Asparagine-Linked Carbohydrate of Penicillium notatum Phospholipase B

Abstract

The asparagine-linked sugar chains of phospholipase B from Penicillium notatum were released by hydrazinolysis. The carbohydrate components were re-N-acetylated and then reductively aminated with 2-aminopyridine. The pyridyl-aminated derivatives were analyzed by size-fractionation and reverse-phase HPLC. The results indicated that the enzyme contains only one kind of oligosaccharide. The carbohydrate was purified by HPLC, and then subjected to NMR and MS analyses for determination of its structure. Based on the results, the structure was determined to be as follows.
Malpha;1-2Manα1_??_Manα1-2Manα1_??_ ⁶₃Manα1_??_ Manα1-2Manα1-2Manα1_??_ ⁶₃Manβ1-4GlcNAcβ1-4GlcNAc