CN115286119A - Microorganism strengthening agent using mineral/biomass as carrier for removing hexavalent chromium and preparation method thereof - Google Patents
Microorganism strengthening agent using mineral/biomass as carrier for removing hexavalent chromium and preparation method thereof Download PDFInfo
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- CN115286119A CN115286119A CN202210857306.3A CN202210857306A CN115286119A CN 115286119 A CN115286119 A CN 115286119A CN 202210857306 A CN202210857306 A CN 202210857306A CN 115286119 A CN115286119 A CN 115286119A
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
- C02F2101/22—Chromium or chromium compounds, e.g. chromates
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Abstract
The invention discloses a microorganism strengthening agent for removing hexavalent chromium by taking minerals/biomass as a carrier and a preparation method thereof, wherein the microorganism strengthening agent is prepared from the following raw materials: clay minerals, waste biomass, microbial feedstocks. The invention combines the waste biomass material and the clay mineral, and adopts the immobilization technology to coat microbial strains, thereby providing long-acting nutrient sources for the microbes, increasing the biomass and improving the capability of removing hexavalent chromium.
Description
Technical Field
The invention belongs to a microorganism strengthening medicament for removing hexavalent chromium by taking minerals/biomasses as carriers and a preparation method thereof, and particularly relates to a microorganism strengthening medicament for removing hexavalent chromium by taking minerals/biomasses as carriers and a preparation method thereof.
Background
The traditional method for treating heavy metal by adopting a flocculating agent, an ion exchange method, an electrochemical method and a biomass adsorbent has the advantages of high cost, narrow application range, inconsistent stability and easy secondary pollution. The microorganism strengthening technology is characterized in that nutrient sources such as a carbon source and a nitrogen source and an electron donor are added, so that the microorganism biomass can be increased, and heavy metals in water and soil can be repaired. Exogenous nutrition addition can assist the growth of microorganisms, but the structure is simpler, the time efficiency is not long, and the soil remediation treatment is easy to be washed by rainwater. Clay minerals are powerful adsorbents and can act as electron donors to provide nutrients to microorganisms.
With the development of industrialization, the heavy metal content in soil and water sources in many chemical plants, industrial gathering areas and the periphery exceeds the standard, serious damage is caused to plants and animals, and finally, the heavy metal enters human bodies in a food chain manner, so that various diseases are caused. The microbial remediation method is an environment-friendly technology. The microorganism stimulator can utilize nutrient substrates, surfactants and organic modifiers added from external sources to strengthen microorganisms.
Disclosure of Invention
The invention aims to provide a microorganism strengthening agent for removing hexavalent chromium by taking minerals/biomasses as carriers and a preparation method thereof, and solves the problem of low efficiency of removing hexavalent chromium by microorganisms in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a microorganism strengthening medicament taking minerals/biomass as carriers for removing hexavalent chromium comprises the following steps:
step 1, placing stenotrophomonas in an LB culture medium to obtain a microbial liquid;
step 2, processing the waste biomass raw material by sodium hydroxide, drying in the sun, smashing and sieving;
step 3, grinding and sieving the clay mineral raw material, modifying the clay mineral, drying and finally sieving;
step 4, preparing a sodium alginate solution, and mixing the raw materials in the step 2 and the step 3 into the sodium alginate solution according to a proportion; sterilizing at 120 ℃ for 15min, cooling to room temperature, adding the microbial liquid obtained in the step 1, dripping the microbial liquid into a 4% calcium chloride solution by using an injector, crosslinking for 8h, and washing with normal saline to obtain immobilized beads;
step 5, gelatinizing starch at 80 ℃ to prepare starch solution, pouring gelatin powder into distilled water to prepare gelatin solution, and mixing the starch solution and the gelatin solution according to the weight ratio of 9:1, sterilizing for 15min at 120 ℃, and placing the immobilized beads obtained in the step 4 in the mixed solution for crosslinking for 12h to obtain the microbial enhancing agent.
Preferably, the concentration of the sodium hydroxide solution in the step 2 is 0.2-2%.
Preferably, the clay mineral raw material in the step 3 is kaolin.
Preferably, the ratio of the waste biomass raw material treated by sodium hydroxide to the modified clay mineral in the step 4 is 3:1.
preferably, the method for modifying the clay mineral in the step 3 is as follows: adding chitosan into acetic acid solution, stirring to completely dissolve, adding clay mineral and prepared chitosan acetic acid solution into distilled water, stirring, adjusting pH of the solution to 10, adding glutaraldehyde solution, crosslinking for 24h, filtering, washing with distilled water until pH is 7, and naturally drying.
Preferably, the amount ratio of the clay mineral to the distilled water is 1g:10ML.
The invention adopts another technical scheme that: a microorganism strengthening agent taking minerals/biomass as a carrier for removing hexavalent chromium is prepared from the following raw materials: modified clay minerals, waste biomass raw materials treated by sodium hydroxide, stenotrophomonas, sodium alginate, calcium chloride, starch solution and gelatin solution.
The invention has the beneficial effects that: the method combines the waste biomass material and the clay mineral, and adopts the immobilization technology to coat microbial strains, thereby providing a long-acting nutrient source for the microbes, increasing the biomass and improving the capability of removing hexavalent chromium; the invention has the characteristics of long effect, low cost, green, changing waste into valuable and treating pollution by waste.
Drawings
FIG. 1 is a schematic flow chart of a method for preparing a microorganism-enhanced medicament for removing hexavalent chromium by using minerals/biomass as carriers according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1
A microorganism strengthening agent taking minerals/biomass as a carrier for removing hexavalent chromium is prepared from the following raw materials: modified clay minerals, waste biomass raw materials treated by sodium hydroxide, stenotrophomonas, sodium alginate, calcium chloride, starch solution and gelatin solution.
As shown in fig. 1, a method for preparing a microorganism-enhanced medicament for removing hexavalent chromium using minerals/biomass as a carrier includes the following steps:
step 1, putting the microbial strains in an LB culture medium for independent culture.
And 2, taking waste biomass as a raw material. Treating straws, peanut shells and fruit peels with 0.2-2% of sodium hydroxide, drying in the sun, crushing, sieving, and mixing according to the proportion of the straws, the peanut shells and the fruit peels being 3:3: 1. and (4) uniformly mixing to obtain the mixed biomass material.
And 3, taking kaolin as a clay mineral raw material, grinding and sieving, and modifying the clay mineral. The method comprises the following specific steps: to 50ml of 2% acetic acid solution, 1g of chitosan was added, and the mixture was stirred with a magnetic stirrer until completely dissolved. Adding 10g of clay mineral and prepared chitosan solution into 100ml of distilled water, stirring for 30min, adjusting the pH of the mixed solution to 10 by using 0.1mol/L sodium hydroxide solution, adding glutaraldehyde solution, crosslinking for 24h, washing with distilled water after suction filtration until the pH is 7, drying and finally sieving.
Step 4, preparing a 4% sodium alginate solution, and mixing the raw materials in the step 2 and the step 3 according to the proportion of 1: 1 (mixed biomass material: modified clay mineral) is mixed into sodium alginate solution in proportion and is mixed evenly by an ultrasonic cleaner; sterilizing at 120 ℃ for 15min, cooling to room temperature, adding 1ml of the microbial solution obtained in the step (1), dripping into a 4% calcium chloride solution by using an injector, crosslinking for 8h, and washing with 0.7-1% normal saline to obtain the immobilized pellet.
Step 5, gelatinizing starch at 80 ℃, preparing a 2% starch solution, pouring gelatin powder into distilled water, preparing a 1% gelatin solution, and mixing the starch solution and the gelatin solution according to a ratio of 9:1, adding 2% glycerol solution, sterilizing at 120 ℃ for 15min, placing the immobilized beads obtained in the step 4 in the mixed solution, and crosslinking for 12h to obtain the microbial strengthening agent.
Example 2
A microorganism strengthening agent taking minerals/biomass as a carrier for removing hexavalent chromium is prepared from the following raw materials: modified clay minerals, waste biomass raw materials treated by sodium hydroxide, stenotrophomonas, sodium alginate, calcium chloride, starch solution and gelatin solution.
As shown in fig. 1, a method for preparing a microbe-strengthening medicament using minerals/biomass as carriers for removing hexavalent chromium comprises the following steps:
step 1, putting the microbial strains in an LB culture medium for independent culture.
And 2, taking waste biomass as a raw material. Treating straws, peanut shells and fruit peels with 0.2-2% of sodium hydroxide, drying in the sun, smashing, and sieving, wherein the ratio of the straws to the peanut shells to the fruit peels is 3:3: 1. and (4) uniformly mixing to obtain the mixed biomass material.
And 3, taking kaolin as a clay mineral raw material, grinding and sieving the kaolin, and modifying the clay mineral. The method comprises the following specific steps: to 50ml of 2% acetic acid solution, 1g of chitosan was added, and the mixture was stirred with a magnetic stirrer until completely dissolved. Adding 10g of clay mineral and the prepared chitosan solution into 100ml of distilled water, stirring for 30min, adjusting the pH of the mixed solution to 10 by using 0.1mol/L sodium hydroxide solution, adding glutaraldehyde solution, crosslinking for 24h, washing by using distilled water after suction filtration until the pH is 7, drying and finally sieving.
Step 4, preparing a 4% sodium alginate solution, and mixing the raw materials in the step 2 and the step 3 according to the ratio of 2: 1 (mixing biomass material: modified clay mineral) is mixed into sodium alginate solution in proportion and is mixed evenly by an ultrasonic cleaner; sterilizing at 120 ℃ for 15min, cooling to room temperature, adding 1ml of the microbial solution obtained in the step (1), dripping into a 4% calcium chloride solution by using an injector, crosslinking for 8h, and washing with 0.7-1% normal saline to obtain the immobilized pellet.
Step 5, gelatinizing starch at 80 ℃ to prepare a 2% starch solution, pouring gelatin powder into distilled water to prepare a 1% gelatin solution, and mixing the two solutions according to the weight ratio of 9:1, adding 2% glycerol solution, sterilizing at 120 ℃ for 15min, placing the immobilized beads obtained in the step 4 in the mixed solution, and crosslinking for 12h to obtain the microbial strengthening agent.
Example 3
A microorganism strengthening agent taking minerals/biomass as a carrier for removing hexavalent chromium is prepared from the following raw materials: modified clay minerals, waste biomass raw materials treated by sodium hydroxide, stenotrophomonas, sodium alginate, calcium chloride, starch solution and gelatin solution.
As shown in fig. 1, a method for preparing a microbe-strengthening medicament using minerals/biomass as carriers for removing hexavalent chromium comprises the following steps:
step 1, putting the microbial strains in an LB culture medium for independent culture.
And 2, taking waste biomass as a raw material. Treating straws, peanut shells and fruit peels with 0.2-2% of sodium hydroxide, drying in the sun, smashing, and sieving, wherein the ratio of the straws to the peanut shells to the fruit peels is 3:3: 1. and (4) uniformly mixing to obtain the mixed biomass material.
And 3, taking kaolin as a clay mineral raw material, grinding and sieving the kaolin, and modifying the clay mineral. The method comprises the following specific steps: to 50ml of 2% acetic acid solution, 1g of chitosan was added, and the mixture was stirred with a magnetic stirrer until completely dissolved. Adding 10g of clay mineral and the prepared chitosan solution into 100ml of distilled water, stirring for 30min, adjusting the pH of the mixed solution to 10 by using 0.1mol/L sodium hydroxide solution, adding glutaraldehyde solution, crosslinking for 24h, washing by using distilled water after suction filtration until the pH is 7, drying and finally sieving.
Step 4, preparing a 4% sodium alginate solution, and mixing the raw materials in the step 2 and the step 3 according to the ratio of 3:1 (mixing biomass material: modified clay mineral) is mixed into sodium alginate solution in proportion and is mixed evenly by an ultrasonic cleaner; sterilizing at 120 ℃ for 15min, cooling to room temperature, adding 1ml of the microbial liquid obtained in the step 1, dripping the microbial liquid into a 4% calcium chloride solution by using an injector, crosslinking for 8h, and washing with 0.7-1% physiological saline to obtain the immobilized pellet.
Step 5, gelatinizing starch at 80 ℃ to prepare a 2% starch solution, pouring gelatin powder into distilled water to prepare a 1% gelatin solution, and mixing the two solutions according to the weight ratio of 9:1, adding 2% glycerol solution, sterilizing at 120 ℃ for 15min, placing the immobilized beads obtained in the step 4 in the mixed solution, and crosslinking for 12h to obtain the microbial strengthening agent.
Experimental example 1
The influence of the microbial strengthening agent prepared in the embodiment 1 on hexavalent chromium pollution in a water body is as follows:
step 1, preparing 50g/L hexavalent chromium mother liquor by using potassium dichromate for later use.
And 2, pouring the mother solution into the sterilized 1L of LB culture solution, and preparing the initial hexavalent chromium concentration of 100 mg/L and 300mg/L. The microbial enhancing agent of example 1 was placed in a 100mL hexavalent chromium conical flask with an initial concentration of 100 mg/L and 300mg/L, and the hexavalent chromium removal rate was measured in parallel with a control of Stenotrophomonas as a pure bacteria and a control of a blank agent without bacteria (except that Stenotrophomonas was not contained, the other components were the same as the microbial enhancing agent of example 1), as shown in table 1.
TABLE 1
As can be seen from Table 1, the removal rate of hexavalent chromium by the microbial enhancing agent is higher than that of Stenotrophomonas (Stenotrophoromonas) by adopting the method of the invention; when the initial concentration of hexavalent chromium is 100 and 300mg/L, the removal rate of the microorganism strengthening medicament to the hexavalent chromium is 88 percent and 59.7 percent respectively, and the removal rate is increased by 4.3 percent and 3.4 percent respectively compared with the removal rate of Stenotrophomonas.
Experimental example 2
The microorganism strengthening agent prepared in the embodiment 2 has the following experimental steps on the influence of hexavalent chromium pollution in a water body:
step 1, preparing 50g/L hexavalent chromium mother liquor by using potassium dichromate for later use.
And 2, pouring the mother solution into the sterilized 1L of LB culture solution, and preparing the initial hexavalent chromium concentration of 100 mg/L and 300mg/L. The removal rate of hexavalent chromium was measured by placing the microbial enhancing agent of example 2 (same as the preparation method of example 1) in a 100mL hexavalent chromium medium Erlenmeyer flask with an initial concentration of 100 mg/L and using Stenotrophomonas as a pure control and a blank agent without added bacteria (same as the microbial enhancing agent of example 1 except that Stenotrophoromonas was not included) as a parallel control, as shown in Table 2.
TABLE 2
As can be seen from Table 2, the removal rate of hexavalent chromium by the microbial enhancing agent is higher than that of Stenotrophomonas (Stenotrophoromonas) by adopting the method of the invention; when the initial concentration of hexavalent chromium is 100 and 300mg/L, the removal rate of the microorganism strengthening medicament to the hexavalent chromium is 95.6 percent and 65.2 percent respectively, and the removal rate is increased by 11.9 percent and 8.9 percent respectively compared with the removal rate of Stenotrophomonas.
Experimental example 3
The microorganism strengthening agent prepared in example 3 has an influence on hexavalent chromium pollution in a water body, and the experimental steps are as follows:
step 1, preparing 50g/L hexavalent chromium mother liquor by using potassium dichromate for later use.
And 2, pouring the mother solution into sterilized 1L of LB culture solution, and preparing the initial hexavalent chromium concentration of 100 mg/L and 300mg/L. The removal rate of hexavalent chromium was measured by placing the microbial enhancing agent of example 3 (same as the preparation method of example 1) in a 100mL hexavalent chromium medium Erlenmeyer flask with an initial concentration of 100 mg/L and using Stenotrophomonas as a pure control and a blank agent without added bacteria (same as the microbial enhancing agent of example 1 except that Stenotrophoromonas was not included) as a parallel control, as shown in Table 3.
TABLE 3
As can be seen from Table 3, the removal rate of hexavalent chromium by the microbial enhancing agent is higher than that of Stenotrophomonas (Stenotrophoromonas) by the method of the present invention; when the initial concentration of hexavalent chromium is 100 and 300mg/L, the removal rate of the microbe strengthening agent to hexavalent chromium is 98 percent and 70.3 percent respectively, and the removal rate is increased by 14.3 percent and 11.1 percent respectively compared with the removal rate of Stenotrophomonas (Stenotrophoromonas).
It can be seen from tables 1 to 3 that the ingredient ratio of example 3 is optimal.
The invention combines the waste biomass material and the clay mineral, and adopts the immobilization technology to coat microbial strains, thereby providing long-acting nutrient sources for the microbes, increasing the biomass and improving the capability of removing hexavalent chromium. The invention has the characteristics of long effect, low cost, green, changing waste into valuable and treating pollution by waste.
While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.
Claims (7)
1. A preparation method of a microorganism strengthening agent taking minerals/biomass as a carrier for removing hexavalent chromium is characterized by comprising the following steps:
step 1, placing stenotrophomonas in an LB culture medium to obtain a microbial liquid;
step 2, processing the waste biomass raw material by sodium hydroxide, drying in the sun, smashing and sieving;
step 3, grinding and sieving the clay mineral raw material, modifying the clay mineral, drying and finally sieving;
step 4, preparing a sodium alginate solution, and mixing the raw materials in the step 2 and the step 3 into the sodium alginate solution in proportion; sterilizing at 120 ℃ for 15min, cooling to room temperature, adding the microbial inoculum obtained in the step 1, dripping into a 4% calcium chloride solution by using an injector, crosslinking for 8h, and washing by using normal saline to obtain immobilized beads;
step 5, gelatinizing starch at 80 ℃ to prepare starch solution, pouring gelatin powder into distilled water to prepare gelatin solution, and mixing the starch solution and the gelatin solution according to the weight ratio of 9:1, sterilizing for 15min at 120 ℃, and placing the immobilized beads obtained in the step 4 in the mixed solution for crosslinking for 12h to obtain the microbial enhancing agent.
2. The method of claim 1, wherein the concentration of sodium hydroxide solution in step 2 is 0.2-2%.
3. The method for preparing a microbe-strengthening chemical using minerals/biomasses as carriers for removing hexavalent chromium according to claim 2, wherein the clay mineral material in the step 3 is kaolin.
4. The method for preparing a microbe-strengthening chemical using minerals/biomasses as carriers for removing hexavalent chromium according to claim 1, wherein the raw material ratio of the waste biomass raw material subjected to the sodium hydroxide treatment in the step 4 to the modified clay minerals is 3:1.
5. the method for preparing a mineral/biomass-based microbial enhancing agent for hexavalent chromium removal according to claim 1, wherein the clay mineral modification in the step 3 is performed by: adding chitosan into acetic acid solution, stirring to completely dissolve, adding clay mineral and prepared chitosan acetic acid solution into distilled water, stirring, adjusting pH of the solution to 10, adding glutaraldehyde solution, crosslinking for 24h, filtering, washing with distilled water until pH is 7, and naturally drying.
6. The method of claim 5, wherein the ratio of the clay mineral to distilled water is 1g:10ML.
7. The mineral/biomass-based microbial enhancing agent for hexavalent chromium removal according to any one of claims 1 to 6, which is prepared from the following raw materials: modified clay minerals, waste biomass raw materials treated by sodium hydroxide, stenotrophomonas, sodium alginate, calcium chloride, starch solution and gelatin solution.
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