Production of Iranian Indigenous Acidithiobacillus ferrooxidans Colony in Modified Solid Media and Investigation Capability in Copper Bioleaching

Acidithiobacillus ferrooxidans, the most widely implicated bacterium in the bioleaching of sulfide ore. Isolation and purification microorganisms occurred by plating and single colony over the solid media. At. ferrooxidans grow readily in liquid media, but it has poor growth on corresponding solid media. To attention of slow growth and economic importance of At. ferrooxidans, selection pure culture solid media is so important. In this work, for isolation of indigenous At. ferrooxidans, PLS (Pregnant Leaching Solution) was added from Sarcheshmeh Copper Mine, in the south part of Iran. PLS inoculated in some liquid and solid media. Isolation of bacterium occurred by single colony. DNA of bacterium was extracted and 16SrDNA gene was investigated by PCR. PCR product was sequenced. Purification by single colony is better by serial dilution method. Among all media, modified 9k solid medium is most favorable for production single colony and serial dilution 102- is best dilution for it. Sequencing results also proved this purified bacteria was At. ferrooxidans. At. ferrooxidans had better growth in ferrous liquid media and had most growth in 9k modified solid media. In addition, At. ferrooxidans had better growth in solid media with agarose and washed agar gelling agent rather than agar.