Assessing Synergism Between Non-Reducing Sugars and Intrinsically Disordered Proteins in Protecting Fragile Biological Materials from Desiccation

Modern society takes full advantage of the wide availability of refrigeration to maintain food, medical supplies (vaccines, plasma), chemicals, and whole cells in a useful state. The cold-/coolchain (CC; as it has become known) ensures that valuable, fragile biological materials (FBM) are stored under conditions that maximize FBM utility. The absolute requirement for uninterrupted CC maintenance to safeguard FBM function underlies the vulnerability of the CC to electrical disruption. Power outage frequently occurs during natural disasters and armed conflict while remote areas remain without access to electricity entirely. Drying FBM while maintaining its serviceability (usually activity or even viability upon rehydration) offers numerous advantages relative to the CC which is expensive, difficult to extend to regions without it, subject to disruption, centrally distributed, and conspicuous by its nature (heavy, bulky, noisy refrigerators/generators and equipment heat signatures at a central locale). The focus of my research is to understand how certain organisms can, as some stage of their life cycle, withstand almost complete loss of water without dying. While in the desiccated state these organisms display extreme resistance to abiotic perturbation and aging. Generally, the attributes cells acquire leading to anhydrobiosis (life with little water) include the accumulation of specific, non-reducing oligosaccharides (trehalose, and sucrose:raffinose admixtures) and some form of intrinsically disordered protein. These macromolecules are thought to: 1) replace water around otherwise desiccation sensitive cellular constituents; 2) vitrify the cellular interior, embedding cellular contents in a glass that severely restricts movement and represses metabolism and; 3) shield sensitive cellular components from reactive oxygen species. The overwhelming body of literature reporting the protective contributions of sugars and intrinsically disordered proteins have focused on one or the other, failing to more closely approximate nature where the two protectants are combined. Neither have many projects examined the consequence of desiccating complex macromolecular assemblages in the presence of the protectants despite reports that sugar recovery from previously desiccated material is considerably reduced by mechanisms not explained by hydrolysis, and suggestions that bound sugar replacing water may become permanently attached. These questions are directly germane to pursuit of an informed utilization of biomimicry to preserve FBM in a useful state through desiccation. We hope to continue our work examining this fascinating aspect of life by replacing our defunct HPLC for sugar detection with a new and functional unit, which is the subject of this request.