CN112047491A - Method for removing phenolic substances in phenol aqueous solution by tyrosinase-metal organic framework compound - Google Patents
Method for removing phenolic substances in phenol aqueous solution by tyrosinase-metal organic framework compound Download PDFInfo
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Abstract
The invention discloses a method for removing phenolic substances in a phenol aqueous solution by a tyrosinase-metal organic framework compound. The tyrosinase-metal organic framework compound is added into a phenol aqueous solution to react with phenols, so that the aim of removing phenol is fulfilled; the tyrosinase-metal organic framework compound is prepared by the following method: and mixing the metal ions or the metal clusters, the organic ligand and the tyrosinase solution, and fixing the tyrosinase on a porous hybrid metal organic framework generated by the coordination of the metal ions or the metal clusters and the organic ligand to obtain the tyrosinase-metal organic framework compound. The tyrosinase-metal organic framework compound disclosed by the invention is simple in preparation process, low in cost, environment-friendly and capable of efficiently removing phenols in water.
Description
Technical Field
The invention relates to the technical field of phenol removal, in particular to a method for removing phenols in a phenol aqueous solution by a tyrosinase-metal organic framework compound.
Background
The phenol pollution in the environment mainly refers to the pollution of phenol compounds to water, and the phenol-containing wastewater is one of industrial wastewater with great harm and wide pollution range in the world at present and is an important source of water pollution in the environment. Even in low concentration, phenolic compounds are pollutants with high toxicity, and phenolic wastewater with high toxicity and difficult degradation can deteriorate the water quality of a water body by being directly discharged without treatment, thereby seriously threatening the health of human beings and other organisms.
At present, the treatment method of phenolic substances in wastewater mainly comprises a chemical method, a physical method and a biological method. The physical treatment method is complex in operation and is mostly only suitable for treating high-concentration phenolic substances; the chemical treatment method has high cost and is easy to cause secondary pollution due to byproducts; compared with the traditional physical and chemical method, the biological treatment method is relatively energy-saving and environment-friendly and has lower cost. The biological method includes a microbial treatment method and an enzyme catalytic treatment method, and the microbial treatment method has the disadvantages of complex process and long period. Enzymatic treatment has proven to be the best method for treating various phenolic substances with different enzymes (e.g. peroxidases, laccases and tyrosinases) under mild conditions. Free tyrosinase has the defects of poor stability, intolerance to extreme conditions, difficulty in recovery and the like, so that the phenol substances are treated by using the immobilized tyrosinase, and the method has the advantages of easiness in reactor operation, convenience in product separation, improvement of reusability of the enzyme and the like.
The current techniques for immobilizing tyrosinase to treat phenolics include carrier-free immobilization techniques and carrier immobilization techniques. Carrier-free immobilization technologies such as Xu and others report that tyrosinase is immobilized in the form of Cross-linked enzyme aggregates (CLEAs) and can effectively remove phenolic pollutants. The carrier immobilization technology such as Seethsram and the like reports that the tyrosinase is covalently immobilized by using silicon dioxide, the clearance rate of 75h after 1mmol/L phenol treatment is only 58%, and the efficiency of eliminating phenolic substances by the immobilization method is low; kourosh Abdollahi et al report that the clearance of treating high-concentration 2500mg/L phenol for 4h is 70% by immobilizing tyrosinase with a cyanuric chloride functionalized magnetic iron oxide nanoparticle, and the method has the defects of complicated preparation process and high cost. Liu and the like fix tyrosinase on a magnetic graphene oxide material, the clearance rate of the magnetic graphene oxide material is 97 percent after the magnetic graphene oxide material is treated by 0.5mmol/L phenol for 4 hours, and the phenol removing efficiency of the method is low. The prior art for treating phenolic substances by immobilized tyrosinase mainly has the defects of complex preparation process of an immobilization method, environmental pollution caused by used chemicals, low efficiency of removing the phenolic substances and the like.
Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a method for removing phenolic substances in a phenol aqueous solution by using a tyrosinase-metal organic framework compound, and aims to solve the problems of complex process and low efficiency of treating the phenolic substances in the prior phenol removal technology.
A method for removing phenols in a phenol aqueous solution by a tyrosinase-metal organic framework compound comprises the following steps:
providing a phenol aqueous solution, wherein the phenol aqueous solution contains phenolic substances;
adding the tyrosinase-metal organic framework compound into the phenol aqueous solution to carry out a dephenolization reaction, so that phenol substances in the phenol aqueous solution can generate ortho-bisphenol through tyrosinase (enzyme-catalyzed) oxidation and further oxidize the ortho-bisphenol to generate quinone substances, and the quinone substances form high molecular polymer precipitates through a series of non-enzyme-catalyzed reactions to realize the removal of the phenol substances in the phenol aqueous solution;
the tyrosinase-metal organic framework compound is prepared by the following method:
and mixing the metal ions or the metal clusters, the organic ligand and the tyrosinase solution, and fixing the tyrosinase on a porous hybrid metal organic framework generated by the coordination of the metal ions or the metal clusters and the organic ligand to obtain the tyrosinase-metal organic framework compound.
The method for removing the phenols in the phenol aqueous solution by the tyrosinase-metal organic framework compound is characterized in that the phenols comprise one or more of phenol, p-alkylphenol (such as p-cresol, nonyl phenol and the like), p-chlorophenol, p-nitrophenol, hydroquinone, 2,4, 6-trichlorophenol, o-nitrophenol, bisphenol A and analogues thereof (such as bisphenol B, E, F, S, Z, AP, AF and the like).
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that the concentration of the phenolic substances in the phenol aqueous solution is 1 mmol/L-25 mmol/L.
The method for removing the phenolic substances in the phenol aqueous solution by the tyrosinase-metal organic framework compound is characterized in that the temperature of the phenol removing reaction is 20-60 ℃.
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that the pH value of the phenol removing reaction is 5.0-8.0.
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that in the phenol removing reaction, the dosage of the tyrosinase-metal organic framework compound is 10-100mg for every 20mL of the phenol aqueous solution.
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that the time of the phenol removing reaction is 1-6 h.
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound, wherein the metal ions comprise: cu2+、Fe3+、Zn2+、Mn2+、Co2+、Cr3+、Ni2+、Ag+、Ca2+One kind of (1).
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that the organic ligand is an azo heterocyclic organic ligand.
The method for removing the phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound is characterized in that the azo heterocyclic organic ligand comprises: 2-methylimidazole, benzimidazole, N-acetylimidazole, 2, 4-dimethylimidazole, 2-undecylimidazole, 1-tritylimidazole, 2-mercapto-1-methylimidazole, 2' -bipyridine, 4' -dimethoxy-2, 2' -bipyridine, 2, 4-diaminopyridine, and 1, 2, 4-triazole.
Has the advantages that: the invention provides an application of a metal organic framework material in treatment of phenolic substances in water. According to the tyrosinase-metal organic framework compound, the metal organic framework grows to form the tyrosinase protective shell, so that the tyrosinase is immobilized, and the immobilized tyrosinase has better capability of treating phenolic substances in water. The tyrosinase-metal organic framework compound can efficiently remove phenols in water, and lays a good foundation for removing phenols in industrial wastewater.
Drawings
FIG. 1 is a schematic diagram of the application of the tyrosinase-metal organic framework composition in the treatment of phenols in water.
FIG. 2 is a graph illustrating the effect of tyrosinase-metal organic framework complex on the efficiency of phenol removal at different concentrations in example 1.
FIG. 3 is a graph illustrating the effect of different temperatures on the clearance of phenol by tyrosinase-metal organic framework complex in example 2 of the present invention.
FIG. 4 is a graph of example 3 of the present invention investigating the effect of different pH on the clearance of phenol by tyrosinase-metal organic framework complex.
FIG. 5 is a graph of example 4 of the present invention investigating the effect of different tyrosinase-metal organic framework complex qualities on phenol removal.
FIG. 6 is a graph illustrating the effect of tyrosinase-metal organic framework complex on the clearance of various phenols in example 5.
FIG. 7 is a graph of the effect of tyrosinase-metal organic framework complex on clearance of various mixed phenols as explored in example 6 of the present invention.
Detailed Description
The present invention provides a method for removing phenols in a phenol aqueous solution, and the present invention is further described in detail below in order to make the objects, technical solutions and effects of the present invention clearer and clearer. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The invention provides a method for removing phenolic substances in a phenol aqueous solution by a tyrosinase-metal organic framework compound, which comprises the following steps:
providing a phenol aqueous solution, wherein the phenol aqueous solution contains phenolic substances;
adding the tyrosinase-metal organic framework compound into the phenol aqueous solution to carry out a dephenolization reaction, so as to remove phenolic substances in the phenol aqueous solution;
the tyrosinase-metal organic framework compound is prepared by the following method:
and mixing the metal ions or the metal clusters, the organic ligand and the tyrosinase solution, and fixing the tyrosinase on a porous hybrid metal organic framework generated by the coordination of the metal ions or the metal clusters and the organic ligand to obtain the tyrosinase-metal organic framework compound.
The invention provides application of a metal organic framework material for fixing tyrosinase to treat phenolic substances in water. The tyrosinase is fixed in a one-pot method mode by mixing metal ions or metal clusters with organic ligands and tyrosinase, and the tyrosinase induces the metal ions or the metal clusters and the organic ligands to form a hybrid porous metal organic framework through coordination, so that the hybrid porous metal organic framework becomes a protective shell of the tyrosinase, and the tyrosinase-metal organic framework compound is obtained. As shown in figure 1, the tyrosinase-metal organic framework compound is added into a water solution containing phenols, the reaction is carried out under the preset reaction condition, the phenols are oxidized by tyrosinase (enzymatic reaction) to generate ortho-bisphenol and further oxidized to generate quinones, and the quinones can generate high molecular polymers through a series of non-enzymatic reactions under natural conditions and are aggregated and precipitated, so that the aim of removing the phenols in water is fulfilled. Moreover, the tyrosinase-metal organic framework compound is simple in preparation process, low in cost and environment-friendly. The tyrosinase-metal organic framework compound is used for phenol removal treatment, and has the advantages of simplicity and convenience in operation, rapidness, high efficiency, mild conditions and the like.
The phenol aqueous solution is a water body containing phenolic substances. The phenolic substance can be mixed phenol consisting of a plurality of phenolic compounds. The phenolic substance can be one or more of phenol and various phenolic substances with substituent groups on the phenol. In one embodiment of the present invention, the phenolic material comprises: one or more of phenol, p-alkyl phenol (such as p-cresol, nonyl phenol, etc.), p-chlorophenol, p-nitrophenol, hydroquinone, 2,4, 6-trichlorophenol, o-nitrophenol, bisphenol a, and the like (such as bisphenol B, E, F, S, Z, AP, AF, etc.). Experiments show that the tyrosinase-metal organic framework compound has good removal effect on phenol, parachlorophenol, para-cresol, nonyl phenol, para-nitrophenol, bisphenol A, hydroquinone and other mixed phenols. The phenolic substances described in the present invention can also include more phenolic substances and are not limited to the above examples.
Experiments show that the removing effect of the tyrosinase-metal organic framework compound on the phenol aqueous solution is reduced along with the increase of the concentration of the phenolic substances, wherein the removing effect is better when the phenolic substances are in low concentration, and the removing effect is limited when the phenolic substances are in high concentration. In one embodiment of the present invention, the concentration of the phenolic substance in the phenol aqueous solution is 1mmol/L to 25 mmol/L. Furthermore, the concentration of the phenolic substances in the phenol aqueous solution is 1 mmol/L-2.5 mmol/L.
In one embodiment of the invention, the dephenolization reaction temperature is 20-60 ℃, or when the aqueous solution of phenol is treated by the tyrosinase-metal organic framework compound, the temperature of the reaction system is 20-60 ℃. When the influence of different temperatures on the phenol removal of the tyrosinase-metal organic framework compound is researched, the phenol removal reaction is found to have the best removal efficiency in the temperature range of 25-30 ℃, and when the temperature is higher than 40 ℃, the enzyme is inactivated due to high temperature, and the removal efficiency is low.
In one embodiment of the invention, the pH of the dephenolization reaction is 5.0-8.0, or the pH of the reaction system of the tyrosinase-metal organic framework compound treatment phenol aqueous solution is 5.0-8.0. Furthermore, the pH value of the phenol removing reaction is 5.0-6.0, the phenol removing effect is good, and when the pH value is higher than 6.5, the removing efficiency is slightly reduced.
In one embodiment of the present invention, the concentration of the phenols in the phenol aqueous solution is 1mmol/L to 25mmol/L, and the amount of the tyrosinase-metal organic framework complex is 0 mg/mL to 5mg/mL, such as 2mg/mL (the ratio of the amount of the tyrosinase-metal organic framework complex to the volume of the phenol aqueous solution).
In the phenol removing reaction, optionally, every 20mL of phenol aqueous solution corresponds to 10-100mg of tyrosinase-metal organic framework compound. In the experiment, the clearance rate of the phenolic substances is increased after 2 hours along with the increase of the dosage of the tyrosinase-metal organic framework compound, and meanwhile, the clearance rate increase amplitude of the phenolic substances is also reduced. In one embodiment of the present invention, in the dephenolization reaction, every 20mL of the phenol aqueous solution is 30mg of the tyrosinase-metal organic framework compound, and the clearance rate of the phenolic substances after 2 hours can reach 96%.
In one embodiment of the invention, the dephenolation reaction time is 1 to 6 hours. In general, for a single phenolic species, the 1h p-cresol clearance was 100%, the 1.5h p-chlorophenol clearance was 100%, and the 2.5h phenol clearance was 100%. For mixed phenol consisting of a plurality of phenolic compounds, the cleaning effect is better at 4-6 h.
Further, the invention also provides a preparation process of the tyrosinase-metal organic framework compound, which comprises the following steps: and mixing the metal ions or the metal clusters, the organic ligand and the tyrosinase solution, and fixing the tyrosinase on a porous hybrid metal organic framework generated by the coordination of the metal ions or the metal clusters and the organic ligand to obtain the tyrosinase-metal organic framework compound.
Wherein the tyrosinase can be derived from animals, plants and microorganisms. For example, the tyrosinase is extracted from plant such as mushroom, and the mushroom can be extracted after being subjected to wall breaking by a wall breaking machine.
The metal organic framework is used as a protective shell of tyrosinase and plays a role in fixing the tyrosinase, and the metal organic framework is a compound of metal ions or metal clusters and organic ligands which are complexed to form a framework structure. In one embodiment of the present invention, the metal ion comprises Cu2+、Fe3+、Zn2+、Mn2+、Co2+、Cr3+、Ni2+、Ag+、Ca2+One kind of (1). In one embodiment of the invention, the organic ligand is an azoheterocyclic organic ligand. Further, the azo heterocyclic organic ligand includes: 2-methylimidazole, benzimidazole, N-acetylimidazole, 2, 4-dimethylimidazole, 2-undecylimidazole, 1-tritylimidazole, 2-mercapto-1-methylimidazole, 2' -bipyridine, 4' -dimethoxy-2, 2' -bipyridine, 2, 4-diaminopyridine, and 1, 2, 4-triazole.
In the process of preparing the tyrosinase-metal organic framework compound, the concentration of the metal ions or metal clusters in a preparation system is 0.1-80 mmol/L, the concentration of the organic ligands in the preparation system is 0.1-640 mmol/L, the volume fraction of the tyrosinase is 50-80% (v/v%), and the preparation temperature is 4-25 ℃.
The technical scheme of the invention is explained by a method corresponding to the application of the metal organic framework material for fixing the phenol substances in the tyrosinase treatment water. The method for fixing the phenolic substances in the tyrosinase treatment water by using the metal organic framework material comprises the following steps:
s1, mixing metal ions or metal clusters, organic ligands and tyrosinase crude enzyme liquid in a mild water environment, and fixing tyrosinase on a porous hybrid metal organic framework generated by coordination of the metal ions or the metal clusters and the organic ligands to obtain a tyrosinase-metal organic framework compound;
s2, adding the tyrosinase-metal organic framework compound into a phenol aqueous solution, and reacting under a preset reaction condition to achieve the purpose of removing phenols in water.
The tyrosinase immobilization method provided by the invention is simple to operate, and the obtained tyrosinase-metal organic framework compound can effectively remove phenolic substances in water.
In the step of S1, the specific preparation method of the tyrosinase-metal organic framework compound is as follows: uniformly mixing the Tyrosinase crude enzyme solution and a metal salt (such as zinc acetate) solution, adding an organic ligand (such as 2-methylimidazole) solution, stirring the total reaction system at 4-25 ℃ for 1-24 h, centrifuging the mixed solution (4 ℃, 8000rpm, 10min) to obtain a precipitate, washing the precipitate for several times by using deionized water until the supernatant is clear and free from enzyme activity, and drying the sample at room temperature in vacuum to constant weight to obtain a Tyrosinase-metal organic framework compound (the compound is called Tyrosinase @ zeolite imidazole ester framework-8, Tyrosinase @ ZIF-8, wherein ZIF ═ zeolitic imidazole framework). Wherein the tyrosinase crude enzyme solution accounts for 50-80% (v/v) of the total system, the final concentration of the metal salt (such as zinc acetate) solution is 0.1-80 mmol/L, and the final concentration of the organic ligand (such as 2-methylimidazole) solution is 0.1-640 mmol/L.
Specifically, the preparation method of the tyrosinase crude enzyme solution comprises the following steps: the mushroom and the phosphate buffer solution are mixed according to the mass (g): and (3) putting the mixture into a homogenizer for mashing, filtering and centrifuging, wherein the volume (mL) ratio is 1-5, and taking the supernatant to obtain the tyrosinase crude enzyme solution.
In the step S2, the tyrosinase-metal organic framework compound is added into a water solution containing phenols, the reaction is carried out under a preset reaction condition, the phenols are oxidized by tyrosinase to generate ortho-bisphenol, the ortho-bisphenol is further oxidized to generate quinone, and the quinone can generate high molecular polymers through a series of non-enzymatic reactions under a natural condition, and the high molecular polymers are aggregated and precipitated, so that the aim of removing the phenols in water is fulfilled.
The predetermined reaction conditions in the step S2 are that the concentration of phenolic substances is 1 mmol/L-25 mmol/L, the reaction temperature of the system is 20-60 ℃, the pH value of the reaction of the system is 5.0-8.0, and the dosage of the tyrosinase-metal organic framework compound is 0-5 mg/mL.
The technical solution of the present invention will be described below by specific examples.
And (3) detecting the content of phenolic substances by HPLC: 2mL of the reaction solution was centrifuged (10000rpm, 1min) from the reaction system, 1mL of the supernatant was collected, and 1mL of the solution to be measured was filtered through a 0.22 μm filter and subjected to HPLC detection.
Detection conditions are as follows: shimadzu high performance liquid chromatograph reverse C18 column (GLSciences, Inc., Japan, InertsilODS-SP, 4.6X 150mmol/L, 5 μm), mobile phase A was ultrapure water (70%), fluidity B was methanol: acetic acid 100: 1(v/v) (30%), a flow rate of 1.0mL/min, a detection wavelength of 270nm, and a column oven temperature of 30 ℃.
Example 1
The influence of the tyrosinase-metal organic framework compound on the removal effect of phenol with different concentrations is researched
Weighing 15mg of tyrosinase-metal organic framework compound into a 50mL centrifuge tube, then respectively reacting with 10mL of phenol solution with concentration gradients of 1mmol/L, 2.5mmol/L, 5mmol/L, 7.5mmol/L, 10mmol/L and 25mmol/L, reacting for 2h in a constant temperature shaking table (30 ℃, 250rpm), sampling, detecting and analyzing by using HPLC, and calculating the clearance rate of different phenol concentrations, wherein each concentration is totally 3 parallel groups. The results are shown in FIG. 2.
From the results, it is known that the removing effect of the tyrosinase-metal organic framework complex on the phenol solution is reduced with the increase of the phenol concentration, the removing effect is better at a low phenol concentration, and the removing effect is limited at a high phenol concentration.
Example 2
The influence of different temperatures on the clearance of phenol by the tyrosinase-metal organic framework compound is explored
Weighing 30mg of tyrosinase-metal organic framework compound into a 50mL centrifuge tube, adding 20mL of 2.5mmol/L phenol solution, then carrying out co-reaction for 2h in a constant-temperature shaking table (250rpm) at different temperatures (20 ℃, 25 ℃, 30 ℃, 40 ℃, 50 ℃ and 60 ℃), sampling every 0.5h, and carrying out HPLC detection analysis and calculation on the clearance rate of phenol at different temperatures, wherein 3 parallel groups are formed at each temperature. The results are shown in FIG. 3.
From the results, it is known that the effect of 20-60 ℃ on the clearance of phenol by the tyrosinase-metal organic framework complex shows that the clearance efficiency is the best in the range of 25-30 ℃, and when the clearance temperature is higher than 40 ℃, the enzyme is inactivated due to high temperature, the clearance efficiency is low, and 30 ℃ is selected as the optimal temperature for the tyrosinase-metal organic framework complex to clear phenol.
Example 3
Exploring the influence of different pH values on the clearance of phenol by the tyrosinase-metal organic framework compound
Weighing 35mg tyrosinase-metal organic framework complex into a 50mL centrifuge tube, adding 20mL phenol solution with 2.5mmol/L and different pH values (5.0, 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0), then co-reacting for 2h in a constant temperature shaking table (30 ℃, 250rpm), sampling every 0.5h, and analyzing and calculating the clearance rate of phenol under different pH values by HPLC detection, wherein each pH value is made into 3 parallel groups. The results are shown in FIG. 4.
From the results, the influence of pH 5.0-8.0 on the clearance of phenol by the tyrosinase-metal organic framework compound shows that the influence of pH on the clearance of phenol is not large in the range of pH 5.0-6.0, the clearance efficiency is slightly reduced when the pH is higher than 6.5, and the pH6.0 is selected as the optimum pH for the clearance of phenol by the tyrosinase-metal organic framework compound.
Example 4
The influence of the quality of different tyrosinase-metal organic framework composites on phenol removal is explored
Weighing tyrosinase-metal organic framework compounds with different masses (5mg, 10mg, 15mg, 20mg, 25mg and 30mg) into a 50mL centrifuge tube, adding 20mL and 2.5mmol/L phenol solution into the centrifuge tube, putting the centrifuge tube into a constant temperature shaking table (30 ℃ and 250rpm) for co-reaction for 2h, sampling every 0.5h, and analyzing and calculating the clearance rate of phenol under the tyrosinase-metal organic framework compounds with different masses by HPLC detection, wherein each mass is totally 3 parallel groups. The results are shown in FIG. 5.
From the results, it can be seen that the phenol removal rate is improved along with the increase of the quality of the tyrosinase-metal organic framework composite, and at 30mg, the phenol removal efficiency reaches 96% after 2h of reaction, and the phenol is almost completely removed.
Example 5
The influence of the tyrosinase-metal organic framework compound on the clearance of different single phenolic substances is researched
Weighing 30mg of tyrosinase-metal organic framework compound into a 50mL centrifuge tube, adding 20mL of phenol solution, 2.5mmol/L of phenol solution, p-cresol solution or p-chlorophenol solution into the centrifuge tube, putting the centrifuge tube and the phenol solution into a constant temperature shaking table (30 ℃, 250rpm) together for co-reaction for 3h, sampling every 0.5h, detecting and analyzing by using HPLC, and calculating the clearance rate of the tyrosinase-metal organic framework compound to different types of phenolic substances, wherein each phenolic substance is made into 3 parallel groups in total. The results are shown in FIG. 6.
From the results, it can be seen that the investigation of the effect of tyrosinase-metal organic framework complex on the removal of the 3 phenols shows that 1h of p-cresol, 1.5h of p-chlorophenol and 2.5h of phenol are completely removed. The tyrosinase-metal organic framework compound has the following efficiency and low removal rate of different types of phenolic substances: p-cresol > p-chlorophenol > phenol, which is related to the substrate specificity of tyrosinase.
Example 6
Exploring the effects of tyrosinase-metal organic framework complex on clearance of various mixed phenols
Weighing 30mg of tyrosinase-metal organic framework compound into a 50mL centrifuge tube, adding 20mL and 2.5mmol/L mixed phenol solution (phenol, p-cresol and p-chlorophenol mixed solution) into the centrifuge tube, putting the centrifuge tube into a constant temperature shaking table (30 ℃, 250rpm) for co-reaction for 10h, sampling at intervals, detecting and analyzing by using HPLC, calculating the clearance rate of the tyrosinase-metal organic framework compound on the mixed phenol substances, and making 3 parallel groups in total, wherein the result is shown in figure 7.
According to the results, the effect of the tyrosinase-metal organic framework compound on removing various mixed phenol solutions shows that p-cresol can be completely removed within 2.5 hours, p-chlorophenol can be completely removed within 6 hours, and the phenol removal rate is 70% after reaction for 10 hours, and the removal is still not complete. The tyrosinase-metal organic framework compound has a lower removing effect on mixed phenols than that of a single phenol substance. Meanwhile, by controlling the reaction time, specific phenolic substances can be removed, and a certain guiding effect is provided for the separation and specific removal of the phenolic substances.
In summary, the invention provides an application of a metal organic framework material in the fixation of tyrosinase to phenolic substances in water, a tyrosinase-metal organic framework compound is synthesized by a one-pot method, a metal ion or a metal cluster and an organic ligand form a hybrid porous metal organic framework through coordination, the hybrid porous metal organic framework becomes a protective shell of the tyrosinase, the fixation of the tyrosinase is realized, and the tyrosinase-metal organic framework compound is obtained. The immobilized tyrosinase has high catalytic performance, and can be used as a catalyst to be added into an aqueous solution containing phenols, so that the phenols in water can be effectively removed. The method for treating the phenolic substances in the water by fixing the tyrosinase by the metal organic framework material has the advantages of simple process, low cost and low requirement on reaction conditions, and has potential application in the aspect of treating the phenolic substances in the industrial phenol-containing sewage.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. A method for removing phenolic substances in a phenol aqueous solution by a tyrosinase-metal organic framework compound is characterized by comprising the following steps:
providing a phenol aqueous solution, wherein the phenol aqueous solution contains phenolic substances;
adding the tyrosinase-metal organic framework compound into the phenol aqueous solution to carry out a dephenolization reaction, so as to remove phenolic substances in the phenol aqueous solution;
the tyrosinase-metal organic framework compound is prepared by the following method:
and mixing the metal ions or the metal clusters, the organic ligand and the tyrosinase solution, and fixing the tyrosinase on a porous hybrid metal organic framework generated by the coordination of the metal ions or the metal clusters and the organic ligand to obtain the tyrosinase-metal organic framework compound.
2. The tyrosinase-metal-organic framework composite removing method for phenols in phenol aqueous solution according to claim 1, wherein the phenols comprise one or more of phenol, p-alkylphenol, p-chlorophenol, bisphenol A, p-nitrophenol, hydroquinone, 2,4, 6-trichlorophenol and o-nitrophenol.
3. The method for removing the phenols in the phenol aqueous solution by using the tyrosinase-metal organic framework compound as claimed in claim 1, wherein the concentration of the phenols in the phenol aqueous solution is 1 mmol/L-25 mmol/L.
4. The method for removing phenols in phenol water solution by tyrosinase-metal organic framework compound according to claim 1, wherein the temperature of the phenol removing reaction is 20-60 ℃.
5. The method for removing phenols in the aqueous solution of phenol by using the tyrosinase-metal organic framework compound as claimed in claim 1, wherein the pH of the phenol removing reaction is 5.0-8.0.
6. The method for removing phenols in aqueous phenol solution with tyrosinase-metal organic framework complex as claimed in claim 1, wherein in the phenol removing reaction, every 20mL of aqueous phenol solution is used in an amount of 10-100mg of tyrosinase-metal organic framework complex.
7. The method for removing phenolic substances in the phenol aqueous solution by using the tyrosinase-metal organic framework compound as claimed in claim 1, wherein the time for the dephenolization reaction is 1-6 h.
8. The method for removing phenols in phenol water solution with tyrosinase-metal organic framework compound as claimed in claim 1, wherein said metal ions comprise Cu2+、Fe3+、Zn2+、Mn2+、Co2+、Cr3+、Ni2+、Ag+、Ca2+One kind of (1).
9. The method for removing phenols in phenol water solution by tyrosinase-metal organic framework compound according to claim 1, wherein the organic ligand is azo heterocyclic organic ligand.
10. The method for removing phenols in phenol water solution by tyrosinase-metal organic framework complex according to claim 9, wherein said azo heterocyclic organic ligand comprises: 2-methylimidazole, benzimidazole, N-acetylimidazole, 2, 4-dimethylimidazole, 2-undecylimidazole, 1-tritylimidazole, 2-mercapto-1-methylimidazole, 2' -bipyridine, 4' -dimethoxy-2, 2' -bipyridine, 2, 4-diaminopyridine, and 1, 2, 4-triazole.
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