• Journal of Plant Biotechnology
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• v.3 no.1
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• pp.13-17
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• 2001

A Chromium (VI)[Cr(VI)] reductase gene from heavy metal reducing bacteria Pseudomonas aeruginosa HP014 was used to transform tobacco plant cells. A chimeric construct containing the Cr(VI) reductase gene was transfered to tobacco leaf disks using an Agrobacteriun tumefaciens binary vector system. From the leaf disks, transformed plantlets were regenerated. Hybridization experiments demonstrated that the Cr(VI) reductase gene was inserted into and expressed in the regenerated plants. The Cr(VI) reduction activity showed that the transgenic plants may be a another possible tool to reduce the pollution of the toxic Cr(VI) in soil.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.672-675
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• 2000

E.coli ATCC 33456 has relatively higher activity of Cr(VI) reduction than other microorganism. The purpose of this research is cloning of Cr(V) reductase from E.coli ATCC 33456. Using colony and southern hybridization, we selected two condidates. Among candidates, pNCR9 is higher Cr(VI) reduction activity than E.coli ATCC 33456. Purified Cr(VI) reductase antibody was reacted at estimated 42Kda protein band of candidate's crude extract on 12% SDS-PAGE. This results showed cloned gene's product is very similar to purified Cr(VI) reductase from E.coli ATCC 33456.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.580-586
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• 2000

A soluble Cr(VI) reductase was purified from the Cr(VI) reducing strain Escherichia coli ATCC33456 by ammonium sulfate fractionation, and chromatographies on Q-Sepharose FF, Cibacron blue 3GA dye affinity, Mono-Q 5/5, and Superdex 200 HR 10/30 columns. The estimated molecular mass of the purified enzyme was 27 kDa on SDS-polyacrylamide gel electrophoresis and 54 kDa on gel filtration, thus indicating a dimeric structure. The isoelectric point of the enzyme was pH 4.85. The optimum reaction pH and storage pH were both 7.0, the optimum reaction temperature was $37^{\circ}C$, and the storage temperature was $4^{\circ}C$. NADH and NADPH both served as electron donors for the reductase, with $V_{max}$ of 68.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 7.6 $\mu$M using HADH, and Vmax of 42.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 14.6 $\muM$ using NADPH. When 1 mM EDTA was added, the Cr(VI) reducing activity increased 4-fold.

• Journal of Microbiology
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• v.43 no.1
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• pp.21-27
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• 2005

A soluble Cr(VI) reductase was purified from the cytoplasm of Escherichia coli ATCC 33456. The molecular mass was estimated to be 84 and 42 kDa by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, indicating a dimeric structure. The pI was 4.66, and optimal enzyme activity was obtained at pH 6.5 and $37^{\circ}C$. The most stable condition existed at pH 7.0. The purified enzyme used both NADPH and NADH as electron donors for Cr(VI) reduction, while NADPH was the better, conferring 61% higher activity than NADH. The $K_m$ values for NADPH and NADH were determined to be 47.5 and 17.2 umol, and the $V_max$ values 322.2 and 130.7 umol Cr(VI) $min^{-1}mg^{-1}$ protein, respectively. The activity was strongly inhibited by N-ethylmalemide, $Ag^{2+},\;Cd^{2+},\;Hg^{2+}$, and $Zn^{2+}$. The antibody against the enzyme showed no immunological cross reaction with those of other Cr(VI) reducing strains.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.547-554
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• 2012

Chromium is generated from several industrial processes. It occurs in different oxidation states, but Cr(III) and Cr(VI) are the most common ones. Cr(VI) is a toxic, soluble environmental contaminant. Some bacteria are able to reduce hexavalent chromium to the insoluble and less toxic Cr(III), and thus chromate bioremediation is of considerable interest. An indigenous chromium-reducing bacterial strain, Rb-2, isolated from a tannery water sample, was identified as Ochrobactrum intermedium, on the basis of 16S rRNA gene sequencing. The influence of factors like temperature of incubation, initial concentration of Cr, mobility of bacteria, and different carbon sources were studied to test the ability of the bacterium to reduce Cr(VI) under variable environmental conditions. The ability of the bacterial strain to reduce hexavalent chromium in artificial and industrial sewage water was evaluated. It was observed that the mechanism of resistance to metal was not due to the change in the permeability barrier of the cell membrane, and the enzyme activity was found to be inductive. Intracellular reduction of Cr(VI) was proven by reductase assay using cell-free extract. Scanning electron microscopy revealed chromium precipitates on bacterial cell surfaces, and transmission electron microscopy showed the outer as well as inner distribution of Cr(VI). This bacterial strain can be useful for Cr(VI) detoxification under a wide range of environmental conditions.