Effect of culture parameters on the heat tolerance and inorganic polyphosphate accumulation by Lacticaseibacillus rhamnosus CRL1505, a multifunctional bacterium.

Lacticaseibacillus rhamnosus CRL1505 can be used in functional products as a probiotic powder (dried live cells) or as a postbiotic intracellular extract containing inorganic polyphosphate as a functional biopolymer. Thus, the aim of this work was to optimize the production of Lr-CRL1505 depending on the target of the functional product (probiotic or postbiotic). For this purpose, the effect of culture parameters (pH, growth phase) on cell viability, heat tolerance and polyphosphate accumulation by Lacticaseibacillus rhamnosus CRL1505 was evaluated. Fermentations at free pH produced less biomass (0.6 log units) than at controlled pH while the growth phase affected both polyphosphate accumulation and cell heat tolerance. Exponential phase cultures showed 4-15 times greater survival rate against heat shock and 49-62% increased polyphosphate level, compared with the stationary phase. Results obtained allowed setting the appropriate culture conditions for the production of this strain according to its potential application, i.e., as live probiotic cells in powder form or postbiotic. In the first case, running fermentations at pH 5.5 and harvesting the cells at the exponential phase are the best conditions for obtaining a high live biomass yield capable of overcoming heat stress. Whereas the postbiotic formulations production requires fermentations at free pH and harvesting the cells in exponential phase to increase the intracellular polyphosphate level as a first step.

PhoB activation in non-limiting phosphate condition by the maintenance of high polyphosphate levels in the stationary phase inhibits biofilm formation in Escherichia coli.

Polyphosphate (polyP) degradation in Escherichia coli stationary phase triggers biofilm formation via the LuxS quorum sensing system. In media containing excess of phosphate (Pi), high polyP levels are maintained in the stationary phase with the consequent inhibition of biofilm formation. The transcriptional-response regulator PhoB, which is activated under Pi limitation, is involved in the inhibition of biofilm formation in several bacterial species. In the current study, we report, for the first time, we believe that E. coli PhoB can be activated in non-limiting Pi conditions, leading to inhibition of biofilm formation. In fact, PhoB was activated when high polyP levels were maintained in the stationary phase, whereas it remained inactive when the polymer was degraded or absent. PhoB activation was mediated by acetyl phosphate with the consequent repression of biofilm formation owing to the downregulation of c-di-GMP synthesis and the inhibition of autoinducer-2 production. These results allowed us to propose a model showing that PhoB is a component in the signal cascade regulating biofilm formation triggered by fluctuations of polyP levels in E. coli cells during stationary phase.

Environmental phosphate differentially affects virulence phenotypes of uropathogenic Escherichia coli isolates causative of prostatitis.

K-12 Escherichia coli cells grown in static media containing a critical phosphate (Pi) concentration ≥25 mM maintained a high polyphosphate (polyP) level in stationary phase, impairing biofilm formation, a phenomenon that is triggered by polyP degradation. Pi concentration in human urine fluctuates according to health state. Here, the influence of environmental Pi concentration on the occurrence of virulence traits in uropathogenic E. coli (UPEC) isolated from acute prostatitis patients was evaluated. After a first screening, 3 isolates were selected according to differential biofilm formation profiles depending on media Pi concentration. For each isolate, biofilm positive and negative conditions were established. Regardless of the isolate, biofilm formation capacity was accompanied with curli and cellulose production and expression of some key virulence factors associated with adhesion. When the selected isolates were grown in their non-biofilm-forming condition, low concentrations of nalidixic acid and ciprofloxacin induced biofilm formation. Interestingly, similar to laboratory strains, polyP degradation induced biofilm formation in the selected isolates. Data demonstrated the complexity of UPEC responses to environmental Pi and the importance of polyP metabolism in the virulence of clinical isolates.