Abstract
Sixty five soil samples and fifteen water samples were collected from different places in which previously explosions were occurred in Iraq. Seven isolates showed ability to utilize 0.1mM trinitrotoluene (TNT) and/or 0.2mM glycerol trinitrate (GTN) as a sole carbon and nitrogen source and one of these isolates showed the highest nitrate reduction which was classified and coded as Pseudomonas sp. SH7. The highest nitrate reductase activity extracted by sonication while optimum conditions for enzyme production in minimal media pH 7 containing 0.25mM GTN at 35oC for 3 days under aerobic condition. Nitrate reductase was purified by 40-60% ammonium sulphate, ion exchange and gel filtration. Nitrate reductase molecular weight determined by SDS-PAGE was 115 kD. The characterization of purified enzyme activity and stability was higher at a pH between 6.5-7.5 and. Maximum activity was at 35oC and stable at 30-40oC for 15 min., while for heat sensitivity 100% activity observed at 45oC for 20 min. Treatment with 200 µM azide and 500 µM cyanide inhibited the activity by 76 and 91% respectively.
Highlights
Life on this planet is based on the continuous cycling of elements
Nitrate reductases can be performed with three different types: the utilization of nitrate as a nitrogen source for growth, the generation of metabolic energy by using nitrate as a terminal electron acceptor, and the dissipation of excess reducing power for redox balancing
Protein determinations: Protein was performed by Bradford, [6] with bovine serum albumin used as standard Optimal conditions for nitrate reductase production: Several factors were studied to determine the optimal conditions for nitrate reductase production from Pseudomonas sp
Summary
In the recent years the massive mobilization of natural resources and the industrial synthesis of chemicals have generated a number of environmental problems as a consequence of the limited incorporation of natural and synthesized molecules into ongoing biological cycles. This is true for xenobiotic compounds, which exhibit structural elements or substituents that are rarely found in natural products. A study focusing on the fluorescent Pseudomonads community showed that 56% of the isolated strains had nar gen only, 51% had the nap gene and 15.5% possessed both nitrate reductase genes [3].
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