Endoglucanase Degradation and Enzyme-Aided Characterization of Cellulose Acetates

Endoglucanases were used as a selective tool to determine the substituent distribution along the chains of partially substituted cellulose acetates. Unsubstituted segments are intensively fragmented while highly substituted segments are only slightly degraded or not affected at all. Two different procedures were developed to perform a preparative separation of the fragments by size exclusion chromatography. One system is based on a direct separation of the fragmentation products using pyridine : water (9 : 1) as an eluent. The isolated fragments can be analysed regarding to their DS values by ATR/FTIR spectroscopy or by hydrolysis and HPLC. The second system is based on deutero-acetylation of endoglucanase fragmented samples. The complete acetylation of all OH groups enables a chromatographic separation in chloroform. Afterwards the isolated fragments can be analysed regarding to their partial DS values by ¹H-NMR spectroscopy.