Storage of oil field-produced waters alters their chemical and microbiological characteristics.

Many oil fields are in remote locations, and the time required for shipment of produced water samples for microbiological examination may be lengthy. No studies have reported on how storage of oil field waters can change their characteristics. Produced water samples from three Alberta oil fields were collected in sterile, industry-approved 4-l epoxy-lined steel cans, sealed with minimal headspace and stored under anoxic conditions for 14 days at either 4 degrees C or room temperature (ca. 21 degrees C). Storage resulted in significant changes in water chemistry, microbial number estimates and/or community response to amendment with nitrate. During room-temperature storage, activity and growth of sulfate-reducing bacteria (and, to a lesser extent, fermenters and methanogens) in the samples led to significant changes in sulfide, acetate and propionate concentrations as well as a significant increase in most probable number estimates, particularly of sulfate-reducing bacteria. Sulfide production during room-temperature storage was likely to be responsible for the altered response to nitrate amendment observed in microcosms containing sulfidogenic samples. Refrigerated storage suppressed sulfate reduction and growth of sulfate-reducing bacteria. However, declines in sulfide concentrations were observed in two of the three samples stored at 4 degrees C, suggesting abiotic losses of sulfide. In one of the samples stored at room temperature, nitrate amendment led to ammonification. These results demonstrate that storage of oil field water samples for 14 days, such as might occur because of lengthy transport times or delays before analysis in the laboratory, can affect microbial numbers and activity as well as water sample chemistry.

Sulfide persistence in oil field waters amended with nitrate and acetate.

Nitrate amendment is normally an effective method for sulfide control in oil field-produced waters. However, this approach has occasionally failed to prevent sulfide accumulation, despite the presence of active nitrate-reducing bacterial populations. Here, we report our study of bulk chemical transformations in microcosms of oil field waters containing nitrate-reducing, sulfide-oxidizing bacteria, but lacking denitrifying heterotrophs. Amendment with combinations of nitrate, acetate, and phosphate altered the microbial sulfur and nitrogen transformations. Elemental sulfur produced by chemotrophic nitrate-reducing bacteria was re-reduced heterotrophically to sulfide. Ammonification, rather than denitrification, was the predominant pathway for nitrate reduction. The application of nitrite led to transient sulfide depletion, possibly due to higher rates of nitrite reduction. The addition of molybdate suppressed both the accumulation of sulfide and the heterotrophic reduction of nitrate. Therefore, sulfidogenesis was likely due to elemental sulfur-reducing heterotrophic bacteria, and the nitrate-reducing microbial community consisted mainly of facultatively chemotrophic microbes. This study describes one set of conditions for continued sulfidogenesis during nitrate reduction, with important implications for nitrate control of sulfide production in oil fields.

X-ray crystal structure of Mycobacterium tuberculosis haloalkane dehalogenase Rv2579.

Haloalkane dehalogenases are enzymes well known to be important in bioremediation; the organisms from which they are produced are able to clean up toxic organohalides from polluted environments. However, besides being found in such contaminated environments, these enzymes have also been found in root or tissue-colonizing bacterial species. The haloalkane dehalogenase Rv2579 from Mycobacterium tuberculosis H37Rv has been cloned, expressed, purified and its crystal structure determined at high resolution (1.2A). In addition, the crystal structure of the enzyme has been determined in complex with the product from the reaction with 1,3-dibromopropane, i.e. 1,3-propanediol and in complex with the classical substrate of haloalkane dehalogenases, 1,2-dichloroethane. The enzyme is a two-domain protein having a catalytic domain of an alpha/beta hydrolase fold and a cap domain. The active site residues and the halide-stabilizing residues have been identified as Asp109, Glu133, His273, Asn39 and Trp110. Its overall structure is similar to those of other known haloalkane dehalogenases. Its mechanism of action involves an SN2 nucleophilic displacement.