• Journal of Microbiology and Biotechnology
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• 제18권11호
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• pp.1747-1754
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• 2008

Acidithiobaeillus ferrooxidans ATCC 23270 is an important model organism for bioleaching and bioremediation studies owing to its diverse metabolic capabilities, whereas lix984n is a widely used extractant. Little is known about the response of cbb genes in A. ferrooxidans to lix984n shock. Thus, to elucidate the response of the $CO_2$ fixation genes in A. ferrooxidans ATCC 23270 to the addition of lix984n, the gene expression of cbb genes was examined using a real-time PCR. Although a natural increase or decrease in the expression of most cbb genes was observed after 5 min of shock with 3% (v/v) lix984n, sdhC and cbbR exhibited quick responses to the shock. Ten min of shock had a greater effect on the cbb gene expression, yet 15 min of shock had a significant effect on the Calvin cycle in A. ferrooxidans ATCC 23270, as the expression of all the cbb genes reached a very high level. Therefore, after a short lix984n shock, a solution of A. ferrooxidans can be re-used for bioleaching.

• Journal of Microbiology and Biotechnology
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• 제18권10호
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• pp.1672-1677
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• 2008

IscR (iron-sulfur cluster regulator) has been reported to be a repressor of the iscRSUA operon, and in vitro transcription reactions have revealed that IscR has a repressive effect on the iscR promoter in the case of [$Fe_{2}S_{2}$] cluster loading. In the present study, the iscR gene from A. ferrooxidans ATCC 23270 was cloned and successfully expressed in Escherichia coli, and then purified by one-step affinity chromatography to homogeneity. The molecular mass of the IscR was 18 kDa by SDS-PAGE. The optical and EPR spectra results for the recombinant IscR confirmed that an iron-sulfur cluster was correctly inserted into the active site of the protein. However, no [$Fe_{2}S_{2}$] cluster was assembled in apoIscR with ferrous iron and sulfide in vitro. Therefore, the [$Fe_{2}S_{2}$] cluster assembly in IscR in vivo would appear to require scaffold proteins and follow the Isc "AUS" pathway.