Bioleaching of Chalcopyrite by a New Strain Leptospirillum ferrodiazotrophum Ksh-L Isolated from a Dump-Bioleaching System of Kashen Copper-Molybdenum Mine

A new strain of Leptospirillum sp. Ksh-L was isolated from a dump-bioleaching system of the Kashen copper-molybdenum mine (South Caucasus). Ksh-L is an obligate chemolithoautotroph, capable of oxidizing ferrous iron (Fe2+). Cells are Gram-negative and vibrio- or spirillum-shaped of a 0.5-3 mu m size. The optimal conditions for the growth are 35 degrees C and pH 1.6-1.8. Cu2+ and Zn2+ have different effects on the oxidizing ability of the Leptospirillum sp. Ksh-L culture depending on the phase of growth and concentration of Fe2+. Under the conditions of gradually increasing the concentration of copper in the medium, during 4-5 successive subculturing experiments, it was possible to obtain an adapted culture of Leptospirillum sp. Ksh-L, capable of growing in the medium in the presence of up to 400 mM Cu2+. A bioleaching experiment indicates that Ksh-L can efficiently oxidize chalcopyrite. However, the bioleaching of copper from chalcopyrite by Leptospirillum ferrodiazotropum Ksh-L increased about 1.8 times in association with At. thiooxidans ATCC 19377. Phylogenetic analysis based on 16S rRNA gene sequences (GenBank ID ON226845) shows that strain Ksh-L forms a single cluster into Group III. The strain possesses 99.59%, 99.52%, and 96.60% sequence similarity with the strains YTW-96-06, YTW-66-06, and Leptospirillum ferrodiazotrophum 5C in Group III, respectively.