Panchal T, Wodzinski R J
Department of Molecular Biology and Microbiology, University of Central Florida, Orlando 32816, USA.
Prep Biochem Biotechnol. 1998 Aug;28(3):201-17. doi: 10.1080/10826069808010136.
The glycosylation patterns of phytase (E C 3.1.3.8) from Aspergillus niger NRRL 3135 with recombinant phytase from A. niger van Tieghem were compared. The following characteristics were studied: size of the whole molecule, type of linkage (N-linked or O-linked oligosaccharide), profile (number of different type of oligosaccharides present), monosaccharide composition, their order, and configuration. The molecular weights of both glycoproteins, after deglycosylation, was approximately 55 kDa as determined by SDS-PAGE. Both glycoproteins were about 35.29% glycosylated (calculations were made on the basis of 85 kDa molecular weight before deglycosylation). Only N-linked oligosaccharides were present. When N-linked oligosaccharides were released and labeled, the same profile was obtained for both glycoproteins. Mannose residues were detected after digestion by combinations of various exoglycosidases. N-acetylneuraminic acid, galactose, and N-acetylglucosamine residues were not detected. Released mannose residues were (alpha 1-2,3,6) linked. The trisaccharide core structure was nonfucosylated for all the oligosaccharides released from both glycoproteins. The only major difference found between the two glycoproteins was the migration pattern of oligosaccharide bands on gel after digestion with alpha-mannosidases. The structure of N-linked oligosaccharides ranged from (Man)5(GlcNAc)2 to (Man)10 (GlcNAc)2.
对黑曲霉NRRL 3135的植酸酶（EC 3.1.3.8）与黑曲霉范蒂根重组植酸酶的糖基化模式进行了比较。研究了以下特征：整个分子的大小、连接类型（N-连接或O-连接寡糖）、谱图（存在的不同类型寡糖的数量）、单糖组成、它们的顺序和构型。经SDS-PAGE测定，两种糖蛋白去糖基化后的分子量约为55 kDa。两种糖蛋白的糖基化程度均约为35.29%（基于去糖基化前85 kDa的分子量进行计算）。仅存在N-连接寡糖。当释放并标记N-连接寡糖时，两种糖蛋白获得相同的谱图。用各种外切糖苷酶组合消化后检测到甘露糖残基。未检测到N-乙酰神经氨酸、半乳糖和N-乙酰葡糖胺残基。释放的甘露糖残基以（α1-2,3,6）连接。从两种糖蛋白释放的所有寡糖的三糖核心结构均无岩藻糖基化。两种糖蛋白之间发现的唯一主要差异是用α-甘露糖苷酶消化后凝胶上寡糖带的迁移模式。N-连接寡糖的结构范围从（Man）5（GlcNAc）2到（Man）10（GlcNAc）2。
