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The effect of LuxR solo EvgA on the spoilage potential of Pseudomonas fluorescens

Dangfeng Wang,Benyou Zhang,Yunting Wang,Likun Ren,Yuqiong Meng,Rui Ma,Shulin Wang,Zhiteng Liu,Yang Jiang,Fangchao Cui,Tingting Li,Jianrong Li

LuxR solos sense exogenous signaling molecules and regulate microbial behaviors, thus facilitating intra- and inter-species communication among bacteria. However, studies on the role of LuxR solos in regulating the spoilage potential of specific spoilage organisms are limited. In this study, we screened a LuxR solo EvgA, derived from Pseudomonas fluorescens PF08. qRT-PCR and molecular docking results indicated that EvgA responds and binds most strongly to exogenous N -3-tetradecanoyl-L-homoserine lactone (C14-HSL) through hydrogen bonds and hydrophobic interactions, with a –CDOCKER ENERGY of 45.87 kcal/mol. Subsequently, the evgA knockout (Δ evgA ) and complementary evgA (Δ evgA -com) mutants of P. fluorescens PF08 were constructed through homologous recombination. Δ evgA mutants displayed diminished eavesdropping to exogenous C14-HSL (by 18.99 ± 2.09%), reduced adhesion ability (by 30.33 ± 0.36%), attenuated biofilm formation (by 15.97 ± 0.14%), and decreased motility (swarming zones by 4.34 ± 0.002 mm and swimming zones by 4.17 ± 0.11 mm). In addition, EvgA deletion reduced spoilage factor production (hydrolysis zone), which included extracellular proteases, lipases, and biogenic amines (by 4.45 ± 0.51 mm, 10.12 ± 0.10 mm, and 14.41 ± 0.57 mm, respectively), thereby diminishing the spoilage of salmon fillets stored at 4 °C. Notably, the addition of exogenous C14-HSL promoted the spoilage ability of all strains. This study emphasizes the important role of LuxR solo EvgA in AHLs-mediated quorum sensing and the spoilage potential of P. fluorescens PF08, which is expected to be a new target for controlling the spoilage of aquatic products.