CN114621907A - Accelerating agent for microbial degradation of polycyclic aromatic hydrocarbon, application thereof and method for microbial degradation of polycyclic aromatic hydrocarbon pollutants - Google Patents
Accelerating agent for microbial degradation of polycyclic aromatic hydrocarbon, application thereof and method for microbial degradation of polycyclic aromatic hydrocarbon pollutants Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 28
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- 238000006731 degradation reaction Methods 0.000 title claims abstract description 20
- 230000000813 microbial effect Effects 0.000 title claims description 14
- 239000003795 chemical substances by application Substances 0.000 title claims description 4
- 244000005700 microbiome Species 0.000 claims abstract description 51
- 230000000593 degrading effect Effects 0.000 claims abstract description 26
- 238000000855 fermentation Methods 0.000 claims abstract description 23
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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Abstract
The invention relates to the field of microorganisms, and discloses an accelerant for degrading polycyclic aromatic hydrocarbons by microorganisms, application thereof and a method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms. The method for degrading the polycyclic aromatic hydrocarbon pollutants by microorganisms comprises the following steps: and inoculating the activated microorganisms into a fermentation medium containing polycyclic aromatic hydrocarbon pollutants and an accelerant for fermentation culture. The invention regulates the cell metabolism of the microorganism by a culture method of adding the accelerant containing at least two micromolecular organic acids, and improves the degradation efficiency of the microorganism on the polycyclic aromatic hydrocarbon pollutants.
Description
Technical Field
The invention relates to the field of microorganisms, and in particular relates to an accelerant for degrading polycyclic aromatic hydrocarbons by microorganisms, application of the accelerant and a method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms.
Background
Polycyclic Aromatic Hydrocarbons (PAHs) are aromatic organic compounds widely existing in the environment and formed by two or more than two fused benzene rings, most of which have chronic toxicity, carcinogenic, teratogenic and mutagenic effects, can be enriched in organisms through food chains, can also be retained in soil and sediments for a long time, and have great harm to human health and ecological environment.
The common artificial treatment methods for the polycyclic aromatic hydrocarbon polluted environment comprise physical repair, chemical repair, biological repair and the like. Physical remediation is mostly used as a pretreatment means, and is a remediation technology for separating PAHs from soil by methods such as extraction, heating and the like, but the remediation effect of the technology is limited; the chemical restoration mainly utilizes strong oxidizing ions and chemical oxidants generated in photochemical action to carry out strong oxidation on polycyclic aromatic hydrocarbon substances so as to convert the polycyclic aromatic hydrocarbon substances into harmless substances, but the chemical restoration has high cost and is easy to cause secondary pollution; bioremediation is an environmental remediation technical means with great application prospect which is rapidly developed in recent years, PAHs efficient degrading strains are mainly screened from polluted environments, and a remediation microbial inoculum is compounded by one or more degrading strains and applied to the polluted environments to realize the degradation of the PAHs, secondary pollution is not generated, and the environmental pollution problem is treated by the bioremediation means, so that the bioremediation method has good application prospect and great potential.
However, the degrading efficiency of the conventional repairing microbial inoculum to PAHs is still not high, and a method capable of promoting the degrading effect of the repairing microbial inoculum to PAHs is urgently needed to be provided.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provide an accelerant for degrading polycyclic aromatic hydrocarbons by microorganisms, application thereof and a method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms.
In order to achieve the above object, the present invention provides an accelerating agent for the microbial degradation of polycyclic aromatic hydrocarbons, which comprises at least two small-molecule organic acids.
Preferably, the small molecule organic acid is selected from citric acid, oxalic acid, glutaric acid, tartaric acid and malic acid.
Preferably, the accelerator comprises citric acid, oxalic acid and glutaric acid.
Preferably, the molar ratio of citric acid, oxalic acid and glutaric acid in the accelerator is 1: 0.5-1.5: 0.5-1.5.
Preferably, the accelerator also contains a surfactant.
Preferably, the surfactant is at least one selected from humic acid, alkyl glycoside, fatty alcohol-polyoxyethylene ether and fatty alcohol-polyoxyethylene ether sulfate, and more preferably humic acid.
Preferably, the molar ratio between the surfactant and the total amount of the small-molecule organic acid is 1: 0.5-3.
The second aspect of the invention provides the application of the accelerant in the microbial degradation of polycyclic aromatic hydrocarbon pollutants.
Preferably, the microorganism is selected from at least one of bacillus subtilis, bacillus cereus, mycobacterium flavum and enterobacter cloacae, more preferably bacillus subtilis.
Preferably, the polycyclic aromatic hydrocarbon contaminant is selected from at least one of phenanthrene, fluoranthene, pyrene and benzopyrene, more preferably phenanthrene.
The third aspect of the invention provides a method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms, which comprises the following steps: inoculating the activated microorganisms into a fermentation medium containing polycyclic aromatic hydrocarbon pollutants and an accelerant for fermentation culture; wherein the accelerant is the accelerant for degrading the polycyclic aromatic hydrocarbon by the microorganisms.
Preferably, the process of microbial activation comprises: inoculating the microorganism into a seed culture medium for seed culture.
Preferably, the conditions of the seed culture include: the temperature is 35-40 ℃, the rotation speed is 150-.
Preferably, the fermentation medium contains 0.8-1.2g/L of ammonium salt, 1.8-2.2g/L of potassium salt, 0.1-0.3g/L of magnesium salt, 0.01-0.03g/L of calcium salt and 0.4-0.5g/L of iron salt.
Preferably, the conditions of the fermentation culture include: the temperature is 35-40 ℃, the rotating speed is 150-.
Preferably, the inoculation amount of the seed solution of the microorganism is 10-50mL, the content of the polycyclic aromatic hydrocarbon pollutant is 0.05-0.25g, and the content of the accelerant is 0.9-1.4g relative to 1000mL of the fermentation medium.
Through the technical scheme, the invention has the beneficial effects that: the accelerant provided by the invention contains at least two kinds of micromolecular organic acids, and when the accelerant is applied to degrading polycyclic aromatic hydrocarbons by microorganisms on the basis of combining different micromolecular organic acids, the cell metabolism of the microorganisms can be effectively regulated, the degradation effect of the microorganisms on the polycyclic aromatic hydrocarbons is promoted, and the degradation efficiency of the microorganisms on polycyclic aromatic hydrocarbon pollutants is further improved.
When the accelerant provided by the invention is used for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms, the accelerant is mild in condition, low-carbon and environment-friendly, low in cost and convenient to operate, and has important significance for degrading polycyclic aromatic hydrocarbon pollutants.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
FIG. 1 shows the growth of Bacillus subtilis ZL09-26 of examples 1, 4, 1 and 2;
FIG. 2 shows the residual phenanthrene content in the fermentation broths obtained in example 1, example 4, comparative example 1 and comparative example 2.
Detailed Description
The following describes in detail specific embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the present invention, are given by way of illustration and explanation only, not limitation.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
The invention provides an accelerant for degrading polycyclic aromatic hydrocarbon by microorganisms, which contains at least two small molecular organic acids.
In the research and development process, the inventor of the invention unexpectedly finds that when the method is applied to degrading polycyclic aromatic hydrocarbons by microorganisms on the basis of combining at least two small molecular organic acids, the cell metabolism of the microorganisms can be effectively regulated, the degradation effect of the microorganisms on the polycyclic aromatic hydrocarbons is promoted, and the degradation efficiency of the microorganisms on polycyclic aromatic hydrocarbon pollutants is further improved.
According to the present invention, preferably, the small molecule organic acid is selected from citric acid, oxalic acid, glutaric acid, tartaric acid and malic acid.
According to the present invention, preferably, the accelerator contains citric acid, oxalic acid and glutaric acid. The above substances are all commercially available.
According to the invention, preferably, the molar ratio of citric acid, oxalic acid and glutaric acid in the accelerator is 1: 0.5-1.5: 0.5-1.5. The inventor finds that under the preferred embodiment, the microbial activity and metabolism can be promoted, and the degradation efficiency of the polycyclic aromatic hydrocarbon pollutants can be improved.
According to the present invention, preferably, the accelerator further contains a surfactant.
According to the present invention, preferably, the surfactant is selected from at least one of humic acid, alkyl glycoside, fatty alcohol-polyoxyethylene ether, and fatty alcohol-polyoxyethylene ether sulfate, and more preferably humic acid.
In the invention, the humic acid is an amorphous polymer compound and is a complex mixture colloid consisting of polymer hydroxycarboxylic acids, the basic macromolecular structure of the humic acid is an aromatic ring and an alicyclic ring, and the ring is connected with functional groups such as carboxyl, hydroxyl, carbonyl, quinonyl, methoxyl and the like.
The above substances are all commercially available.
According to the present invention, preferably, the molar ratio between the surfactant and the total amount of the small organic acid is 1: 0.5-3. The inventor finds that under the preferred embodiment, the activity and metabolism of microorganisms can be further promoted, and the degradation efficiency of the polycyclic aromatic hydrocarbon pollutants is improved.
The invention provides application of the accelerant in degrading polycyclic aromatic hydrocarbon pollutants through microorganisms.
According to the present invention, the microorganism may be any one of strains capable of degrading polycyclic aromatic hydrocarbons in the art. Preferably, the microorganism is selected from at least one of bacillus subtilis, bacillus cereus, mycobacterium flavum and enterobacter cloacae, more preferably bacillus subtilis. Illustratively, the invention adopts bacillus subtilis ZL09-26, and the preservation information of the strain is shown in Chinese patent with the publication number CN107937321A, and the preservation number is GDMCC No: 60293.
according to the present invention, preferably, the polycyclic aromatic hydrocarbon contaminants are selected from at least one of phenanthrene, fluoranthene, pyrene and benzopyrene, more preferably phenanthrene.
In a third aspect, the invention provides a method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms, which comprises the following steps: inoculating the activated microorganisms into a fermentation medium containing polycyclic aromatic hydrocarbon pollutants and an accelerant for fermentation culture; wherein the accelerant is the accelerant for degrading the polycyclic aromatic hydrocarbon by the microorganisms.
According to the present invention, the microorganism activation process may employ strain activation methods that are conventional in the art, including but not limited to solid plate culture, liquid culture. Preferably, the process of microbial activation comprises: inoculating the microorganism into a seed culture medium for seed culture.
The method of seed culture according to the present invention is not particularly limited as long as the microorganism can be activated and proliferated by the method, and preferably, the conditions of seed culture include: the temperature is 35-40 deg.C, specifically 35 deg.C, 36 deg.C, 37 deg.C, 38 deg.C, 39 deg.C, 40 deg.C, or any value between the above two values; the rotation speed is 150-200rpm, specifically can be 150rpm, 160rpm, 170rpm, 180rpm, 190rpm, 200rpm, or any value between the two values; the pH is 7.5-8.5, specifically 7.5, 8.0, 8.5, or any value between the two values; the time is 10-12h, specifically 10h, 11h, 12h, or any value between the two values. The seed medium may be a medium conventionally used in the art, and for example, may be an LB liquid medium (0.8-1 wt% peptone, 0.5-0.8 wt% yeast extract, 1-1.5 wt% sodium chloride).
In the present invention, the fermentation medium may be a medium commonly used in the art, and preferably, a liquid medium of inorganic salts is used. The inorganic salt liquid culture medium comprises ammonium salt 0.8-1.2g/L, potassium salt 1.8-2.2g/L, magnesium salt 0.1-0.3g/L, calcium salt 0.01-0.03g/L, and iron salt 0.4-0.5g/L, specifically ammonium nitrate 0.9-1.1g/L, potassium dihydrogen phosphate 0.4-0.6g/L, potassium dihydrogen phosphate 1.4-1.6g/L, magnesium sulfate heptahydrate 0.1-0.3g/L, anhydrous calcium chloride 0.01-0.03g/L, and ferric chloride 0.4-0.5 g/L. The inventors have found that in this preferred embodiment, the efficiency of degradation of polycyclic aromatic hydrocarbon contaminants by microorganisms can be increased.
The method of the present invention for fermentation culture is not particularly limited as long as the microorganism can be proliferated in a large amount by the method, and preferably, the conditions of the fermentation culture include: the temperature is 35-40 deg.C, specifically 35 deg.C, 36 deg.C, 37 deg.C, 38 deg.C, 39 deg.C, 40 deg.C, or any value between the above two values; the rotation speed is 150-200rpm, specifically can be 150rpm, 160rpm, 170rpm, 180rpm, 190rpm, 200rpm, or any value between the two values; the time is not less than 12 h. The inventors have found that in this preferred embodiment, the growth rate of the microorganisms can be increased, thereby increasing the efficiency of degradation of the polycyclic aromatic hydrocarbon contaminants.
According to the invention, preferably, the inoculation amount of the seed liquid of the microorganism is 10-50mL, the content of the polycyclic aromatic hydrocarbon pollutant is 0.05-0.25g, and the content of the accelerant is 0.9-1.4g relative to 1000mL of the fermentation medium. The inventors have found that in this preferred embodiment, the efficiency of degradation of polycyclic aromatic hydrocarbon contaminants by microorganisms can be increased.
According to a particularly preferred embodiment of the present invention, a method for the microbial degradation of polycyclic aromatic hydrocarbon contaminants comprises the steps of:
(1) inoculating bacillus subtilis ZL09-26 into an LB liquid culture medium according to the inoculation amount of 0.5-2% by volume, and culturing for 10-12h under the conditions that the temperature is 35-40 ℃, the rotation speed is 150-;
(2) inoculating the seed liquid of the bacillus subtilis ZL09-26 into an inorganic salt liquid culture medium containing 50-250mg/L phenanthrene and 900-1400mg/L promoter according to the inoculation amount of 1-5 volume percent, and culturing for at least 1d under the conditions that the temperature is 35-40 ℃ and the rotating speed is 150-200 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid is 1: 0.5-1.5: 0.5-1.5, the molar ratio of humic acid to the total amount of citric acid, oxalic acid and glutaric acid is 1: 0.5-3;
the LB liquid medium comprises the following components: 0.8-1 wt.% peptone, 0.5-0.8 wt.% yeast extract, 1-1.5 wt.% sodium chloride;
the inorganic salt liquid culture medium comprises the following components: 0.9-1.1g/L of ammonium nitrate, 0.4-0.6g/L of monopotassium phosphate, 1.4-1.6g/L of dipotassium phosphate, 0.1-0.3g/L of magnesium sulfate heptahydrate, 0.01-0.03g/L of anhydrous calcium chloride and 0.4-0.5g/L of ferric trichloride.
In the above-mentioned particularly preferred embodiment, when the accelerant containing at least two kinds of small-molecular organic acids is used for the microbial degradation of polycyclic aromatic hydrocarbon pollutants, the conditions are mild, the carbon is low, the environment is protected, the cost is low, the operation is convenient, and the production cost is low.
The present invention will be described in detail below by way of examples.
In the following examples, humic acid was obtained from Shanghai Merlian under the model number H811077 (the content of fulvic acid is 90% or more); unless otherwise specified, all experimental materials used were purchased from conventional biochemical reagent stores.
In the following examples, the microorganism used is Bacillus subtilis ZL09-26, and the preservation information of the strain is disclosed in Chinese patent with publication No. CN107937321A, with the preservation number being GDMCC No: 60293.
the formula of the LB liquid culture medium is as follows: 10g of peptone, 5g of yeast extract and 10g of sodium chloride, and distilled water till the volume is 1L, pH and 7.0.
The formula of the inorganic salt liquid culture medium is as follows: 1.0g of ammonium nitrate, 0.5g of monopotassium phosphate, 1.5g of dipotassium phosphate, 0.2g of magnesium sulfate heptahydrate, 0.02g of anhydrous calcium chloride and 0.5g of ferric trichloride, and the volume of distilled water is adjusted to 1L, pH to 8.0.
Example 1
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 37 ℃ temperature, 180rpm rotation speed and pH of 8.0 to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed liquid of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 200mg/L phenanthrene and 1135mg/L accelerant according to the inoculation amount of 2 volume percent, and culturing for 3d under the conditions that the temperature is 37 ℃ and the rotating speed is 180 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid to the humic acid is 1: 1: 1: 1.
example 2
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 10h under the conditions of the temperature of 35 ℃, the rotation speed of 150rpm and the pH value of 8.0 to obtain a seed solution of the bacillus subtilis ZL 09-26;
(2) inoculating seed solution of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 50mg/L phenanthrene and 908mg/L accelerant according to the inoculation amount of 1 volume percent, and culturing for 3d under the conditions that the temperature is 35 ℃ and the rotating speed is 150 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid to the humic acid is 1: 0.5: 0.5: 1.2.
example 3
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 40 ℃ of temperature, 200rpm of rotation speed and 8.0 of pH to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating the seed solution of bacillus subtilis ZL09-26 into an inorganic salt liquid culture medium containing 250mg/L phenanthrene and 1362mg/L accelerant according to the inoculation amount of 3 volume percent, and culturing for 3d under the conditions that the temperature is 40 ℃ and the rotating speed is 200 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid to the humic acid is 1: 1.5: 1.5: 8.
example 4
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 37 ℃ of temperature, 180rpm of rotation speed and 8.0 of pH value to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed liquid of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 200mg/L phenanthrene and 1135mg/L accelerant according to the inoculation amount of 2 volume percent, and culturing for 3d under the conditions that the temperature is 37 ℃ and the rotating speed is 180 rpm;
wherein the accelerant contains citric acid, oxalic acid and glutaric acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid is 1: 1: 1.
example 5
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 0.8 volume percent, and culturing for 12h under the conditions that the temperature is 37 ℃, the rotation speed is 180rpm and the pH is 8.0 to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed liquid of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 200mg/L phenanthrene and 1135mg/L accelerant according to the inoculation amount of 2%, and culturing for 3d under the conditions that the temperature is 37 ℃ and the rotating speed is 180 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, wherein the molar ratio of the citric acid to the oxalic acid to the glutaric acid to the humic acid is 1: 1: 1: 8.
example 6
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 37 ℃ of temperature, 180rpm of rotation speed and 8.0 of pH value to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed liquid of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 200mg/L phenanthrene and 227mg/L accelerant according to the inoculation amount of 2 volume percent, and culturing for 3d under the conditions that the temperature is 37 ℃ and the rotating speed is 180 rpm;
the accelerant contains citric acid, oxalic acid, glutaric acid and humic acid, and the molar ratio of the citric acid to the oxalic acid to the glutaric acid to the humic acid is 1: 1: 1: 4.
comparative example 1
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 37 ℃ of temperature, 180rpm of rotation speed and 8.0 of pH value to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed solution of Bacillus subtilis ZL09-26 in an inoculum size of 2 vol% in inorganic salt liquid culture medium containing 200mg/L phenanthrene, and culturing at 37 deg.C and 180rpm for 3 d.
Comparative example 2
(1) Inoculating bacillus subtilis ZL09-26 into LB liquid culture medium according to the inoculation amount of 2 volume percent, and culturing for 12h under the conditions of 37 ℃ of temperature, 180rpm of rotation speed and 8.0 of pH value to obtain a seed solution of bacillus subtilis ZL 09-26;
(2) inoculating seed liquid of bacillus subtilis ZL09-26 into inorganic salt liquid culture medium containing 200mg/L phenanthrene and 1135mg/L accelerant according to the inoculation amount of 2 volume percent, and culturing for 3d under the conditions that the temperature is 37 ℃ and the rotating speed is 180 rpm;
wherein the promoter contains only humic acid.
Test example 1
The growth of Bacillus subtilis ZL09-26 in examples 1, 4, 1 and 2 was measured using a microplate reader (SpectraMax M3, USA) at an absorption wavelength of 600 nm.
The growth of Bacillus subtilis ZL09-26 in the fermentation liquid obtained in example 1, example 4, comparative example 1 and comparative example 2 is shown in figure 1, and as can be seen from figure 1, compared with comparative example 1 and comparative example 2, the accelerant containing citric acid, oxalic acid, glutaric acid and humic acid is added in example 1, the accelerant containing citric acid, oxalic acid and glutaric acid is added in example 4, and the growth of microbial cells is promoted and the activity of microorganisms is improved after the accelerant is added in example 1 and example 4.
Test example 2
Examples 1 and 4 were collected,Comparative example 1 and comparative example 2 the fermentation broths obtained by culturing 1d, 2d and 3d were subjected to gas chromatography-mass spectrometry (GC-MS, Trace GC2000 DSQ, Agilent, USA) using a DB-5MS column (30 m.times.0.25 mm.times.0.25 μm) to determine the content of phenanthrene remaining in the fermentation broths. Nitrogen was used as a carrier gas, the column temperature was initially 80 ℃ for 25 ℃ min-1The rate of (A) is reduced to 200 ℃ for 10 min-1Cooling to 300 deg.C, and keeping the temperature at 300 deg.C for 6 min.
The content of the phenanthrene remaining in the fermentation broth obtained in example 1, example 4, comparative example 1 and comparative example 2 is shown in fig. 2, and the results of the efficiency of bacillus subtilis ZL09-26 in example 1, example 4, comparative example 1 and comparative example 2 in degrading phenanthrene are shown in table 1.
TABLE 1
As is apparent from the results in Table 1, the efficiency of degrading phenanthrene by Bacillus subtilis ZL09-26 in examples 1 and 4 is superior to that of Bacillus subtilis ZL09-26 in comparative examples 1 and 2.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (10)
1. An accelerant for degrading polycyclic aromatic hydrocarbon by microorganisms is characterized in that the accelerant contains at least two small molecular organic acids.
2. The enhancer of claim 1, wherein the small molecule organic acid is selected from the group consisting of citric acid, oxalic acid, glutaric acid, tartaric acid, and malic acid;
preferably, the accelerator comprises citric acid, oxalic acid and glutaric acid;
preferably, the molar ratio of citric acid, oxalic acid and glutaric acid in the accelerator is 1: 0.5-1.5: 0.5-1.5.
3. An accelerator according to claim 1 or 2, further comprising a surfactant;
preferably, the surfactant is selected from at least one of humic acid, alkyl glycoside, fatty alcohol-polyoxyethylene ether and fatty alcohol-polyoxyethylene ether sulfate, and is more preferably humic acid;
preferably, the molar ratio between the surfactant and the total amount of the small-molecule organic acid is 1: 0.5-3.
4. Use of an accelerating agent as defined in any one of claims 1 to 3 for the microbial degradation of polycyclic aromatic hydrocarbon contaminants.
5. The use according to claim 4, wherein the microorganism is selected from at least one of Bacillus subtilis, Bacillus cereus, Mycobacterium flavum and Enterobacter cloacae, preferably Bacillus subtilis;
preferably, the polycyclic aromatic hydrocarbon contaminant is selected from at least one of phenanthrene, fluoranthene, pyrene and benzopyrene, more preferably phenanthrene.
6. A method for degrading polycyclic aromatic hydrocarbon pollutants by microorganisms is characterized by comprising the following steps: inoculating the activated microorganisms into a fermentation medium containing polycyclic aromatic hydrocarbon pollutants and an accelerant for fermentation culture; wherein the accelerator is the accelerator according to any one of claims 1 to 3.
7. The method of claim 6, wherein the process of microbial activation comprises: inoculating the microorganism into a seed culture medium for seed culture;
preferably, the conditions of the seed culture include: the temperature is 35-40 ℃, the rotation speed is 150-.
8. The method of claim 6 or 7, wherein the fermentation medium comprises 0.8-1.2g/L ammonium salt, 1.8-2.2g/L potassium salt, 0.1-0.3g/L magnesium salt, 0.01-0.03g/L calcium salt, and 0.4-0.5g/L iron salt.
9. The method of claim 6 or 7, wherein the conditions of the fermentation culture comprise: the temperature is 35-40 ℃, the rotating speed is 150-.
10. The method of claim 6 or 7, wherein the inoculation amount of the seed solution of the microorganism is 10-50mL, the content of the polycyclic aromatic hydrocarbon pollutant is 0.05-0.25g, and the content of the promoter is 0.9-1.4g, relative to 1000mL of the fermentation medium.
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| LEI ZHANG ET AL.: "How humic acid and Tween80 improve the phenanthrene biodegradation efficiency: Insight from cellular characteristics and quantitative proteomics", JOURNAL OF HAZARDOUS MATERIALS, vol. 421, 21 July 2021 (2021-07-21), pages 3, XP086823758, DOI: 10.1016/j.jhazmat.2021.126685 * |
| LEI ZHANG ET AL.: "Root exuded low-molecular-weight organic acids affected the phenanthrene degrader differently: A multi-omics study", JOURNAL OF HAZARDOUS MATERIALS, vol. 414, 31 December 2021 (2021-12-31), pages 2 * |
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