UTILIZATION OF MICROORGANISM FOR CYANIDE BIOREMEDIATION

BESS A. TULIO

BACHELOR OF SCIENCE IN BIOLOGY

UNIVERSITY OF THE PHILIPPINES MINDANAO

APRIL 2017

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 Cyanide is a process used to recover valuable metals through a leaching process.
Accounting for 90 % of world production, cyanide leaching becomes the viable lixivant for gold
recovery (Mudder, 1999; Mudder and Botz, 2001). The reagent availability, cost, effectiveness
and environmental compatibility paved way to most active lixivant for gold and silver extraction
(McNulty, 2001).
Gold is extracted from the ore where there is very low concentration. It is separated from
the other minerals found inside the ore. Sodium cyanide (NaCN) is added to make gold soluble
and the cyanide ion forms a complex ion with the gold, [Au (CN)2]. Ore is then crushed and ground
leaving behind the gold which are too large to react with cyanide. Gold ore contains other metals
which require more treatments. However, the treated gold is added with sodium cyanide giving a
reaction (4Au + 8CN- + O2 + 2H2O ⇌ 4[Au (CN)2] - + 4OHIn) which makes gold soluble, this
process is called leaching. Since, gold is now soluble, it can move through membrane while the
remnants of ore. Sodium cyanide has lime added to it with pH 10-11 ensuring the cyanide ion is
not converted to hydrogen cyanide. Slurry is treated with activated carbon or zinc to extract the
gold, as cementation needs zinc electrode in carbon paste to directly immerse in the gold cyanide
solution this leaves the remaining cyanide in the tailings or leach pads which are naturally degraded
(“Extraction of gold using cyanide”, 2015).
In line with this, mining industry releases several billion pounds of toxic wastewaters
(Luque-Almagro et al., 2016). This wastewater contains cyanide which however seeps into the
groundwater, in the case of small-scale mining, cyanide runoff from the mountains. Sometimes,
disposal of these toxic wastewater involves throwing it to the nearby river or stream making the
community at stake also risking bodies of water and the species around it.
However, negative environmental impacts of cyanide steered for United Nations Environment
Program (UNEP) international code for management in gold mining (Mudder 2002, Mudder &
Chadwick 2001).
In Philippines, small-scale mining is widely used and account for most of the livelihood of the
Indigenous People and farmers especially those in rural mountainous areas, this give them access
to easy, faster and bigger income compared to traditional farming. According to Simeon (2017)
from The Philippine Star, local small-scale mining sector, accounts for 60 percent of the country’s
total gold production. Small scale mining also digs up holes in mountains that caused most of the
landslides, cyanide disposal in the mountain streams brought upon polluted drinking water to the
affected community nearby.
Cyanide is one of the most toxic chemicals produced on a large scale for its use in the mining,
electroplating, printed circuit board manufacturing, steel and chemical industries. Consequently,
these industries, discharge large quantities of cyanide-containing liquid wastes (Knowles 1976)
Toxicological Profile for Cyanide (July 2006) classified Cyanide belonging to a chemical
group consisting of one atom of carbon connected to one atom of nitrogen by three molecular
bonds (C≡N). Moreover, Cyanides are compounds consists of combination of two or more atom
of carbon connected containing a cyanide group (CN). Cyanides are both naturally occurring and
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man-made. An insight of these are small amounts of cyanide found in plants such as almonds,
millet sprouts, lima beans, soy, spinach, bamboo shoots, and cassava roots, there are certain
bacteria, fungi that can produce or degrade cyanide. In large doses like breathing, drinking or
eating, cyanide poisoning occurs, where there is rapid, deep breathing and shortness of breath,
followed by convulsions (seizures) and loss of consciousness. However, exposure of cyanide
through skin take slower uptake into the body and can produce sores especially contact with
compounds such as hydrogen cyanide or cyanide salts. Breathing large doses of hydrogen cyanide
experienced difficulty in breathing, chest pain, vomiting, blood changes, headaches, and
enlargement of the thyroid gland.
Toxic liquid residues is treated to minimize the health and environmental risk and
bioremediation of cyanide by microorganisms from tailings and wastewaters is a proven and viable
alternative to chemical and physical treatment processes.
Microbial treatment process involves cyanide molecule degradation through an aerobic
biological process in the presence of microorganisms. Cyanide also degrades biologically through
anaerobic processes but time consuming. Biological treatment of cyanide has been shown a viable
and robust process for destroying cyanide in the mining areas (Ackil, 2003).
The use of biological treatment for removal of residual cyanide from leach pads was first
examined in the early twentieth century and was first commercially demonstrated in the gold
mining industry at the Homestake Gold Mine, USA in the middle 1980s (Mudder & Whitlock
1984).
First application was the addition of nutrients to recycled process solutions on the heap
leach pads. In this process, only phosphate was added as a limiting nutrient to the recycled process
solutions to enhance and accelerate the natural degradation of cyanide within the water and process
solution. The process involves isolating and identifying resident species of bacteria with the ability
to use or transform cyanide into non-toxic components, such as carbonate and ammonia
(Thompson et al. 1994).
Ability to degrade simple and inorganic cyanides has been exhibited by both eukaryotes
and prokaryotes from a diverse range of taxa across a wide range of metabolic pathways (Baxter
and Cummings 2006).
Degradation of cyanide pathways involved include hydrolytic, oxidative, reductive,
substitution and transfer reactions (Ebbs, 2004) depending on the microorganism’s requirement
and conditions.
Given et al. and Mudder et al. (1998) states that the aerobic and nutrient rich environment
promotes the growth of the microbial population capable of uptake, conversion, sorption, and
precipitation of thiocyanate, cyanide, ammonia, nitrate, sulfate, and metals. Some of the organisms
known to oxidize cyanide include species of the genera Actinomyces, Alcaligenes, Arthrobacter,
Bacillus, Micrococcus, Neisseria, Paracoccus, Pseudomonas, and Thiobacillus,

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 One of the organism used in microbial treatment are cyanotrophic microorganisms which
can be used for biodegradation cyanurated wastes, most specifically alkaliphilic bacterium
Pseudomonas pseudoalcaligenes strain CECT5344 uses cyanide and its derivatives as a nitrogen
source for growth, making biodegradation of cyanide possible (Luque-Almagro et al., 2016)
Other promising bacteria such as Fusarium spp., can catabolize free cyanide (Knowles and
Bunch 1986), and their growth can be increased in the presence of cyanide (Timonim 1941).
Selected strains of Trichoderma also degrades cyanide (Ezzi and Lynch 2002, 2005; Ezzi et al.
2003).
A mixed culture mainly dominated by Bacillus sp. (Bacillus safensis, Bacillus lichenformis
and Bacillus tequilensis) discovered to be able to tolerate and degrade high concentrations of F-
CN. Cyanide biodegradation was successful even at elevated concentrations, with subsequent
production of ammonium. However, the presence of ammonium was found to have an inhibitory
effect on cyanide degradation as the microorganisms preferred utilizing the ammonium over
cyanide, thus reducing the efficacy of the cyanide biodegradation. Activation/deactivation of
metabolic pathway depending on the substrate that is present was the species mechanism used
(Mekuto, L., Jackson, V.A., and Ntwampe, S.K.O., 2013).
Fungal species also degrade cyanide through the action of cyanide hydratases, a specialized
group of nitrilases which hydrolyze cyanide to formamide (Basile, 2008). Stemphylium loti
mycelia induced cyanide hydrtase, a type of enzyme that converts cyanide to formamide.
(Brinkman & Kuhn, 1975). As well as, immobilized mycelia of Gloeocercospora sorghi (Fry &
Munch, 1975) obtained very high activities of cyanide degradation.
Different variety of taxa exploit cyanides, as a mechanism to avoid predation or to inhibit
competitors which somehow led to the evolution in many organisms of enzymes that catalyze
degradation of a range of cyanide compounds. Species expressing pathways involved in cyanide
degradation paved way to microbial applications of bacteria and fungi in the biodegradation of
cyanide contamination (Baxter, J and Cummings, S., 2006).
Gold Mining’s cyanide wastes can rapidly be degraded by utilization of microorganism.
Bacteria naturally degrades both complex and free cyanide. According to Akcil (2003), the metals
that are freed in the process are either absorbed by the biofilm or are precipitated out of solution
but rate of metal-cyanide complexes (Cu, Ni, Zn and Fe) degradation is directly proportional to
their chemical stability.
Mining industry applied Pseudomonas, Achromobacter, Flavobacterium, Nocardia,
Bdellovibrio, Mycobacterium, and true nitrifiers, Nitrosomonas and Nitrobacter as a biological
treatment. However, microbial nitrilases showed most promising because they do not require any
special conditions or cofactors for activity. Of these nitrilases, the cyanide hydratases of fungi are
more active than the cyanide dihydratases of bacteria. Additionally, fungal nitrilases are also active
at higher pH values than the cyanide dihydratases (Basile, 2008).

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 Utilization of microorganism of cyanide bioremediation in tailing solutions and other
mining related wastewaters is a proven alternative to the traditional chemical and physical
processes (Akcil, 2003). Moreover, the ability of a biological system to be engineered to handle
large flows and high cyanide levels make biological treatment even more valuable compared to
chemical and physical process which are quite expensive and advance to operate.
Microbial detoxification of cyanide in mine waste waters has an advantage over
conventional chemical methods because of its low treatment cost, complete detoxification and its
natural non-toxic products (Adams, et.al, 2001, Thompson et al., 1994).
Bioremediation now is proven to be a reliable alternative in cleaning polluted water and
degrading mining’s toxic wastes. Stimulating naturally existing indigenous microorganism not
only open doors to be able to bioengineer these microorganism to enhance and rapidly degrade
pollutants but also inexpensive which can be used by local miners and are natural reliable
alternative process.

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