Cyanide Degradation Research Articles

Genome mining of cyanide-degrading nitrilases from filamentous fungi

A variety of fungal species are known to degrade cyanide through the action of cyanide hydratases, a specialized subset of nitrilases which hydrolyze cyanide to formamide. In this paper, we report on two previously unknown and uncharacterized cyanide hydratases from Neurospora crassa and Aspergillus nidulans. Recombinant forms of four cyanide hydratases from N. crassa, A. nidulans, Gibberella zeae, and Gloeocercospora sorghi were prepared after their genes were cloned with N-terminal hexahistidine purification tags, expressed in Escherichia coli, and purified using immobilized metal affinity chromatography. These enzymes were compared according to their relative specific activity, pH activity profiles, thermal stability, and ability to remediate cyanide contaminated waste water from silver and copper electroplating baths. Although all four were similar, the N. crassa cyanide hydratase (CHT) has the greatest thermal stability and widest pH range of >50% activity. N. crassa also demonstrated the highest rate of cyanide degradation in the presence of both heavy metals. The CHT of A. nidulans has the highest reaction rate of the four fungal nitrilases evaluated in this work. These data will help determine optimization procedures for the possible use of these enzymes in the bioremediation of cyanide-containing waste. Similar to known plant pathogenic fungi, both N. crassa and A. nidulans were induced to express CHT by growth in the presence of KCN.

Biodegradation of cyanide containing effluents by Scenedesmus obliquus

Biological degradation of cyanide has been shown a viable and robust process for degrading cyanide in mining process wastewaters. Several algal cultures can effectively degrade cyanide as carbon and/or nitrogen source for their growth. In this study, cyanide effluent degradation by Scenedesmus obliquus was examined. Gold mill effluents containing WAD cyanide concentration of 77.9 mg/L was fed to batch unit to examine the ability of S. obliquus for degrading cyanide. Cyanide was reduced down to 6 mg/L in 77 h. Microbial growth and metal uptake of Zn, Fe and Cu was examined during cyanide degradation. The cells well adapted to high pH and the effluent contained cyanide and the metals. It is important that Zn level reduced down 50%, of the starting concentration. pH was kept at 10.3 to prevent loss of cyanide as HCN, due its volatile nature. The bio treatment process was considered to be successful in degrading cyanide in the mine process water.

Copper (II) phthalocyanines immobilized on alumina and encapsulated inside zeolite-X and their applications in photocatalytic degradation of cyanide: A comparative study

Abstract Copper phthalocyanine (CuPc) complexes either deposited on alumina (CuPc/Al); synthesized by sol–gel, or encapsulated inside zeolite-X (CuPc/ZX) while in situ synthesis were characterized by X-ray diffraction (XRD), DSC, FT-IR, diffuse reflectance UV–vis, dynamic light scattering and N2 adsorption techniques. These materials were tested for photocatalytic oxidation of cyanide in comparison with ZX and CuPc free complex. Unlike the case on Al, CuPc was accommodated inside ZX, as confirmed from lattice parameter increment (from 25.101 of ZX to 25.250 A of CuPc/ZX) and decreasing both SBET and pore volume (705 m2/g, 0.550 cm3/g) as compared to the neat zeolite (837 m2/g, 0.684 cm3/g). Bending modes vibrations of δC–H showed bands at 1041 and 970 cm−1 for CuPc/Al where CuPc/ZX did not, suggesting that these bonds are formed while CuPc vibrating on the former. Contrarily, they are restricted in the latter proposing accommodation of the complex inside zeolite cages as emphasized from ΔS values that showed marked decrease in disorderness (−29 J/mol K) compared with CuPc (−13 J/mol K). In accordance, Q band revealed a red shift from 614 to 632 nm. A very strong photo-oxidation capability of oxidizing CN− was exhibited for the encapsulated complex (CuPc/ZX; 100% removal in 1 h; rate constant = 0.1 min−1) comprehending the importance of electron transfer reactions within ZX to external surfaces, and the zeolite framework that seems participating efficiently in the electron transfer process.

Generation of hydrogen peroxide and removal of cyanide from solutions using ultrasonic waves

The generation of hydrogen peroxide and the decomposition of free cyanide by ultrasonic waves were studied and the statistical analysis of the results for significance was performed using Ergun's test (essentially One-way Analysis of Variance (ANOVA) for gradients). Effects of the ultrasonic intensity, the external addition of hydrogen peroxide, aeration, temperature and pH on the rate and extent of formation/accumulation of hydrogen peroxide were demonstrated. The generation of H 2O 2 was found to increase with increasing the ultrasonic intensity (9–114 W/cm 2), which also controls the accumulation of H 2O 2 in solution (400 ml). External addition of H 2O 2 or high temperatures (>30–50°C) appeared to suppress the production of H 2O 2 in water. Formation of H 2O 2 tends to be adversely affected by the increase in alkalinity (pH 4–10.5) whilst effects of air-saturation prior to the ultrasonic irradiation and aeration during the ultrasonic irradiation were shown to be statistically insignificant. The results have also shown that a high ultrasonic power input is required for the degradation of cyanide ([CN] 0: 20 mg/l, 200 ml) to become significant (i.e. 25% reduction in cyanide level at 114 W/cm 2). The removal of cyanide by ultrasonic irradiation appeared to be substantially enhanced with the aid of additives (NaCl and CCl 4) with the complete removal of cyanide in the presence of 24 g/l CCl 4. These findings suggest that ultrasonic treatment could be used more suitably for the treatment of the effluents containing low concentrations of cyanide and the addition of NaCl and CCl 4 is essential to improve its effectiveness.

FENTON REACTION IN THE PRESENCE OF HUMATES. TREATMENT OF HIGHLY CONTAMINATED WASTEWATER AT NEUTRAL pH

This paper presents the results of a treatability study conducted on highly contaminated wastewater using Fenton reaction at neutral pH, in the presence of humic substances. It has been found that addition of humates significantly accelerates the degradation of cyanide, thiocyanide, phenols and arsenic by ferrous iron and hydrogen peroxide. Only negligible amounts of contaminants of concern were removed when humates were added without the Fenton reagent. The mechanism of the humates effect is discussed. It is suggested that the combination of Fenton reagent with humates could provide a convenient and economical way for treatment of highly contaminated water at neutral pH.

Efficient biodegradation of cyanide and ferrocyanide by Na-alginate beads immobilized with fungal cells of Trichoderma koningii

Cyanide or metal cyanide contaminations have become serious environmental and food-health problems. A fungal mutant of Trichoderma koningii, TkA8, constructed by restriction enzyme-mediated integration, has been verified to have a high cyanide degradation ability in our previous study. In this study, the mutant cells were entrapped in sodium-alginate (Na-alginate) immobilization beads to degrade cyanide and ferrocyanide in a liquid mineral medium. The results showed that the fungus in immobilization beads consisting of 3% Na-alginate and 3% CaCl2 could degrade cyanide more efficiently than a nonimmobilized fungal culture. For maximum degradation efficiency, the optimal ratio of Na-alginate and wet fungal biomass was 20:1 (m/m) and the initial pH was 6.5. In comparison, cell immobilization took at least 3 and 8 days earlier, respectively, to completely degrade cyanide and ferrocyanide. In addition, we showed that the immobilized beads could be easily recovered from the medium and reused for up to 5 batches without significant losses of fungal remediation abilities. The results of this study provide a promising alternative method for the large-scale remediation of soil or water systems from cyanide contamination.

New approach to optimize operational conditions for the biological treatment of a high-strength thiocyanate and ammonium waste: pH as key factor

Biological treatment of coke and steel-processing wastewaters has to satisfy both industrial economic needs and environmental protection regulations. Nevertheless, as some of the pollutants contained in these waters or produced during the treatment are highly toxic, an effective and safe treatment has proved to be difficult to obtain. This paper reports the study of a biological method for the treatment of wastewaters containing free cyanide, thiocyanate and ammonium (NH 4). Laboratory-scale activated-sludge reactors were fed with a synthetic solution reproducing a steel-processing industrial wastewater and inoculated with the same industrial bacterial seeding used on-site (Ecosynergie Inc.). The results demonstrated that free cyanide and thiocyanate were efficiently degraded. Nevertheless, thiocyanate degradation and nitrification processes were actually inhibited by the free ammonia form (NH 3) in place of the ionized NH 4 form (NH 4 +) currently dosed and often unproperly named “ammonia” [IUPAC, 1997. In: McNaught, A.D., Wilkinson, A. (compilers). Compendium of Chemical Terminology. Royal Society of Chemistry, Cambridge, UK]. Optimum degradation rates were obtained for very narrow ranges of ammonia nitrogen (NH 3-N) concentrations. This result can be explained by the role of pH, which mainly controls the NH 3/NH 4 equilibrium. Pollutants and NH 3 concentrations influenced degradation rates of main pollutants. This influence was determined and expressed through elementary equations. Although the Michaelis–Menten equation could have been used to describe thiocyanate degradation, a Haldane-inhibition model was used to satisfactorily describe cyanide degradation. On the other hand, a slightly modified Haldane model was applied to describe both NH 4 oxidation using NH 3-N as substrate and thiocyanate degradation using NH 3-N as inhibitor. These findings emphasize the role of pH on degradation rates and allow one to optimize operational conditions in the biological treatment of coke and steel industrial wastewaters.

EPR and kinetic investigation of free cyanide oxidation by photocatalysis and ozonation

Oxidation of free cyanide in aqueous suspensions of three commercial TiO2 specimens, with different anatase crystal size, has been carried out in a batch photoreactor by simultaneously applying ozonation and photocatalysis. Dissolved ozone participates both in homogeneous and catalytic reactions with cyanide; the extents of these two processes are comparable to that of the photodegradation with oxygen. The reactivity results are well described by the Langmuir-Hinshelwood kinetic model, providing the values of the kinetic and equilibrium adsorption constants for the catalytic and photocatalytic reactions contributing to cyanide oxidation. The cyanide concentration decreases faster with time for catalysts with increasing anatase crystal size, being more marked under UV irradiation. EPR studies on gaseous ozone adsorption on the three samples in the dark have shown stronger ozone interactions with Ti4+ and O2− ions of the samples with largest anatase crystal size, leading to the formation of significant signals of Ti3+ and s O−−O2 radicals than with the anatase with the lowest crystal size, where ozone was mainly adsorbed on water molecular arrangements covering its surface. The hampering of the ozone and/or cyanide adsorption by the water molecular arrangements covering the surface of the catalyst with the lowest crystal size would justify the low cyanide degradation rate observed for this sample.

Sol-gel synthesis, characterization and photocatalytic activity of mixed oxide ZnO-Fe2O3

Synthesis by sol-gel method of mixed semiconductor ZnO-Fe2O3 was carried out using inorganic salts as precursors. The powders were synthesized at different pH (neutral and basic) in order to study its influence on the properties of the catalysts. These materials were characterized by thermal analysis TGA-DTA, X-ray Diffraction, FTIR spectroscopy, scanning electron microscopy and X-ray energy dispersive spectroscopy. The specific surface area of the solids was calculated from nitrogen adsorption isotherms by BET method. The band gap energy (E g ) of the catalysts was obtained from it UV-Vis spectra (diffuse reflectance) when the crystalline phase was formed under thermal treatment at 350°C by 3 h. The incorporation of Fe2O3 to ZnO semiconductor gives as a result a lower E g value for zinc oxide. Catalytic activity of the materials was evaluated using as test reaction the photocatalytic degradation of potassium cyanide in aqueous dissolution. The catalyst ZnO-Fe2O3 synthesized at pH 7 turned out to be superior than sol-gel and commercial ZnO under similar conditions.

Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI)

REMI technique was used to construct mutants with improved cyanide-degradation ability from biocontrol fungus Trichoderma koningii strain T30. The plasmid pV2 transformation was confirmed by PCR and Southern blot analysis. Out of 21 transformants, 15 single-copied transformants (71.4%) were found. To compare enzyme activities of rhodanese and cyanide hydratase, T. atroviride T23, T. harzianum T21 and their transformants constructed by REMI previously were also included. Transformants TkB6 (0.173 μmols thiocyanate formed min −1 mg protein −1) from T30 and TaK1 (0.174 μmols thiocyanate formed min −1 mg protein −1) from T23 showed higher rhodanese activity than other transformants and their wild strains. TkA9 (5.53 μmols formamide formed h −1 mg protein −1) from T30 and Th64 (5.35 μmols formamide formed h −1 mg protein −1) from T21 had higher cyanide hydratase activity than other transformants and their wild strains.

Photocatalytic degradation of pollutants from Elcogas IGCC power station effluents

The aim of this work is to improve the quality of water effluents coming from Elcogas IGCC power station (Puertollano, Spain) with the purpose of fulfilling future more demanding normative, using heterogeneous photocatalytic oxidation processes (UV/H 2O 2/TiO 2 or ZnO). The efficiency of photocatalytic degradation for the different catalysts (TiO 2 and ZnO) was determined from the analysis of the following parameters: cyanides, formates and ammonia content. In a first stage, the influence of two parameters (initial concentration of H 2O 2 and amount of catalyst) on the degradation kinetics of cyanides and formates was studied based on a factorial experimental design. pH was always kept in a value >9.5 to avoid gaseous HCN formation. The degradation of cyanides and formates was found to follow pseudo-first order kinetics. Experimental kinetic constants were fitted using neural networks (NNs). The mathematical model reproduces experimental data within 90% of confidence and allows the simulation of the process for any value of parameters in the experimental range studied. Moreover, a measure of the saliency of the input variables was made based upon the connection weights of the neural networks, allowing the analysis of the relative relevance of each variable with respect to the others. Results showed that the photocatalytic process was effective, being the degradation rate of cyanides about five times higher when compared to removal of formates. Finally, the effect of lowering pH on the degradation of formates was evaluated after complete cyanides destruction was reached (10 min of reaction). Under the optimum conditions (pH 5.2, [H 2O 2] = 40 g/l; [TiO 2] = 2 g/l), 100% of cyanides and 92% of initial NH 3 concentration are degraded after 10 min, whereas 35 min are needed to degrade 98% of formates.

Modelling anaerobic digestion acclimatisation to a biodegradable toxicant: application to cyanide

The observed acclimatisation to biodegradable toxicants in anaerobic cassava wastewater treatment is explained by modelling anaerobic cyanide degradation. A complete degradation pathway is proposed for cyanide. Cyanide degradation is modelled as enzymatic hydrolysis to formate and ammonia. Ammonia is added to the inorganic nitrogen content of the digester while formate is degraded by the hydrogenotrophic methanogens. Cyanide irreversible enzyme inhibition is modelled as an inhibition factor to acetate uptake processes. Cyanide irreversible toxicity is modelled as a decay factor to the acetate degraders. Cyanide as well as added phosphorus buffer solution were considered in the chemical equilibrium calculations of pH. The observed reversible effect after acclimatisation of sludge is modelled by a population shift between two aceticlastic methanogens that have different tolerance to cyanide toxicity. The proposed pathway is added to the IWA Anaerobic Digestion Model no.1 (ADM1). The ADM1 model with the designed extension is validated by an experiment using three lab-scale upflow anaerobic sludge bed reactors which were exposed to different cyanide loadings.

Biodegradation of Cyanide by a White Rot Fungus, Trametes versicolor

The cyanide degradation abilities of three white rot fungi, Trametes versicolor ATCC 200801, Phanerochaete chrysosporium ME 496 and Pleurotus sajor-caju, were examined. T. versicolor was the most effective with 0.35 g dry cell/100 ml degrading 2 mM KCN (130 mg/l) over 42 h, at 30 degrees C, pH 10.5 with stirring at 150 rpm.

The Impact of Bioaugmentation on Metal Cyanide Degradation and Soil Bacteria Community Structure

Metal cyanides are significant contaminants of many soils found at the site of former industrial activity. In this study we isolated bacteria capable of degrading ferric ferrocyanide and K2Ni(CN)4. One of these bacteria a Rhodococcus spp. was subsequently used to bioaugment a minimal medium broth, spiked with K2Ni(CN)4, containing 1 g of either an uncontaminated topsoil or a former coke works site soil. Degradation of the K2Ni(CN)4 was observed in both soils, however, bioaugmentation did not significantly impact the rate or degree of K2Ni(CN)4 removal. Statistical analysis of denaturing gradient gel electrophoresis profiles showed that the topsoil bacterial community had a higher biodiversity, and its structure was not significantly affected by either K2Ni(CN)4 or bioaugmentation. In contrast, profiles from the coke works site indicated significant changes in the bacterial community in response to these additions. Moreover, in both soils although bioaugmentation did not affect rates of biodegradation the Rhodococcus spp. did become established in the communities in broths containing both top and coke works soil. We conclude that bacterial communities from contaminated soils with low biodiversity are much more readily perturbed through interventions such as contamination events or bioaugmentation treatments and discuss the implications of these findings for bioremediation studies.

The current and future applications of microorganism in the bioremediation of cyanide contamination

Inorganic cyanide and nitrile compounds are distributed widely in the environment, chiefly as a result of anthropogenic activity but also through cyanide synthesis by a range of organisms including higher plants, fungi and bacteria. The major source of cyanide in soil and water is through the discharge of effluents containing a variety of inorganic cyanide and nitriles. Here the fate of cyanide compounds in soil and water is reviewed, identifying those factors that affect their persistence and which determine whether they are amenable to biological degradation. The exploitation of cyanides by a variety of taxa, as a mechanism to avoid predation or to inhibit competitors has led to the evolution in many organisms of enzymes that catalyse degradation of a range of cyanide compounds. Microorganisms expressing pathways involved in cyanide degradation are briefly reviewed and the current applications of bacteria and fungi in the biodegradation of cyanide contamination in the field are discussed. Finally, recent advances that offer an insight into the potential of microbial systems for the bioremediation of cyanide compounds under a range of environmental conditions are identified, and the future potential of these technologies for the treatment of cyanide pollution is discussed.

Cyanide metabolism of Pseudomonas pseudoalcaligenes CECT5344: role of siderophores

Cyanide is one of the most potent and toxic chemicals produced by industry. The jewelry industry of Córdoba (Spain) generates a wastewater (residue) that contains free cyanide, as well as large amounts of cyano-metal complexes. Cyanide is highly toxic to living systems because it forms very stable complexes with transition metals that are essential for protein function. In spite of its extreme toxicity, some organisms have acquired mechanisms to avoid cyanide poisoning. The biological assimilation of cyanide needs the concurrence of three separate processes: (i) a cyanide-insensitive respiratory chain, (ii) a system for iron acquisition (siderophores) and (iii) a cyanide assimilation pathway. Siderophores are low-molecular-mass compounds (600-1500 Da) that scavenge iron (Fe(3+)) ions (usually with extremely high affinity) from the environment under iron-limiting conditions. There are two main classes of siderophores: catechol and hydroxamate types. The catechol-type siderophores chelate ferric ion via a hydroxy group, whereas the hydroxamate-type siderophores bind iron via a carbonyl group with the adjacent nitrogen. In the presence of cyanide, bacterial proliferation requires this specific metal uptake system because siderophores are able to break down cyano-metal complexes. Pseudomonas pseudoalcaligenes CECT5344 is able to use free cyanide or cyano-metal complexes as nitrogen source. A proteomic approach was used for the isolation and identification, in this strain, of a protein that was induced in the presence of cyanide, namely CN0, that is involved in siderophore biosynthesis in response to cyanide. An overview of bacterial cyanide degradation pathways and the involvement of siderophores in this process are presented.

Cyanide utilization in Pseudomonas fluorescens NCIMB 11764 involves a putative siderophore

Cyanide utilization in P. fluorescens NCIMB 11764 requires the induction of cyanide oxygenase. The enzyme is induced during growth on cyanide as the sole nitrogen source or when cyanide is added to stationary-phase cells grown on limiting ammonia, however, enzyme induction and cyanide degradation were found to occur non-concomitantly. Cyanide removal by chloramphenicol-treated cells and a mutant strain (JL102) defective in cyanide oxygenase were also observed. We now report a non-enzymatic mechanism for cyanide biotransformation by an iron-chelating species identified as a putative siderophore. Partially purified siderophore preparations removed cyanide at initial rates as high as 7.6 mmol min−1 mg−1. The reaction product was further shown to be enzymatically oxidized to NH3 (and CO2) and also supported growth. The results indicate that both cyanide oxygenase and a putative siderophore component are involved in cyanide utilization.

The distribution and natural degradation of cyanide in goldmine tailings and polluted soil in arid and semiarid areas

Farmlands and rivers have been seriously polluted by cyanide from a goldmine tailings dam that collapsed in early spring of 1995 in Yining County, Xinjiang Autonomous Region of China. The cyanide distribution in the polluted farmland and the abandoned tailings dam was studied, three and 4 years after the accident occurred. The results indicated that natural degradation of cyanide in soils is slower than in natural water bodies. The cyanide transport in the soil section is similar to freely soluble salts. In arid and semiarid areas, cyanide can be highly enriched in the salt crust in which the concentration is even higher than the fresh tailings debris though cyanide has decomposed for 4 years. In the polluted farmland, the sticky layer in the soil section can highly adsorb and enrich cyanide so it can partly prevent cyanide transfer to groundwater. According to the characteristics of cyanide natural degradation in soil, the measures for prevention and cure of soil polluted by goldmine tailing dam collapse have been discussed.

Heterogeneous catalytic degradation of cyanide using copper-impregnated pumice and hydrogen peroxide

The main objective of this research was to investigate the oxidative destruction of free cyanide with hydrogen peroxide and copper-impregnated pumice as a heterogeneous catalyst. Original or copper-impregnated pumices added alone were not effective adsorbents of negatively charged cyanide ions due to incompatible surface interactions. Peroxide and original pumices added together were also ineffective in removing cyanide. However, for all of the three natural pumices tested with various particle size fractions, the use of copper-impregnated pumices and peroxide together significantly enhanced both the initial rate and extent of cyanide removal. Although copper-impregnated specific surface area was the major factor affecting the rate and extent of cyanide destruction for a particular pumice source with similar surface chemistries, the type of surface chemistry (i.e., specific functional groups) within different pumice sources also appears to be a very important factor. Lower rates and extents of cyanide removals were observed at pH 11 compared to pH 8 probably because of the negative impacts of alkaline conditions in terms of scavenging peroxide and forming more negatively charged pumice surfaces. Both the initial rate and ultimate extent of cyanide removals were generally higher at a temperature of 20 °C compared with those found at 10 °C. The use of copper-impregnated pumice as a light, cheap, readily available, natural, and porous heterogeneous catalyst either in completely mixed/suspended or fixed-bed reactor configurations may be an effective treatment technology for cyanide removal from solution. This new approach may minimize downstream metal removal problems experienced in conventional cyanide oxidation technologies.

Biodegradation of cyanide by Trichoderma spp. and Fusarium spp.

The rate of degradation of cyanide by certain strains of the Trichoderma spp. was evaluated. For comparison two Fusarium spp., which had previously been demonstrated to degrade metallocyanides were also studied. Studies were carried out to assess the rate of degradation using cyanide as the sole source of carbon or in the presence of glucose. Biodegradation was observed in flask cultures using cyanide as the sole carbon source. Strong evidence of cyanide biodegradation and co-metabolism emerged from studies with flask cultures where glucose was provided as a co-substrate. The rate of degradation of 2000 ppm CN − was enhanced almost three times in the presence of glucose. A concomitant increase in fungal biomass was also observed in all the strains over the experimental period. Growth yield calculations performed provided values that were comparable to those reported in literature for one-carbon substrates.