Nakajima T, Tamari K, Matsuda K
J Biochem. 1977 Dec;82(6):1647-55. doi: 10.1093/oxfordjournals.jbchem.a131860.
A purified proteo-heterolgycan, [alpha]D + 72.5 degrees, was isolated from Piricularia oryzae, a pathogenic fungus of rice blast disease (Imochi-byo), by means of hot citrate buffer extraction, cetavlon fractionation, and DEAE-Sephade chromatography. It was found to be homogeneous by electrophoresis and by analytical ultracentrifugation to have an s value 6.1 and to contain 91% (w/w) of carbohydrate, which consists of D-mannose, D-glucose, and D-galactose in a molar ratio of 6:2:1. Partial acid hydrolysis and methylation analysis of the carbohydrate moiety of the proteo-heteroglycan indicate that the molecule is composed of mannan, the side chain terminals of which are partially modified with D-glucopyranose and D-galactofuranose. Enzymatic hydrolysis with bacterial alpha-D-mannanase has been shown to remove most of the side chains from the heteroglycan, leaving an (1-6) linked mannan back-bone with a small amount of side chains, the terminals of which must be modified with D-glucopyranose or D-galactofuranose. The carbohydrate to protein linkage of the proteo-heteroglycan was shown by alkaline beta-elimination, to be mannosyl serine or mannosyl threonine.
通过热柠檬酸盐缓冲液提取、西曲溴铵分级分离和DEAE - 葡聚糖凝胶色谱法,从稻瘟病(稻热病)的致病真菌稻梨孢中分离出一种比旋光度[α]D + 72.5°的纯化蛋白聚糖杂多糖。经电泳和分析超速离心表明其具有均一性,沉降系数s为6.1,且含有91%(w/w)的碳水化合物,该碳水化合物由摩尔比为6:2:1的D - 甘露糖、D - 葡萄糖和D - 半乳糖组成。对该蛋白聚糖杂多糖碳水化合物部分进行部分酸水解和甲基化分析表明,该分子由甘露聚糖组成,其侧链末端部分被D - 吡喃葡萄糖和D - 呋喃半乳糖修饰。已证明用细菌α - D - 甘露聚糖酶进行酶解可从杂多糖中去除大部分侧链,留下具有少量侧链的(1→6)连接的甘露聚糖主链,其末端必定被D - 吡喃葡萄糖或D - 呋喃半乳糖修饰。通过碱性β - 消除反应表明,该蛋白聚糖杂多糖的碳水化合物与蛋白质的连接方式为甘露糖基丝氨酸或甘露糖基苏氨酸。