Use of resorufin-labelled N-glycopeptide in a high-performance liquid chromatography assay to monitor endoglycosidase activities during cultivation of Flavobacterium meningosepticum.
Peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase F (PNGase F) and endo-beta-N-acetyl glucosaminidase F (Endo F) activities were monitored during cultivation of Flavobacterium 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.
Bourgerie
S
S
Institute de Biotechnologie, Université de Limoges, France.
Karamanos
Y
Y
Berger
S
S
Julien
R
R
D000581:Amidohydrolases / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005417:Flavobacterium / Q000201:enzymology* / Q000254:growth & development; D006020:Glycopeptides / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism*; D008969:Molecular Sequence Data; D010078:Oxazines; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
D006020:Glycopeptides; D010078:Oxazines; C014180:resorufin; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase