CN114084967B - Lucid ganoderma fermented Shuanghuanglian mushroom dreg for repairing polycyclic aromatic hydrocarbon benzo [ a ] pyrene polluted water environment and application thereof - Google Patents
Lucid ganoderma fermented Shuanghuanglian mushroom dreg for repairing polycyclic aromatic hydrocarbon benzo [ a ] pyrene polluted water environment and application thereof Download PDFInfo
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- CN114084967B CN114084967B CN202111336921.1A CN202111336921A CN114084967B CN 114084967 B CN114084967 B CN 114084967B CN 202111336921 A CN202111336921 A CN 202111336921A CN 114084967 B CN114084967 B CN 114084967B
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- aromatic hydrocarbon
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- FMMWHPNWAFZXNH-UHFFFAOYSA-N Benz[a]pyrene Chemical compound C1=C2C3=CC=CC=C3C=C(C=C3)C2=C2C3=CC=CC2=C1 FMMWHPNWAFZXNH-UHFFFAOYSA-N 0.000 title claims abstract description 122
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 title claims abstract description 63
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 60
- 241000222336 Ganoderma Species 0.000 title claims description 8
- 235000001674 Agaricus brunnescens Nutrition 0.000 title description 3
- 239000009254 shuang-huang-lian Substances 0.000 title description 3
- 240000008397 Ganoderma lucidum Species 0.000 claims abstract description 73
- 235000001637 Ganoderma lucidum Nutrition 0.000 claims abstract description 43
- 241000233866 Fungi Species 0.000 claims abstract description 36
- 102000004190 Enzymes Human genes 0.000 claims abstract description 30
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 8
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- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 229910052698 phosphorus Inorganic materials 0.000 claims description 6
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- 240000006023 Trichosanthes kirilowii Species 0.000 description 2
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- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 2
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- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 description 1
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- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- DRKYTUDHOKREMS-UHFFFAOYSA-N ent 27,313 Chemical group ClC1=C(Cl)C2(Cl)C3CCC(Cl)C3C1(Cl)C2(Cl)Cl DRKYTUDHOKREMS-UHFFFAOYSA-N 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- C—CHEMISTRY; METALLURGY
- 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
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
-
- C—CHEMISTRY; METALLURGY
- 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
-
- 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
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
- C02F2101/327—Polyaromatic Hydrocarbons [PAH's]
Abstract
The invention relates to a ganoderma lucidum fermentation double coptis fungus dreg for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene and application thereof, which are characterized in that the metabolic activity of polycyclic aromatic hydrocarbon degrading bacteria in the environment is improved by influencing the survival and degradation performance of microorganisms in the environment, the polycyclic aromatic hydrocarbon is degraded, then the water environment polluted by polycyclic aromatic hydrocarbon BaP is repaired by extracellular enzyme and adsorption capacity contained in the fungus dreg, the problem of protecting the water environment is effectively realized, ganoderma lucidum seed liquid is inoculated in a culture medium of the double coptis fungus dreg, the ganoderma lucidum is cultured, the rest culture medium residue is ganoderma lucidum fungus dreg, the ganoderma lucidum fungus dreg is added into a water body polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene at the pH of 4.0-8.5 and the temperature of 20-50 ℃ by 5-10 percent of volume.
Description
Technical Field
The invention relates to the technical field of water environment protection, in particular to ganoderma lucidum double coptis chinensis fermentation fungus residues for repairing a water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene and application thereof.
Background
Polycyclic aromatic hydrocarbons (Polycyclic aromatic hydrocarbons, PAHs) are a very typical class of organic pollutants consisting of two or more carbon and hydrogen atoms, and are highly hydrophobic, stable and highly intractable, with toxicity, mutability and carcinogenicity severely threatening the health of humans. In the past 30 years, various methods and technologies are applied to repair of PAHs pollution, such as physical methods, chemical oxidation and biological repair, and currently, research on a polycyclic aromatic hydrocarbon removal method in water environment has achieved certain effects at home and abroad. Among the conventional control measures effective in the treatment are adsorption method, bioremediation method, photolysis method, advanced oxidation method, etc.
Benzo [ a ] pyrene (BaP) is a polycyclic aromatic hydrocarbon composed of five benzene rings, and is one of the most carcinogenic polycyclic aromatic hydrocarbons. It has very low water solubility, vapor pressure and high octanol water partition coefficient, and BaP tends to be non-aqueous and low in microbial utilization, so BaP is more likely to accumulate in the environment. It is known from a large number of references that PAHs are in practical use
The removal process is mainly a biological method or a physical method, wherein the biological method is considered as an economic and effective method for degrading the polycyclic aromatic hydrocarbon in the polluted environment by utilizing microorganisms, but researches find that the polycyclic aromatic hydrocarbon with more than four rings has strong antimicrobial activity degradation capability, and exogenous microorganisms are influenced by the polluted environment factors (such as temperature, pH value, oxygen content, nutrient content and the like) to cause larger degradation effect difference in different media. It is widely believed that pH is the primary factor affecting the efficiency of bioremediation of polycyclic aromatic hydrocarbons. Since the optimal pH values for different species of microorganisms are different, the pH values affect the growth and metabolic activity of the microorganisms. In addition, the temperature has a critical influence on the bioremediation system of the polycyclic aromatic hydrocarbon. In a proper temperature range, the activity of the microorganism increases with the increase of temperature, because it can promote the metabolism of the microorganism and the activity of enzymes, thereby accelerating the bioremediation process of polycyclic aromatic hydrocarbon. In view of the limitation of microorganism reinforced repair, the invention utilizes the combination of biological method and physical method to achieve the effect of improving the degradation of polycyclic aromatic hydrocarbon according to the defects of the prior art. The fungus dreg is a waste culture medium after fruiting of edible fungi, china is a main production country of the edible fungi, and accounts for 75% of the total world yield. The fungus dregs contain various nutrients including organic matters, nitrogen, phosphorus and potassium, and various microorganisms (such as bacillus, bacillus brevis, arthrobacter and microbacterium), and the nutrients in the fungus dregs can be used for environmental improvement, such as changing the pH value and the nutrient content in the environment, and the like, and all the nutrients can promote the degradation of polycyclic aromatic hydrocarbon, so that the fungus dregs have the dual advantages of biological stimulation and biological enhancement. The biological stimulation means that nitrogen sources and phosphorus sources are added to activate microorganisms which are stored in polluted environment, so that the degradation effect of the polycyclic aromatic hydrocarbon is improved; the bio-enhancement refers to a method for degrading PAHs by adding exogenous polycyclic aromatic hydrocarbon degrading bacteria on the basis of biological stimulation, and in addition, various extracellular enzymes including lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (LaC) are secreted in the growth and development process of the edible fungi, and the extracellular enzymes accumulate in bacterial residues and can degrade organic pollutants.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide ganoderma lucidum fermentation double coptis fungus dreg for repairing the water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene and application thereof, and the method can improve the metabolic activity of polycyclic aromatic hydrocarbon degrading bacteria in the environment by influencing the survival and degradation performance of microorganisms in the environment, degrade the polycyclic aromatic hydrocarbon, and repair the water environment polluted by polycyclic aromatic hydrocarbon BaP by extracellular enzyme and adsorption capacity contained in the fungus dreg, thereby effectively realizing the protection of the water environment.
According to the technical scheme, ganoderma lucidum fermentation double coptis fungus residues for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene are prepared by inoculating ganoderma lucidum seed liquid into double coptis fungus residue culture medium, culturing, collecting ganoderma lucidum, and obtaining the rest culture medium residues, wherein the ganoderma lucidum fungus residues contain lignin-decomposing enzyme, organic matters, nitrogen, phosphorus, potassium and various microorganisms, the lignin-decomposing enzyme can secrete lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (LaC) of various extracellular enzymes, and the extracellular enzymes have good degradation effects on various organic pollutants, in particular to polycyclic aromatic hydrocarbon benzo [ a ] pyrene;
the ganoderma lucidum solid fermentation medium is prepared from 450g of rhizoma coptidis dregs, 50g of corn flour, 15g of bran, 5g of gypsum, 5g of sucrose, 0.06g of monopotassium phosphate, 0.06g of magnesium sulfate and 0.06g of vitamin B;
the medicinal residue of the double coptis is medicinal residue remained after decoction of a composition consisting of baical skullcap root, honeysuckle and weeping forsythiae capsule (known technology);
the ganoderma lucidum dreg is added into a water body polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene in a weight volume of 5-10% under the conditions of an acid-base environment with the pH value of 4.0-8.5 and the temperature of 20-50 ℃, the weight volume is calculated in g, the volume is calculated in mL, the survival and degradation performance of microorganisms for degrading the polycyclic aromatic hydrocarbon benzo [ a ] pyrene can be effectively improved, the polycyclic aromatic hydrocarbon benzo [ a ] pyrene in the water body with the polycyclic aromatic hydrocarbon benzo [ a ] pyrene pollution concentration of 50mg/L is removed, the ganoderma lucidum dreg shows better degradation effect especially under the conditions of the pH value of slightly acidic of 4.5-5.5 and the lower temperature of 20-25 ℃, the degradation rate of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene reaches more than 90% after 7 days, and the application of the ganoderma lucidum dreg in the water environment restoration of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene pollution is realized.
The invention has the advantages of abundant raw materials, easy obtainment of fungus residues, convenient use and good effect, can effectively solve the problem of water environment restoration polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene, and is a great innovation for restoring the water body polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene, and has huge economic and social benefits.
Detailed Description
The following describes the embodiments of the present invention in detail with reference to specific cases.
The invention relates to ganoderma lucidum fungus dreg for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene, which is obtained by taking Shuanghuanglian herb dreg as a base material and fermenting residues obtained after ganoderma lucidum collection by the following method:
(1) Screening of fungus residues
And detecting the activity of lignin-decomposing enzyme on the collected ganoderma lucidum fungus residues, oyster mushroom fungus residues and mushroom fungus residues, and screening out fungus residues with higher lignin-decomposing enzyme for subsequent degradation tests. 10.0g of the bacterial residues are weighed into a 250mL conical flask, 25mL of distilled water is added, and then the conical flask is put into a shaking table for shake culture at 37 ℃ and 180rpm for 1h. After shaking for 1h, 1mL of fungus dreg liquid is taken in a centrifuge tube, the centrifuge tube is centrifuged at 12000rpm for 5min, and the supernatant is taken to detect lignin enzyme activity;
through detection, the laccase activity of the ganoderma lucidum fungus dreg is 468.2U/g, the manganese peroxidase activity is 699.52U/g, and compared with other fungus dreg, the lignin decomposition enzyme activity of the ganoderma lucidum fungus dreg is higher, and the enzyme activity is higher than that reported in the prior literature.
Laccase activity detection: the laccase activity is determined by adopting an ABTS method, the reaction system is 1mL, 0.5mL of tartaric acid buffer solution with pH of 4.0, 0.38mL of distilled water, 10 mu L of fermentation enzyme solution and 100 mu L of ABTS are contained, water bath is carried out at 30 ℃ for 1min, the reaction is carried out for 30s, the light absorption value is determined at 420nm, and each group is repeated for 3 times. 1 enzyme activity unit is defined as the amount of enzyme required to oxidize 1 mu mol abts per minute under the current reaction conditions.
Determination of the enzymatic Activity of manganese peroxidase: the manganese peroxidase kit produced by Solarbio was used, and the measurement reagent and the sample were sequentially added according to the instructions of the kit, and the absorbance was measured at 465nm with a spectrophotometer. 1 enzyme activity unit is defined as the amount of enzyme required to oxidize 1nmol of guaiacol per gram of sample per minute, as one enzyme activity unit.
(2) Culturing Ganoderma lucidum seed solution
Inoculating Ganoderma strain into seed culture medium, and shake culturing at 30deg.C and 180rmp with constant temperature shaking table; punching a ganoderma lucidum flat plate into a round hole with the diameter of 0.5cm, inoculating the round hole into a liquid ganoderma lucidum seed culture medium, and culturing for 3 days at the temperature of 30 ℃ and at the speed of 180 rpm;
the seed culture medium is as follows: 30g of corn flour, 15g of bean cake powder and 5g of bran, adding water, boiling for 1h, filtering by 8 layers of gauze, adding 20g of glucose, 0.2g of magnesium sulfate, 1g of monopotassium phosphate and 5g of yeast powder, fixing the volume to 1000mL, and sterilizing at 115 ℃ for 20min;
(3) Solid fermentation of ganoderma lucidum
Adding solid fermentation culture medium of Ganoderma into fungus bags, wherein each Ganoderma bag has weight of 500g, adding water to volume water content of 60%, sterilizing at 121deg.C for 2 hr, and naturally cooling;
ganoderma lucidum solid fermentation culture medium: 450g of double coptis root dregs, 50g of corn meal, 15g of bran, 5g of gypsum, 5g of sucrose, 0.06g of monopotassium phosphate, 0.06g of magnesium sulfate and 0.06g of vitamin B;
inoculating ganoderma lucidum seed liquid into ganoderma lucidum fungus bags, wherein the inoculation amount is 2% of the weight volume of a culture medium, the weight volume is calculated in terms of g, the liquid is calculated in terms of mL, after inoculation is completed, the liquid is transferred into a darkroom to generate hyphae, the fungus bags full of hyphae are transferred into an illumination incubator to perform ganoderma lucidum fruiting management, the temperature of the incubator is kept at 26-28 ℃, the humidity is 85-90%, the fungus handles are used for keeping out wind (without ventilation) in growth period, ventilation is performed in time when the ganoderma lucidum is sliced, the illumination intensity is 180-220Lx, ganoderma lucidum is harvested in the slicing period of 450-550Lx, and the solid fermentation culture medium residues in the fungus bags after ganoderma lucidum harvesting are ganoderma lucidum fungus residues.
The application of the ganoderma lucidum fungus dreg for repairing the water environment polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene, which is obtained by the method, in repairing the water environment polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene, wherein the initial pollution concentration of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene is 50mg/L, the water environment is an acid-base environment with pH of 4.0-8.5, and the temperature is 20-50 ℃;
through tests, the ganoderma lucidum fungus residues contain lignin-decomposing enzymes, organic matters, nitrogen, phosphorus, potassium, various microorganisms and the like, wherein the lignin-decomposing enzymes comprise lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (LaC), and the extracellular enzymes have proved to have good degradation effects on various organic pollutants.
The ganoderma lucidum fungus dreg can remove polycyclic aromatic hydrocarbon benzo [ a ] pyrene under acidic and alkaline water environment and different temperature conditions, wherein the pH value of the acid-base environment is 4.0-8.5, and the different temperature is 20-50 ℃.
The method is characterized in that ganoderma lucidum residues are subjected to liquid chromatography detection after 5 days degradation under the conditions of different acid-base water environments (pH 4.0-8.5) and culture temperatures of 37 ℃, the initial concentration of benzo [ a ] pyrene is 50mg/L, the removal rate of benzo [ a ] pyrene is 60% -80%, the initial concentration of benzo [ a ] pyrene is 50mg/L under the culture conditions of different temperatures (20-50 ℃) and pH6.0, and the removal rate of benzo [ a ] pyrene is more than 90% after 5 days degradation.
The ganoderma lucidum fungus dreg can efficiently degrade high molecular weight polycyclic aromatic hydrocarbon benzo [ a ] pyrene under acidic and alkaline environments and different temperature conditions, wherein the acidic and alkaline environments have pH of 4.0-8.5, and the different temperatures are 20-50 ℃, so that the bioremediation of the high molecular weight polycyclic aromatic hydrocarbon benzo [ a ] pyrene in water environment is effectively solved; the ganoderma lucidum fungus dreg is a culture medium waste after fruiting of edible fungi, contains various nutrient substances including organic matters, nitrogen, phosphorus and potassium, and various microorganisms (such as bacillus, bacillus brevis, arthrobacterium and microbacterium) which can promote the degradation of polycyclic aromatic hydrocarbon, and has the triple advantages of adsorption, biological stimulation and biological reinforcement; in addition, various extracellular enzymes are secreted in the growth and development process of the edible fungi, including lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (LaC), and the extracellular enzymes accumulate in fungus residues, and can degrade organic pollutants, such as polycyclic aromatic hydrocarbon, pentachlorophenol, heptachloro, trichosanthes kirilowii and the like; finally, the ganoderma lucidum dreg is different from the traditional culture medium formula, the culture medium substrate used by the ganoderma lucidum dreg is the double coptis chinensis dreg, and the dreg is rich in crude proteins, crude fibers, amino acids and other nutrient substances and insufficiently extracted medicinal active ingredients, so that a material basis can be provided for the growth of microorganisms in the dreg substrate. And the experiment shows that the method has very good beneficial technical effects, and related data are as follows:
1. under different pH conditions, the degradation experiment of the ganoderma lucidum fungus residues on benzo [ a ] pyrene in water environment:
phosphate buffer solutions with different pH values (4.0-8.5) are prepared. Preparing benzo [ a ] pyrene standard substance mother liquor with the concentration of 1g/L by using acetone, adding 1mL of benzo [ a ] pyrene mother liquor into a 100mL bottle with a stopper, volatilizing the acetone at room temperature, adding 20mL of phosphate buffer solution with different pH values after volatilizing the acetone, then adding 1.5g of ganoderma lucidum fungus residues, and placing the ganoderma lucidum fungus residues in a constant-temperature shaking table with the temperature of 30 ℃ and 180rpm for degradation experiments, repeating each treatment for 3 times, and detecting the content of benzo [ a ] pyrene after 5 days of culture. Percent removal = (control concentration-degradation concentration)/control concentration x 100%. The Biolog method determines the functional diversity of microorganisms in contaminated water.
The pretreatment method of the liquid phase sample comprises the following steps: the extract phase is extracted by using chromatographic grade normal hexane, 10mL of normal hexane is added into each sample for extraction, vortex oscillation is carried out for 10min, after the sample is layered and stabilized, the upper organic phase is added for analysis by anhydrous sodium sulfate.
HPLC analysis: and (3) measuring the PAHs content by adopting an Agilent LC-1200 type high performance liquid chromatograph. The sample injection amount is 20 mu L, the separation column is ZORBAX SB-C18 column (0.46×150mm, agilent), the column temperature is 30deg.C, the ultraviolet detection wavelength is 254nm, the mobile phase is acetonitrile and water (volume ratio is 80%: 20%), and the flow rate is 2.0mL min -1 。
The results showed that the removal rate of benzo [ a ] pyrene was 85% or more after 5 days of liquid phase detection (averaging). Biolog
The result shows that the microbial metabolism activity in the polluted water is obviously improved after the bacterial residues are added.
2. Degradation experiment of ganoderma lucidum fungus residues on benzo [ a ] pyrene in water environment under different temperature conditions:
phosphate buffer solution of pH6.0 was prepared. Preparing benzo [ a ] pyrene standard substance mother liquor with the concentration of 1g/L by acetone, adding 1mL of benzo [ a ] pyrene mother liquor into a 100mL bottle with a stopper, volatilizing the acetone at room temperature, adding 20mL of phosphate buffer solution with the pH of 6.0 after volatilizing the acetone, then adding 1.5g of ganoderma lucidum fungus residues, comparing the ganoderma lucidum fungus residues with benzo [ a ] pyrene aqueous solution without the ganoderma lucidum fungus residues, placing the benzo [ a ] pyrene aqueous solution into a constant temperature shaking table with different temperatures (20-50 ℃), carrying out degradation experiments at 180rpm, repeating each treatment for 3 times, and detecting the content of benzo [ a ] pyrene after 5 days of culture. Percent removal = (control concentration-degradation concentration)/control concentration x 100%. The Biolog method determines the functional diversity of microorganisms in contaminated water.
The pretreatment method of the liquid phase sample comprises the following steps: the extract phase is extracted by using chromatographic grade n-hexane, 10mL of n-hexane is added into each sample for extraction, vortex oscillation is carried out for 10min, after the sample is layered and stabilized, the upper organic phase is added for analysis by taking a certain amount of anhydrous sodium sulfate.
HPLC analysis: the present study uses Agilent LC-1200 type high performance liquid chromatograph to determine PAHs content. The sample injection amount is 20 mu L, the separation column is ZORBAX SB-C18 column (0.46×150mm, agilent), the column temperature is 30deg.C, the ultraviolet detection wavelength is 254nm, the mobile phase is acetonitrile and water (volume ratio is 80%: 20%), and the flow rate is 2.0mL min -1 。
The results showed that the removal rate of benzo [ a ] pyrene was 90% or more after 5 days of liquid phase detection (averaging). The ganoderma lucidum fungus residues have good removal effect on high molecular weight polycyclic aromatic hydrocarbon under different temperature conditions, and the Biolog results show that the microbial metabolism activity in the polluted water is obviously improved after the fungus residues are added.
The invention obtains the same or similar effect as the experiment through repeated experiments, and the experiments are not detailed and listed one by one, and the experiments show that the ganoderma lucidum fungus residues can effectively remove the high molecular weight polycyclic aromatic hydrocarbon benzo [ a ] pyrene in the aqueous solution. The bacteria residue has potential application value in the aspect of repairing high molecular weight polycyclic aromatic hydrocarbon under the water environment condition, can effectively solve the problem of repairing the water environment under the conditions that the pH value of the water body is pH4.0-8.5 and the temperature is 20-50 ℃ and the pollution concentration of polycyclic aromatic hydrocarbon benzo [ a ] pyrene is 50mg/L, and realizes the protection of the water environment, and compared with the prior art, the invention has the following advantages:
firstly, the ganoderma lucidum is fermented by using the double coptis chinensis dregs, the green recycling of the high added value of the ganoderma lucidum is realized by converting the traditional Chinese medicine dregs by microorganisms, waste is changed into valuable, and technical support is provided for modernization and health sustainable development of the traditional Chinese medicine industry.
And secondly, after the ganoderma lucidum is collected, the ganoderma lucidum fungus residues are utilized to effectively degrade benzo [ a ] pyrene in the water environment, repair of polluted water is realized, waste materials are changed into valuable materials, and a green circular economy mode is realized.
In addition, the fungus residues contain a large amount of edible fungus mycelium residues, extracellular enzymes, polysaccharides, crude fibers degraded by biological enzymes, lignin which is not completely utilized and other components, so that the fungus residues are high-quality biomass resources. The nutritional ingredients are even higher than the original culture materials, and the environmental conditions can be optimized and improved, so that the metabolic activity of the original polycyclic aromatic hydrocarbon degrading bacteria in the environment is improved, and the degradation efficiency is further improved. Meanwhile, mycelium in the fungus dreg can secrete hormone substances and a series of extracellular enzymes for degrading lignocellulose, including laccase, lignin peroxidase, cellulase, xylanase and the like, during the growth and development process, and the extracellular enzymes can degrade organic pollutants, such as polycyclic aromatic hydrocarbon, pentachlorophenol, heptachloride, trichosanthes kirilowii and the like.
Meanwhile, the ganoderma lucidum fungus dreg itself can absorb certain benzo [ a ] pyrene pollutant, has triple functions of absorption, biological reinforcement (degradation) and biological stimulation, is a great innovation on the repair of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene pollution, and shows better degradation effect on the benzo [ a ] pyrene under the conditions of meta-acidity and lower temperature, the removal rate of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene can reach more than 90%, and provides powerful technical support for the repair of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene polluted water body, effectively protects the water environment and has huge economic and social benefits.
Claims (2)
1. A preparation method of ganoderma lucidum fungus dreg for repairing water environment polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene is characterized in that ganoderma lucidum seed liquid is inoculated into ganoderma lucidum solid fermentation culture medium for culturing, ganoderma lucidum is collected, and the rest ganoderma lucidum solid fermentation culture medium residue is ganoderma lucidum fungus dreg which contains lignin decomposing enzyme, organic matters, nitrogen, phosphorus and potassium and various microorganisms; the water environment is an acid-base environment with pH value of 4.0-8.5, the temperature is 20-50 ℃, and the initial pollution concentration of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene polluted water body is 50mg/L;
the preparation method comprises the following steps:
(1) Culturing ganoderma lucidum seed liquid:
inoculating Ganoderma strain into seed culture medium, and shake culturing at 30deg.C and 180rmp with constant temperature shaking table; punching a ganoderma lucidum flat plate into a round hole with the diameter of 0.5cm, inoculating the round hole into a liquid ganoderma lucidum seed culture medium, and culturing for 3 days at the temperature of 30 ℃ and at the speed of 180 rpm;
the ganoderma lucidum seed culture medium is as follows: 30g of corn flour, 15g of bean cake powder and 5g of bran, adding water, boiling for 1h, filtering by 8 layers of gauze, adding 20g of glucose, 0.2g of magnesium sulfate, 1g of monopotassium phosphate and 5g of yeast powder, fixing the volume to 1000mL, and sterilizing at 115 ℃ for 20min;
(2) Solid fermentation of ganoderma lucidum
Adding solid fermentation culture medium of Ganoderma into fungus bags, wherein each Ganoderma bag has weight of 500g, adding water to volume water content of 60%, sterilizing at 121deg.C for 2 hr, and naturally cooling;
the ganoderma lucidum solid fermentation medium is prepared from 450g of rhizoma coptidis dregs, 50g of corn flour, 15g of bran, 5g of gypsum, 5g of sucrose, 0.06g of monopotassium phosphate, 0.06g of magnesium sulfate and 0.06g of vitamin B;
inoculating ganoderma lucidum seed liquid into ganoderma lucidum fungus bags, wherein the inoculation amount is 2% of the weight volume of a culture medium, the weight volume is calculated in terms of g, the liquid is calculated in terms of mL, after inoculation is completed, the liquid is transferred into a darkroom to generate hyphae, the fungus bags full of hyphae are transferred into an illumination incubator to perform fruiting management, the temperature of the incubator is kept at 26-28 ℃, the humidity is 85-90%, the growth period of a stipe is kept away from wind, ventilation is performed in time when the ganoderma lucidum is sliced, the illumination intensity is 180-220Lx, the slicing period is 450-550Lx, ganoderma lucidum is harvested in 28-32 days, and solid fermentation culture medium residues in the fungus bags after ganoderma lucidum harvesting are ganoderma lucidum fungus residues.
2. The application of the ganoderma lucidum fungus dreg prepared by the method in the restoration of the water environment polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene, wherein the initial pollution concentration of the water body polluted by the polycyclic aromatic hydrocarbon benzo [ a ] pyrene is 50mg/L, the water environment is an acidic environment with pH of 4.0-5.5, and the temperature is 20-25 ℃; the method is characterized in that the method is added into a water body polluted by polycyclic aromatic hydrocarbon benzo [ a ] pyrene by 5-10% of weight volume, wherein the weight volume refers to the weight calculated in g and the volume calculated in mL, the survival and degradation performance of microorganisms for degrading the polycyclic aromatic hydrocarbon benzo [ a ] pyrene can be effectively improved, the polycyclic aromatic hydrocarbon benzo [ a ] pyrene in the water body with the polycyclic aromatic hydrocarbon benzo [ a ] pyrene pollution concentration of 50mg/L is removed, the degradation rate of the benzo [ a ] pyrene reaches more than 90% after 7 days, and the restoration of the polycyclic aromatic hydrocarbon benzo [ a ] pyrene pollution water environment is realized.
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