Effect of sorbitol and glycerol on the stability of trypsin and difference between their stabilization effects in the various solvents

Abstract

The effect of glycerol and sorbitol on the stability of porcine pancreas trypsin was investigated in this work. Molecular dynamics simulation and thermostability results showed that trypsin has two flexible regions, and polyols (sorbitol and glycerol) stabilize the enzyme by decreasing the flexibility of these regions. Radial distribution function results exhibited that sorbitol and glycerol were excluded from the first water layer of the enzyme, therefore decrease the flexibility of the regions by preferential exclusion. Also, results showed that the stabilization effect of sorbitol is more than glycerol. This observation could be because of the larger decrease in the fluctuations of trypsin in the presence of sorbitol. We also examined the role of solvent's hydrophobicity in enzyme stabilization by sorbitol and glycerol. To do so, the thermostability of trypsin was evaluated in the presence of solvents with different hydrophobicity (methanol, ethanol, isopropanol and n-propanol) in addition to the polyols. Our results depicted that glycerol is a better stabilizer than sorbitol in the presence of hydrophobic solvents (n-propanol), whereas sorbitol is a better stabilizer than glycerol in the presence of hydrophilic solvents (methanol).

Substrate scope of a dehydrogenase from Sphingomonas species A1 and its potential application in the synthesis of rare sugars and sugar derivatives

“…A). Although the theoretical V max was not reached here, as this value only reflects the activity without any product present, SpsADH seemed to be stabilized by sorbitol, as it has also been reported for various other proteins (Brennan et al ., ; Khajehzadeh et al ., ; Pazhang et al ., ). This allowed SpsADH to be active for an extended time period of approximately 230 h, resulting in this high yield.…”

Enhanced trypsin thermostability in Pichia pastoris through truncating the flexible region

“…In many instances, a high thermostability of trypsin is usually required in some industrial and biotechnological applications. Some methods, such as immobilization, the addition of polyol and enzymatic glycation, have been used for improving the thermostability of trypsin [ 6 , 27 , 28 ]. In the present study, we improved the thermostability of trypsin by truncating the flexible region.…”

How to Lengthen the Long-Term Stability of Enzyme Membranes: Trends and Strategies

“…Cipolatti et al prepared lipase-immobilized PEGylated polyurethane particles and showed a stabilizing effect for oil ethanolysis [36]; lipase was adsorbed on the polymer, and the protection of the PEG derivative against the oil environment was effective on the enzyme. Pazhang et al showed the stabilizing effect of sorbitol and glycerol on trypsin in organic solvents [37]. As a result, immobilized enzymes were stabilized by the mechanism, wherein amphiphilic molecules are not required to bind covalently.…”