Purification process of peroxidase enzyme from
In this study, loaded Luffa sponge membrane forms were modified with ZnO, Fe3O4, ZnO/Fe3O4 nanoparticles (NPs) to remove of Direct Blue 15 (DB15), which is a carcinogenic azo dye in aqueous solution. ZnO and Fe3O4 NPs were synthesized using purified peroxidase enzymes from Euphorbia amygdaloides using green synthesis method. Adsorption of DB15 azo dyes was separately studied with membrane forms (LS-pure, LS-ZnO, LS-Fe3O4, and LS-ZnO/Fe3O4). Optimum contact time, optimum pH, optimum temperature, optimum dye concentration, and optimum LS amount were found as 45 min, pH 8.0, 20°C, 200 mg/L, and 0.025 g in line with the optimization studies, respectively. The obtained membrane forms were characterized using SEM, FT-IR, and XRD techniques. According to obtained results, NPs loaded LS membrane forms are promising in removal of DB15 from textile wastewater contaminated water.
- Luffa sponge
- Direct Blue 15
There are more than 3000 different dyes available and half of them belong to the azo dyes compounds class . Azo dyes are the most frequently used dyes in textile industry and are characterized by the presence of one or more azo linkages (─N═N─), usually in number of one or four, linked to phenyl and naphthyl radicals, which are usually replaced with some combinations of functional groups including: amino (─NH2), chlorine (─Cl), hydroxyl (─OH), methyl (─CH3), nitro (─NO2), sulfonic acid (─SO3H), and sodium salts (─SO3Na) [2, 3, 4]. These compounds can lead to significant ecological problems because of the creation of carcinogenic or mutagenic compounds [5, 6, 7]. Several azo dyes have been described to lead human bladder cancer, splenic sarcomas, and hepatocarcinoma, because azo dye reduction in the intestinal tract release aromatic amines which are absorbed by the intestine and excreted in the urine . The acute toxicity of azo dyes, with respect to the criteria of the European Union for the sorting of unsafe substances, is low and the values of LD50 are 250–2000 mg/kg body weight [9, 10].
The textile dyes can be removed by using physical, chemical, and biological methods . Nevertheless, most of these methods, which simply accumulate or concentrate the dyes, and trigger secondary contamination, resulted in the extreme usage of chemical materials [11, 12].
Because of nanoparticles’ features arising from size effect, nanotechnology has emerged in many scientific and industrial fields [13, 14]. It involves studies of measurement, modeling, and manipulation of substance in nanoscale. Nanoremediation is economic and has improved overall efficiency of fragmentation process. Potential catalytic activity of Au, Ag, Pd, Mg, Cu, Zn, and Fe nanoparticles have been reported for degradation of some aqueous cationic and anionic dyes [14, 15, 16, 17, 18]. Researchers studied degradation of Methyl Orange, Sunset Yellow, Acid Blue A azo dyes using zero valent iron nanoparticles (NZVI) with diameters between 20 and 110 nm. Methyl Orange, Sunset Yellow, Acid Red A were removed using solution prepared with 2 g NZVI rate of 79.9, 98.9, and 98.8, respectively .
However, nanoparticles are left in the ecosystem after their use in the removal of environmental contaminants. Thus, nanoparticles immobilized on a support material are to the fore for environmental remediation [20, 21]. LS is eco-friendly, cost effective, easy to use matrix material successfully used as a biotechnological tool for variety of systems, purposes and applications. LS immobilized cell systems have efficiently studied toward biofilm development for remediation of domestic and industrial wastewater containing toxic metal, paint, chlorinated compounds [22, 32].
In this study, ZnO and Fe3O4 nanoparticles were obtained by catalyzing using purified peroxidase enzymes from
2. Materials and methods
Direct Blue 15 (CAS no: 2429-74-5), FeCl2, ZnCI2, and other chemicals were purchased from Sigma-Aldrich. Euphorbia (
2.2. Green synthesis of Fe3O4 and ZnO nanoparticles
2.2.1. Collection of plant sample and preparation of plants extract
2.2.2. Partial purification of the peroxidase enzyme with ammonium sulfate precipitation
Prepared Euphorbia (
2.2.3. Peroxidase enzyme activity test
Determination of peroxidase activity was made by substrate of 1 mM 2,2′-azino-bis(3-ethylbenzthiazoline-sulfonic acid) diammonium salt (ABST) prepared in 0.1 M phosphate buffer at pH 6. For this purpose, 2.8 mL ABST was transferred to a test tube, and then the reaction mixture was formed by the addition 100 μL of 80% enzyme and 100 μL of 3.2 mM H2O2 solution into the test tube. The change in absorbance was monitored at 412 nm using UV–Visible spectrophotometer at 1 min intervals for 3 min. Blank test tube was prepared using distilled water instead of enzyme in the reaction mixture.
2.2.4. Synthesis of Fe3O4 and ZnO nanoparticles
100 μL of purified peroxidase enzyme from Euphorbia (
2.2.5. Characterization of Fe3O4 and ZnO nanoparticles
Synthesized Fe3O4 and ZnO NPs were characterized by scanning at range of 200–1000 nm by using UV-Vis spectrophotometer (Epoch nanodrop spectrophotometer). Determination of topography for Fe3O4 and ZnO nanoparticles was performed by SEM (Scanning Electron Microscope). In addition, XRD analysis (X-ray diffraction analysis) and FT-IR (Fourier transform infrared spectroscopy) were performed for Fe3O4 and ZnO NPs.
Contact time, pH, temperature, and metal ion concentration were determined for the purpose of optimization synthesized Fe3O4 and ZnO NPs. For determination of the optimum contact time, samples were spectrophotometrically measured between 0 and 240 min with 3 min intervals. Synthesis of Fe3O4 and ZnO NPs was performed in sodium phosphate buffer at pH 2.0–3.0, sodium acetate buffer at pH 4.0–6.0, sodium phosphate buffer at pH 7.0–8.0 and sodium carbonate buffer at pH 9.0–11.0 and the values of absorbance were measured. pH was adjusted by using 0.1 N HCl and 0.1 N NaOH. Synthesis of NPs was separately carried out from 10° to 90°C, respectively, and changes in absorbants of the samples were measured. Synthesis of NPs was performed by using related solution at 0.5, 1, 3, 5, and 7 mM and the absorbance of samples was measured. All measurements were performed by UV–VIS spectrophotometer and deionized water was used for blank sample.
2.3. Preparation of LS material, immobilization of nanoparticles procedure
Dried LS material was made into small pieces and was autoclaved for 20 min to soften the fibrous structure. Then, it was transformed into dough using blender. It was incubated for 4 h with 1 N NaOH at 80°C. Then, the fibers were collected and were thoroughly washed with distilled water until NaOH is resolved. About 0.1% hypochlorite was used for decoloration of washed fibers and then, they were washed with distilled water. Fibers with the length of 10–50 μm were collected and were dispersed with distilled water to form a suspension form. The suspension was filtered under aseptic conditions using filter paper and obtained LS fibers were dried on filter paper at 40°C for 4 h . Immobilization was performed by treatment solutions containing Fe3O4 and ZnO NPs with LS which was pretreated in ultrasonic bath for 1 h. Then, the obtained membrane forms (LS-pure, LS-ZnO, LS-Fe3O4, LS-ZnO/Fe3O4) was dried in oven for 2 h.
2.4. Azo dye remediation
The prepared membranes were used for decolorization of DB15 solution which was prepared in the laboratory. Synthetic wastewater was prepared by dissolving DB15 dye. A calibration curve was prepared in the range 0–40 ng/cm3 of DB15. The reaction mixture was prepared by adding membrane forms and in flasks containing 50 mL volume DB15 dye solution. The samples were taken out from the flasks periodically with a micropipette and were centrifuged at 5000 rpm for 10 min. The supernatant solutions were filtered with 0.45 mm filters. Then, the concentration of DB15 was measured with a UV–VIS spectrophotometer at λ = 596 nm. Scanning electron microscopy (SEM) was used to examine the surface of the adsorbents before and after dye adsorption (JEOL JSM-6400 SEM) and FTIR, XRD were performed for dye adsorption. Optimum contact time, pH, temperature, concentration of dye to determine optimal conditions for the decolorization of DB15 azo dye were analyzed using UV-Visible spectrophotometer.
The amounts of the dyes adsorbed onto LS-pure, LS-ZnO, LS-Fe3O4, and LS-ZnO/Fe3O4 (
2.5. Adsorption isotherms
Plots of ln(
3. Results and discussion
3.1. Partial obtaining peroxidase enzyme from
Euphorbia amygdaloides plant
The data obtained in the purification process of peroxidase enzyme are given in Table 1.
|Enzyme fraction||Volume (mL)||Activity (EU/mL)||Total activity (EU) 103/%||Protein (mg protein) (mL)||Specific activity (EU/mg)||Purification coefficient (EU/mg)|
|Crude extract||50||236.4 ± 1.0||11.82/100||3.82×102± 0.7||0.62||—|
|(NH4)2SO4 (60–80%)||20||174.3 ± 1.02||3.49/29.5||1.88 ± 0.16||92.71||149.54|
There are many studies on the purification of peroxidase enzyme in the literature. Plants such as wheat seeds, barley and wheat, soybeans, fava beans, sorghum, watermelon seeds, red beets, cotton, pearl millet seedlings, Asian rice, lettuce, wild radish, and pearl barley hybrids were used for the purification of peroxidase enzyme [25, 26, 27, 28, 29].
Peroxidase enzyme was purified by ammonium sulfate precipitation from
3.2. Characterization of Fe3O4 and ZnO NPs
The results of the optimization studies for the green synthesis of nanoparticles are given in Table 2.
|Nanoparticle||Wavelength (nm)||Contact time (h)||pH||Temperature (°C)||Metal ion concentration (mM)|
No research has been found in the literature on the green synthesis of Fe3O4 and ZnO NPs from the
SEM (scanning electron microscope) basically works on the basis of obtaining the images of the surface morphology which are scanned with the help of electrons. The electrons sourced from tungsten tip are sent to the surface to be scanned. After that, the emitted electrons are captured by the detector and the image is formed. SEM analysis images of ZnO and Fe3O4 nanoparticles in this study taken in this way are shown in Figure 2. The SEM image recorded for the Fe3O4 nanoparticle structure resembled a dust particle, suggesting that it may be a soft structure. It has been determined that these nanoparticle structures obtained by the surface characterization process have an average size of 30–80 nm. The SEM image for ZnO nanoparticle was taken at 5 μm. In this image, the nanoparticle structure exhibits a wavier surface. Generally, the liquid containing peroxidase enzyme prevents the formation of ZnO nanoparticle formations composed of dust and flake-like structures. However, it appears that the powder and flake structures combine in some regions and exhibit a wavy appearance after this enzyme liquid is evaporated. After the surface characterization analysis of ZnO nanoparticles, it was found that these structures vary between 60 and 80 nm on average.
The Fourier Transform Infrared Spectrophotometer (FTIR) is used to determine the bond formation between the elements present in the structure to be measured. In this direction, the structural formation that the sample possesses can be understood by measuring the vibrations of the bond occurrences in the structure at certain frequencies. This helps in the determination of the functional groups in the material being measured. The FT-IR spectrums of ZnO and Fe3O4 nanoparticles are given in Figure 3.
When looking at Figure 3A, it is possible to observe the absorbance at 510–564 cm−1, which is indicative of Zn-O band formation in this analysis which was carried out to detect the biomolecule by revealing the stabilization ability and bandwidth of the metal nanoparticles synthesized by green synthesis . Geetha et al.  synthesized ZnO nanoparticles by green synthesis using the
In the FT-IR spectrum of the Fe3O4 nanoparticle structure shown in Figure 3B, there are oscillations of bond structures that oxygen forms with iron. It is known that this nanoparticle structure oscillates between 200 and 650 cm−1. In this direction, as shown in the graph, the Fe3O4 nanoparticle structure obtained by green synthesis exhibited oscillations indicating specific bonds between iron and oxygen elements between 256 cm and 636 cm−1.
XRD (X-ray diffraction) method was used for the analysis of crystallized structures of nanoparticles used in the study. In this method, since the diffraction pattern to be produced by each structure will be different, the planar structure of the elements arranged symmetrically or periodically can be determined. The graphs obtained by XRD analysis of ZnO and Fe3O4 nanoparticles are given in Figure 4.
Plots of 100, 002, 101, 102, 110, 103, 200, 112, 201, 004, and 202 were determined in the XRD analysis graph to show the crystallized structure of the ZnO nanoparticle. ZnO nanoparticle structures are located at 2θ angles of 31.77, 34.40, 36.22, 47.61, 56.58, 62.85, 66.41, 67.93, 69.08, 72.54, and 76.85° corresponding to the plane distances of the atoms present in the structure of the nanoparticle. In this case, the XRD chart confirms that the nanoparticle we are analyzing is a ZnO nanoparticle.
Nearly all of the characterization studies for ZnO nanoparticles synthesized by different methods in the literature are discussed according to XRD analysis results. In the analyses, the planes are generally observed at the highest peak 101 with the planes of 100, 002, 101, 102, 110, 103, 112 [30, 39]. In parallel with these studies in the XRD spectrum we obtained in our study, the 101 plane has the highest peak.
The peaks obtained in the XRD spectrum of the Fe3O4 nanoparticles reveal that the desired nanoparticle structure is obtained in the green synthesis process carried out. 2θ values specific to this nanoparticle were determined as 30, 33, 44, 53, 56, and 62°.The distances between the planes determined in this direction are 220, 311, 400, 422, 511, and 440 respectively. In the light of this information, it is determined that the Fe3O4 nanoparticle structure is in a spherical crystal structure.
3.3. Immobilization efficiency
The data obtained for the immobilization of nanoparticles on the LS are given in Table 3.
|Nanoparticle solution||Wavelength (nm)||Absorbance before immobilization||Absorbance after immobilization||% immobilization|
3.4. Adsorption studies
Analytical methods used for quantitative analysis require calibration. Calibration is a process for accurately determining the relationship between the signal measured at the output of any device and the concentration of the material causing the signal. The curve obtained is a straight line. Since the calibration curve R2 value is 0.9941, the slope is assumed to be 0.0094.
As seen in Figure 5, the DB15 azo dye had value of about qe = 80 mg/L with all membrane forms in the first 15 min. Measurements at 30, 45, and 60 min resulted in 80–100 mg/L qe. However, since the highest values were noted at 45 min, optimum contact time for this azo dye was accepted as 45 min. ZnO and Fe3O4 NPs have been used to study the remodeling of many azo dyes [40, 41]. The LS-ZnO membrane form provided slightly lower adsorption compared to the LS-Fe3O4 NPs membrane form. The highest adsorption was obtained with LS-ZnO/Fe3O4 NPs membrane form.
Maximum adsorption peaks were observed at pH 8.0 in the spectrophotometric measurements performed on pH optimizations for the adsorption of DB15 azo dye solution at 50 mg/L concentration with the formed membrane forms. According to adsorption data obtained with LS-ZnO NPs and LS-Fe3O4 NPs membrane forms in pH 7.0 medium, pure LS membrane form provides a more effective adsorption in this pH environment. However, at pH 7.0, the same adsorption data were observed for LS-ZnO/Fe3O4 NPs and pure LS membrane forms (Figure 5).
In some studies performed onto Fenton process, a high decolorization rate was achieved for DB15 azo dye in highly acidic media such as pH 3.0 and 4.0 [42, 43]. Providing maximum yield in alkaline environment such as pH 8.0 near neutral with the formed membrane forms is advantageous in terms of operating.
The adsorption of the DB15 azo dye solution with membrane form exhibited the highest adsorption peaks at 20°C. However, when the temperature was gradually increased above 20°C, the adsorption with membrane forms showed an inverse proportion and gradually decreased. This result is quite advantageous in terms of the industrial application because the approximate temperature of 20°C is accepted as the optimum value.
In the work for the degradation of DB15 azo dye using copper hydroxide nitrate as a catalyst by wet peroxide oxidation, it has been reported that degradation activity of 85 and 90% of this dye is obtained after 10 and 60 min at 60°C . In another study, remediation of DB15 azo dye was studied with Fenton reaction, and in this study, the degradation efficiency of the system was increased in parallel with the temperature increasing from 20 to 40°C . The temperatures in these studies are very high and cause extra energy consumption and therefore financial loss. In this respect, the membrane forms proposed in our work offer advantages at 20°C with effective performance.
When Figure 6 is examined, the adsorption of concentration of 200 mg/L DB15 azo dye solution prepared with the membrane forms had the highest value. The adsorption of the DB15 azo dye solution prepared at the concentration of 200 mg/L with LS-ZnO NPs, LS-Fe3O4 NPs, and LS-ZnO/Fe3O4 NPs membrane forms showed the highest values were measured according to the adsorption of azo dye solutions prepared at other concentrations (10, 25, 50, and 100 mg/L). The use of nanoparticles has affected adsorption quite positively. Pure LS membrane form showed very low efficiency in dye adsorption compared to nanoparticle loaded membrane forms.
Adsorption values were read close to each other in the adsorption study of the DB15 azo dye solution with membrane forms formed with LS quantities of 0.025, 0.05, 0.1, 0.3, and 0.5 g. In the adsorption study of azo dye solution with nanoparticle-loaded membrane forms formed with all LS quantities used in the experiment, higher adsorption values were obtained compared to the pure LS membrane form used in the same amount. The highest percentage of recovery was obtained with 0.025 g LS (Figure 6). Kesraoui et al.  conducted biosorption of the Alpasit Blue with LS. In this study, maximum efficiency was obtained with 1 g LS fibrils after 2 h in pH 2.0 medium with 20mg/L concentration dye. In our study, the highest yield was achieved with an adsorbent amount of 0.025 g. In addition, nanoparticle loading has made this more efficient.
3.5. Characterization studies of dye absorption with prepared nanoparticle loaded membrane forms
SEM images for the adsorption of DB15 azo dye with the membrane forms obtained in the study are given in Figure 7. SEM images were taken at a size of 10 μm at 8000× magnification. It is seen that the DB15 azo dye has drained the membrane forms like a cover.
As shown in Figure 8, membrane forms formed with LS material exhibit significant peaks, especially at permeability of 1000 cm−1. At the same time, certain peaks were observed at about 500 and around 3350 cm−1. In fact, FT-IR bands specific to the cellulose structure of LS seen here. C─H bands and ─OH groups are found at around 3350 and 2800–2900 cm−1 [46, 47]. In the adsorption of DB15 azo dye with LS membrane forms, FT-IR bands were observed especially for Fe3O4 nanoparticle-loaded membrane forms. ZnO NPs loaded membrane had higher permeability to dye adsorption than other membrane types.
The XRD spectrum of the LS membrane form showed peaks at 2θ = 15, 20, and 38 areas. The peak intensity is above 30,000 in the area 2θ = 20. However, in the XRD spectrum of DB15 azo dye adsorption with this membrane form, the 2θ = 20 area shifted to 2θ = 22 and the peak intensity approached 60.000 in this area. In addition, additional peak was observed at 2θ = 35, 38, 43, and 50 areas. Nanoparticle-loaded membrane forms exhibited significant changes in XRD spectra when DB15 azo dye adsorbed with these membrane forms. These membrane forms exhibited very low XRD peaks compared to the pure LS membrane form, but they exhibited very high XRD peaks especially in the 2θ = 15 and 20 areas after dye adsorption. A distinctive feature of ZnO and Fe3O4 NPs in the adsorption of this azo dye was not observed in the XRD spectrum. The values were very close to each other (Figure 8) [47, 48, 49].
3.6. Langmuir and Freundlich adsorption isotherm studies
The following Langmuir isotherm equation is used to plot Langmuir adsorption isotherm graphs for the adsorption of DB15 azo dye with the membrane forms in this study. The correlation between and
Using the following Freundlich adsorption isotherm equation, Freundlich adsorption isotherm graph for the adsorption of DB15 azo dyes of the working membrane forms was drawn. This graph showing the relation between log qe and log Ce was given in Figure 9.
The Langmuir adsorption isotherm is generally used to describe the maximum adsorption capacity of an adsorbent. qm and b values were calculated from the above equation. b is a constant related to adsorption net enthalpy (L/mg), and qm is the amount of adsorbed material (mg/g) in the unit weight of the adsorbent to form a single layer at the surface. In the Langmuir isotherm study, the highest qm value was obtained LS-ZnO/Fe3O4 membrane form with 274.6 mg/g and the lowest qm value was obtained with pure LS membrane form with 45.0 mg/g. The highest b value was achieved with the pure LS membrane form with 1.186 L/mg and the LS-ZnO/Fe3O4 NPs membrane form with a minimum b value of 0.06 L/mg. High correlation coefficient R2 (0.9605) was provided with Langmuir model, the linear form application for LS-Fe3O4. This indicates that the sorption system of Langmuir isotherm provides a good model for this membrane form (Table 4).
|KF (mg/g) (L/mg)1/n||6.85||1.84||1.53||1.36×10−4|
Freundlich isotherm model is an empirical relationship that defines the adsorption of solubles from a liquid to a solid surface and assumes that there are different areas with several adsorption energies. Freundlich constants are related to the sorption capacity of the adsorbent (mg/g) and adsorption energy. In the Freundlich model, KF and n are constants that show the adsorption capacity and intensity, respectively. High KF and n values indicate high adsorption capacity and magnitude of n value is an indication of the suitability of adsorption. The LS-ZnO NPs membrane form with 2.65 values of n had a good adsorption capacity relative to the Freundlich isotherm. The pure LS membrane form is quite advantageous according to R2 (0.9885) (Table 4).
3.7. Reaction kinetics of first and second order
The adsorption kinetics of DB15 azo dye with LS, LS-ZnO NPs, LS-Fe3O4 NPs, LS-ZnO/Fe3O4 NPs membrane forms and were investigated against 10, 25, 50, 100, and 200 mg/L concentrations of dye solutions. To determine the adsorption constants, first order conformity to reaction kinetics was investigated. For this purpose, the time-dependence of ln(
|Adsorbent||Initial dye conc.||qeexp (mg/g)||First order||Second order|
|k1 (L/min)||qecal (mg/g)||R2||k2 (g/mg min)||qecal (mg/g)||R2|
|LS||10||8.8||0.034||0.845||0.9954||5.77 × 10−4||16.53||0.9751|
|25||12.6||0.030||0.978||0.9977||2.27 × 10−3||20.70||0.9955|
|50||33.6||0.0.28||1.296||0.9873||6.8 × 10−5||96.15||0.6882|
|LS||10||9.1||0.053||0.809||0.9974||2.63 × 10−3||12.03||0.9702|
|25||17.1||0.038||1.071||0.9986||1.03 × 10−3||26.46||0.9923|
|50||49.65||0.049||1.395||0.9744||8.78 × 10−5||95.24||0.9547|
|LS-Fe3O4 NPs||10||8.6||0.055||0.949||0.925||2.03 × 10−3||13.14||0.9912|
|25||22.7||0.049||1.113||0.9983||1.45 × 10−3||27.25||0.991|
|50||44.3||0.054||1.414||0.9603||1.48 × 10−4||76.33||0.9654|
|LS-ZnO/Fe3O4 NPs||10||8.9||0.047||0.839||0.9821||2.83 × 10−3||12.76||0.9971|
|25||24.0||0.039||1.075||0.9494||1.23 × 10−3||29.94||0.9537|
|50||48.6||0.054||1.400||0.9715||2.54 × 10−4||67.57||0.9876|
The k1 constant is calculated using the first order reaction equation given below:
Then, its suitability of second-order reaction kinetics was investigated to calculate the adsorption constants of DB15 azo dye. (t/qt) depicting the time dependence graphs were plotted and R2 values for 10, 25, and 50 mg/L concentrations of DB15 azo dyes were calculated (Table 5).
The second-order reaction equation is used to calculate the k2 constant and all calculated values (qeexperimental, qecalculated, k1, k2, R2) for adsorption of azo dye DB15 are shown in Table 5.
When Table 5 is examined, there is high difference between values of qeexperimental and qecalculated in the reaction kinetics of the first- and second-order in dye adsorption with pure LS membrane form. Furthermore, when the R2 values are examined, it is clear that the second order is inadequate. There is lower difference than the other concentration between values of qeexperimental and qecalculated in reaction kinetics of the second order in adsorption of azo dye at a concentration of 10 mg/L with the LS-ZnO NPs membrane form. However, when we take into account the R2 values, there is a compatibility with the first-order reaction kinetics. We see that the adsorption of azo dye at 25 mg/L concentration with Fe3O4 NPs-loaded membrane forms is more appropriate for the second-order reaction kinetics in terms of R2 values.
3.8. Calculation of thermodynamic parameters
Plots of LnKL against 1/T obtained in adsorption experiments with membrane forms of DB15 azo dye are given in Figure 10.
Values for ΔGo Gibbs free energy, ΔH° enthalpy change and ΔS° entropy thermodynamic parameters for membrane forms used in the adsorption of DB15 azo dyes are given in Table 6. ΔG° values decreased as the temperature increases in adsorption of DB15 azo dye with all membrane forms. This shows an increasing tendency in the feasibility and spontaneity of DB15 azo dye adsorption. The fact that ΔG° has negative values means that the adsorption of DB15 azo dye is spontaneously. The negative values of ΔH° confirm the exothermic structure of the adsorption process. Therefore, the adsorption of DB15 azo dye with membranes formed by the use of LS and nanoparticle is a natural chemical. Positive values of ΔS° indicate increasing disorder and randomness at the solid solution interface of the adsorbent and DB15 azo dye . This was observed in membrane forms immobilized with Fe3O4 NPs (Table 6).
|∆G° (kJ/mol K)||∆H° (kJ/mol)||∆S° (kJ/mol)|
|LS-ZnO NPs 20°C||−19855.73||−346.62||−68.95|
|LS-ZnO NPs 25°C||−20200.48|
|LS-ZnO NPs 30°C||−20545.23|
|LS-Fe3O4 NPs 20°C||−58727.95||−941.5||203.65|
|LS-Fe3O4 NPs 25°C||−59746.20|
|LS-Fe3O4 NPs 30°C||−60764.45|
|LS-ZnO/Fe3O4 NPs 20°C||−86188.31||−1386.46||298.89|
|LS-ZnO/Fe3O4 NPs 25°C||−87682.76|
|LS-ZnO/Fe3O4 NPs 30°C||−89177.21|
In this study on remediation, the possibilities offered by the environment are evaluated. ZnO and Fe3O4 NPs were produced by green synthesis with catalyzed peroxidase enzyme partially purified from
LS is a natural plant that can grow in many countries, can be used for many purposes, and has recently undergone a lot of research. In this study, nanoparticles were immobilized successfully on this material. In this way, it is aimed to prevent the nanoparticles accumulation in the environment and the creation of a separate pollution. Adsorption of DB15, a carcinogenic azo dye, was studied with nanoparticle-loaded membrane forms. Optimization, characterization, kinetic, thermodynamic studies demonstrated effectiveness of the membrane forms used in dye adsorption. For this reason, we can easily say that this work will be a source for commercialized membrane systems in the future.
This research was performed under the project numbered 115Z810 and supported by the Scientific and Technical Research Council of Turkey (TUBITAK). The authors acknowledge the support of TUBITAK, Turkey for this work.
Majcen-Le Marechal A, Slokar YM, Taufer T. Decoloration of chlorotriazine reactive azo dyes with H2O2/UV. Dyes and Pigments. 1997; 33:281-298
Shaul GM, Lieberman RJ, Dempsey CR, Dostal KA. Treatability of water soluble azo dyes by the activated sludge process. Proceedings of the Industrial Wastes Symposia WPCF. 1986:1-18
Gomes JR. Estrutura e Propriedades dos Corantes. Braga, Portugal: Barbosa e Xavier Lda; 2001
Pereira L, Alves M. Dyes—Environmental impact and remediation. In: Malik A, Grohmann E, editors. Environmental Protection Strategies for Sustainable Development, Strategies for Sustainability. Netherlands: Springer; 2012. pp. 111-154
Balan DSL. Biodegradação e toxicidade de efluentes têxteis. Revista Brasileira de Técnicos Têxteis - ABTT. 2009; 1(1):16-18
Gupta VK, Khamparia S, Tyagi I, Jaspal D, Malviya A. Decolorization of mixture of dyes: A critical review. Global Journal of Environmental Science and Management. 2015; 1:71-94
Campos-Takaki GM, Vilar Junior JC, Cavalcanti DL, Alves da Silva CA, Andrade RFS. Decolorization of black B azo dye by Pseudomonas aeruginosa. International Journal of Current Microbiology and Applied Sciences. 2015; 4(7):720-728
Gunasekaran P, Puvaneswari N, Muthukrishnan J. Toxicity assessment and microbial degradation of azo dyes. Indian Journal of Experimental Biology. 2006; 44:618-626
Clarke EA, Anliker R. Organic dyes and pigment. In: Hutzinger O, editor. The Handbook of Environmental Chemistry. 3. A. Anthropogenic Compounds. United States: Springer-Verlag; 1980. pp. 1-215
Ventura-Camargo BC, Marin-Morales MA. Azo dyes: Characterization and toxicity—A review. Textiles and Light Industrial Science and Technology (TLIST). 2013; 2(2):85-103
Robinson T, Mcmullan G, Marchant R, Nigam P. Remediation of dyes in textile effluent: A critical review on current treatment technologies with a proposed alternative. Bioresource Technology. 2001; 77:247-255
Jadhav JP, Parshetti GK, Kalme SD, Govindwar SP. Decolourization of azo dye methyl red by Saccharomyces cervisiaeMTCC 463. Chemosphere. 2007; 68:394-400
Karn B, Kuiken T, Otto M. Nanotechnology and in situ remediation: A review of the benefits and potential risks. Environmental Health Perspectives. 2009; 117:1823-1831
Nam S, Tratnyek PG. Reduction of azo dyes with zero-valent iron. Water Research. 2000; 34:1837-1845
Gupta N, Singh HP, Sharma RK. Metal nanoparticles with high catalytic activity in degradation of methyl orange: An electron relay effect. Journal of Molecular Catalysis A: Chemical. 2011; 335:248-252
Wang JQ, Liu YH, Chen MW, Louzguine-Luzgin DV, Inoue A, Perepezko JH. Excellent capability in degrading azo dyes by MgZn-based metallicglass powders. Scientific Reports. 2012; 2:418-423
Vidhu VK, Philip D. Catalytic degradation of organic dyes using biosynthesized silver nanoparticles. Micron. 2014; 56:54-62
Philip D, MeenaKumari M. Degradation of environment pollutant dyes using phytosynthesized metal nanocatalysts. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2015; 135:632-638
Rahman N, Abedin Z, Hossain MA. Rapid degradation of azo dyes using nano-scale zero valent iron. American Journal of Environmental Sciences. 2014; 10(2):157-163
Adams LK, Lyon DY, Alvarez PJJ. Comparative ecotoxicity of nanoscale TiO2, SiO2 and ZnO water suspensions. Water Research. 2006; 40:3527-3532
Lovern SB, Strickler JR, Klaper R. Behavioral and physiological changes in Daphnia magnawhen exposed to nanoparticle suspensions (titanium dioxide, nano-C60 and C60HxC70Hx). Environmental Science & Technology. 2007; 41(12):4465-4470
Saeed A, Iqbal M. Loofa (Luffa cylindrica) sponge: Review of development of the biomatrix as a tool for biotechnological applications. Biotechnology Progress. 2013; 29:573-600
Güngör AA, Demir N, Demir Y. Purification of peroxidase from latex of euphorbia ( Euphorbia amygdaloides) and investigation of kinetic properties. Asian Journal of Chemistry. 2008; 20(1):477-482
Su CH, Liu DZ, Jiang PL, Chien MY, Sheu MT, Huang YY, Chen MH. Dried fruit of the Luffa spongeas a source of chitin for applications as skin substitutes. BioMed Research International. 2014; 2014:1-9 [article ID 458287]
Converso DA, Fernandez ME. Peroxidase isozymes from wheat germ: Purification and properties. Phytochemistry. 1995; 40:1341-1345
Billaud C, Louarme L, Nicolas J. Comparison of peroxidases from barley kernel ( Hordeum vulgareL.) and wheat germ ( Triticum aestivumL.): Isolation and preliminary characterization. Journal of Food Biochemistry. 1999; 23:145-172
Chen Z, Mabrouk PA. Isolation and purification of soybean peroxidase from “Montsew” Chinese soybeans. National Undergraduate Research Clearinghouse; 2000. pp. 2-7
Dicko MH, Gruppen H, Hilhorst R, Voragen AG, Berkel WJV. Biochemical characterization of the major sorghum grain peroxidase. FEBS Journal. 2006; 273:2293-2307
Suzuki T, Honda Y, Mukasa Y, Kim SJ. Characterization of peroxidase in buckwheat seed. Phytochemistry. 2006; 67:219-224
Geetha MS, Nagabhushana H, Shivananjaiah HN. Green mediated synthesis and characterization of ZnO using Euphorbia Millilatex as fuel. IJSR. 2016; 5(4):158-163
Nasrollahzadeh M, Sajadi SM. Preparation of Pd/Fe3O4 nanoparticles by use of Euphorbia stracheyi Boissroot extract: A magnetically recoverable catalyst for one-pot reductive amination of aldehydes at room temperature. Journal of Colloid and Interface Science. 2016; 464:147-152
Slman AA. Antibacterial activity of ZnO nanoparticle on some gram-positive and gram-negative bacteria. Iraq. Journal de Physique. 2012; 10(18):5-10
Ng JCY, Cheung WH, McKay G. Equilibrium studies for the sorption of lead from effluents using chitosan. Chemosphere. 2003; 52:1021-1030
Sharma P, Sreenivas K, Rao KV. Analysis of ultraviolet photoconductivity in ZnO films prepared by unbalanced magnetron sputtering. JJAP. 2003; 93:3963-3970
Rahman OU, Mohapatra SC, Ahmad S. Fe3O4 inverse spinal super paramagnetic nanoparticles. Materials Chemistry and Physics. 2012; 132:196-202
Nagarajan S, Kuppusamy KA. Extracellular synthesis of zinc oxide nanoparticle using seaweeds of gulf of Mannar, India. Journal of Nanbiotechnology. 2013; 11:39
Manouchehr N, Mehriana A, Reza L, Rostami MH. The optimum conditions for synthesis of Fe3O4/ZnO core/shell magnetic nanoparticles for photodegradation of phenol. IJEHSE. 2014; 12(21):1-6
Gnanasangeetha D, Sarala TD. Benign ZnO nanoparticle as a practical adsorbent for removal of As3+ embedded on activated silica using Ocimum Sanctum. Discovery. 2014; 16(46):33-41
Davar F, Majedi A, Mirzaei A. Green synthesis of ZnO nanoparticles and its application in the degradation of some dyes. Journal of the American Ceramic Society. 2015; 98(6):1739-1746
Rasool K, Lee DS. Effect of ZnO nanoparticles on biodegradation and biotransformation of co-substrate and sulphonated azo dye in anaerobic biological sulfate reduction processes. International Biodeterioration & Biodegradation. 2016; 109:150-156
Sharma ACD, Sun Q, Li J, Wang Y, Suanon F, Yang J, Yu CP. Decolorization of azo dye methyl red by suspended and co-immobilized bacterial cells with mediators anthraquinone-2,6-disulfonate and Fe3O4 nanoparticles. International Biodeterioration & Biodegradation. 2016; 112:88-97
Sun JH, Shi SH, Lee YF, Sun SP. Fenton oxidative decolorization of the azo dye direct blue 15 in aqueous solution. Chemical Engineering Journal. 2009; 155(3):680-683
Weng CH, Lin YT, Chang CK, Liu N. Decolourization of direct blue 15 by Fenton/ultrasonic process using a zero-valent iron aggregate catalyst. Ultrasonics Sonochemistry. 2013; 20(3):970-977
Zhan Y, Zhou X, Fu B, Chen Y. Catalytic wet peroxide oxidation of azo dye (direct blue 15) using solvothermally synthesized copper hydroxide nitrate as catalyst. Journal of Hazardous Materials. 2011; 187(1-3):348-354
Kesraoui A, Moussa A, Ben Ali G, Seffen M. Biosorption of Alpacide blue from aqueous solution by lignocellulosic biomass: Luffa cylindricafibers. Environmental Science and Pollution Research. 2016; 23(16):15832-15840
Siquera G, Bras J, Follain N, Belbekhouche S, Marais S, Dufresne A. Thermal and mechanical properties of bionanocomposites reinforced by Luffa cylindricacellulose nanocrystals. Carbohydrate Polymers. 2013; 91:711-717
Tong Y, Zhao S, Ma J, Wang L, Zhang Y, Gao Y, Xie YM. Improving cracking and drying shrinkage properties of cement mortar by adding chemically treated luffa fibres. Construction and Building Materials. 2014; 71:327-333
Chen C, Luo Wen J, Tong Q. Elemental analysis, chemical composition, cellulose crystallinity, and FT-IR spectra of Toona sinensiswood. Abrégé en. Monatshefte fuer Chemie. 2014; 145(1):175-185
Kalkan E, Nadaroglu H, Celebi N, Tozsin G. Removal of textile dye reactive black 5 from aqueous solution by adsorption on laccase-modified silicafume. Desalination and Water Treatment. 2014; 52(31-33):6122-6134