Oxidative dehalogenation and mineralization of polychlorinated biphenyls by a resuscitated strain Streptococcus sp. SPC0.

Most functional microorganisms cannot be cultivated due to entering a viable but non-culturable (VBNC) state, which limits the characterization and application of polychlorinated biphenyl (PCB)-degrading strains. Resuscitating VBNC bacteria could provide huge candidates for obtaining high-efficient PCB degraders. However, limited studies have focused on the ability of resuscitated strains for PCBs degradation. In the present study, whole-genome analysis of a resuscitated strain SPC0, and its performances in degradation of three prevalent PCB congeners (PCBs 18, 52 and 77) were investigated. The results indicate that the strain SPC0 belonged to the genus Streptococcus, possessed the degradation potential for aromatic xenobiotics. The SPC0 could effectively degrade PCBs 18 and 52, but exhibited lower degradation efficiency of PCB 77. Degradation of PCBs 18 and 52 could be fitted well by zero-order model, whereas the fittest model for PCB 77 degradation was pseudo second-order kinetics. The bph genes expression, chloride ions release and degradation metabolites identification, suggest that SPC0 possessed the capability of oxidative dehalogenation and mineralization of PCBs. Interestingly, SPC0 can degrade PCBs via the bph-encoded biphenyl pathway, and further mineralize metabolite dichlorobenzoate via protocatechuate pathway. This study is the first to show that a strain belonging to genus Streptococcus possessed PCB-degrading capability, which uncovered the powerful potential of resuscitated strains for bioremediation of PCB-contaminated sites.

Enhancement of polychlorinated biphenyl biodegradation by resuscitation promoting factor (Rpf) and Rpf-responsive bacterial community.

The activities of indigenous bacterial communities in polychlorinated biphenyls (PCBs) contaminated environments is closely related to the efficiency of bioremediation processes. Using resuscitation promoting factor (Rpf) from Micrococcus luteus is a promising method for resuscitation and stimulation of functional bacterial populations under stressful conditions. This study aims to use the Rpf to accelerate the biodegradation of Aroclor 1242, and explore putative PCB degraders which were resuscitated by Rpf addition. The Rpf-responsive bacterial populations were investigated using culture-dependent and culture-independent approaches, respectively. The results confirm that Rpf was capable of enhancing PCB degradation of enriched cultures from PCB-contaminated soils, and improving the activities of cultures with low tolerance to PCBs. High-throughput 16S rRNA analysis displays that the Rpf greatly altered the composition and abundance of bacterial populations in the phylum Proteobacteria. Identification of the resuscitated strains further suggests that the Rpf-responsive population was mostly represented by Sphingomonas and Pseudomonas, which are most likely the key PCB-degraders for enhanced biodegradation of PCBs.

Aerobic degradation of 3,3',4,4'-tetrachlorobiphenyl by a resuscitated strain Castellaniella sp. SPC4: Kinetics model and pathway for biodegradation.

Resuscitated strains which were obtained by addition of resuscitation promoting factor (Rpf) could provide a vast majority of microbial source for obtaining highly efficient polychlorinated biphenyl (PCB)-degrading bacteria. In this study, the Castellaniella sp. strain SPC4 which was resuscitated by Rpf addition showed the highest efficiency in degradation of 3,3',4,4'-tetrachlorobiphenyl (PCB 77) among the resuscitated and non-resuscitated isolates. Further investigations on the PCB 77 degradation capability of the resuscitated strain SPC4 showed that SPC4 could efficiently degrade PCB 77 with maximum degradation rate (qmax) of 0.066/h at about 20 mg/L of PCB 77. The maximum growth rate on PCB 77 was 2.663 × 107 CFU/(mL·h) (0.024/h). The most suitable model of Edward demonstrated that the SPC4 could achieve qmax of 0.9315/h, with substrate-affinity of 11.33 mg/L and substrate-inhibition constants of 11.41 mg/L. Meanwhile, the presence of bphA gene expression and chlorine ions release, together with the identification of metabolites, confirmed that the bph-encoded biphenyl pathway was involved in PCB 77 mineralization by SPC4. This report is the first to demonstrate aerobic degradation of PCB 77 by the resuscitated strain Castellaniella sp. SPC4, indicating excellent potential for PCB bioremediation.