Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying

Abstract

The microorganisms Escherichia coli DH5␣ and Bacillus thuringiensis HD-1 show an increased tolerance to freeze-drying when dried in the presence of the disaccharides trehalose and sucrose. When the bacteria were dried with 100 mM trehalose, 70% of the E. coli and 57% of the B. thuringiensis organisms survived, compared with 56 and 44%, respectively, when they were dried with sucrose. Only 8% of the E. coli and 14% of the B. thuringiensis organisms survived drying without the sugars. Fourier transform infrared spectroscopy was used to investigate the role of membrane phase transitions in the survival of the organisms during drying and rehydration. Both E. coli and B. thuringiensis showed an increase of 30 to 40؇C in the temperature of their phospholipid phase transition when dried without the sugars, while phase transition temperatures of those dried with the sugars remained near those of the hydrated cells. A Fourier transform infrared spectroscopy microscope made it possible to investigate the effects of drying on the protein structure in the intact cells. The amide II peak shifts from 1,543 cm ؊1 in the hydrated cells to about 1,533 cm ؊1 in the cells dried without sugar. There is no shift in the amide II peak when the cells are dried with trehalose or sucrose. We attribute the increased survival to the sugars' ability to lower the membrane phase transition temperature and to protect protein structure in the dry state. In addition to increasing the immediate survival of both species, the addition of trehalose protected the cells from the adverse effects of exposure to light and air while dry. E. coli dried with trehalose and exposed to light and air for 4 h had an increase in CFU of between 2,000 and 4,000 times the number obtained with E. coli dried without trehalose. B. thuringiensis showed an increase in CFU of 150% in samples dried with trehalose compared with samples dried without trehalose. The cells dried with sucrose did not show an increased tolerance to exposure following drying.

Trehalose‐phosphate synthase of Mycobacterium tuberculosis

“…The charged sugars were then eluted with a 0-0.3 M linear gradient of NH 4 HCO 3. As shown in profile A, a sharp symmetrical peak of radioactivity emerged at 0.1-0.15 M NH 4 HCO 3 and fractions [23][24][25][26][27][28][29][30][31][32] containing radioactivity were pooled and concentrated several times with triethylamine to remove the NH 4 HCO 3 . In profile B, the charged compound was treated with alkaline phosphatase and subjected to paper chromatography in solvent A.…”

Effects of Moisture Content on the Storage Stability of Dried Lipoplex Formulations

“…15,16 It has been conclusively demonstrated that sugars (e.g., sucrose, trehalose) are able to preserve macromolecules during drying. 17 The mechanism by which sugars exert their protective effect is thought to involve both glass formation 18,19 and hydrogen bonding, 20 although the validity of these mechanisms is still debated. 21 Regardless of the precise mechanism responsible for stabilization, the role of water as both a reactant and plasticizer suggests that progressively greater stability should be observed at lower moisture contents.…”

Hydrophilins in the filamentous fungus Neosartorya fischeri (Aspergillus fischeri) have protective activity against several types of microbial water stress

“…Compatible solutes consist of sugars, polyols, amino acids and derivatives thereof. They stabilize cellular structures and restore the osmotic balance without interfering with the functioning of biomolecules (Leslie et al, 1995;Shen et al, 1999;Billi et al, 2000). Protective proteins bind to and structurally stabilize other proteins, membranes and RNA (Warner et al, 2004).…”