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Purification and Characterization of Intracellular Chromate Reductase from Pseudomonas putida Isolated from Soil | ||
| Al-Mustansiriyah Journal of Science | ||
| Article 1, Volume 19, Issue 2, 2008, Pages 1-14 | ||
| Author | ||
| Sahira N. Muslim | ||
| Abstract | ||
| Pseudomonas putida isolates were obtained at 12(60%) isolates out of 20 polluted soil samples from different locations in AL-Dora strainer. The isolate that gave higher reduction activity was chosen to purify chromate reductase enzyme through four stages of purification including(Ultracentrifugation, ammonium sulfate precipitation, ion exchange chromatography by DEAE- sepharose Cl-6B and gel filtration chromatography by sephadex G-75) with 77.2-fold purification and 45.8% recovery. The purified chromate reductase was characterized: the molecular weight was about 49000 daltons. The temperature and pH optima for chromate reductase were 80°C and 5, respectively. The reductase activity remained virtually unaltered upto 30 min of exposure to 50°C; even exposure to 80°C did not immediately inactivate the enzyme. Cationic metal Cd2+ had inhibitory effect on reduction activity more than Cu2+ and Zn2+, while the other metals such as Ni2+,Mg2+,Ca2+ and Mn2+ had no effect on the activity. Therefore, this research leads to increase benefit in using chromate reductase as an important strategy for bioremediation and detoxification of chromate as environmental pollutant. | ||
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