229. Epub 2013 Mar 5.

DOI:10.1002/jobm.201200229

Abstract

Aqueous extracts of basidiomycete fungi were screened for the presence of lectins by hemagglutination (HA) assays with mouse, rabbit, and sheep red blood cells. From mycelia and/or fruiting bodies, 23 extracts were prepared; 15 extracts exhibited HA activity towards mouse erythrocytes, with specific activities ranging from 12 to 440 lectin units (LU) mg(-1) protein. In HA inhibition assays, 43 carbohydrates including mono-, di-, tri-, tetrasaccharides, glycoproteins, and polysaccharides were tested as haptens, to determine the saccharide-binding specificities of the lectins. A novel lectin with specificity towards N-acetyl-glucosamine was purified from mycelia of Punctularia atropurpurascens using affinity chromatography on chitosan-Sepharose. The lectin has a subunit molecular mass of 67 kDa determined by SDS-PAGE and a pI of 5.0.

摘要

采用血凝（HA）试验，以小鼠、兔和绵羊的红细胞为研究对象，筛选了担子菌真菌的水提物中是否存在凝集素。从菌丝体和/或子实体中制备了 23 种提取物；其中 15 种提取物对小鼠红细胞具有 HA 活性，其比活性范围为 12 至 440 个凝集素单位（LU）/mg（-1）蛋白。在 HA 抑制试验中，测试了 43 种碳水化合物，包括单糖、二糖、三糖、四糖、糖蛋白和多糖，作为半抗原，以确定凝集素的糖结合特异性。从暗紫丝膜菌的菌丝体中，通过壳聚糖-琼脂糖亲和层析，从暗紫丝膜菌的菌丝体中纯化出一种对 N-乙酰葡萄糖胺具有特异性的新型凝集素。通过 SDS-PAGE 测定，该凝集素的亚基分子量为 67 kDa，等电点为 5.0。