Kopecká M, Fleet G H, Phaff H J
Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
J Struct Biol. 1995 Mar-Apr;114(2):140-52. doi: 10.1006/jsbi.1995.1013.
The ultrastructure of isolated cell walls of Schizosaccharomyces pombe was studied by electron microscopy after treatment with the following purified enzymes: endo-beta-(1-->3)-glucanase, endo-beta-(-->6)-glucanase, and endo-alpha-(1-->3)-glucanase produced by Bacillus circulans; exo-beta-(1-->3)-glucanase and endo-beta-(1-->3)-glucanase produced by Schizosaccharomyces japonicus var. versatilis. The exo-beta-(1-->3)-glucanase had no detectable effect on the walls, but amorphous wall material was removed by action of the endo-beta-(1-->3)- and endo-beta-(1-->6)-glucanases of B. circulans to reveal a wall component consisting of densely interwoven microfibrils. The fibrils were hydrolyzed by treatment with the Schiz. japonicus endo-beta-(1-->3)-glucanase followed by B. circulans endo-alpha-(1-->3)-glucanase--suggesting that they were composed of -beta-(1-->3)-linked glucan and alpha-(1-->3)-linked glucan. The presence of a fibrillar component in untreated walls was evident after negative staining.
用以下纯化酶处理后,通过电子显微镜研究了粟酒裂殖酵母分离细胞壁的超微结构:环状芽孢杆菌产生的内切-β-(1→3)-葡聚糖酶、内切-β-(→6)-葡聚糖酶和内切-α-(1→3)-葡聚糖酶;日本裂殖酵母变种产生的外切-β-(1→3)-葡聚糖酶和内切-β-(1→3)-葡聚糖酶。外切-β-(1→3)-葡聚糖酶对细胞壁没有可检测到的作用,但环状芽孢杆菌的内切-β-(1→3)-和内切-β-(1→6)-葡聚糖酶作用去除了无定形壁物质,从而揭示出一种由紧密交织的微纤维组成的壁成分。用日本裂殖酵母内切-β-(1→3)-葡聚糖酶处理,然后用环状芽孢杆菌内切-α-(1→3)-葡聚糖酶处理,微纤维被水解,这表明它们由β-(1→3)-连接的葡聚糖和α-(1→3)-连接的葡聚糖组成。负染色后,未处理的细胞壁中纤维状成分的存在很明显。