Harz H, Burgdorf K, Höltje J V
Max-Planck-Institut für Entwicklungsbiologie, Abt. Biochemie, Tübingen, Federal Republic of Germany.
Anal Biochem. 1990 Oct;190(1):120-8. doi: 10.1016/0003-2697(90)90144-x.
The length distribution of the glycan strands in the murein (peptidoglycan) sacculus of Escherichia coli has been analyzed after solubilization of the murein by complete digestion with human serum amidase. The glycan strands released were separated according to length by reversed-phase HPLC on wide-pore Nucleosil 300 C18 material at 50 degrees C, employing a convex gradient from 5 to 11% acetonitrile. The length of the fractionated glycan strands, which carry a nonreducing 1,6-anhydromuramic acid as a natural end group, was calculated from the ratio of total to nonreducing terminal muramic acid residues. This was possible after complete hydrolysis of the isolated glycan strands by muramidase followed by separation of the released nonreducing and reducing di- and tetrasaccharides by reversed-phase HPLC on Hypersil C18. The method established allows the separation of the glycan strands of murein, a poly-GlcNAc(beta 1-4)MurNAc-polysaccharide, up to a degree of polymerization of approximately 60. The predominant lengths of the glycan strands were 5 to 10 GlcNAc(beta 1-4)MurNAc disaccharide units.
用人类血清酰胺酶完全消化使大肠杆菌的胞壁质（肽聚糖）囊溶解后，分析了胞壁质中聚糖链的长度分布。在50℃下，采用5%至11%乙腈的凸形梯度，在宽孔Nucleosil 300 C18材料上通过反相高效液相色谱法（HPLC）根据长度分离释放的聚糖链。带有非还原性1,6 - 脱水 Muramic 酸作为天然末端基团的分级分离的聚糖链的长度，是根据总 Muramic 酸残基与非还原性末端 Muramic 酸残基的比例计算得出的。在通过溶菌酶将分离的聚糖链完全水解后，再通过在Hypersil C18上的反相HPLC分离释放的非还原性和还原性二糖及四糖，这才得以实现上述计算。所建立的方法能够分离胞壁质的聚糖链，即一种聚 - GlcNAc(β1 - 4)MurNAc - 多糖，聚合度可达约60。聚糖链的主要长度为5至10个GlcNAc(β1 - 4)MurNAc二糖单位。
