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Campylobacter jejuni  is a common gastrointestinal pathogen that colonizes food animals; it is transmitted via fecal contamination of food, and infections in immune-compromised people are more likely to result in serious long-term illness. Environmental phosphate is likely an important sensor of environmental fitness and the ability to obtain extracellular phosphate is central to the bacteria's core metabolic responses. PhoX is the sole alkaline phosphatase in  C. jejuni , a substrate of the TAT transport system. Alkaline phosphatases mediate the hydrolytic removal of inorganic phosphate (Pi) from phospho-organic compounds and thereby contribute significantly to the polyphosphate kinase 1 ( ppk1 ) mediated formation of poly P, a molecule that regulates bacterial response to stresses and virulence. Similarly, deletion of the  tatC  gene, a key component of the TAT system, results in diverse phenotypes in  C. jejuni  including reduced stress tolerance and  in vivo  colonization. Therefore, here we investigated the contribution of  phoX  in poly P synthesis and in TAT-system mediated responses. The  phoX  deletion mutant showed significant decrease (P<0.05) in poly P accumulation in stationary phase compared to the wild-type, suggesting that PhoX is a major contributor to the inorganic phosphate pool in the cell which is essential for poly P synthesis. The  phoX  deletion is sufficient for a nutrient stress defect similar to the defect previously described for the Δ tatC  mutant. Additionally, the  phoX  deletion mutant has increased resistance to certain antimicrobials. The Δ phoX  mutant was also moderately defective in invasion and intracellular survival within human intestinal epithelial cells as well as in chicken colonization. Further, the Δ phoX  mutant produced increased biofilm that can be rescued with 1 mM inorganic phosphate. The qRT-PCR of the Δ phoX  mutant revealed transcriptional changes that suggest potential mechanisms for the increased biofilm phenotype.

Contribution of TAT System Translocated PhoX to Campylobacter jejuni Phosphate Metabolism and Resilience to Environmental Stresses