Microbial community shifts as a response to efficient degradation of chlorobenzene under hypoxic conditions

Limitations in the availability of oxygen restrict aerobic biodegradation of chloroaromatic compounds in groundwater ecosystems. In this context the activity of ring-cleaving chlorocatechol dioxygenases (CC12O) is crucial for effective mineralization. Previously we demonstrated that oxygen-related enzyme characteristics of CC12O can vary widely among the Proteobacteria (Balcke et al. submitted). Here, we investigated how strains with different ability to transform intermediary 3-chlorocatechol integrate into biodegradation of chlorobenzene (CB) under low or high oxygen availability. Pseudomonas veronii UFZ B549 and Acidovorax facilis UFZ B530, which had differing oxygen affinities for CC12O, were mixed together at different proportions (20:80; 80:20), and compared for degradation of chlorobenzene under oxic (215 mu M O2) and hypoxic (11 mu M O2) conditions. Changes in community composition in binary mixed cultures were determined and compared with an indigenous groundwater community, cultivated under comparable conditions. Community shifts were determined by FISH (fluorescent in situ hybridization) in our model system and SSCP (single stranded conformation polymorphism) fingerprinting in the groundwater community, as well as by analysis of respiratory quinones of taxonomic value. Hypoxia led to enrichment of Acidovoracae in the groundwater and binary cultures. Under hypoxic conditions cis,cis-2-chloromuconate released to the medium by A. facilis allowed for concomitant growth of P. veronii, although its low-affinity type CC12O would not imply growth on CB. Vice versa, increasing abundance of P. veronii induced intermediary 3-chlorocatechol accumulation, which was reduced by growth of A. facilis. Thus, reduced oxygen availability caused syntrophic rather than competitive interactions.