CN113578279B - Wastewater treatment method based on beta-lactoglobulin - Google Patents
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Abstract
The invention provides a wastewater treatment method based on beta-lactoglobulin, and relates to the field of water treatment. The processing method comprises the following steps: adding beta-lactoglobulin into wastewater, adjusting the pH value of the wastewater to 2-5, heating the wastewater, generating amyloid protein fibers in situ in the wastewater to adsorb harmful ions in the wastewater, and removing the amyloid protein fibers adsorbing the harmful ions in the wastewater through solid-liquid separation. The invention introduces beta-lactoglobulin into the waste water containing harmful ions to prepare amyloid fiber in situ, the generated amyloid fiber has abundant amino acid groups and can combine with various heavy metal ions or arsenic and other metalloids through metal coordination and the like, thereby realizing the effect of synchronously removing the harmful ions in the waste water while preparing the amyloid fiber. The beta-lactoglobulin comes from nature, has low cost, and achieves the purposes of no pollution and resource saving. The method can be used for treating various ions such as heavy metals of nickel, copper, lead, mercury, metalloid arsenic and the like.
Description
Technical Field
The invention relates to the field of water treatment, in particular to a wastewater treatment method based on beta-lactoglobulin.
Background
With the rapid development of various industries such as electronics, semiconductors, metallurgy, mining and the like, the pollution caused by harmful substances such as heavy metals and the like is also concerned widely, and especially in industrial accidents, the discharge of industrial wastewater containing the harmful substances such as heavy metals and the like can cause more serious pollution. The long-term exposure to toxic heavy metals or arsenic, etc. can threaten human health and ecological environment, and the biological accumulation effect can generate carcinogenic effect on human body. In industrial wastewater, electroplating and acid mine wastewater contains heavy metals such as nickel, copper and the like, and has certain acidity. At present, for heavy metalsThe ion/arsenic removal method mainly comprises precipitation, ion exchange electrochemistry and the like, but the method has certain limitation in treating the acidic wastewater under the condition of low pH value. For example, chemical precipitation methods require precipitation of harmful ions by adjusting the pH of the solution to an alkaline range or adding large amounts of drugs; while in the process of ion exchange and adsorption, the H concentration is high + A competitive protonation effect is exhibited, affecting the removal of harmful ions to some extent. Therefore, the method for treating the waste water containing harmful ions without pollution becomes an important research direction in the field of water treatment.
Beta-lactoglobulin (beta-LG), a protein mainly derived from mammalian milk, is composed of 162 amino acids and has two disulfide bonds and a free thiol group. Beta-lactoglobulin is heated at high temperature under the condition of low pH value, and amyloid protein fibers can be synthesized by changing the internal folding structure of the beta-lactoglobulin. One foreign english-language document, "Amyloid-carbon hybrids for undivided water purification", discloses an Amyloid fiber-carbon hybrid membrane for water purification, which is obtained by mixing an activated carbon dispersion and a β -lactoglobulin fiber solution and then vacuum-filtering, and can be used for removing heavy metal ions and radioactive wastes in water. Although the purpose of green and pollution-free can be achieved, the treatment method is complex to operate, and the removal rate of the heavy metal ions by using the amyloid protein fibers alone is low, such as the removal rate of mercury ions is 7.38 mug/mg, and the removal rate of lead ions is 38.80 mug/mg. Therefore, the research on a wastewater treatment method which is simple and convenient to operate and has a good harmful ion removal effect becomes a technical problem to be solved in the field.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the defects of complex operation and low harmful ion removal rate of the treatment method of the waste water containing harmful ions in the prior art, thereby providing the treatment method of the waste water for preparing the amyloid protein fibers in situ by utilizing the beta-lactoglobulin.
The invention provides a wastewater treatment method based on beta-lactoglobulin, which comprises the following steps:
adding beta-lactoglobulin into wastewater, adjusting the pH value of the wastewater to 2-5, heating the wastewater, generating amyloid protein fibers in situ in the wastewater to adsorb harmful ions in the wastewater, and removing the amyloid protein fibers adsorbing the harmful ions in the wastewater through solid-liquid separation.
Furthermore, the dosage of the beta-lactoglobulin is 1-20 g/L, preferably 1-10 g/L, based on the volume of the wastewater.
Further, the heat treatment is performed at 70 to 90 ℃ for 4 to 6 hours, preferably for 5 hours.
Further, the solid-liquid separation adopts a centrifugal method and/or a membrane filtration method.
Further, the centrifugation is carried out for 5-10min under the condition of 7000-12000 rpm.
Further, the membrane filtration selects 0.22 μm or 0.45 μm organic filter membrane or filter head for treatment.
Further, adding 1-6 mol/L HCl to adjust the pH value of the wastewater.
Further, the beta-lactoglobulin is obtained by purification, and the purification method comprises the following steps:
(1) Dissolving beta-lactoglobulin to be purified in ultrapure water, and centrifuging;
(2) Taking the supernatant obtained by centrifugation in the step (1), and adjusting the pH value;
(3) Filtering to remove insoluble protein;
(4) Dialyzing to remove ions;
(5) Adjusting the pH value of the solution, and freeze-drying to obtain the purified beta-lactoglobulin.
Further, in the step (1), the beta-lactoglobulin to be purified is dissolved in ultrapure water at a concentration of 5 to 10% by mass fraction; the centrifugation is carried out for 5-10min under the conditions of 4 ℃ and 7000-12000 rpm;
in the step (2), adjusting the pH value of the supernatant to 2 by using 1-6 mol/L HCl solution;
in step (3), removing insoluble proteins using a filter;
in the step (4), a 3-8 kDa dialysis bag is used for removing ions until the pH value of the solution reaches 4-5;
in the step (5), the pH value of the solution is adjusted to 2-5.
Further, the wastewater contains at least one harmful ion of nickel ions, copper ions, lead ions, mercury ions and trivalent arsenic.
The technical scheme of the invention has the following advantages:
1. the invention introduces beta-lactoglobulin (beta-LG) into the wastewater containing harmful ions to prepare the amyloid fiber in situ, and the generated amyloid fiber has rich amino acid groups and can be combined with various heavy metal ions or arsenic and the like through metal coordination and the like, thereby realizing the effect of synchronously removing the harmful ions in the wastewater while preparing the amyloid fiber. Specifically, in the process of preparing the amyloid protein fiber serving as the adsorbent in situ, the secondary structure of the protein is changed, wherein the proportion of the folded structure is increased, the proportion of the irregular coiled structure is reduced, harmful ions preferentially occupy adsorption active sites, and the removal efficiency of the amyloid protein fiber is improved.
2. The beta-LG comes from nature, has low cost and achieves the aims of no pollution and resource saving.
3. The in-situ preparation of the amyloid protein fiber only needs to add the beta-LG into the wastewater and carry out a little treatment, and the method for treating the wastewater is simple and easy to operate. And experiments show that compared with the method of directly adding the prepared amyloid protein fibers into the wastewater, the method for preparing the amyloid protein fibers in situ in the wastewater has higher removal rate of harmful ions. That is, the method provided by the invention can be used for preparing the amyloid protein fibers and adsorbing harmful ions at the same time, so that the preparation steps are saved and the better treatment effect is realized.
4. The method for treating the wastewater based on the beta-lactoglobulin is particularly suitable for the acidic wastewater containing heavy metal ions or arsenic, can fully utilize the low pH characteristic of the acidic wastewater, does not need to prepare amyloid fibers in advance, does not generate a strong acid solution for preparing the amyloid fibers, namely in-situ synthesis can avoid pollution caused by acidic waste liquid generated in the process of preparing the amyloid fibers, saves the water treatment cost, and fully utilizes the value of the acidic wastewater to solve the problem of harmful ion pollution under the condition of low pH value. The method provided by the invention has wide application scene and is not limited by the low pH value of the acidic wastewater.
5. The water treatment method for preparing the amyloid protein fibers in situ in the wastewater can be used for treating various heavy metal ions such as nickel, copper, lead, mercury and the like, and has a good treatment effect on arsenic; meanwhile, the method can also be applied to the removal and recovery of metals such as gold, cadmium and the like, and has good removal effect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a scanning electron micrograph of amyloid fibrils formed in situ in wastewater according to example 1 of the present invention;
fig. 2 is a scanning electron micrograph of amyloid fiber prepared in comparative example 1 of the present invention.
Detailed Description
The following examples are provided to better understand the present invention, not to limit the best mode, and not to limit the content and protection scope of the present invention, and any product that is the same or similar to the present invention and is obtained by combining the present invention with other features of the prior art and the present invention falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The raw materials or equipment used are all conventional products which can be obtained commercially, including but not limited to the raw materials or equipment used in the examples of the present application.
Example 1
This example provides a nickel-containing ion (Ni) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Dissolving beta-lactoglobulin (sigma) to be purified into ultrapure water according to the concentration of 10% by mass fraction, centrifuging for 5min at the temperature of 4 ℃ and the rotation speed of 12000rpm, taking the supernatant obtained by centrifugation, adjusting the pH value of the supernatant to 2 by using an HCl solution with the concentration of 6mol/L, removing insoluble protein by using a Jinteng 250mL vacuum filter, removing ions in the protein by using a 7kDa dialysis bag (Viskease MD 25) until the pH value of the solution reaches 4-5, adjusting the pH value of the solution to 2 again, and freeze-drying to obtain the purified beta-lactoglobulin.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 100mg/LNi 2+ Adjusting the pH value of the wastewater to 2, adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Ni in the wastewater by using amyloid protein fibers generated in situ 2 + Filtering the wastewater by a 0.45 mu m filter head to remove Ni adsorbed in the wastewater 2+ Amyloid fibrils.
Example 2
This example provides a copper-containing ion (Cu) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 100mg/L Cu 2+ Adjusting the pH value of the wastewater to 2, adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of the wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Cu in the wastewater by using amyloid protein fibers generated in situ in the wastewater 2 + Filtering the wastewater by a 0.45 mu m filter head to remove Cu adsorbed in the wastewater 2+ Amyloid fibrils.
Example 3
This example provides a lead-containing ion (Pb) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fiber
Will contain 100mg/LPb 2+ Adjusting the pH value of the wastewater to 2, adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of the wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Pb in the wastewater by using amyloid protein fibers generated in situ in the wastewater 2 + Filtering the waste water by a 0.45 mu m filter head to remove Pb adsorbed in the waste water 2+ Amyloid fibrils.
Example 4
The present example provides a mercury ion (Hg) containing solution 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 100mg/L Hg 2+ Adjusting the pH value of the wastewater to 2, adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Hg in the wastewater by using amyloid protein fibers generated in situ in the wastewater 2 + Filtering the waste water by a 0.45 mu m filter head to remove adsorbed Hg in the waste water 2+ Amyloid fibrils.
Example 5
The embodiment provides a method for treating arsenic (III) -containing wastewater, which comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and arsenic adsorption of amyloid protein fibers
Adjusting the pH value of wastewater containing 100mg/LAs (III) to 2, adding 0.01g of purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5h, generating amyloid fibers in situ in the wastewater to adsorb As (III) in the wastewater, and filtering through a 0.45-micron filter head to remove the amyloid fibers adsorbing As (III) in the wastewater.
Example 6
This example provides a copper ion (Cu) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 100mg/L Cu 2+ Adjusting the pH value of the wastewater to 5, adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Cu in the wastewater by using amyloid fibers generated in situ in the wastewater 2 + Filtering the wastewater by a 0.45 mu m filter head to remove Cu adsorbed in the wastewater 2+ Amyloid fibrils.
Example 7
This example provides a copper ion (Cu) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 1000mg/L Cu 2+ Adjusting the pH value of the wastewater to 2, adding 0.1g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Cu in the wastewater by using amyloid fibers generated in situ in the wastewater 2 + Filtering the wastewater by a 0.45 mu m filter head to remove Cu adsorbed in the wastewater 2+ Amyloid fibrils.
Example 8
This example provides a copper ion (Cu) 2+ ) The wastewater treatment method comprises the following steps:
(1) Purification of beta-lactoglobulin
Refer to step (1) of example 1.
(2) In situ preparation and heavy metal ion adsorption of amyloid protein fibers
Will contain 100mg/L Cu 2+ Adjusting the pH value of the wastewater to 2, and adding 0.01g of the purified beta-lactoglobulin obtained in the step (1) into 10mL of wastewaterHeating in 70 deg.C water bath for 5 hr to adsorb Cu in the wastewater by in-situ generation of amyloid protein fiber in the wastewater 2 + Filtering the wastewater by a 0.45 mu m filter head to remove Cu adsorbed in the wastewater 2+ Amyloid fibrils.
Comparative example 1
This comparative example provides a nickel ion (Ni) 2+ ) The wastewater treatment method comprises the following steps:
(1) Preparation of amyloid fibrils
Dissolving 0.01g of purified beta-LG (purification method in reference to example 1) in 10mL of ultrapure water, adjusting the pH value of the solution to 2, heating in a water bath at 90 ℃ for 5h, centrifuging at 4 ℃ and 12000rpm for 10min, and freeze-drying the precipitate to obtain the amyloid fiber.
(2) Adsorption of heavy metal ions
Will contain 100mg/LNi 2+ Adjusting the pH value of the wastewater to 2, adding 0.01g of the amyloid protein fibers obtained in the step (1) into 10mL of wastewater, heating in a water bath at 90 ℃ for 5 hours, and adsorbing Ni in the wastewater by the added amyloid protein fibers 2+ Filtering the wastewater by a 0.45 mu m filter head to remove Ni adsorbed in the wastewater 2+ Amyloid fibrils.
Comparative example 2
This comparative example provides a copper ion (Cu) 2+ ) A method for treating wastewater, which is different from comparative example 1 only in that the object to be treated is replaced with a wastewater containing 100mg/L of Cu 2+ The waste water of (2).
Comparative example 3
This comparative example provides a lead ion (Pb) 2+ ) A method for treating wastewater, which comprises, with reference to comparative example 1, only replacing the object to be treated with a wastewater containing 100mg/LPb 2+ The wastewater of (2).
Comparative example 4
This comparative example provides a mercury ion (Hg) containing solution 2+ ) A method for treating wastewater, which is different from comparative example 1 only in that the object to be treated is replaced with a wastewater containing 100mg/LHg 2+ The waste water of (2).
Comparative example 5
This comparative example provides a method for treating As (iii) -containing wastewater, which is different from comparative example 1 only in that the object of treatment is replaced with wastewater containing 100mg/LAs (iii).
Comparative example 6
This comparative example provides a copper ion (Cu) 2+ ) A method for treating wastewater, which is different from comparative example 1 in that the object to be treated is replaced with a wastewater containing 100mg/L Cu 2+ The pH value of the wastewater is adjusted to 5.
Comparative example 7
This comparative example provides a copper ion (Cu) 2+ ) A method for treating wastewater, which is different from comparative example 1 in that the object to be treated is replaced with a wastewater containing 1000mg/L of Cu 2+ The amount of the added amyloid protein fiber in the wastewater of (1) is 0.1g.
Comparative example 8
This comparative example provides a copper ion (Cu) 2+ ) A method for treating wastewater, which is different from comparative example 1 in that the object to be treated is replaced with a wastewater containing 100mg/L of Cu 2+ The water bath heating temperature of the wastewater of (1) is 70 ℃.
Examples of the experiments
In order to verify the removal effect of the wastewater treatment method based on the beta-lactoglobulin provided by the invention on heavy metal ions/arsenic, the wastewater treated according to the methods in examples 1 to 8 and comparative examples 1 to 8 is respectively collected, the concentration of corresponding harmful ions (Ni, cu, pb, hg, as) in the treated wastewater is measured by adopting an inductively coupled plasma emission method (Agilent 710 ICP-OES), and the adsorption capacity is calculated by the following calculation method:
wherein Q e (mg/g) is the adsorption capacity of harmful ions, C 0 (mg/L) and C e (mg/L) is the concentration before harmful ion adsorption and after adsorption equilibrium is reached, V (mL) is the volume of the reaction solution, and m (g) is the mass of the adsorbent. The results are shown in Table 1.
TABLE 1 Effect of treating harmful ions in examples 1 to 8 and comparative examples 1 to 8
As shown in table 1, the methods of preparing amyloid protein fibers in situ in wastewater (examples 1 to 8) have higher treatment efficiency for harmful ions and better treatment effect than the methods of directly throwing prepared amyloid protein fibers into wastewater (comparative examples 1 to 8). The method provided by the invention can be used for containing Ni 2+ 、Cu 2+ 、Pb 2+ 、Hg 2+ And the As (III) wastewater treatment is simple to operate, green and pollution-free.
As shown in fig. 1 and 2, the present inventors also observed the amyloid fibrils produced in situ in the wastewater of example 1 and the amyloid fibrils prepared in comparative example 1, and analyzed the causes of the difference in morphology between them: in the case of in situ preparation, the presence of heavy metal ions interferes with the pathway of amyloid fiber conversion from the original protein, preferentially occupies binding sites during the uncoiling process of the fiber conversion (e.g., exposed binding sites cys-121 during uncoiling), and also serves as a site of action for fiber formation, so that the proportion of the generated fiber secondary structure is changed, for example, the folding structure is increased from 30.89% to 34.36% of the original folding structure, the proportion of the random coil secondary structure is reduced from 29.17% to 27.6%, the length of the formed fiber is reduced compared with the original fiber, and the morphology is also different.
Therefore, based on different conditions for forming the amyloid fiber, the length and the shape of the fiber obtained by in-situ preparation and ex-situ preparation are different, experiments show that the treatment efficiency of the amyloid fiber prepared in situ is obviously increased, and compared with ex-situ preparation, the method can avoid pollution caused by acidic waste liquid generated in the process of preparing the amyloid fiber, and has important application value.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. This need not be, nor should it be exhaustive of all embodiments. And obvious variations or modifications derived therefrom are intended to be within the scope of the invention.
Claims (9)
1. Based onβ-a method for the treatment of lactoglobulin wastewater, characterized in that it comprises the following steps:
will be provided withβ-adding lactoglobulin into wastewater, adjusting the pH value of the wastewater to 2-5, generating amyloid protein fibers in situ in the wastewater after heating treatment to adsorb harmful ions in the wastewater, and removing the amyloid protein fibers adsorbing the harmful ions in the wastewater through solid-liquid separation, wherein the heating treatment is to heat the wastewater for 4-6 hours at 70-90 ℃, and the wastewater contains at least one harmful ion of nickel ions, copper ions, lead ions, mercury ions and arsenic.
2. The method according to claim 1β-a method for treating a wastewater containing lactoglobulin, characterized in that said wastewater is treated by the volume of said wastewaterβThe dosage of lactoglobulin is 1 to 20 g/L.
3. The base of claim 1β-a method for treating wastewater containing lactoglobulin, characterized in that the solid-liquid separation is performed by centrifugation and/or membrane filtration.
4. The method according to claim 3βA method for treating wastewater of lactoglobulin, which is characterized in that the centrifugation is carried out for 5 to 10min under the condition of 7000 to 12000 rpm.
5. The base of claim 3βA method for treating lactoglobulin-containing wastewater, characterized in that the membrane filtration is performed using a 0.22 μm or 0.45 μm organic filter membrane or filter head.
6. The method according to claim 1βA wastewater treatment method of lactoglobulin, which is characterized in that 1 to 6mol/L HCl is added to regulate wastewaterThe pH value of (1).
7. The method according to claim 1β-a method for treating a wastewater containing lactoglobulin, characterized in thatβ-lactoglobulin is purified, said purification process comprising:
(1) To be purifiedβ-lactoglobulin is dissolved in ultrapure water and centrifuged;
(2) Taking the supernatant obtained by centrifugation in the step (1), and adjusting the pH value;
(3) Filtering to remove insoluble protein;
(4) Dialyzing to remove ions;
(5) Adjusting the pH of the solution, and freeze-drying to obtain purified productβLactoglobulin.
8. The method according to claim 7βA process for the treatment of lactoglobulin-containing waste water,
in step (1), the compound to be purifiedβ-dissolving lactoglobulin in 5 to 10% by mass in ultrapure water; the centrifugation is carried out for 5 to 10min at the temperature of 4 ℃ and the rotating speed of 7000 to 12000 rpm;
in the step (2), adjusting the pH value of the supernatant to 2 by using 1 to 6mol/L HCl solution;
in step (3), removing insoluble proteins using a filter;
in the step (4), removing ions by using a 3 to 8kDa dialysis bag until the pH value of the solution reaches 4 to 5;
in the step (5), the pH value of the solution is adjusted to 2 to 5.
9. The method according to claim 1β-a method for treating wastewater containing lactoglobulin, characterized in that arsenic contained in the wastewater is trivalent arsenic.
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| CL2015001254A1 (en) * | 2015-05-08 | 2015-09-11 | Cyra Consultora Limitada | Chelating arsenic composition. |
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