X-Message-Number: 24297
Date: Sat, 26 Jun 2004 08:43:51 -0700 (PDT)
From: Doug Skrecky <>
Subject: Glycine betaine as a cryoprotectant

J Microbiol Methods. 2004 Jul;58(1):31-8.
Glycine betaine as a cryoprotectant for prokaryotes.
Osmoprotectants are low molecular weight, hydrophilic, nontoxic molecules
that assist a cell under osmotic stress to stabilize its concentration of
internal solutes. These properties are similar to compounds used as
cryoprotectants for the preservation of prokaryotic cells during
freezing. This study tested the ability of a common compatible solute,
glycine betaine (GB), to act as a cryoprotectant. In a series of
freeze-drying studies using a variety of prokaryotes, GB performed as
well, or better than,two commonly used cryoprotectants, sucrose/bovine
serum albumin (S/BSA) and trehalose/dextran (T/D). GB did especially well
maintaining cell viability after long-term storage (simulated equivalent
of 20 years) for microorganisms like Neisseria gonorrhoeae and
Streptococcus pneumoniae. GB was tested for its ability to preserve
members of the genus Acidothiobacillus, a difficult genus to preserve. For
two strains of Acidithiobacillus ferrooxidans that were preserved using
liquid drying, GB performed as well as S/BSA. Results were more mixed for
two strains of Acidithiobacillus thiooxidans; one strain could be
preserved with S/BSA but not GB, the other strain gave low recoveries with
both cryoprotectants. GB also proved to be a useful cryoprotectant for
liquid nitrogen preservation yielding equivalent results to the
cryopreservative, glycerol for halophilic archaea, and neutrophilic
Fe-oxidizing bacteria. These results indicate that GB is a simple and
useful cryoprotectant that works for a wide range of prokaryotic organisms
under different cryopreservation regimens.

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