Bioleaching of low-grade copper sulfide ore by extremely thermoacidophilic consortia at 70℃in column reactors

The effects of introducing M.sedula derivatives having different Cu^2+-resistance on bioleaching capacity of a defined consortium(consisting of A.brierleyi DSM1651 and M.hakonensis HO1-1)were studied in column reactors at 70℃.Introducing M.sedula copA mutant,a copper sensitive derivative,only had negligible effects on bioleaching.While introducing M.sedula ARS50-2,a Cu^2+resistant strain,substantially consolidated bioleaching process,with 27.77%more copper recovered after 58 d of bioleaching.Addition of M.sedula ARS50-2 likely enhanced the sulfur oxidation capacity of consortium after the 24th day under the Cu^2+stress.The majority of extreme thermoacidophiles were attached on minerals surface as indicated by quantitative PCR(qPCR)data.Successions of microbial community of extremely thermoacidophilic consortia that attached on surface of minerals were different from those in leachate.M.hakonensis HO1-1 was the dominant species attached on minerals surface in each column reactor throughout bioleaching process.The sessile M.sedula ARS50-2 remained as a major species till the 34th day.A.brierleyi DSM1651 was the most abundant planktonic species in leachate of each column reactor.These results highlight that higher Cu^2+-resistance is a beneficial trait for extreme thermoacidophiles to process copper minerals.

STUDY ON BIOLEACHING OF PRIMARY CHALCOPYRITE ORE WITH THERMOACIDOPHLIC ARCHAE

A high temperature-tolerating thermoacidophilic archae （TA） was isolated from water samples collected from a hot sulfur-containing spring in the Yunnan Province, China, and was used in bioleaching experiments of a low-grade chalcopyrite ore. The TA grow at temperatures ranging from 40 to 80℃, with 65℃ being the optimum temperature, and at pH values of l.5 to 4.0, with an optimum pH value of 2.0. The bioleaching experiments of the chalcvpyrite ore were conducted in both laboratory batch bioreactors and leaching columns. The results obtained from the bioreactor experiments showed that the TA bioleaching rate of copper reached 97% for a 12-day leaching period, while the bioleaching rate was 32.43% for thiobacillus ferrooxidans （Tf） leaching for the same leaching time. In the case of column leaching, tests of a two-phase leaching （196 days）, that is, a two-month （56 days） Tf leaching in the first phase, followed by a 140-day TA leaching in the second phase were performed. The average leaching rate of copper achieved for the 140-day TA leaching was 195mg/（L.d）, while for the control experiments, it was as low as 78mg/（L .d） for the Tf leaching, indicating that the TA possesses a more powerful oxidizing ability to the chalcopyrite than Tf Therefore, it is suggested that the two-phase leaching process be applied to .for the heap leaching operations, whereas, the TA can be used in the second phase when the temperature inside the heap has increased, and the primary copper sulfide minerals have already been partially oxidized with Tf beforehand in the first phase.

Purification and Characterization of β-Glucosidase from a Newly Isolated Strain Tolypocladium cylindrosporum Syzx4

A higher β-glucosidase-producing strain was isolated and identified as Tolypocladium cylindrosporum syzx4 based on its morphology and internal transcribed spacer（ITS） rDNA gene sequence.The present study is to ferment,purify and characterize a β-glucosidase from T.cylindrosporum gams.The enzyme was purified to homogeneity by sulfate precipitataion,diethylaminoethyl cellulose anion exchange chromatography and Sephadex G-100 gel filtration with a 9.47-fold increase in specific activity and a recovery of 12.27%.The molecular weight（Mw） of the β-glucosidase was estimated to be 58600 by SDS-PAGE,which is much lower than that of the enzyme from other fungi.The β-glucosidase shows high activity over a wide range of temperature from 35℃ to 70℃ and the optimum pH is approximately 2.4.Then the kinetic parameters Km（0.85 mmol/L） and vmax[85.23 mmol/（L·s）] of the β-glucosidase were studied,respectively with high affinity p-nitrophenyl-β-D-glucopyranoside（p-NPG） as the substrate,inhibition constant Ki（39.5 mmol/L） with the tolerance of glucose.Although β-glucosidases have been purified and characterized from several other sources,T.cylindrosporum β-glucosidase with the unique optimal pH and higher tolerance of glucose distinguished from others makes the β-glucosidase a potential application in simultaneous saccharification and fermentation（SSF）.