Protecting Enzymes from Stress-Induced Inactivation

Abstract

The pharmaceutical and chemical industries depend on additives to protect enzymes and other proteins against stresses that accompany their manufacture, transport, and storage. Common stresses include vacuum-drying, freeze-thawing, and freeze-drying. The additives include sugars, compatible osmolytes, amino acids, synthetic polymers, and both globular and disordered proteins. Scores of studies have been published on protection, but the data have never been analyzed systematically. To spur efforts to understand the sources of protection and ultimately develop more effective formulations, we review ideas about the mechanisms of protection, survey the literature searching for patterns of protection, and then compare the ideas to the data.

Optimization of Protective Agents for The Freeze-Drying of Paenibacillus polymyxa Kp10 as a Potential Biofungicide

“…Sucrose was reported to replace water around polar residues in macromolecular, thus stabilizing cell membranes and proteins during desiccation [35]. Sugars from disaccharides could preserve cell structure by hydrogen bond formation that maintains tertiary protein structure when water molecules are absent [36]. Meanwhile, skim milk rich in protein is capable of preventing cells from injury caused by extracellular ice formation during the freezing stage by providing a protective coat to the cells [32].…”

“…In this study, a combination of skim milk, sucrose, and lactose gave a significant effect on cell viability after the freeze-drying process by maintaining maximum cell viability. water molecules are absent [36]. Meanwhile, skim milk rich in protein is capable of preventing cells from injury caused by extracellular ice formation during the freezing stage by providing a protective coat to the cells [32].…”

Toxicity and Immunogenicity of a Tardigrade Cytosolic Abundant Heat Soluble Protein in Mice

“…In addition to vaccines, there are more than 200 FDA-approved protein- and peptide-based therapeutics on the market, including treatments for type 2 diabetes, breast cancer, and prostate cancer ( Fosgerau and Hoffmann, 2015 ; Usmani et al., 2017 ). To stabilize proteins and increase shelf life, sugars, polymers, amino acids, globular proteins, and osmolytes have been investigated as excipients ( Piszkiewicz and Pielak, 2019 ). Even in the presence of excipients, proteins must be kept at low temperatures.…”

“…Even in the presence of excipients, proteins must be kept at low temperatures. Lyophilization and desiccation can increase the shelf life of some proteins but may lead to problems such as aggregation and denaturation ( Piszkiewicz and Pielak, 2019 ).…”

Exploring Enzymatic Activity in Multiparameter Space: Cosolvents, Macromolecular Crowders and Pressure

“…Obviously, such extremolytes, i. e. compatible cosolutes that are found in extremophiles, may be of particular use. They accumulate to high concentrations in cells in response to diverse environmental stresses, such as heat, cold, osmotic pressure, desiccation, and high hydrostatic pressure without interfering with cellular metabolism and allow their hosts to survive harsh ecological conditions [28,29,34] . Here, we explored the effects of urea and TMAO, which are also widely used in protein folding studies.…”