• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. Oct 1995; 61(10): 3592–3597.
PMCID: PMC167656

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

Abstract

The microorganisms Escherichia coli DH5 alpha 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 degrees 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.(ABSTRACT TRUNCATED AT 250 WORDS)

Full Text

The Full Text of this article is available as a PDF (246K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Blok MC, van der Neut-Kok EC, van Deenen LL, de Gier J. The effect of chain length and lipid phase transitions on the selective permeability properties of liposomes. Biochim Biophys Acta. 1975 Oct 6;406(2):187–196. [PubMed]
  • Carpenter JF, Arakawa T, Crowe JH. Interactions of stabilizing additives with proteins during freeze-thawing and freeze-drying. Dev Biol Stand. 1992;74:225–239. [PubMed]
  • Carpenter JF, Crowe JH. An infrared spectroscopic study of the interactions of carbohydrates with dried proteins. Biochemistry. 1989 May 2;28(9):3916–3922. [PubMed]
  • Carpenter JF, Crowe LM, Crowe JH. Stabilization of phosphofructokinase with sugars during freeze-drying: characterization of enhanced protection in the presence of divalent cations. Biochim Biophys Acta. 1987 Jan 20;923(1):109–115. [PubMed]
  • Carpenter JF, Martin B, Crowe LM, Crowe JH. Stabilization of phosphofructokinase during air-drying with sugars and sugar/transition metal mixtures. Cryobiology. 1987 Oct;24(5):455–464. [PubMed]
  • Crowe JH, Crowe LM, Carpenter JF, Aurell Wistrom C. Stabilization of dry phospholipid bilayers and proteins by sugars. Biochem J. 1987 Feb 15;242(1):1–10. [PMC free article] [PubMed]
  • Crowe JH, Crowe LM, Carpenter JF, Rudolph AS, Wistrom CA, Spargo BJ, Anchordoguy TJ. Interactions of sugars with membranes. Biochim Biophys Acta. 1988 Jun 9;947(2):367–384. [PubMed]
  • Crowe JH, Crowe LM, Jackson SA. Preservation of structural and functional activity in lyophilized sarcoplasmic reticulum. Arch Biochem Biophys. 1983 Feb 1;220(2):477–484. [PubMed]
  • Crowe JH, Hoekstra FA, Crowe LM. Membrane phase transitions are responsible for imbibitional damage in dry pollen. Proc Natl Acad Sci U S A. 1989 Jan;86(2):520–523. [PMC free article] [PubMed]
  • Crowe JH, Hoekstra FA, Crowe LM, Anchordoguy TJ, Drobnis E. Lipid phase transitions measured in intact cells with Fourier transform infrared spectroscopy. Cryobiology. 1989 Feb;26(1):76–84. [PubMed]
  • Crowe LM, Crowe JH. Hydration-dependent hexagonal phase lipid in a biological membrane. Arch Biochem Biophys. 1982 Sep;217(2):582–587. [PubMed]
  • Crowe LM, Crowe JH, Rudolph A, Womersley C, Appel L. Preservation of freeze-dried liposomes by trehalose. Arch Biochem Biophys. 1985 Oct;242(1):240–247. [PubMed]
  • Gordon-Kamm WJ, Steponkus PL. Lamellar-to-hexagonalII phase transitions in the plasma membrane of isolated protoplasts after freeze-induced dehydration. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6373–6377. [PMC free article] [PubMed]
  • Haris PI, Chapman D. Does Fourier-transform infrared spectroscopy provide useful information on protein structures? Trends Biochem Sci. 1992 Sep;17(9):328–333. [PubMed]
  • Hoekstra FA, Crowe JH, Crowe LM. Effect of Sucrose on Phase Behavior of Membranes in Intact Pollen of Typha latifolia L., as Measured with Fourier Transform Infrared Spectroscopy. Plant Physiol. 1991 Nov;97(3):1073–1079. [PMC free article] [PubMed]
  • Israeli E, Shaffer BT, Lighthart B. Protection of freeze-dried Escherichia coli by trehalose upon exposure to environmental conditions. Cryobiology. 1993 Oct;30(5):519–523. [PubMed]
  • Koster KL, Webb MS, Bryant G, Lynch DV. Interactions between soluble sugars and POPC (1-palmitoyl-2-oleoylphosphatidylcholine) during dehydration: vitrification of sugars alters the phase behavior of the phospholipid. Biochim Biophys Acta. 1994 Jul 13;1193(1):143–150. [PubMed]
  • Leslie SB, Teter SA, Crowe LM, Crowe JH. Trehalose lowers membrane phase transitions in dry yeast cells. Biochim Biophys Acta. 1994 Jun 1;1192(1):7–13. [PubMed]
  • MacKenzie AP. Comparative studies on the freeze-drying survival of various bacteria: gram type, suspending medium, and freezing rate. Dev Biol Stand. 1976 Oct;36:263–277. [PubMed]
  • Prestrelski SJ, Byler DM, Liebman MN. Comparison of various molecular forms of bovine trypsin: correlation of infrared spectra with X-ray crystal structures. Biochemistry. 1991 Jan 8;30(1):133–143. [PubMed]
  • Rudolph AS, Crowe JH. Membrane stabilization during freezing: the role of two natural cryoprotectants, trehalose and proline. Cryobiology. 1985 Aug;22(4):367–377. [PubMed]
  • Stambuk BU, Crowe JH, Crowe LM, Panek AD, de Araujo PS. A dependable method for the synthesis of [14C]trehalose. Anal Biochem. 1993 Jul;212(1):150–153. [PubMed]
  • Susi H, Byler DM. Protein structure by Fourier transform infrared spectroscopy: second derivative spectra. Biochem Biophys Res Commun. 1983 Aug 30;115(1):391–397. [PubMed]
  • Wharton CW. Infra-red and Raman spectroscopic studies of enzyme structure and function. Biochem J. 1986 Jan 1;233(1):25–36. [PMC free article] [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...