Production of ectoine by Vreelandella boliviensis using non-aseptic repeated-batch and continuous cultivations in an air-lift bioreactor

Ectoine, an osmolyte produced by various microorganisms, has numerous commercial applications. Vreelandella boliviensis (formerly called Halomonas boliviensis) generates high ectoine concentrations, i.e., 78.6 g/L. This study investigated three cultivation strategies for ectoine production in a non-aseptic air-lift bioreactor. The first strategy was performed in a repeated-batch mode with 5% (w/v) NaCl to induce cell growth, followed by the addition of solid NaCl to a final concentration of 12.5% (w/v) to prompt ectoine production. A maximum dry cell weight of 13.8 g/L at 46.5 h, a maximum ectoine concentration of 1.37 g/L at 37.5 h, and a maximum volumetric productivity of 0.93 g/L/d at 34.5 h were reached. The second strategy employed a three-step repeated-batch cultivation method. In the first step, cells were grown at the optimum salt concentration, harvested by centrifugation, and cultivated in a replenished medium for the second step. In the third step, the cells were harvested again and grown in a fresh medium containing 12.5% (w/v) NaCl. This strategy improved dry cell weight to 32 g/L, ectoine concentration to 4.37 g/L, and productivity to 1.76 g/L/day at 60 h of cultivation. The third strategy consisted of continuous cultivations that were investigated using different NaCl concentrations. The highest ectoine concentration of 2.83 g/L and productivity of 3.49 g/L/d were obtained with 8.5% (w/v) NaCl at a dilution rate of 0.05 (1/h). This study is the first to report ectoine production by V. boliviensis in continuous air-lift bioreactors under non-aseptic conditions.