SOLID-STATE CP MASS C-13-NMR SPECTROSCOPY - A SENSITIVE METHOD TO MONITOR ENZYMATIC-HYDROLYSIS OF CHITIN/

The time-course hydrolysis of colloidal chitin by the chitinase comple x isolated from Myrothecium verrucaria was monitored using solution an d solid-state C-13-NMR spectroscopy. The solution NMR studies showed t he presence of N-acetylglucosamine (GlcNAc) as the sole product of hyd rolysis. Solid-state C-13 CP/MASS studies, on the other hand, indicate d the presence of high molecular weight oligomers as well as GlcNAc. T he linewidth of the C-1 carbon of the oligomers obtained after hydroly sis is found to be less than that of the unhydrolysed sample. The line widths calculated from the spin-spin relaxation times (T-2) of colloid al chitin and its products of hydrolysis were in the restricted range of 40-50 Hz, compared with the observed linewidths of 143-123 Hz. Peak area measurement on monomer to polymer/oligomer indicated an initial slow formation of the monomer, GlcNAc. From the NMR data, the involvem ent of endo-enzymes in the initial phase of hydrolysis is suggested.