Draft Genome Sequencing of Three Glutaraldehyde-Tolerant Bacteria from Produced Water from Hydraulic Fracturing.

Here, we report the draft genome sequence of three glutaraldehyde-resistant isolates from produced water from hydraulic fracturing operations. The three strains were identified as Marinobacter sp. strain G11, Halomonas sp. strain G15, and Bacillus sp. strain G16. The genome sequences of these isolates will provide insights into biocide resistance in hydraulic fracturing operations.

Chronic effects of lead exposure on topsmelt fish (Atherinops affinis): Influence of salinity, organism age, and relative sensitivity to other marine species.

The aim of the present study was to determine the influence of salinity and organism age on the chronic toxicity of waterborne lead (Pb) to Atherinops affinis and to compare the relative Pb sensitivity of A. affinis with other marine species. Chronic Pb exposure experiments were conducted in a water flow-through testing system. Survival, standard length, dry weight, and tissue Pb concentration were measured and lethal concentrations (LCs), effect concentrations (ECs), and bioconcentration factors (BCFs) were calculated. In general, increasing salinity and organism age decreased Pb toxicity. The LC50s for larval fish at 14 and 28 ppt salinity were 15.1 and 79.8 µg/L dissolved Pb, respectively; whereas, the LC50 for juvenile fish was 167.6 µg/L dissolved Pb at 28 ppt salinity. Using standard length data, the EC10 values for larval fish were 16.4 and 82.4 µg/L dissolved Pb at 14 and 28 ppt salinity, respectively. The dry weight EC25s for low and high salinity were 15.6 and 61.84 µg/L dissolved Pb, respectively. The BCFs were higher with the lower salinity study (1703) in comparison to the higher salinity study (654). Results of Pb speciation calculation showed higher fraction of Pb2+ in water with lower salinity, explaining the higher observed toxicity of Pb in lower salinity water than higher salinity water. Atherinops affinis is more sensitive to Pb than several other marine species. Evidence of abnormal swimming and skeletal deformities were observed in Pb exposure treatments. Results of the present study are useful for marine biotic ligand modeling and support ecological risk assessment and deriving Pb environmental quality criteria for marine environments. Environ Toxicol Chem 2018;37:2705-2713. © 2018 SETAC.

Silica gel-encapsulated AtzA biocatalyst for atrazine biodegradation.

Encapsulation of recombinant Escherichia coli cells expressing a biocatalyst has the potential to produce stable, long-lasting enzyme activity that can be used for numerous applications. The current study describes the use of this technology with recombinant E. coli cells expressing the atrazine-dechlorinating enzyme AtzA in a silica/polymer porous gel. This novel recombinant enzyme-based method utilizes both adsorption and degradation to remove atrazine from water. A combination of silica nanoparticles (Ludox TM40), alkoxides, and an organic polymer was used to synthesize a porous gel. Gel curing temperatures of 23 or 45 °C were used either to maintain cell viability or to render the cells non-viable, respectively. The enzymatic activity of the encapsulated viable and non-viable cells was high and extremely stable over the time period analyzed. At room temperature, the encapsulated non-viable cells maintained a specific activity between (0.44 ± 0.06) µmol/g/min and (0.66 ± 0.12) µmol/g/min for up to 4 months, comparing well with free, viable cell-specific activities (0.61 ± 0.04 µmol/g/min). Gels cured at 45 °C had excellent structural rigidity and contained few viable cells, making these gels potentially compatible with water treatment facility applications. When encapsulated, non-viable cells were assayed at 4 °C, the activity increased threefold over free cells, potentially due to differences in lipid membranes as shown by FTIR spectroscopy and electron microscopy.

X-ray structure and mutational analysis of the atrazine Chlorohydrolase TrzN.

Atrazine chlorohydrolase, TrzN (triazine hydrolase or atrazine chlorohydrolase 2), initiates bacterial metabolism of the herbicide atrazine by hydrolytic displacement of a chlorine substituent from the s-triazine ring. The present study describes crystal structures and reactivity of wild-type and active site mutant TrzN enzymes. The homodimer native enzyme structure, solved to 1.40 Å resolution, is a (ßα)(8) barrel, characteristic of members of the amidohydrolase superfamily. TrzN uniquely positions threonine 325 in place of a conserved aspartate that ligates the metal in most mononuclear amidohydrolases superfamily members. The threonine side chain oxygen atom is 3.3 Å from the zinc atom and 2.6 Å from the oxygen atom of zinc-coordinated water. Mutation of the threonine to a serine resulted in a 12-fold decrease in k(cat)/K(m), largely due to k(cat), whereas the T325D and T325E mutants had immeasurable activity. The structure and kinetics of TrzN are reminiscent of carbonic anhydrase, which uses a threonine to assist in positioning water for reaction with carbon dioxide. An isosteric substitution in the active site glutamate, E241Q, showed a large diminution in activity with ametryn, no detectable activity with atratone, and a 10-fold decrease with atrazine, when compared with wild-type TrzN. Activity with the E241Q mutant was nearly constant from pH 6.0 to 10.0, consistent with the loss of a proton-donating group. Structures for TrzN-E241Q were solved with bound ametryn and atratone to 1.93 and 1.64 Å resolution, respectively. Both structure and kinetic determinations suggest that the Glu(241) side chain provides a proton to N-1 of the s-triazine substrate to facilitate nucleophilic displacement at the adjacent C-2.