Use of resorufin-labelled N-glycopeptide in a high-performance liquid chromatography assay to monitor endoglycosidase activities during cultivation ofFlavobacterium meningosepticum

Peptide-N⁴-(N-acetyl--glucosaminyl) asparagine amidase F (PNGase F) and endo--N-acetyl glucosaminidase F (Endo F) activities were monitored during cultivation ofFlavobacterium meningosepticum using a new fluorescence-HPLC procedure based on a commercially available substrate. The PNGase F activity reached a maximum level at the end of the log phase and remained constant during the stationary phase, while Endo F continuously increased until late stationary phase. PNGase F obtained at the end of the log phase was less contaminated by other proteins compared with late stationary phase.Abbreviations Con A concanavalin A - Endo F endo--N-acetyl glucosaminidase F (EC 3.2.1.96) - GlcNAc N-acetylglucosamine - PNGase F peptide-N⁴-(N-acetyl--glucosaminyl) asparagine amidase F (EC 3.5.1.52).