Activation of N-acyl-homoserine lactone-mediated quorum sensing system improves long-term preservation of anammox microorganisms by vacuum lyophilization.

The long-term preservation of anaerobic ammonium oxidation (anammox) microorganisms via vacuum lyophilization process would help commercialize the technique. In this study, vacuum lyophilization was evaluated for the cost-effective long-term preservation of such microorganisms. Skim milk was found to be the most effective cryoprotectant for maintaining the physiological properties (heme c, EPS, and the PN/PS ratio) of anammox microorganisms. Conversely, the vacuum lyophilization technique was shown to cause serious damage to the quorum sensing (QS) system of anammox, so that anammox activity was not adequately recovered afterwards. To overcome this limitation, activation of the AHL-mediated QS system were applied to the vacuum lyophilization process. Endogenous (i.e., fresh anammox sludge of 10%) and exogenous (i.e., C6-HSL of 60 mg/L) QS autoinducers significantly increased anammox activity to 88.2 ± 12.2 and 130.0 ± 12.2 mgTN/gVSS/d, respectively, after 56 d of reactivation. In addition, nitrogen removal potentials were estimated to be 123.5 and 87.5 gTN/m/d, respectively. The effect of the exogenous QS autoinducer on anammox reactivation was reconfirmed through the comparison experiment. The results of this study will be greatly significant to this field since they improve the feasibility of the once-underestimated vacuum lyophilization technique.