Cyanide bioremediation: the potential of engineered nitrilases

The cyanide-degrading nitrilases are of notable interest for their potential to remediate cyanide contaminated waste streams, especially as generated in the gold mining, pharmaceutical, and electroplating industries. This review provides a brief overview of cyanide remediation in general but with a particular focus on the cyanide-degrading nitrilases. These are of special interest as the hydrolysis reaction does not require secondary substrates or cofactors, making these enzymes particularly good candidates for industrial remediation processes. The genetic approaches that have been used to date for engineering improved enzymes are described; however, recent structural insights provide a promising new approach.

     

Precipitation of gold(I) cyanide from dicyanoaurate solutions by Acinetobacter RFB1

Summary The precipitation of gold(I) cyanide by removal of one cyanide group from dicyanoaurate by Acinetobacter RFB1 is reported. In the absence of essential salts but in the presence of ferrous ions, precipitation of gold(I) cyanide could be achieved without growth of the microorganism. Maximal precipitation was also dependent on the level of dissolved oxygen in the reaction media. Silver cyanide and other metal cyanide salts could not be precipitated as effectively as gold(I) cyanide. This system may find application in the separation of gold from other metal cyanide complexes in mining eluants.Offprint requests to: I. Finnegan     

Environmental impacts of gold mining in Essakane site of Burkina Faso

In this study, environmental impacts on air, water, and soil pollution caused by the exploit of Essakane gold mine, which is located in North Eastern part of Burkina Faso were investigated. Analyses on drinking water were made in the laboratory to determine the concentration of essential chemicals used in gold mining. Dust fallouts have been measured to assess the level of air pollution. The results showed that exploiting of gold mine directly or indirectly contributes to air pollution in Essakane district. The use of chemicals such as cyanide (industrial gold mining) and mercury (artisanal gold mining) to obtain the gold from the ore constitutes a potential risk for the ecosystem, the local population's health, and livestock production. The results also showed that there is a significant degradation of natural landscape and topography of the soil by open-pits mining (industrial mine) and holes dug by artisanal miners.     

Microbial destruction of cyanide wastes in gold mining: process review

Microbial destruction of cyanide and its related compounds is one of the most important biotechnologies to emerge in the last two decades for treating process and tailings solutions at precious metals mining operations. Hundreds of plant and microbial species (bacteria, fungi and algae) can detoxify cyanide quickly to environmentally acceptable levels and into less harmful by-products. Full-scale bacterial processes have been used effectively for many years in commercial applications in North America. Several species of bacteria can convert cyanide under both aerobic and anaerobic conditions using it as a primary source of nitrogen and carbon. Other organisms are capable of oxidizing the cyanide related compounds of thiocyanate and ammonia under varying conditions of pH, temperature, nutrient levels, oxygen, and metal concentrations. This paper presents an overview of the destruction of cyanide in mining related solutions by microbial processes.     

High Mercury Levels in the Indigenous Population of the Yaigojé Apaporis National Natural Park,Colombian Amazon

Biological Trace Element Research - Mercury (Hg) use in artisanal gold mining in the Colombian Amazon is widespread, and little is known about the exposure on local indigenous people. The aim of...     

Bacterial cyanide degradation is under review: Pseudomonas pseudoalcaligenes CECT5344, a case of an alkaliphilic cyanotroph

There are thousands of areas in the U.S.A. and Europe contaminated with cyanide-containing wastes as a consequence of a large number of industrial activities such as gold mining, steel and aluminium manufacturing, electroplating and nitrile pesticides used in agriculture. Chemical treatments to remove cyanide are expensive and generate other toxic products. By contrast, cyanide biodegradation constitutes an appropriate alternative treatment. In the present review we provide an overview of how cells deal in the presence of the poison cyanide that irreversible binds to metals causing, among other things, iron-deprivation conditions outside the cell and metalloenzymes inhibition inside the cell. In this sense, several systems must be present in a cyanotrophic organism, including a siderophore-based acquisition mechanism, a cyanide-insensitive respiratory system and a cyanide degradation/assimilation pathway. The alkaliphilic autochthonous bacterium Pseudomonas pseudocaligenes CECT5344 presents all these requirements with the production of siderophores, a cyanide-insensitive bd-related cytochrome [Cio (cyanide-insensitive oxidase)] and a cyanide assimilation pathway that generates ammonium, which is further incorporated into organic nitrogen.     

1H, 13C NMR, and electronic absorption spectroscopic studies of the interaction of cyanide with aurothiomalate

The reaction between cyanide and aurothiomalate (Autm) has been studied by 1H and 13C NMR spectroscopy and by uv spectroscopy. At cyanide:Autm ratios greater than or equal to 2, aurocyanide, [Au(CN)2]-, is the sole product but was also produced at lower ratios. Two intermediates were also identified. These were a mixed ligand complex, [tmAuCN]-, which accounted for over 80% of the gold at a ratio of cyanide to Autm of 1, and a bisthiomalato complex, [Autm2]-, which accounted for 6.8% of the total gold at this ratio of cyanide to Autm. The formation of these complexes may be significant in the antiarthritic activity of Autm since cyanide is produced by potential target cells such as polymorphonuclear leukocytes.     

Detoxification of cyanide using titanium dioxide and hydrocyclone sparger with chlorine dioxide

Abstract

The extraction of gold and silver from minerals and concentrates with cyanide is an important hydrometallurgy process that has been studied for more than 120 years. This technology, which consists of the dissolutions of the precious metals in cyanide solutions, followed by the recovery of the values by cementation, activated carbon or ion exchange resin. Most of the wastes in the industrial effluents’ milling are known to contain high contents of free cyanide as well as metallic cyanide complexes, which give them a high degree of toxicity. Appropriate methods for the treatment of cyanide solutions include cyanide destruction by oxidation using a photoelectrocatalytic detoxification technique with titanium dioxide microelectrodes. This is one of the most innovative ways for the treatment of wastewater containing cyanide. Another is the use of chlorine dioxide (ClO₂) with a gas-sparged hydrocyclone as the reactor. The results show that photodegradation of cyanide was 93% in 30 minutes using a 450 W halogen lamp, and in the case of ClO₂ the destruction of cyanides was 99% in 1 minute. In both cases, excellent performances can be achieved with the high capacity of these technologies.     

Bioremediation of cyanide‐containing wastes: The potential of systems and synthetic biology for cleaning up the toxic leftovers from mining

Synthetic biology could harness the ability of microorganisms to use highly toxic cyanide compounds for growth applied to bioremediation of cyanide‐contaminated mining wastes and areas. Subject Categories: Biotechnology & Synthetic Biology, Evolution & Ecology, Metabolism

Cyanide is a highly toxic chemical produced in large amounts by the mining and jewellery industries, steel manufacturing, coal coking, food processing and chemical synthesis (Luque‐Almagro et al, 2011). The mining industry uses so‐called cyanide leaching to extract gold and other precious metals from ores, which leaves large amounts of cyanide‐containing liquid wastes with arsenic, mercury, lead, copper, zinc and sulphuric acid as cocontaminants.Although these techniques are very efficient, they still produce about one million tonnes of toxic wastewaters each year, which are usually stored in artificial ponds that are prone to leaching or dam breaks and pose a major threat to the environment and human health (Luque‐Almagro et al, 2016). In 2000, a dam burst in Baia Mare, Romania, caused one of the worst environmental disasters in Europe. Liquid waste from a gold mining operation containing about 100 tonnes of cyanide spilled into the Somes River and eventually reached the Danube, killing up to 80% of wildlife in the affected areas. A more recent spill was caused by a blast furnace at Burns Harbor, IN, USA, which released 2,400 kg of ammonia and 260 kg of cyanide at concentrations more than 1,000 times over the legal limit into Calumet River and Lake Michigan, severely affecting wildlife. Notwithstanding the enormous damage such major spills cause, industrial activities that continuously release small amounts of waste are similarly dangerous for human and environmental health.The European Parliament, as part of its General Union Environment Action Programme, has called for a ban on cyanide in mining activities to protect water resources and ecosystems against pollution. Although several EU member states have joined this initiative, there is still no binding legislation. Similarly, there are no general laws in the USA to prevent cyanide spills, and former administration even authorized the use of cyanide for control predators in agriculture.     

Life Cycle Inventories of Gold Artisanal and Small‐Scale Mining Activities in Peru

No life cycle assessment (LCA) of artisanal and small‐scale mining activities (A&Sma) has been identified as of today, and there are limited studies about large‐scale mining and alluvial mining. The A&Sma are relevant economic sectors in countries with large reserves of mineral resources. Gold is the most representative metal mined with these practices and is used not only in jewelry but also in several electronics appliances. South America accounted for 17% of the total worldwide gold extraction in 2005; A&Sma occurred mostly in Colombia, Peru, and Brazil. The aim of this study is to estimate environmental indicators using methodologies for life cycle inventories (LCIs) in one of the two largest producers of gold through A&Sma in South America, Peru, and to discuss possible indicators for A&Sma in South America. Different functional units were used for each case study, as gold with different concentrations was produced and it was not possible to collect data for downstream processes for both bases. The product systems start in the mining and end with the gold production. Data were collected in two mining sites and, later on, related to the functional units. The results showed the amount of energy and water consumed as well as mercury used and released, carbon dioxide (CO₂) emissions, and solid wastes for each type of gold produced.     

Recent advances in microbial mining

Microbial mining of copper sulphide ores, has been practiced on an industrial scale since the late 1950s. Since then, advances in microbial mining and the role of microorganisms involved in solubilization of metals have assumed commerical importance. The fact that bioleaching processes save energy, have a minimum pollution potential and are able to yield value-added by-products make these processes invaluable. The metal extraction processes using microorganisms, which are currently in active use, concern copper and uranium bioleaching. Biobeneficiation is also applied at an industrial scale for recovery of gold from arsenopyrites. The developments in these processes during the last 15 years, with particular reference to developing nations, are reviewed. Information gathered on molecular genetics of these microorganisms should lead to a better understanding and control of microbial leaching processes. Areas still needing research to sustain economic expansion of microbial mining techniques are indicated.The author is with the Agharkar Research Institute, Agarkar Road, Pune 411 004, India     

Bacterial oxidation of propane

Abstract Much recent work in the field of biohydrometallurgy has been directed to the study of bio-oxidation of gold ores by acidophilic iron and sulfur oxidizing microorganisms. This work has been done worldwide and has resulted in several pilot plant and commercial scale operations for gold ore bio-oxidation. Bioleaching of gold by metabolic products of microorganisms has received less attention, but also offers opportunities for industrial application, especially if future regulations restrict the use of cyanide. This paper reviews recent progress in the use of microorganisms tooxidize the sulfidic matrix in refractory gold ores (bio-oxidation) and to solubilize elemental gold (bioleaching).     

Maintenance ofRhodotorula rubra isolated from liquid samples of gold mine effluents

TheRhodotorula rubra strain isolated from waste waters of a gold mining plant has demonstrated the ability to grow in the presence of cyanide. The maintenance of this strain in complex organic media leads to a loss of this ability. To preserve the cyanide resistance ofR. rubra we tested the following maintenance methods: subculturing in sterile distilled water, freezing at −20, −40, −70°C, liquid nitrogen freezing and the paper replica method. The ability to grow in the presence of cyanide was preserved and a higher viability level was observed for cells maintained frozen at −70°C, in liquid nitrogen and by the paper replica method. Preservation in distilled water resulted in the lowest viability after twelve months of storage.     

Biogenic production of cyanide and its application to gold recovery

Chromobacterium violaceum is a cyanogenic (cyanide-producing) microorganism. Cyanide is used on an industrial scale to complex and recover gold from ores or concentrates of ores bearing the precious metal. A potentially useful approach in gold mining operations could be to produce cyanide biologically in relatively small quantities at the ore surface. In this study, C. violaceum grown in nutrient broth formed a biofilm and could complex and solubilize 100% of the gold on glass test slides within 4–7 days. Approximately 50% of the cyanide-recoverable gold could be mobilized from a biooxidized sulfidic-ore concentrate. Complexation of cyanide in solution by gold appeared to have a beneficial effect on cell growth — viable cell counts were nearly two orders of magnitude greater in the presence of gold-coated slides or biooxidized ore substrates than in their absence. C. violaceum was cyanogenic when grown in alternative feedstocks. When grown in a mineral salt solution supplemented with 13.3% v/v swine fecal material (SFM), cells exhibited pigmentation and suspended cell concentrations comparable to cultures grown in nutrient broth. Glycine supplements stimulated production of cyanide in 13.3% v/v SFM. In contrast, glycine was inhibitory when added at the time of inoculation in the more concentrated SFM, decreasing cell numbers and reducing ultimate bulk-solution cyanide concentrations. However, aeration and addition of glycine to stationary phase cells grown on 13.3% v/v SFM anaerobically resulted in rapid production and high concentrations (up to 38 mg l⁻¹) of cyanide. This indicates that biogenesis of cyanide may be supported in remote areas using locally produced and inexpensive agricultural feedstocks in place of commercial media. Journal of Industrial Microbiology & Biotechnology (2001) 26, 134–139. Received 06 June 2000/ Accepted in revised form 30 September 2000     

Natural attenuation potential of cyanide via microbial activity in mine tailings

Biological removal by indigenous microflora of cyanide, contained in old (6-9 years) and fresh tailings (3 months), was studied in order to assess its natural attenuation potential via biodegradation. To investigate the presence of indigenous microflora in tailings, total heterotrophic and cyanide resistant bacteria were counted using the spread-plate method. The free cyanide mineralization potential was estimated using K14CN in the presence of various unlabeled cyanide concentrations (0, 5, and 10 mg CN/kg). The biodegradation of cyanide contained initially in the samples was also investigated by monitoring formate, formamide, ammonia and total cyanide (CNT) concentrations over 111 days. The enumeration of total heterotrophic and cyanide-resistant bacteria in old tailings showed an average population of 105 cfu/g. However, no growth was detected in fresh tailings. Nevertheless, cyanide mineralization tests indicated the presence, in both old and fresh tailings, of a cyanide-degrading microflora. In old tailings, maximum mineralization percentages of free cyanide ranging from 85% to 100% were obtained after 65 days at all concentrations tested. A mineralization percentage of 83% after 170 days was also observed in fresh tailings. No decrease of total cyanide concentration in old tailings was observed when the biodegradation of endogenous cyanide was tested whereas a significant decrease was recorded in fresh tailings after 96 days. The presence of strong metal-cyanide complexes resistant to biodegradation could explain the absence of biodegradation in old tailings. This study demonstrated the presence of an indigenous free cyanide-degrading microflora in both old and fresh tailings, and suggests that natural attenuation of cyanide in gold mine tailings is likely to occur via microbial activity.     

Transport of Ferrocyanide by Two Eucalypt Species and Sorghum

The wastes from some industrial processes and the tailings from gold mining contain elevated concentrations of cyanide, which reacts with iron in the media to form iron cyanide complexes. This research examined the transport and possible metabolism of ferrocyanide by two native Australian trees, blue mallee and sugar gum, and by sorghum. Hydroponic studies using ¹⁵N-labeled ferrocyanide showed that both tree species transported ferrocyanide into roots and displayed significant increases in ¹⁵N enrichment and concentration with no evidence of phytotoxicity. A subsequent experiment with blue mallee and membrane-transport inhibitors showed that ¹⁵N enrichment was significantly inhibited in the presence of the protonophore carbonyl cyanide m-chlorophenylhydrazone, suggesting that ferrocyanide uptake is mediated partly by H⁺-symporters. A study of the time dependence of ¹⁵N translocation showed a rapid equilibration of ¹⁵N from ferrocyanide in the root of blue mallee, accompanied by a slow increase in shoot ¹⁵N, suggestive of the metabolism of ferrocyanide in plant roots. A similar experiment with sorghum showed a more rapid translocation of ¹⁵N, suggesting that the transport and/or metabolism of ferrocyanide by roots of this species may differ. The results offer additional incentive for the use of these species as vegetative cover over cyanidation wastes and for cyanide phytoremediation.     

Influence of small-scale gold mining on French Guiana streams: Are diatom assemblages valid disturbance sensors?

The ongoing gold-rush in French Guiana could cause severe disturbance to ecosystems. Although illegal, small gold mining sites are rapidly expanding. Few studies have attempted to measure the consequences of the increased gold mining on the biota of small forest streams, and to date no study has dealt with primary producers. Here we measured the response of diatom assemblages to gold mining in ten sites differently affected by the mining activity (i.e., reference, formerly gold-mined and currently exploited). Our results showed that both taxonomic and functional structure of the diatom assemblages were influenced by the intensity of gold mining activity. A significant relationship between soil erosion and diatom motility ability has been demonstrated. These findings show that diatom assemblages are sensitive to gold mining disturbance and suggest that diatom communities may be used as sensors of the environmental stress caused by small-scale gold mining activities.     

Production of rhodanese by bacteria present in bio-oxidation plants used to recover gold from arsenopyrite concentrates

Considerably larger quantities of cyanide are required to solubilize gold following the bio-oxidation of gold-bearing ores compared with oxidation by physical-chemical processes. A possible cause of this excessive cyanide consumption is the presence of the enzyme rhodanese. Rhodanese activities were determined for the bacteria most commonly encountered in bio-oxidation tanks. Activities of between 6.4 and 8.2 micromol SCN min(-1) mg protein(-1) were obtained for crude enzyme extracts of Thiobacillus ferrooxidans, Thiobacillus thiooxidans and Thiobacillus caldus, but no rhodanese activity was detected in Leptospirillum ferrooxidans. Rhodanese activities 2-2.5-fold higher were found in the total mixed cell mass from a bio-oxidation plant. T. ferrooxidans synthesized rhodanese irrespective of whether it was grown on iron or sulphur. With a PCR-based detection technique, only L. ferrooxidans and T. caldus cells were detected in the bio-oxidation tanks. As no rhodanese activity was associated with L. ferrooxidans, it was concluded that T. caldus was responsible for all of the rhodanese activity. Production of rhodanese by T. caldus in batch culture was growth phase-dependent and highest during early stationary phase. Although the sulphur-oxidizing bacteria were clearly able to convert cyanide to thiocyanate, it is unlikely that this rhodanese activity is responsible for the excessive cyanide wastage at the high pH values associated with the gold solubilization process.     

Development of a plant uptake model for cyanide

A model for cyanide species uptake by willow (Salix eriocephala L. var. Michaux) was developed to interpret data from hydroponic experiments quantitatively. While the potential for cyanide phytoremediation has been demonstrated modeling will aid in determining plant processes that contribute to cyanide transport and metabolism in willow and will target specific physiological parameters for field-scale phytoremediation design and optimization. The objective of the model development was to gain insight into the relative role of different processes with respect to dissolved free and iron-complexed cyanide transport and assimilation in plants and to determine rates at which these processes occur within the willow plant under the experimental conditions. A physiologically-based model describing plant uptake, transport, and metabolism of cyanide species was developed to reflect the processes that influence the movement of cyanide into and throughout the plant. Plant compartmentalization (root, stem, and leaf) corresponded to the level of detail in the data collected via hydroponic experiments. Inclusion of more detailed intra- and intercellular processes would create a model inconsistent with the macroscale nature of the data. Mass balances around each compartment were developed via kinetic representations for the mass transfer processes and were combined to form a model describing the fate of cyanide species within plant-water systems.     

Biological cyanide destruction mediated by microorganisms

Many microorganisms have an inherent capacity to degrade the toxic organic compounds that enter the environment as a result of pollution and natural activities. Significant degradation of these compounds may take many years and it is frequently necessary to consider methods that can accelerate this process. There have been several demonstrations of enhanced biological degradation of toxic wastes, both in the laboratory and under field conditions. The prospects for enhanced biological cyanide degradation are reviewed. Compared with bench-scale processes, there are very few reports of field-scale processes for cyanide bioremediation. The implementation of such field-scale degradation requires inputs from biology, hydrology, geology, chemistry and civil engineering. A conceptual framework is emerging that can be adapted to develop new processes for bioremediation of toxic organic wastes. In terms of cyanide biodegradation, this framework incorporates identification of microbes, determination of the optimal conditions for degradation, establishment of the metabolic pathways involved in cyanide degradation, identification and localization of the genes involved, identification of suitable microbial strains for practical application and development of practical engineering processes. The present review addresses the progress that has been made in each of these aspects of cyanide biodegradation. It also examines the existing field applications of biological cyanide degradation and makes recommendations for future research.Dr S.K. Dubey is and Dr D.S. Holmes was with the Department of Biology, Clarkson University, Potsdam, NY 13699, USA. Dr D.S. Holmes is now affiliated with Centro de Estudios Cientifigos de Santiago, Av. Presedente Errazuriz 3132, Casilla 16443, Santiago 9, Chile.