Trehalose 6-phosphate

In some of our own work, we have developed formulations to lyophilize and preserve bacteria for extended periods of time at relatively high temperatures (Conrad et al., 2000 Ekdawi-Sever and de Pablo, 2003). Some of our data are shovm in Figure 9.2, where it can be seen that freeze-dried samples of Lactobacillus acidophilus in trehalose-phosphate mixtures retain more than 60% of their viability after 3 months of storage at relatively high temperatures. In contrast, commercial processes and formulations lead to complete loss of viability after a few days of storage. 

Viability of Lactobacillus acidophilus during storage at 37°C as a function of storage time. The cells were lyophilized in a solution of trehalose and phosphate ions. The symbols show results for cells lyophilized without additives (inverted triangles), with trehalose only (diamonds), with a trehalose-borate mixture (bullets), and with a trehalose-phosphate mixture (triangles). 

Analysis of the enzymically synthesized product by chemical and enzymic methods, as well as by optical rotation, showed that it was identical with the trehalose phosphate isolated from the fermentation products of dry yeast. ... 

Trehalose phosphate is synthesized from G-6-P and UDPG by trehalose phosphate synthetase and converted to trehalose by a phosphatase. 

According to the conditions, hexose monophosphate and trehalose phosphate also accumulate in varying amounts. Separation of the fructose diphosphate from the monophosphate can be effected by precipitating the diphosphate with neutral lead acetate or barium ions, followed by precipitation of the monophosphate from the supernatant liquid with basic lead acetate or barium ions in 50% or stronger ethanol (199). The separation of the different monophosphates was carried out by fractional crystallization (200). Chromatographic methods have also been used (Chapter XI). 

Some salts of sugar phosphates may be obtained in cr) talline form, as in the case of barium glucose 6-phosphate and barium trehalose phosphate. Salts of organic bases usually tend to crystallize more easily. Those which have been found more useful are cyclohexylamine, brucine, strychnine and benzylamine. 

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