• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.39-44
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• 2010

Acidithiobacillus caldus is an acidophilic, chemolithotrophic bacterium that plays an important role in bioleaching. Gene transformation into A. caldus is difficult, and only the conjugation method was reported successful, which was a relatively sophisticated method. In this research, electrotransformation of A. caldus species was achieved for the first time using A. caldus Y-3 and plasmid pJRD215. Transformants were confirmed by colony PCR specific to the str gene on pJRD215, and the recovery of the plasmid from the presumptive transformants. Optimizations were made and the transformation efficiency was increased from 0.8 to $3.6{\times}10^4$ transformants/${\mu}g$ plasmid DNA. The developed electrotransformation method was convenient in introducing foreign genes into A. caldus.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.162-167
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• 2007

A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-hostrange IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-hostrange IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The $Km^r\;and\;Sm^r$ selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.12
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• pp.1126-1133
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• 2010

A two-stage biofilter was constructed and utilized to determine the removal efficiency when treating dynamic loading of a mixture of odorous compounds including benzene, toluene, p-xylene, ammonia and hydrogen sulfide. A yeast strain, Candida tropicalis, and a sulfur oxidizing bacterial (SOB) strain, Acidithiobacillus caldus sp., were immobilized in polyurethane media and packed in the two-stage biofilter. The experiment of dynamic loading variation was composed of (1) stepwise loading variation of all the odorous compounds (total EC test), (2) stepwise loading variation of each odorous compound, and (3) intermittent loading variation with 2-day-off and 3-day-on. The total EC test showed that the maximum elimination capacity was $61\;g/m^3/hr$ for total VOCs, and 5.2 and $9.1\;g/m^3/hr$ for ammonia and hydrogen, respectively. In addition, the inhibition between VOCs was observed when the loading of each individual VOC was varied. Especially the stepwise increase in toluene loading resulted in decreases of benzene and p-xylene removal efficiencies about 30% and 25%, respectively. However, the inhibition between organic and inorganic compounds was not observed. The intermittent loading variation with 2-day-off and 3-day-on showed that greater than 95% of the overall removal efficiency was restored in two days after the loading resumed. Consequently, the two-stage biofilter packed with immobilized microorganisms showed advantages over conventional biofilters for the simultaneous treatment of the mixture of organic and inorganic odorous compounds.