Transport kinetics of ectoine, an osmolyte produced by Brevibacterium epidermis.

Brevibacterium epidermis DSM 20659 is a halotolerant Gram-positive bacterium which can synthesize the osmolyte, ectoine, but prefers to take it up from its environment. The present study revealed that B. epidermis is equipped with at least one transport system for ectoine, with a maximal transport velocity of 15.7 +/- 4.3 nmol/g CDW.min. The transport requires energy (ATP) and is completely inhibited by the proton uncoupler, CCCP. The ectoine uptake system is constitutively expressed at a basal level of activity and its activity is immediately 10-fold increased by hyper-osmotic stress. Initial uptake rates are not influenced by the intensity of the hyper-osmotic shock but the duration of the increased activity of the uptake system could be directly related to the osmotic strength of the assay solution. Competition assays indicate that betaine, but not proline, is also transported by the ectoine uptake system.

Dynamics and optimal conditions of intracellular ectoine accumulation in Brevibacterium sp.

The optimal conditions for the intracellular synthesis of ectoine were determined in a halotolerant Brevibacterium sp. The size of the intracellular ectoine pool in the bacterial cells is shown to depend on the external salt concentrations, type of carbon source and aeration level. In erlenmeyer flasks a maximum concentration of intracellular ectoine of about 0.9 g/l was obtained. Under controlled aeration in a 1.5 l fermentor this level could be increased to 1.2 g/l. Consecutive cell transfers to media with increasingly higher salt concentrations enabled us to reach even higher levels, up to 1.6 g/l on erlenmeyer scale. The ectoine synthesis takes place immediately after the osmotic upshock. Within one generation time, the new corresponding specific intracellular ectoine concentration is reached.