Pressure-Enhanced Enzymes Application Notes

Pressure-Enhanced Enzymes Application Notes

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Proteolysis-PrEP: Optimization of Pressure Cycling Conditions for Efficient Digestion of Native IgG

Introduction
PCT has been proven to accelerate enzymatic protein digestion and the positive effect of PCT on digestion by both trypsin and Lys-C is well established [1-4]. Additionally, the enhancing effects of PCT on the activity of enzymes including Proteinase K, PNGase F, chymotrypsin, and lysozyme have been reported [5-8]. High hydrostatic pressure accelerates protein digestion via a combination of two mechanisms.

Guidelines for Protocol Development:

Introduction
Pressure-enhanced proteolytic digestion exploits the ability of high hydrostatic pressure to promote protein denaturation and the access of proteolytic enzymes to their target sites. Pressure denaturation is fundamentally different from thermal denaturation as it occurs by virtue of hydration of hydrophobic residues and by water saturation of protein substrate cavities normally inaccessible to solvent. Pressure denaturation is more efficient for hydrophobic proteins, while some hydrophilic soluble proteins are reported to retain relatively compact conformation, even when saturated by water molecules [1-6].

Proteolysis-PrEP: Pressure-Accelerated Activity of Elastase Enzyme

Introduction
The positive effect of Pressure Cycling Technology (PCT) on trypsin digestion is well established [1-4], and has been shown to result in improved sequence coverage, higher recovery and significantly reduced digestion times. Additionally, the enhancing effect of PCT on the activity of several other enzymes, including Lys-C, Proteinase K, PNGase F, chymotrypsin, Glu-C, thermolysin, and lysozyme has been reported [5-12].

Proteolysis-PrEP: Pressure-Accelerated Protein Digestion by Thermolysin Enzyme

Introduction
The positive effect of Pressure Cycling Technology (PCT) on digestion with trypsin is well established [1-4], and has been shown to result in improved sequence coverage, higher peptide intensities and significantly reduced digestion times. Additionally, the enhancing effect of PCT on the activity of several other enzymes, including Lys-C, Proteinase K, PNGase F, chymotrypsin, Glu-C and lysozyme has been reported [5-11].

Pressure Cycling-Accelerated Digestion with Trypsin: Digestion of Native Proteins in Whole Tissue Lysate

Introduction
The positive effect of Pressure Cycling Technology (PCT) on digestion with trypsin is well established [1-4], and has been shown to result in improved sequence coverage, higher peptide intensities and significantly reduced digestion times. Additionally, the enhancing effect of PCT on the activity of several other enzymes, including Lys-C, chymotrypsin, Glu-C, thermolysin, Proteinase K, and lysozyme, has been reported [5-11].