Accumulation of Ectoines By Halophilic Bacteria Isolated from Fermented Shrimp Paste: An Adaptation Mechanism to Salinity, Temperature, and pH Stress

Shrimp paste is a traditional fermented food produced by many Asian countries. Bacteria play important roles in the shrimp paste fermentation process. In order to survive under the low water activity (A(w)) conditions caused by the high salt concentration, the bacteria need to employ a special adaptation strategy. This study found that most halophilic bacteria isolated from shrimp paste accumulated ectoines (ectoine and hydroxyectoine) as protective osmotic agents. Five isolated bacteria, including three high ectoine producers and two high hydroxyectoine producers, were selected for further study. Based on their morphological and biochemical characteristics and 16S rRNA gene sequences, the five strains were classified into three genera: Salinivibrio (strains M7 and M316), Salimicrobium (strains M31 and M69), and Vibrio (strain M92). The accumulation of ectoines by Salimicrobium species is reported here for the first time. The effects of salinity, incubation temperature, and initial pH on the growth rate and accumulation of ectoines by the five strains were investigated. The results revealed that the bacterial growth rate was inhibited while the accumulation of ectoines by the five selected strains was triggered by an increase in the external salinity, incubation temperature, or initial pH. In addition, a high concentration of ectoine only (21.2 wt%) was produced by strain M316 at the optimum salinity and temperature, and under pressure of a high initial pH value. To the best of our knowledge, this is the first report demonstrating that the production of ectoines by bacterial strains can be enhanced by increasing the pH of the culture medium to induce pH stress. This finding suggests a new ectoine producer and fermentation strategy that may help to improve the production of ectoines in the future.