snr-1 Gene Is Required for Nitrate Reduction in Pseudomonas aeruginosa PAO1

ABSTRACT Pseudomonas aeruginosa is able to use nitrate for both assimilation and anaerobic respiration. One set of genes, designatedsnr (for “shared nitrate reduction”), have been recently cloned and partially characterized. In this study, we demonstrate that the snr-1 gene encodes a predicted 52.5-kDa protein that is 82% similar to a unique cytochromec of Desulfomonile tiedjei DCB-1. Importantly, the Snr-1 protein sequence of P. aeruginosa differed from that of the cytochrome c of D. tiedjeiprimarily in the first 25 amino acids, which are required for membrane attachment in D. tiedjei. In P. aeruginosa, the Snr-1 protein hydropathy profile indicates that it is a soluble protein. An isogenic snr-1::Gm insertional mutant was unable to grow aerobically with nitrate as a sole nitrogen source or anaerobically with nitrate as an electron acceptor. Complementation of the snr-1::Gm mutant with thesnr-1 gene restored the wild-type phenotypes. Interestingly, anaerobic growth rates were significantly higher in thesnr-1 mutant harboring a multicopy plasmid containingsnr-1. In contrast, aerobic growth rates of the restored mutant using nitrate as the sole nitrogen source were similar to those of the wild type. Transcriptional lacZ fusions demonstrated that snr-1 was not regulated by molybdate, oxygen, or nitrate.

Adhesion of Biodegradative Anaerobic Bacteria to Solid Surfaces

ABSTRACT In order to exploit the ability of anaerobic bacteria to degrade certain contaminants for bioremediation of polluted subsurface environments, we need to understand the mechanisms by which such bacteria partition between aqueous and solid phases, as well as the environmental conditions that influence partitioning. We studied four strictly anaerobic bacteria, Desulfomonile tiedjei,Syntrophomonas wolfei, Syntrophobacter wolinii, and Desulfovibrio sp. strain G11, which theoretically together can constitute a tetrachloroethylene- and trichloroethylene-dechlorinating consortium. Adhesion of these organisms was evaluated by microscopic determination of the numbers of cells that attached to glass coverslips exposed to cell suspensions under anaerobic conditions. We studied the effects of the growth phase of the organisms on adhesion, as well as the influence of electrostatic and hydrophobic properties of the substratum. Results indicate thatS. wolfei adheres in considerably higher numbers to glass surfaces than the other three organisms. Starvation greatly decreases adhesion of S. wolfei and Desulfovibrio sp. strain G11 but seems to have less of an effect on the adhesion of the other bacteria. The presence of Fe3+ on the substratum, which would be electropositive, significantly increased the adhesion ofS. wolfei, whereas the presence of silicon hydrophobic groups decreased the numbers of attached cells of all species. Measurements of transport of cells through hydrophobic-interaction and electrostatic-interaction columns indicated that all four species had negatively charged cell surfaces and that D. tiedjei andDesulfovibrio sp. strain G11 possessed some hydrophobic cell surface properties. These findings are an early step toward understanding the dynamic attachment of anaerobic bacteria in anoxic environments.

Degradation of chlorinated aromatic compounds in UASB reactors

Data on anaerobic degradation of chloroaromatic compounds in Upflow Anaerobic Sludge Blanket Reactors (UASB-reactor) are presented and compared. Special attention is given to the metabolic pathways for degradation of chlorinated phenols by granular sludge. Results indicate that PCP can be degraded in UASB-reactors via stepwise dechlorination to phenol. Phenol will subsequently be converted to benzoate before ring cleavage. Dechlorination proceeds via different pathways dependent upon the inocula used. Results are further presented on the design of special metabolic pathways in granules which do not possess this activity using the dechlorinating organism, Desulfomonile tiedjei. Additionally, it is shown that it is possible to immobilize Dechlorosporium hafniense, a newly isolated dechlorinating anaerobe, into granular sludge, thereby introducing an ability not previously present in the granules.