Heterologous expression of AHL lactonase AiiK by Lactobacillus casei MCJ[DELA]1 with great quorum quenching ability against Aeromonas hydrophila AH-1 and AH-4

Nowadays, microbial infections have caused increasing economic losses in aquaculture industry and deteriorated worldwide environments. Many of these infections are caused by opportunistic pathogens through cell-density mediated quorum sensing (QS). The disruption of QS, known as quorum quenching (QQ), is an effective and promising way to prevent and control pathogens, driving it be the potential bio-control agents. In our previous studies, AHL lactonase AiiK was identified with many characteristics, and constitutive expression vector pELX1 was constructed to express heterologous proteins in Lactobacillus casei MCJ[DELA]1 (L. casei MCJ[DELA]1). In this study, recombinant strain pELCW-aiiK/L. casei MCJ[DELA]1 (LcAiiK) and wild-type Aeromonas hydrophila (A. hydrophila) were co-cultured to test the QQ ability of LcAiiK against A. hydrophila. A cell wall-associated expression vector pELCW for L. casei MCJ[DELA]1 was constructed. Localization assays revealed that the expressed AiiK was anchored at the surface layer of LcAiiK via vector pELCW-aiiK. LcAiiK (OD.sub.600 = 0.5) degraded 24.13 [mu]M of C.sub.6-HSL at 2 h, 40.99 [mu]M of C.sub.6-HSL at 12 h, and 46.63 [mu]M of C.sub.6-HSL at 24 h. Over 50% LcAiiK cells maintained the pELCW-aiiK plasmid after 15 generations of cultivation without erythromycin. Furthermore, LcAiiK inhibited the swimming motility, extracellular proteolytic activity, haemolytic activity and biofilm formation of A. hydrophila AH-1 and AH-4. The AHL lactonase AiiK is firstly and constitutively expressed at the surface layer of L. casei MCJ[DELA]1. LcAiiK displayed considerable AHL lactonase activity and great QQ abilities against A. hydrophila AH-1 and AH-4 by attenuating their QS processes instead of killing them. Therefore, the LcAiiK can be exploited as an anti-pathogenic drug or a bio-control agent to control the AHL-mediated QS of pathogenic bacteria.