Stabilization and Preservation of Lactobacillus acidophilus in Saccharide Matrices

• Published in: Cryobiology 2000-08, Vol.41 (1), p.17-24
• Main Authors: Conrad, Paul B., Miller, Danforth P., Cielenski, Peter R., de Pablo, Juan J.
• Format: Article
• Language: English
• Subjects: Action of physical and chemical agents on bacteria; Ammonium Hydroxide; Bacteriology; Biological and medical sciences; borate; Borates - pharmacology; Buffers; Citric Acid - pharmacology; Cryopreservation; Cryoprotective Agents - pharmacology; Desiccation - methods; Freeze Drying; Fundamental and applied biological sciences. Psychology; glass transition; Hydrogen-Ion Concentration; Hydroxides - pharmacology; Lactic Acid - pharmacology; lactic acid bacteria; Lactobacillus acidophilus; Lactobacillus acidophilus - drug effects; Lactobacillus acidophilus - growth & development; Lactobacillus acidophilus - metabolism; lyophilization; Microbiology; Solutions; trehalose; Trehalose - pharmacology; Vacuum
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1006/cryo.2000.2260

Lyophilization and vacuum- or spray-drying are some of the most useful techniques for preserving foods, agricultural products, and pharmaceuticals. Biological materials, however, can be irreversibly damaged during these treatments. Therefore, it is essential to design protective agents to preserve protein activity and cell viability. In this paper we examine the use of α,α-trehalose–borate systems as protectants for Lactobacillus acidophilus during freeze- and vacuum-drying. Trehalose was found to be an effective protectant for freeze-dried and vacuum-dried samples, and it is equivalent to a protective formulation which is in current industrial use. It is known from our previous work on enzymes that the presence of borate can dramatically enhance the protective ability of trehalose. In this work, the addition of trehalose–borate to bacterial concentrate greatly improves the recovery of viable cells after storage. This improvement was seen in freeze-dried samples stored at 37°C as well as for vacuum-dried samples held at room temperature. A tailored buffering strategy was tested to counteract the high pH resulting from the addition of borate to the mixture. Use of citric or lactic acids in combination with ammonium hydroxide gave a protectant solution with high pH (resulting in effective crosslinking between trehalose and borate) but a dry product with reduced pH upon rehydration (conducive to cell survival). These results raise exciting possibilities for protection of more labile prokaryotic species as well as simple eukaryotes.