CN114807287A - Method for promoting microbial degradation of weathered coal to produce humic acid - Google Patents
Method for promoting microbial degradation of weathered coal to produce humic acid Download PDFInfo
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- 239000003245 coal Substances 0.000 title claims abstract description 74
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 239000004021 humic acid Substances 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 44
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 24
- 230000015556 catabolic process Effects 0.000 title claims abstract description 22
- 230000001737 promoting effect Effects 0.000 title claims abstract description 18
- 230000000813 microbial effect Effects 0.000 title claims abstract description 17
- 241000222393 Phanerochaete chrysosporium Species 0.000 claims abstract description 30
- 239000002736 nonionic surfactant Substances 0.000 claims abstract description 24
- 241000222355 Trametes versicolor Species 0.000 claims abstract description 23
- 239000001963 growth medium Substances 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 15
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 12
- 229920000053 polysorbate 80 Polymers 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 7
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 7
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 5
- 244000061456 Solanum tuberosum Species 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 229930003270 Vitamin B Natural products 0.000 claims description 3
- 235000019156 vitamin B Nutrition 0.000 claims description 3
- 239000011720 vitamin B Substances 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 abstract description 10
- 230000001580 bacterial effect Effects 0.000 abstract description 5
- 239000002689 soil Substances 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 230000035558 fertility Effects 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 16
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- 108010054320 Lignin peroxidase Proteins 0.000 description 7
- 230000009286 beneficial effect Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000009630 liquid culture Methods 0.000 description 5
- 108010029541 Laccase Proteins 0.000 description 4
- 108010059896 Manganese peroxidase Proteins 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 239000004094 surface-active agent Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000222354 Trametes Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004939 coking Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- PTKRHFQQMJPPJN-UHFFFAOYSA-N dipotassium;oxido-(oxido(dioxo)chromio)oxy-dioxochromium;sulfuric acid Chemical compound [K+].[K+].OS(O)(=O)=O.[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O PTKRHFQQMJPPJN-UHFFFAOYSA-N 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- FQENQNTWSFEDLI-UHFFFAOYSA-J sodium diphosphate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]P([O-])(=O)OP([O-])([O-])=O FQENQNTWSFEDLI-UHFFFAOYSA-J 0.000 description 1
- 229940048086 sodium pyrophosphate Drugs 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 235000019818 tetrasodium diphosphate Nutrition 0.000 description 1
- 239000001577 tetrasodium phosphonato phosphate Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/14—Fungi; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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 discloses a method for promoting microbial degradation of weathered coal to produce humic acid, which comprises the following steps: inoculating the microorganisms to a PDA culture medium containing weathered coal and a nonionic surfactant for culture; wherein the microorganism is trametes versicolor and/or Phanerochaete chrysosporium. The invention utilizes trametes versicolor and/or phanerochaete chrysosporium to degrade weathered coal, and adds nonionic surfactant in the degradation process, so as to cooperate with the bacterial strain to degrade weathered coal to produce humic acid. The method for extracting humic acid by efficiently degrading weathered coal by using microorganisms can increase the utilization efficiency of the weathered coal humic acid to the maximum extent on the basis of economy and environmental protection, increase the soil fertility and improve the soil property.
Description
Technical Field
The invention relates to the technical field of comprehensive utilization of weathered coal, in particular to a method for promoting microbial degradation of weathered coal to produce humic acid.
Background
The weathered coal mainly refers to coal with shallow buried depth, has high probability of contacting with the external environment, and generates a series of physical and chemical changes after the weathered coal is contacted with weather phenomena such as rain, snow, frost and the like for a long time, thereby leading to the generation of a large amount of humic acid in the metamorphic coal, and the metamorphic coal has the characteristics of high brittleness, easy fragmentation, deteriorated mechanical performance, poor coking property, cohesiveness and other indexes, low heat value and the like. The reserves of weathered coal on the earth are 80 million tons and 50 million tons in Shanxi and Nemontage two provinces (regions), and the reserves are more in Xinjiang, Heilongjiang, Jiangxi, Yunnan, Sichuan, Henan, Gansu, Guizhou provinces and other provinces, but can not be directly used as fire coal, thereby causing a great deal of resource waste. Some coal in some regions has a deep weathering degree, the utilization efficiency of humic acid of weathered coal is improved to a certain extent by some physical and chemical methods, but the environmental pollution is inevitably generated while related target products are obtained, and in addition, the physical and chemical conversion modes are used or the cost is higher, the requirements on conversion conditions are strict, or the conversion rate is lower. Therefore, a method for producing humic acid from weathered coal, which is efficient, simple and convenient and does not produce secondary pollution, is urgently needed to be researched.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims to provide a method for promoting microbial degradation of weathered coal to produce humic acid, so as to solve the problems of high cost, secondary pollution and the like when the weathered coal is used for extracting the humic acid.
The technical scheme for solving the technical problems is as follows: the method for promoting the microbial degradation of weathered coal to produce humic acid comprises the following steps:
inoculating the microorganisms to a PDA culture medium containing weathered coal and a nonionic surfactant for culture; wherein the microorganism is trametes versicolor and/or Phanerochaete chrysosporium.
The invention has the beneficial effects that: the phanerochaete chrysosporium and the trametes versicolor can generate a plurality of enzymes with strong degradation capability on lignin substances in the growth process, for example, the phanerochaete chrysosporium can generate lignin peroxidase and manganese peroxidase, the trametes versicolor has strong capability of generating lignin peroxidase and laccase, weathered coal contains lignin-like structures, and the phanerochaete chrysosporium can be used for degrading weathered coal to generate humic acid. However, the enzymes for degrading lignin substances produced by Phanerochaete chrysosporium are influenced by various factors, the PDA culture medium contains nutrients required by the growth of the Phanerochaete chrysosporium, weathered coal can also provide part of carbon sources for the Phanerochaete chrysosporium, and the nonionic surfactant can promote the degradation of the weathered coal by the enzymes, so that more humic acid is produced, the efficiency of extracting the humic acid by degrading the weathered coal is effectively improved, and meanwhile, the method is simple to operate, low in cost and free of secondary pollution.
On the basis of the technical scheme, the invention can be further improved as follows:
further, the nonionic surfactant is tween 80 or tween 20.
The beneficial effect of adopting the further technical scheme is as follows: the Tween 80 or Tween 20 can improve the permeability of cell membranes of Coriolus versicolor and Phanerochaete chrysosporium, so that enzyme solution in cells can be secreted out through the cell membranes, thereby improving the production of lignin peroxidase and laccase in Coriolus versicolor, and improving the production of lignin peroxidase and manganese peroxidase in Phanerochaete chrysosporium, and especially in the process of degrading weathered coal, the enzymes can stably maintain a high enzyme activity state for a long time.
Further, the volume ratio of the nonionic surfactant to the medium is 0.8 to 1.2:100, preferably 1: 100.
The beneficial effect of adopting the further technical scheme is as follows: the addition amount of the nonionic surfactant cannot be too much or too little, and the production of lignin peroxidase, laccase and manganese peroxidase in the Coriolus versicolor and Phanerochaete chrysosporium can be effectively improved only within a specific dosage range, so that the efficiency of degrading weathered coal is promoted.
Further, the weight volume ratio of the weathered coal to the PDA culture medium is (8-15) g: 100ml, preferably 10 g: 100 ml.
Further, the weathered coal is pretreated when being added into the PDA culture medium, and the specific process is as follows: pulverizing weathered coal, and sieving with 0.2-0.5mm sieve.
The beneficial effect of adopting the further technical scheme is as follows: the pretreatment of the weathered coal can affect the degradation effect of microorganisms on the weathered coal, when the weathered coal is crushed and sieved by a sieve of 0.2-0.5mm, the particle size of the weathered coal can be effectively contacted with the microorganisms, and simultaneously, under the action of a nonionic surfactant, after enzyme generated by the microorganisms is fully contacted with the weathered coal, the weathered coal can be effectively degraded to form humic acid.
Further, the PDA culture medium comprises the following components: 1L of potato leachate, 20g of glucose and KH 2 PO 4 3g、MgSO 4 ·7H 2 1.5g of O and 10.01-0.03 g of vitamin B, and the pH is natural.
The beneficial effect of adopting the further technical scheme is as follows: the PDA culture medium contains nutrients for promoting growth of trametes versicolor and Phanerochaete chrysosporium, such as potato leachate for providing carbon source, nitrogen source, water and inorganic salt for trametes versicolor and Phanerochaete chrysosporium, glucose for providing carbon source for trametes versicolor and Phanerochaete chrysosporium, and KH 2 PO 4 3g、MgSO 4 ·7H 2 O represents an inorganic salt, etc.
Further, the culture conditions were: the temperature is 25-30 ℃, the rotating speed is 120-160r/min, the time is 20-40 days, the preferable culture temperature is 28 ℃, the rotating speed is 150r/min, and the time is 30 days.
The beneficial effect of adopting the further technical scheme is as follows: in order to promote the growth of the bacteria, the full contact with the weathered coal and the good growth of the bacteria in the culture process, shaking table culture can be carried out, the rotating speed is 120-160r/min, the culture temperature in the culture process is 25-30 ℃, the microorganisms can rapidly grow under the condition of the culture time, the microorganisms are continuously contacted with the weathered coal from the beginning of growth, the non-ionic surfactant continuously promotes the production of enzymes in the microorganisms in the growth process, more enzymes continuously permeate the cell membranes and are fully contacted with the weathered coal to degrade the weathered coal to produce humic acid, the produced humic acid is increased along with the increase of the culture time, and the yield of the humic acid is highest when the culture time reaches 30 days.
The invention has the following beneficial effects:
the invention utilizes trametes versicolor and/or phanerochaete chrysosporium to degrade weathered coal, and adds a nonionic surfactant in the degradation process, wherein the nonionic surfactant can enable enzyme solution in cells to be secreted out through cell membranes, thereby improving the production of lignin peroxidase and laccase in the tramete versicolor and improving the production of the lignin peroxidase and manganese peroxidase in the phanerochaete chrysosporium, and particularly, the enzymes can stably maintain a high enzyme activity state for a long time in the process of degrading weathered coal. The weathered coal contains lignin-like structural substances, and the enzymes can carry out enzymolysis on the substances to generate humic acid. The method for extracting humic acid by efficiently degrading weathered coal by using microorganisms can increase the utilization efficiency of the weathered coal humic acid to the maximum extent on the basis of economy and environmental protection, increase the soil fertility and improve the soil property.
Detailed Description
The following examples are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The phanerochaete chrysosporium and trametes versicolor used in the invention are purchased strains, and the specific process is as follows:
example 1:
a method for promoting microbial degradation of weathered coal to produce humic acid comprises the following steps:
(1) activated strain
Inoculating Phanerochaete chrysosporium into a PDA solid culture medium for activated culture; wherein, the PDA solid culture medium is prepared from the following components: 1L of potato extract, 20g of glucose and KH 2 PO 4 3g、MgSO 4 ·7H 2 1.5g of O, 10.02g of vitamin B and 20g of agar, and the pH is natural.
(2) Preparation of liquid culture Medium
The liquid culture medium, namely the PDA liquid culture medium, is prepared from the following components: 1L of potato extract, 20g of glucose and KH 2 PO 4 3g、MgSO 4 ·7H 2 O1.5 g, vitamin B10.02g, and natural pH.
(3) Pouring the prepared PDA liquid culture medium into a 250ml triangular flask, filling 100ml of PDA liquid culture medium in each flask, then adding 10ml of Tween 80 and 10g of weathered coal, uniformly mixing, and placing at 121 ℃ for autoclaving for 21 min; wherein, the weathered coal is pretreated before being added, and the concrete process is as follows: the weathered coal was crushed through a 0.25mm sieve.
(3) Cooling the sterilized culture medium, dividing strains on the solid culture medium into 0.5cm by 0.5cm blocks with uniform sizes on an aseptic operating platform, inoculating 5 blocks into each triangular flask for culturing, wherein the culture conditions are as follows: the culture temperature is 28 ℃, the rotation speed is 150r/min, and 10ml of culture is respectively taken to detect the content of humic acid when the culture is cultured for 1 st, 10 th, 20 th and 30 th days.
Example 2:
example 2 is different from example 1 in that the added strain is trametes versicolor, and the rest of the procedure is the same as example 1.
Example 3:
example 3 is different from example 1 in that tween 20 was added as a nonionic surfactant, and the rest of the procedure was the same as in example 1.
Example 4:
example 4 was different from example 2 in that tween 20 was added as a nonionic surfactant, and the rest of the procedure was the same as in example 2.
Example 5:
example 5 is different from example 1 in that the added strains are Phanerochaete chrysosporium and trametes versicolor, the addition amount is half of each, and the rest process is the same as example 1.
Comparative example 1:
comparative example 1 is different from example 1 in that tween 80 was not added and the rest of the procedure was the same as in example 1.
Comparative example 2:
comparative example 2 is different from example 2 in that tween 80 was not added and the rest of the procedure was the same as in example 2.
Comparative example 3:
comparative example 3 is different from example 5 in that the added fungus is white rot fungus, and the rest of the procedure is the same as example 5.
Comparative example 4:
comparative example 4 is different from example 5 in that the added bacteria are white rot fungi and phanerochaete chrysosporium, each in half, and the rest of the procedure is the same as example 5.
Comparative example 5:
comparative example 5 differs from example 5 in that the surfactant added is glycerol and the rest of the procedure is the same as in example 5.
The detection process of the content of humic acid is as follows: transferring 10ml of the culture taken out into a 250ml volumetric flask, then adding 100ml of sodium pyrophosphate aqueous alkali, taking out in a boiling water bath for 60min, adding 1% sodium hydroxide solution into the volumetric flask, cooling to a constant volume of 250ml, filtering and pumping 10ml of leaching solution, evaporating to dryness in an oil bath, then adding 10ml of potassium dichromate sulfuric acid solution, boiling in an oil bath kettle at the temperature of 170-. The calculation formula of the content of the humic acid refers to a soil humic acid detection formula.
The culture obtained in examples 1 to 5 and comparative examples 1 to 5 were subjected to humic acid content measurement, and the results were:
| day 1 | Day 10 | Day 20 | Day 30 | |
| Example 1 | 4.56％ | 7.35％ | 12.89％ | 19.79％ |
| Example 2 | 4.51％ | 6.21％ | 10.78％ | 16.24％ |
| Example 3 | 4.54％ | 7.20％ | 12.17％ | 19.32％ |
| Example 4 | 4.45％ | 6.01％ | 10.53％ | 15.88％ |
| Example 5 | 4.55％ | 10.56％ | 18.65％ | 28.31％ |
| Comparative example 1 | 4.50％ | 6.12％ | 8.25％ | 11.93％ |
| Comparative example 2 | 4.42％ | 6.08％ | 8.13％ | 12.76％ |
| Comparative example 3 | 4.48％ | 5.86％ | 7.56％ | 10.51％ |
| Comparative example 4 | 4.59％ | 8.87％ | 14.15％ | 20.16％ |
| Comparative example 5 | 4.21％ | 8.91％ | 15.36％ | 22.25％ |
As can be seen from the above table, when only Phanerochaete chrysosporium is added without adding the nonionic surfactant, the humic acid content in 10ml of the bacterial liquid is increased from the initial 4.50% to 11.93%, and the humic acid content is increased by 7.43%. And the bacterial strain is changed into trametes versicolor, when the nonionic surfactant is not added, the content of humic acid in 10ml of bacterial liquid is increased from the initial 4.42 percent to 12.76 percent, and the content of humic acid is increased by 8.34 percent. From this, it can be seen that the two microorganisms have a certain effect on the degradation of weathered coal, but there is no significant difference.
When the Phanerochaete chrysosporium and the nonionic surfactant are added, if the surfactant is Tween 80, the content of humic acid in 10ml of the bacterial liquid is increased from the initial 4.56% to 19.79%, and the content of humic acid is increased by 15.23%. Compared with the method without adding Tween 80, the method has the advantages that the content of humic acid is obviously improved, and therefore, the surfactant has obvious influence on degradation of weathered coal by Phanerochaete chrysosporium. When the strain is changed to trametes versicolor, under the combined action of the strain and Tween 80, the content of humic acid in 10ml of the strain is increased from the initial 4.51% to 16.24%, and the content of humic acid is increased by 11.73%. From this, it is known that the nonionic surfactant has a greater influence on Phanerochaete chrysosporium in degrading decomposed coal to produce humic acid than on trametes versicolor.
When the Phanerochaete chrysosporium and the trametes versicolor and the nonionic surfactant such as Tween 80 degrade the weathered coal to produce the humic acid, the effect of the combination of the two strains on the nonionic surfactant is more remarkable than that of the combination of the two strains on the weathered coal to degrade the weathered coal, but when the nonionic surfactant is changed from Tween 80 or Tween 20 to other surfactants such as glycerol, the yield of the humic acid is reduced, which indicates that only the Tween 80 or the Tween 20 can generate the synergistic effect of degrading the weathered coal to produce the humic acid with the Phanerochaete chrysosporium and the trametes versicolor. In addition, when the strain is replaced by Phanerochaete chrysosporium and trametes versicolor, the yield of humic acid is far lower than the effects of Phanerochaete chrysosporium and trametes versicolor.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A method for promoting microbial degradation of weathered coal to produce humic acid is characterized by comprising the following steps:
inoculating the microorganisms to a PDA culture medium containing weathered coal and a nonionic surfactant for culture; wherein the microorganism is trametes versicolor and/or phanerochaete chrysosporium.
2. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 1 wherein the non-ionic surfactant is tween 80 or tween 20.
3. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 2 wherein the volume ratio of the non-ionic surfactant to the culture medium is 0.8-1.2: 100.
4. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 3 wherein the volume ratio of the non-ionic surfactant to the culture medium is 1: 100.
5. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 1, wherein the weight volume ratio of weathered coal to PDA culture medium is (8-15) g: 100 ml.
6. The method for promoting the microbial degradation of weathered coal to produce humic acid according to claim 1, wherein the weathered coal is pretreated when being added into the PDA culture medium, and the method comprises the following specific processes: pulverizing weathered coal, and sieving with 0.2-0.5mm sieve.
7. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 1 wherein the PDA culture medium comprises the following components: 1L of potato extract, 20g of glucose and KH 2 PO 4 3g、MgSO 4 ·7H 2 1.5g of O and 10.01-0.03 g of vitamin B, and the pH is natural.
8. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 1, wherein the culture conditions are as follows: the temperature is 25-30 ℃, the rotating speed is 120-160r/min, and the time is 20-40 days.
9. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 8, wherein the culture temperature is 28 ℃ and the rotation speed is 150 r/min.
10. The method for promoting microbial degradation of weathered coal to produce humic acid according to claim 8 wherein the culturing period is 30 days.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101372424A (en) * | 2008-09-12 | 2009-02-25 | 黑龙江省宏达生物工程有限公司 | Method for preparing humic acid bio-fertilizer from lignite |
| CN103168619A (en) * | 2013-03-04 | 2013-06-26 | 武汉理工大学 | Cultural method for improving degradation performance of phanerochaete chrysosporium |
| CN111961595A (en) * | 2020-08-25 | 2020-11-20 | 中南林业科技大学 | Coriolus versicolor trametes capable of efficiently decoloring multiple textile dyes |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101372424A (en) * | 2008-09-12 | 2009-02-25 | 黑龙江省宏达生物工程有限公司 | Method for preparing humic acid bio-fertilizer from lignite |
| CN103168619A (en) * | 2013-03-04 | 2013-06-26 | 武汉理工大学 | Cultural method for improving degradation performance of phanerochaete chrysosporium |
| CN111961595A (en) * | 2020-08-25 | 2020-11-20 | 中南林业科技大学 | Coriolus versicolor trametes capable of efficiently decoloring multiple textile dyes |
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Application publication date: 20220729 |