Extracellular Secretion Of Glycolytic Housekeeping Enzyme In Edwardsiella Tarda

Glycolytic enzymes is housekeeping enzymes of organism and they usually play a physiological and biochemical role in the cytoplasm.Research has shown that glycolytic enzymes are secreted to extracellular region playing a function except glycolysis in some bacteria.Edwardsiella tarda is a wide host bacteria that it can infect a variety of freshwater and seawater fishes.This study were focus on secretion of the related glycolytic housekeeping enzymes in E.tarda.Using western blot and ELISA,it was found that five glycolytic enzymes are widespread in the extracellular protein of 50 strains of Edwardsiella.The contents have a certain relationship with species though the various secretions of five enzymes in different strains are not consistent with the virulence of pathogens.Later,we proved that the secretions of these five glycolytic enzymes in E.tarda are not closely related to the secretion systems of T3SS,T4SS,T6SS and Tat.We also found that these five glycolytic enzymes exist in the periplasmic space of E.tarda EIB202,which suggested that these enzmyes are translocated from the cytoplasm to the extracellular region through a two-step way.Moreover,these five enzymes located on the surface of E.tarda EIB202 are demonstrated to has a role during the infection of the pathogen.Finally,based on the His-tag,GST-tag and MBP-tag pull downs and Flag-tag Co-IP,two proteins(90 and 19 kDa)were obtained which might have interactions with FBA.With further analysis,the possible secretion pathway of FBA will be elucidated.The aboved results showed that these different glycolytic enzymes in E.tarda has a correlation function of the infection.FBA interacting protein screening laid a good foundation for the moonlighting function researches and secretion mechanism exploration of these glycolytic enzymes.