Leaching of Copper by Fungi

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Jorge July 18, 2019

Bacterial and fungal populations were observed in samples taken from the White Pine
Copper Company tailing pond and mine, and isolates were prepared of the dominant
species. The isolates were then screened for solubilization of copper from tailing, ore,
and concentrate during incubation in a carbon and nitrogen supplying medium. Certain
Penicillium fungi solubilized significant amounts of copper under neutral to slightly
acidic conditions. Leaching apparently was accomplished by release into the medium of
metabolites which chelated with the copper. The process was found to be highly
dependent upon the medium and the concentration of copper in the source material.

Methods

The White Pine tailing disposal area covers about 1700 acres adjacent to the plant; the
holding pond takes up about 150 acres. The samples were sealed in sterile one-half pint
glass jars and refrigerated at 4° C. until they could be plated within the following 24
hours. After

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vigorous shaking of the sample jar, one milliliter of fluid and suspended solids was
pipetted into a sterile petri-dish and 25 milliliters of agar medium at 48° C. was added.
The contents were mixed by swirling, and after the mixture hardened the dish was
stored at room temperature and exposed to light for several days. Agar media of the
following compositions were used:

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lthough the original objective was to study biological processes for leaching copper from
tailing and other wastes, it was found necessary to use ores and concentrates of higher
copper content in the work so as to have leaching effects that were significantly greater
than the experimental error White Pine flotation plant feed and concentrate were
obtained for the study, also native copper jig concentrate was obtained from the Ahmeek
mill of Calumet & Hecla, Inc.

A primary screen of the pure cultures was carried out to determine their activity on the
copper-bearing materials. Czapek’s broth was used as medium for the fungi and
nutrient broth for the bacteria. One gram of the copper-bearing material was sterilized
in a 125 millimeter flask and 50 milliliters of sterile broth was then added. Triplicate
flasks were inoculated with a fungus or a bacterium taken from the isolates by the sterile
loop technique, and a control flask was prepared in the same way without fungus or
bacterium. The flasks were then stored for 15 – 21 days at room temperature.

After the standard procedure had been set up, the process variables considered to be
significant were investigated in an effort to maximize leaching of copper. Various
sources of carbon for the fungi were tried by substituting other sugars for glucose in the
Czapek’s broth. Sources of nitrogen were investigated by substituting the individual
amino acids for sodium nitrate in the Czapek’s broth; saturated aqueous solutions of
these amino acids were made, sterile filtered, and then added to each flask. Sodium
citrate was tried as an agent for reducing the toxicity of copper on the fungi. It was
found that concentrations up to one gram per liter in the medium did not solubilize
significant amounts of copper in control flasks but that it did significantly increase the
amount of copper solubilized in the inoculated flasks.

Rod mill feed, flotation concentrate, and jig concentrate were leached with nitric acid to
compare copper yields with those from fungal leaching. Concentrated nitric acid was
added to a flask containing one-half gram of the ore-bearing material in 50 milliliters of
distilled water until the desired initial pH was reached; the initial pH was varied from 7
to 1. The flasks were agitated on the gyratory shaker for five days, and then the amount
of copper in solution was determined and the final pH was measured.

Results and Discussion

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The pond is a unique habitat with respect to ecological adversities. The foremost
adversity is the high pH (approximately 10) which is caused by basic gangue minerals
and small amounts of alkaline salts in the ore and by addition of lime for sedimentation
of tailing particles. The second adversity is murkiness of the water which affects light
penetration, thereby reducing photosynthetic organisms. A third adversity is the low
temperature, which was found to be 14 – 15°C. in October and May and 20 – 22°c. in
the summer. During the winter months, the holding pond is frozen over. The overflow
water from the pond contains no copper in solution detectable by colorimetric test.

The most common bacteria isolated in the pond were of the genera Bacillus,
Brevibacterium, and Micrococcus. Bacterial population counts varied greatly; in
general, the mud samples contained the largest population with a maximum of 74,000
cells per milliliter, the bottom water samples were the next highest with up to 17,000
cells per milliliter, while the surface and middle water samples were usually low with
counts of 2790 and 200 cells per milliliter, respectively.

The increase in solubilization was much the same with both the chalcocite flotation
concentrate and the native copper jig concentrate. The peak at 50 percent copper might
be attributed to various factors including toxicity, effect of scouring by sand particles
during agitation and others.

The improvement in yield of copper as a result of standardization of procedures and

optimization of test conditions is apparent; also the improvement in reproducibility is
apparent in the markedly lower coefficient of variation for the standard

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