Introduction
Pressure-induced denaturation of substrate proteins leads to better access of enzymes to previously inaccessible, or poorly accessible, target sites, resulting in improved and accelerated digestion. Here we report that PNGase F deglycosylation of native proteins is accelerated when the reactions are carried out under pressure.
PNGase F is a glycosidase that cleaves N-linked high mannose, hybrid, and complex oligosaccharides. We tested PNGase F activity under high pressure conditions using two model glycoproteins, human alpha1 acid glycoprotein (α1AGP), and bovine fetuin.

High Hydrostatic Pressure in the Barocycler System Pressure Cycling Technology (PCT) uses hydrostatic pressure cycles to control biomolecular interactions and to accelerate certain enzymatic reactions. The samples in the pressure chamber are exposed to rapid pressurization from atmosphere (14.7 psi) to levels as high as 45,000psi (45kpsi), or higher in some applications.
The system is comprised of a Barocycler instrument, such as the 2320EXT, and high pressure-resistant sample containers such as PCT MicroTubes. Up to 16 MicroTubes at a time can be processed in the 2320EXT.