CN109880647B

CN109880647B - Low-rank coal microbial grading degradation method

Info

Publication number: CN109880647B
Application number: CN201910232033.1A
Authority: CN
Inventors: 李建涛; 刘向荣; 蔡会武; 杨再文; 赵顺省; 杨杰; 石晨; 康红丽
Original assignee: Xian University of Science and Technology; Shangluo University
Current assignee: Xian University of Science and Technology; Shangluo University
Priority date: 2019-03-26
Filing date: 2019-03-26
Publication date: 2021-02-05
Anticipated expiration: 2039-03-26
Also published as: CN109880647A

Abstract

The invention discloses a method for degrading low-rank coal by microorganism in a grading way, which comprises the following steps: firstly, preparing low-rank coal into coal powder; secondly, carrying out photo-oxidation on the coal powder to obtain photo-oxidized coal powder; thirdly, adding the photo-oxidation coal powder into a liquid culture medium inoculated with streptomyces viridocystoris for primary degradation; fourthly, adding the sterilized first-stage degraded coal residues into a liquid culture medium inoculated with pseudomonas putida for second-stage degradation; and fifthly, adding the sterilized second-stage degraded coal residues into a liquid culture medium inoculated with phanerochaete chrysosporium to carry out third-stage degradation. The invention adopts streptomyces viridosporus, pseudomonas putida and phanerochaete chrysosporium to sequentially degrade the low-rank coal in a grading way, and utilizes various active substances such as alkali, enzyme, chelating agent, surfactant and the like generated by the metabolism of the three bacteria to perform synergistic action on different degradation active points in the structure of the low-rank coal, thereby improving the degradation rate of the low-rank coal and creating basic conditions for further clean and efficient utilization of the low-rank coal.

Description

Low-rank coal microbial grading degradation method

Technical Field

The invention belongs to the technical field of coal microbial conversion, and particularly relates to a method for degrading low-rank coal by microorganism in a grading manner.

Background

Coal is known as one of the most important energy sources, Chinese coal reserves are third in the world, and coal yield and consumption are always the first in the world. In the process of mining and utilizing coal, along with the preemptive mining, processing and utilizing of high-quality high-rank coal, the proportion of low-rank coal in the coal reserves is larger and larger. At present, more than 50% of low-rank coal in China is used for power generation, and then coking, gasification, liquefaction, direct combustion heating and the like are carried out. The utilization process of the low-rank coal has the following problems:

(1) the power generation efficiency is low; the low-rank coal has the characteristics of low heat value, high moisture and the like, so that the heat efficiency is low;

(2) the processing conditions are harsh; the processing modes of gasification, liquefaction, pyrolysis, coking and the like of low-rank coal generally need high temperature and high pressure, and have high requirements on equipment;

(3) the combustion pollution is large; n, S, heavy metal elements and ash content in the low-rank coal are high, and the low-rank coal can generate a large amount of pollutants such as smoke dust, nitrogen oxides, sulfur oxides and heavy metal compounds besides carbon dioxide and hydrocarbons in the combustion process.

The direct utilization of low-rank coal as fuel is increasingly unfavorable for the development of the current society, so the clean and efficient processing and utilization of the low-rank coal is an inevitable way for the sustainable development of coal.

The microbial degradation of coal is the latest technology after the liquefaction and gasification processing technology of coal, and is a new field relating to the subjects of microbiology, biochemistry, enzymology, molecular biology, separation engineering, coal chemistry, mineral processing and the like. The microbial degradation of the coal only needs to be carried out at room temperature and normal pressure, and has the advantages of mild reaction conditions, simple requirements on reaction equipment, low energy consumption, high product utilization rate and the like. The development prospect is bright in the aspect of clean and efficient utilization of coal, particularly low-rank coal.

Since 80 s of the last century scientists discovered that microorganisms can degrade coal, the microbial degradation of coal, as a new technology for coal processing, has achieved certain results through research and development for forty years, but with the progress of research, has discovered a number of problems, such as: (1) the degradation efficiency of the microorganism on the coal is low; (2) the research on the bacterium-coal matching rule is insufficient, efficient degrading bacteria are lacked, the universality of the degrading bacteria is lacked, and the like; (3) the composition and structure of coal are complex, so that microbial degradation products of the coal are difficult to separate and utilize; (4) the mechanism of microbial degradation of coal is not yet clear. The above problems are all restricting the industrialization of coal microbial degradation technology to different degrees, wherein the low degradation rate of coal by microbes is the most basic and critical problem. Because the degradation rate is too low, the degradation products are less, so that the research on the application and industrialization of the degradation products is hindered, and the subsequent research is influenced; the research of the mechanism is the key point, and the clear mechanism can guide the microbial degradation process of the coal from an objective rule, so that reasonable and scientific measures are made to improve the microbial degradation rate of the coal and design the composition of degradation products of the coal; the separation and the efficient utilization of the degradation products are the purposes, and the wider the application of the products is, the larger the additional value is, the research on the coal microbial degradation technology by people can be promoted in turn. Therefore, the above problems are urgently needed to be researched and broken through systematically.

The microbial degradation mechanism of the coal is as follows according to the proposed sequence: an enzymatic mechanism of action, an alkaline mechanism of action, a chelating agent mechanism of action, a surfactant mechanism of action, and an ABCDE mechanism of action. The ABCDE mechanism of action points out the functional group structure sites in the macromolecular structure of coal which are acted on by enzymes, alkali, chelating agents and surfactants respectively, and is proposed on the basis of the previous mechanisms, which think that the process of microbial degradation of coal is the result of the combined action of multiple mechanisms. Often a single strain is not capable of producing many coal-degrading active substances (enzymes, bases, chelating agents, surfactants, etc.) that degrade coal. At present, researchers mostly use single strains to carry out microbial degradation on coal, so the degradation rate is often low, even if the matching of individual strains and certain coal is good (active substances for degrading coal generated by the strains are exactly corresponding to degradation active points which are abundantly existed in the structure of certain coal), the strains are not always high in degradation rate on coal with different structure functional groups, namely the strains lack universality. Therefore, the method for researching and improving the coal microbial degradation rate by taking the coal microbial degradation mechanism as a starting point is the premise and the basis of high-efficiency clean utilization of coal, particularly low-rank coal.

Disclosure of Invention

The invention aims to solve the technical problem of providing a method for degrading low-rank coal by microorganism in a grading manner aiming at the defects of the prior art. The method sequentially degrades low-rank coal by streptomyces viridosporus, pseudomonas putida and phanerochaete chrysosporium in a grading manner, utilizes various active substances such as alkali, enzyme, chelating agent and surfactant generated by metabolism of the three bacteria respectively, and has synergistic effect on different degradation active points in the structure of the low-rank coal, so that the degradation rate of the low-rank coal is improved, the types and the number of products such as fine chemicals and liquid fuels in degradation solutions at all levels are improved, and basic conditions are created for further clean and efficient utilization of the low-rank coal.

In order to solve the technical problems, the technical scheme provided by the invention is as follows: the method for degrading the low-rank coal by the microorganism in a grading manner is characterized by comprising the following steps of:

step one, crushing and drying low-rank coal, and then sequentially grinding and screening to obtain coal powder with the particle size of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the pulverized coal obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized pulverized coal;

step three, adding the photo-oxidation coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridosporus, placing the liquid culture medium into an incubator for primary degradation, then filtering a primary degradation product, respectively collecting a primary degradation liquid and primary degradation coal residues, and then sterilizing the primary degradation coal residues;

step four, adding the sterilized first-stage degraded coal residues in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for second-stage degradation, filtering a second-stage degradation product, respectively collecting a second-stage degradation liquid and second-stage degraded coal residues, and then sterilizing the second-stage degraded coal residues;

and step five, adding the second-level degraded coal residues after sterilization in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium in an incubator for third-level degradation, filtering the third-level degradation products, and respectively collecting third-level degradation liquid and third-level degraded coal residues.

In the first step, the coal powder with the granularity of-0.15 mm +0.075mm is coal powder with the granularity of more than or equal to 0.075mm and less than 0.15 mm.

The method comprises the steps of crushing and screening low-rank coal to obtain coal powder, carrying out photo-oxidation pretreatment on the coal powder to obtain photo-oxidized coal powder, increasing the oxygen content in the coal powder so that the coal powder is more easily degraded by microorganisms, and then sequentially carrying out primary degradation of streptomyces viridospori (actinomycetes), secondary degradation of pseudomonas putida (bacteria) and tertiary degradation of phanerochaete chrysosporium (fungi) on the photo-oxidized coal powder. The mechanism of the first-stage degradation of actinomycete streptomyces viridochromogenes is an alkali action and a surfactant action, a group with stronger acidity in the photo-oxidation coal powder acts under the weaker alkali action in the first-stage degradation process, and the hydrophilicity of the photo-oxidation coal powder is enhanced under the surfactant action, so that the first-stage degradation process can be smoothly carried out; the secondary degradation mechanism of the pseudomonas putida comprises an alkali action, a surfactant action and a chelating agent action, the alkali action in the secondary degradation process is stronger, groups with weaker acidity in the primary degraded coal residues can be further reacted, the primary degradation process consumes the groups with stronger acidity in the photo-oxidized coal powder, so that alkaline substances generated in the secondary degradation process only react with the groups with weaker acidity, the groups with stronger acidity are prevented from being consumed, the alkaline degradation efficiency is improved, meanwhile, the hydrophilicity of the primary degraded coal residues is further enhanced by the substances with the surfactant property generated in the secondary degradation process, the full contact between the degraded coal active substances generated in the secondary degradation process and the degradable active sites in the primary degraded coal residues is facilitated, the secondary degradation efficiency is improved, and the chelating agent action enables metal ions playing a 'bridging' role in the primary degraded coal residues to be removed, macromolecular structures in the first-stage degraded coal residues collapse and depolymerize to a certain degree, so that further exposure of degradable active sites is promoted, and subsequent degradation is facilitated; the three-time degradation mechanism of the fungus Phanerochaete chrysosporium is an enzyme action, an alkali action and a surfactant action, the enzyme action in the three-stage degradation process is strong, so that the second-stage degraded coal residues are reacted and degraded under the action of enzyme generated in the three-stage degradation process, alkaline substances generated in the three-stage degradation process are further reacted with acid groups left in the second-stage degraded coal residues after the previous two-stage degradation, and meanwhile, the surfactant action enhances the hydrophilicity of the second-stage degraded coal residues and improves the efficiency of the three-stage degradation. Under the action of different microorganisms, various degradation products with different types and structures are contained in each level of degradation liquid obtained by three-level degradation, and fine chemicals, liquid fuels and other products can be obtained through further separation, so that high-efficiency utilization of low-rank coal is realized.

The sufficient and efficient degradation of the low-rank coal by the microorganisms is the premise and the basis of breakthrough of the microbial technology in the aspect of clean and efficient utilization of the low-rank coal. The invention utilizes three microorganisms to generate various active substances for degrading coal, such as alkali, enzyme, chelating agent, surfactant and the like, according to different action mechanisms of different microorganisms on the low-rank coal, the low-rank coal is degraded step by step, the later degradation is carried out on the basis of the former degradation, and the former degradation provides a foundation for the later degradation, so that the later degradation is easier to carry out, and simultaneously the synergistic action among the active substances is activated, the action on different degradation active points in the structure of the low-rank coal is jointly carried out, the succession rule of microorganism groups in the natural degradation process of the low-rank coal is met, thereby improving the degradation rate of the low-rank coal, improving the types and the quantity of products such as fine chemicals, liquid fuels and the like in various levels of degradation liquid, and laying a solid foundation for further clean and efficient utilization of the low-rank coal.

The rotary bed photochemical reactor disclosed in the utility model with the publication number of ZL201621380305.0 is adopted in the rotary bed photochemical reactor.

The method for degrading the low-rank coal in the microbial classification manner is characterized in that the conditions of the photo-oxidation pretreatment in the step two are as follows: the coal feeding amount is 20g/L calculated by the mass of the coal powder added into the unit volume of the rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min. And determining and obtaining the optimal process condition of the photo-oxidation pretreatment by a single-factor test and an orthogonal test variance analysis method. Under the optimal technological condition of the photo-oxidation pretreatment, the rotary bed photochemical reactor has the best photo-oxidation pretreatment effect on the pulverized coal, so that the oxygen content in the photo-oxidation pulverized coal is greatly improved, the degradation rate of microorganisms on the photo-oxidation pulverized coal in the subsequent grading degradation process is further improved, and the method is safer and more environment-friendly compared with a common nitric acid oxidation method.

The method for degrading the low-rank coal in the microbial classification manner is characterized in that the conditions of the first-level degradation in the third step are as follows: 9.50g/L of the addition amount of the photo-oxidative pulverized coal in the liquid culture medium inoculated with the streptomyces viridis, 180mL/L of the inoculation amount of the streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis, 160r/min of oscillation frequency of an incubator, 10d of culture time and 28 ℃ of culture temperature; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is not less than 3.0 multiplied by 10⁵one/mL.

The method for microbial graded degradation of low-rank coal is characterized in that the conditions of secondary degradation in the fourth step are as follows: the adding amount of the sterilized first-stage degraded coal residues in the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L of the mass of the photo-oxidized coal powder in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria solution in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria liquid is not less than 8.0 multiplied by 10⁵one/mL.

The method for microbial graded degradation of the low-rank coal is characterized in that the conditions of the third-level degradation in the fifth step are as follows: the addition amount of the second-stage degraded coal residue after sterilization in the liquid culture medium inoculated with the phanerochaete chrysosporium is 13.00g/L of the mass of the photo-oxidized coal powder in the first-stage degradation process, and the liquid of the phanerochaete chrysosporiumThe inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration in the phanerochaete chrysosporium mother liquor is not less than 2.0 multiplied by 10⁵one/mL.

The three degradation bacteria are cultured, growth curves of the three degradation bacteria are respectively drawn by combining a thallus dry weight method, the lag phase, the exponential growth phase, the stationary phase and the decay phase of various growth processes of the three degradation bacteria are respectively determined, and then the optimal degradation process conditions of all levels corresponding to the three degradation bacteria are respectively determined by a single-factor test and a response surface method. The degradation of the low-rank coal is carried out in sequence under the optimal degradation process conditions of all levels corresponding to the three degradation bacteria, and the degradation process of all levels is guaranteed to have higher degradation rate to degradation substrates, so that the total degradation rate of the low-rank coal is improved, the types and the number of products such as fine chemicals, liquid fuels and the like in all levels of degradation liquid are further improved, and the high-efficiency utilization of the low-rank coal is further realized on the basis.

Because the structures of the coal types (mainly lignite) belonging to the category of low-rank coal have similarity and can be expressed by a unified coal structure model, and the structures of the coal residues are not greatly changed compared with the structures of the corresponding low-rank coal after the microbial action, the types of substances contained in degradation liquid obtained by degrading the low-rank coal and the corresponding coal residues are not greatly different on the premise that the degradation process and the action bacteria are the same, and only the concentrations of the substances are different. That is, the concentration of the substance in the degradation liquid having a large degradation rate is large, the absorbance of the degradation liquid to be measured is large, the concentration of the substance in the degradation liquid having a small degradation rate is small, the absorbance of the degradation liquid to be measured is small, therefore, a determined relationship exists between the degradation rate of the low-rank coal and the corresponding coal residues and the absorbance of the degradation liquid, the three equations are correspondingly established through experimental research, and are suitable for degradation liquid obtained by corresponding microbial degradation of the low-rank coal and degradation liquid obtained by corresponding microbial degradation of coal residues obtained by other microbial degradation.

The method for degrading the low-rank coal in the microbial grading manner is characterized in that the absorbance of the first-level degradation liquid obtained in the third step at 450nm is measured by adopting a spectrophotometry method, and then the measured absorbance is brought into a relation equation between the first-level degradation rate of the streptomyces viridis degrading and photooxidizing the low-rank coal and the absorbance of the first-level degradation liquid, and the first-level degradation rate is calculated; the relation equation of the first-stage degradation rate of the streptomyces viridocystoris for degrading the photooxidation low-rank coal and the absorbance of the first-stage degradation liquid is as follows: eta₁＝0.02466+0.07453Y₁With a goodness-of-fit determination coefficient of R₁ ²0.98392, where eta₁First order degradation rate, Y₁The absorbance of the first-order degradation liquid is shown.

The method for degrading the low-rank coal in the microbial grading manner is characterized in that the absorbance of the secondary degradation liquid obtained in the fourth step at 450nm is measured by adopting a spectrophotometry method, and then the absorbance is brought into a relation equation between the secondary degradation rate of the pseudomonas putida degrading and photooxidizing the low-rank coal and the absorbance of the secondary degradation liquid, and the secondary degradation rate is obtained through calculation; the relation equation of the secondary degradation rate of the pseudomonas putida degrading and photo-oxidizing low-rank coal and the absorbance of the secondary degradation liquid is as follows: eta₂＝0.02919+0.06412Y₂With a goodness-of-fit determination coefficient of R₂ ²0.99075, where eta₂Is a second level of degradation, Y₂The absorbance of the second-order degradation liquid is shown.

The method for degrading the low-rank coal in the microbial grading manner is characterized in that the absorbance of the third-level degradation liquid obtained in the fifth step at 450nm is measured by adopting a spectrophotometry method, and then the absorbance is brought into a relation equation between the third-level degradation rate of the phanerochaete chrysosporium degradation photooxidation low-rank coal and the absorbance of the third-level degradation liquid, and the third-level degradation rate is obtained through calculation; the relation equation of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium for degrading the photo-oxidized low-rank coal is as follows: eta₃＝0.02336+0.08945Y₃With a goodness-of-fit determination coefficient of R₃ ²＝0.97836，Wherein eta₃Three-stage degradation rate, Y₃The absorbance of the third-order degradation liquid is shown.

And (3) measuring at 450nm by a spectrophotometry method to respectively obtain a relation equation between each level of degradation rate and absorbance of the corresponding degradation liquid in the three-level degradation process. The degradation rate and the absorbance are combined for use, a relational equation between the degradation rate of the microorganisms degrading the photooxidation low-rank coal and the absorbance of degradation liquid is established, and the accuracy of the degradation rate is improved, so that various levels of degradation rates can be quickly obtained according to the relational equation in the application process of the method, the method is convenient and efficient, and the degradation processes can be adjusted and controlled according to the degradation rate results of various levels, so that the degradation rate of the low-rank coal is improved.

The method for degrading the low-rank coal in the microbial classification manner is characterized in that the sterilization conditions in the third step and the fourth step are as follows: sterilizing at 121 deg.C for 20 min. The sterilization conditions can effectively kill streptomyces viridisporus remained in the first-stage degraded coal residues and pseudomonas putida remained in the second-stage degraded coal residues, thereby avoiding the influence of the streptomyces viridisporus in the first-stage degraded coal residues on the second-stage degradation process and the influence of the pseudomonas putida in the second-stage degraded coal residues on the third-stage degradation process, enhancing the degradation effects of the second-stage degradation and the third-stage degradation, and further improving the degradation rate of low-rank coal.

Compared with the prior art, the invention has the following advantages:

1. the invention sequentially degrades the low-rank coal by streptomyces viridosporus, pseudomonas putida and phanerochaete chrysosporium in a grading way, utilizes various active substances such as alkali, enzyme, chelating agent, surfactant and the like generated by the metabolism of the three bacteria according to different action mechanisms of the three bacteria on the low-rank coal, and has synergistic effect on different degradation active points in the structure of the low-rank coal, thereby improving the degradation rate of the low-rank coal, improving the types and contents of fine chemicals, liquid fuels and other products in various levels of degradation liquid, and laying a foundation for further clean and efficient utilization of the low-rank coal.

2. The method adopts the rotary bed photochemical reactor to carry out the photo-oxidation treatment on the pulverized coal prepared from the low-rank coal, increases the oxygen content in the photo-oxidation pulverized coal, improves the microbial degradation activity in the photo-oxidation pulverized coal, further improves the degradation rate of the photo-oxidation pulverized coal, and is simple, safe and environment-friendly.

3. The invention respectively carries out graded degradation under the optimal process condition of degrading the photo-oxidation coal dust by each grade of microorganism, ensures that degradation substrates have higher degradation rate in each grade of degradation process, thereby improving the total degradation rate of low-rank coal, further improving the types and the number of products such as fine chemicals, liquid fuels and the like in each grade of degradation liquid, and being beneficial to realizing the high-efficiency utilization of the low-rank coal.

4. The invention establishes a relation equation between the degradation rate in the degradation process of each level and the absorbance of the degradation liquid at 450nm, so that the absorbance A450 of the degradation liquid and the corresponding degradation rate can be converted quickly according to the relation equation, and the method is simple, direct, convenient and efficient.

5. The degradation liquid obtained by degrading the low-rank coal by microorganism grading contains various compounds, can be further separated to obtain various valuable compounds, and is used in the fields of clean liquid fuels, medicaments and industrial raw materials.

The technical solution of the present invention is further described in detail by the accompanying drawings and examples.

Drawings

FIG. 1 is a linear fitting graph of the degradation rate of Streptomyces viridis degrading photooxidation low-rank coal and the degradation liquid absorbance relationship.

FIG. 2 is a linear fitting graph of the relationship between the degradation rate of the pseudomonas putida degrading and photooxidizing low-rank coal and the absorbance of degradation liquid.

FIG. 3 is a linear fitting graph of the degradation rate of Phanerochaete chrysosporium degrading photooxidation low-rank coal and the degradation liquid absorbance relationship.

Detailed Description

Streptomyces viridochromosporus (Streptomyces viridosporus), Pseudomonas putida (Pseudomonas putida) and Phanerochaete chrysosporium (Phanerochaete chrysosporium) used in the invention are purchased from China general microbiological culture Collection center and are numbered as Streptomyces viridochromospora (4.1770), Pseudomonas putida (1.1820) and Phanerochaete chrysosporium (3.7212).

The culture medium adopted by the Streptomyces viridosporus of the invention is a Gao's No. one culture medium, and the composition of the culture medium is as follows: soluble starch 20g, KNO₃ 1g，K₂HPO₄ 0.5g，MgSO₄·7H₂O 0.5g，NaCl0.5g，FeSO₄·7H₂0.01g of O, (15 g of agar in a solid culture medium), 1000mL of distilled water and 7.4-7.6 of pH. Firstly reviving the preserved Streptomyces viridosporus, adopting a liquid culture medium to culture the Streptomyces viridosporus to the third generation, and culturing the Streptomyces viridosporus to the third generation 3d with the concentration of not less than 3.0 multiplied by 10⁵And (3) taking the streptomyces viridosporus bacterial liquid per mL as a mother bacterial liquid for inoculation to obtain a liquid culture medium inoculated with the streptomyces viridosporus for primary degradation and independent degradation of low-rank coal.

The culture medium adopted by the Pseudomonas putida (Pseudomonas putida) is an LB culture medium, and the composition of the culture medium is as follows: 10g of peptone, 5g of yeast powder, 10g of NaCl, (15 g of agar in a solid culture medium), 1000mL of distilled water and pH 7.4-7.6. Reviving the preserved Pseudomonas putida (Pseudomonas putida), culturing in liquid culture medium to the third generation, and culturing to the third generation 2d with bacteria concentration of 8.0 × 10⁵And (3) inoculating the pseudomonas putida bacterial liquid serving as mother bacterial liquid to obtain a liquid culture medium inoculated with the pseudomonas putida for secondary degradation and independent degradation of low-order coal.

The culture medium adopted by the Phanerochaete chrysosporium (Phanerochaete chrysosporium) of the invention is an improved martin culture medium, and the culture medium comprises the following components: 5g of peptone, 2g of yeast powder, 20g of glucose, 1g of dipotassium phosphate, 0.5g of magnesium sulfate, (15 g of agar added into a solid culture medium), 1000mL of distilled water and pH of 6.2-6.6. Reviving preserved Phanerochaete chrysosporium, culturing in liquid culture medium to the third generation, culturing to the third generation 2d with spore concentration not less than 2.0 × 10⁵And (3) inoculating the individual/mL phanerochaete chrysosporium bacterial liquid serving as a mother bacterial liquid to obtain a liquid culture medium inoculated with the phanerochaete chrysosporium for three-stage degradation and independent degradation of low-order coal.

Generally, the absorbance of the microbial degradation liquid at 450nm or the degradation rate of the microbes to the substrate coal is used as an index for evaluating the coal degradation effect, and the degradation rate of the microbes for degrading the low-rank coal is calculated according to the following formula:

in the formula (a), η is the degradation rate (%), m₀The mass (g) of the low-rank coal is m₁The mass (g) of coal residue obtained for degradation, A₀Ash mass (g) in low rank coal, A₁The mass (g) of ash in the coal residue obtained by degradation.

The degradation effect is evaluated by absorbance, the operation is simple, but subsequent calculation is needed, the degradation effect is evaluated by the degradation rate, the operation is complex, the degradation rate of the microorganisms to the low-rank coal is related to the granularity of the low-rank coal, and the smaller the granularity of the low-rank coal is, the higher the degradation rate of the microorganisms to the low-rank coal is. When the particle size of the low-rank coal is larger, the coal residue obtained by degradation can be fully separated from microbial thalli, so that the degradation rate is accurately calculated by using the formula (a); when the particle size of the low-rank coal is too small, the coal residue obtained by microbial degradation is not completely separated from microbial thalli, so that the parameter values in the formula (a) are inaccurate, and the degradation rate obtained according to the formula (a) has a large error. Therefore, the two indexes are combined for use, a relational equation of the degradation rate of the microbially degraded and photo-oxidized low-rank coal and the absorbance of the degradation liquid is established according to the corresponding relational rule of the degradation liquid obtained by the microbial degradation of the low-rank coal with larger granularity and the degradation rate calculated by the formula (a), and then the applicability analysis is carried out on the relational equation. The method for obtaining the degradation rate by adopting the relational equation is simple and easy to implement, the degradation solution is obtained by only carrying out simple centrifugal operation on the degradation solution, the absorbance value of the degradation solution is measured, the coal residue obtained by degradation is not required to be completely separated from microbial cells, the operation difficulty is reduced, and the accuracy of the degradation rate is greatly improved.

(1) Establishment of relation equation between degradation rate of streptomyces viridosporus degrading photooxidation low-rank coal and degradation liquid absorbance

Degrading photo-oxidation endosymbiont lignite (GSH) with the granularity of-1.7 mm +1mm (namely the granularity is more than or equal to 1mm and less than 1.7mm), -1mm +0.7mm (namely the granularity is more than or equal to 0.7mm and less than 1mm), -0.7mm +0.5mm (namely the granularity is more than or equal to 0.5mm and less than 0.7mm), -0.5mm +0.25mm (namely the granularity is more than or equal to 0.25mm and less than 0.5mm), -0.25mm +0.15m (namely the granularity is more than or equal to 0.15mm and less than 0.25mm), -0.15mm +0.075mm (namely the granularity is more than or equal to 0.075mm and less than 0.045mm), and-0.045 mm (namely the granularity is less than 0.045mm) by adopting streptomyces viridis, three groups of parallel experiments are set for the degradation of photooxidation inner Mongolian lignite (GSLH) with each granularity, and the degradation conditions are as follows: the coal adding amount is 9.50g/L in terms of the mass of photooxidation endosymbiont lignite (GSLH) in a unit volume of liquid culture medium of streptomyces viridis, the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.1 multiplied by 10⁵one/mL.

After degradation, respectively centrifuging degradation products for 15min under the condition of 10000r/min, filtering obtained supernate, performing secondary filtration through a filter membrane of 0.22 mu m to obtain secondary filtrate, taking deionized water as reference, determining the absorbance A450 of the secondary filtrate at 450nm by adopting a TU-1900 type spectrophotometer, and taking the absorbance average value of three parallel experiments of photooxidation inner Mongolian lignite (GSLH) of each granularity as the value of A450 under the granularity; and then, carrying out multiple times of washing on precipitates obtained by centrifuging the degradation products of the photooxidation endosymbiont lignite (GSLH) with the granularity of-1.7 mm +1mm, -1mm +0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm, -0.25mm +0.15m, removing thalli, drying at 60 ℃ to constant weight, calculating the degradation rate corresponding to the five-granularity photooxidation endosymbiont lignite (GSLH) according to a formula (a), and taking the average degradation rate of three groups of parallel experiments of the photooxidation endosymbiont lignite (GSLH) with each granularity as the degradation rate under the granularity.

According to the results of experimental data, the larger size of the degraded Streptomyces viridis is (namely-1.7 mm +1mm,-1mm +0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm, -0.25mm +0.15m) and a linear fitting graph of degradation rate and absorbance relationship of the photooxidation inner Mongolian lignite (GSLH), as shown in figure 1, the obtained equation of the relationship between the degradation rate and the absorbance of degradation liquid of the streptomyces viridis degradation photooxidation low-rank coal is as follows: eta₁₀＝0.02466+0.07453Y₁₀With a goodness-of-fit determination coefficient of R₁₀ ²0.98392, good fitting degree and high reliability, wherein eta₁₀For the degradation rate, Y₁₀The absorbance of the degradation solution is shown. The A450 values (2.693, 2.587 and 2.145 respectively) of photo-oxidized inner Mongolian lignite (GSLH) with the grain sizes of-0.15 +0.075mm, -0.075+0.045mm and-0.045 mm are used for obtaining the corresponding degradation rate eta₁₀Respectively as follows: 22.54%, 21.68% and 18.45%.

(2) Applicability analysis of relation equation between degradation rate of streptomyces viridocystoris for degrading photooxidation low-rank coal and absorbance of degradation liquid

Respectively degrading photo-oxidized Yunnan Zhaotong lignite (GZTH), photo-oxidized muddy mountain lignite (GHYH) and photo-oxidized inner Mongolian Yuanbao mountain lignite (GYBH) by adopting streptomyces viridisporus, wherein the granularity of each photo-oxidized low-rank coal is divided into-1.7 mm +1mm, -1mm +0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm and-0.25 mm +0.15m, three groups of parallel experiments are set for the degradation of each granularity of the photo-oxidized low-rank coal, and the degradation conditions are as follows: the coal adding amount is 9.50g/L of the mass of the photooxidation low-order coal in a liquid culture medium of streptomyces viridis per unit volume, the inoculation amount of streptomyces viridis mother liquor in the culture medium of streptomyces viridis liquid is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.1 multiplied by 10⁵one/mL.

After degradation, respectively centrifuging the degradation products for 15min under the condition of 10000r/min, filtering the obtained supernatant, performing secondary filtration through a filter membrane of 0.22 mu m to obtain secondary filtrate, taking deionized water as reference, measuring the absorbance A450 of the secondary filtrate at 450nm by adopting a TU-1900 type spectrophotometer, and taking the absorbance average value of three parallel experiments of photooxidation low-rank coal of each granularity as the A450 value of the granularity; then, for each granularity of the photo-oxidation low-rank coal degradation product, the product is separatedWashing the sediment obtained from the core for multiple times, removing thalli, drying at 60 ℃ to constant weight, calculating the degradation rates corresponding to the five-granularity photooxidation low-rank coal according to a formula (a), and taking the average value of the degradation rates of three groups of parallel experiments of the photooxidation low-rank coal with each granularity as the degradation rate under the granularity; then the A450 value of the photooxidation low-rank coal with each granularity is substituted into a relation equation eta of degradation rate and degradation liquid absorbance of streptomyces viridis for degrading the photooxidation low-rank coal₁₀＝0.02466+0.07453Y₁₀The corresponding degradation rate is obtained as a predicted value (eta)_Prediction) And the degradation rate (. eta.) calculated by the formula (a)_{Practice of}) Comparative studies were conducted and the results are shown in table 1 below.

TABLE 1 analysis result of the applicability of the equation of the relationship between the degradation rate of Streptomyces viridogriseus degraded photooxidation low-rank coal and the absorbance of the degraded solution

As can be seen from Table 1, the degradation rates eta of the three types of photo-oxidized low-rank coals with different particle sizes are obtained according to the relation equation between the degradation rate of the streptomyces viridosporus in degrading the photo-oxidized low-rank coals and the absorbance of the degradation liquid_PredictionDegradation rate eta obtained by the formula (a)_{Practice of}The relative error between the degradation rate and the degradation solution absorbance equation is small, so that the degradation rate and the degradation solution absorbance equation of the streptomyces viridosporus degrading the photooxidation low-rank coal have good applicability to different photooxidation low-rank coals, and the method can be used for degrading the first-stage degradation solution absorbance (Y) of the photooxidation low-rank coal by the streptomyces viridosporus under the first-stage degradation process condition of the invention₁) And first order degradation rate (. eta.)₁) Conversion between, i.e. expressed as η₁＝0.02466+0.07453Y₁With a goodness-of-fit determination coefficient of R₁ ²＝0.98392。

(3) Establishment of relation equation between degradation rate of pseudomonas putida degrading and photo-oxidizing low-rank coal and absorbance of degradation liquid

Pseudomonas putida is adopted to respectively degrade photo-oxidation endosymbiont lignite (GSH) with the granularity of-1.7 mm +1mm (namely the granularity is more than or equal to 1mm and less than 1.7mm), -1mm +0.7mm (namely the granularity is more than or equal to 0.7mm and less than 0.7mm), -0.5mm +0.25mm (namely the granularity is more than or equal to 0.25mm and less than 0.5mm), -0.25mm +0.15m (namely the granularity is more than or equal to 0.15mm and less than 0.25mm), -0.15mm +0.075mm (namely the granularity is more than or equal to 0.075mm and less than 0.15mm), -0.075mm +0.045mm (namely the granularity is more than or equal to 0.045mm) and-0.045 mm (namely the granularity is less than 0.045mm), three groups of parallel experiments are set for the degradation of photooxidation inner Mongolian lignite (GSLH) with each granularity, and the degradation conditions are as follows: the coal adding amount is 13.00g/L of the mass of the photo-oxidation coal powder in the liquid culture medium of the pseudomonas putida in unit volume, the inoculation amount of a pseudomonas putida mother bacteria liquid in the culture medium of the pseudomonas putida liquid is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵one/mL.

According to the results of experimental data, the larger particle size of the pseudomonas putida degradation (namely-1.7 mm +1mm, -1mm +0.7mm, -0.7mm +0.5mm and-0) is drawn5mm +0.25mm, -0.25mm +0.15m) photo-oxidation inner Mongolia lignite (GSLH) degradation rate and absorbance relation linear fitting graph, as shown in figure 2, the obtained pseudomonas putida degradation photo-oxidation low-rank coal degradation rate and degradation liquid absorbance relation equation is as follows: eta₂₀＝0.02919+0.06412Y₂₀With a goodness-of-fit determination coefficient of R₂₀ ²0.99075, good fitting degree and high reliability, wherein eta₂₀For the degradation rate, Y₂₀The absorbance of the degradation solution is shown. The A450 values (5.875, 5.321 and 4.827 respectively) of photo-oxidized inner Mongolian lignite (GSLH) with the grain sizes of-0.15 +0.075mm, -0.075+0.045mm and-0.045 mm are used for obtaining the corresponding degradation rate eta₂₀Respectively as follows: 40.59%, 37.04% and 33.87%.

(4) Applicability analysis of relation equation between degradation rate of pseudomonas putida degrading and photo-oxidizing low-rank coal and absorbance of degradation liquid

Adopting pseudomonas putida to degrade photo-oxidized Yunnan Zhaotong lignite (GZTH), photo-oxidized Shanxi muddy source lignite (GHYH) and photo-oxidized inner Mongolian Yuanshan lignite (GYBH) respectively, wherein the granularity of each photo-oxidized low-rank coal is divided into-1.7 mm +1mm, -1mm +0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm and-0.25 mm +0.15m, three groups of parallel experiments are set for the degradation of each granularity of the photo-oxidized low-rank coal, and the degradation conditions are as follows: the coal adding amount is 13.00g/L of the mass of the photo-oxidation coal powder in the liquid culture medium of the pseudomonas putida per unit volume, the inoculation amount of a pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵one/mL.

After degradation, respectively centrifuging the degradation products for 15min under the condition of 10000r/min, filtering the obtained supernatant, performing secondary filtration through a filter membrane of 0.22 mu m to obtain secondary filtrate, taking deionized water as reference, measuring the absorbance A450 of the secondary filtrate at 450nm by adopting a TU-1900 type spectrophotometer, and taking the absorbance average value of three parallel experiments of photooxidation low-rank coal of each granularity as the A450 value of the granularity; then obtaining the product of photo-oxidation low-rank coal degradation of each granularity by centrifugationWashing the precipitate for multiple times, removing thalli, drying at 60 ℃ to constant weight, calculating the degradation rate corresponding to the five-granularity photooxidation low-rank coal according to a formula (a), and taking the average value of the degradation rates of three groups of parallel experiments of the photooxidation low-rank coal with each granularity as the degradation rate under the granularity; then the A450 value of the photooxidation low-rank coal with each granularity is substituted into a relation equation eta of the degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of degradation liquid₂₀＝0.02919+0.06412Y₂₀The corresponding degradation rate is obtained as a predicted value (eta)_Prediction) And the degradation rate (. eta.) calculated by the formula (a)_{Practice of}) Comparative studies were conducted and the results are shown in table 2 below.

Table 2 applicability analysis results of relation equation between degradation rate of pseudomonas putida degrading and photo-oxidizing low-rank coal and absorbance of degradation liquid

As can be seen from Table 2, the degradation rates eta of the three types of photo-oxidized low-rank coals with different particle sizes are obtained according to the equation of the relationship between the degradation rate of the pseudomonas putida degraded photo-oxidized low-rank coal and the absorbance of the degradation liquid_PredictionDegradation rate eta obtained by the formula (a)_{Practice of}The relative error between the degradation rate and the degradation solution absorbance equation is small, which shows that the degradation rate and the degradation solution absorbance equation of the pseudomonas putida degraded photooxidation low-rank coal have good applicability to different photooxidation low-rank coals, and the degradation solution absorbance (Y) of the pseudomonas putida degraded photooxidation low-rank coal can be used under the secondary degradation process condition of the invention₂) And second order degradation rate (. eta.)₂) The conversion between. I.e. as η₂＝0.02919+0.06412Y₂With a goodness-of-fit determination coefficient of R₂ ²＝0.99075。

(5) Establishment of relation equation between degradation rate of phanerochaete chrysosporium for degrading photo-oxidized low-rank coal and absorbance of degradation liquid

Phanerochaete chrysosporium is adopted to respectively degrade the photo-oxidation endospore lignite with the granularity of-1.7 mm +1mm (namely the granularity is more than or equal to 1mm and less than 1.7mm), -1mm +0.7mm (namely the granularity is more than or equal to 0.7mm and less than 1mm), -0.7mm +0.5mm (namely the granularity is more than or equal to 0.5mm and less than 0.7mm), -0.5mm +0.25mm (namely the granularity is more than or equal to 0.25mm and less than 0.5mm), -0.25mm +0.15m (namely the granularity is more than or equal to 0.075mm and less than 0.15mm), -0.075mm +0.045mm (namely the granularity is more than or equal to 0.045mm and less than 0.045mm), three groups of parallel experiments are set for the degradation of photooxidation inner Mongolian lignite (GSLH) with each granularity, and the degradation conditions are as follows: the coal adding amount is 13.00g/L of the mass of the photo-oxidation coal powder in a unit volume of a liquid culture medium of the phanerochaete chrysosporium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.3 multiplied by 10⁵one/mL.

According to the results of experimental data, the larger degradation particle sizes (i.e., -1.7mm +1mm, -1 mm) of Phanerochaete chrysosporium are plotted+0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm, -0.25mm +0.15m) of the degradation rate and absorbance relationship linear fitting graph of the photooxidation inner Mongolia lignite (GSLH), as shown in figure 3, the obtained degradation rate and degradation solution absorbance relationship equation of the phanerochaete chrysosporium degradation photooxidation low-order coal is as follows: eta₃₀＝0.02336+0.08945Y₃₀The goodness-of-fit determination coefficient is R₃₀ ²0.97836, good fitting degree and high reliability, wherein eta₃₀For the degradation rate, Y₃₀The absorbance of the degradation solution is shown. The A450 values (5.228, 4.409 and 3.511 respectively) of photo-oxidized inner Mongolian lignite (GSLH) with the grain sizes of-0.15 +0.075mm, -0.075+0.045mm and-0.045 mm are used for obtaining the corresponding degradation rate eta₃₀Respectively as follows: 48.09%, 40.95% and 33.12%.

(6) Applicability analysis of relation equation between degradation rate of phanerochaete chrysosporium for degrading photo-oxidized low-rank coal and absorbance of degradation liquid

The method comprises the steps of respectively degrading photo-oxidized Yunnan Zhaotong lignite (GZTH), photo-oxidized Shanxi muddy source lignite (GHYH) and photo-oxidized inner Mongolian Yuanbao mountain lignite (GYBH) by adopting Phanerochaete chrysosporium, wherein the granularity of each photo-oxidized low-rank coal is divided into-1.7 mm +1mm, -1mm +0.7mm, -0.7mm +0.5mm, -0.5mm +0.25mm and-0.25 mm +0.15m, three groups of parallel experiments are set for the degradation of each granularity of the photo-oxidized low-rank coal, and the degradation conditions are as follows: the coal adding amount is 13.00g/L of the mass of the photo-oxidation coal powder in a unit volume of a liquid culture medium of the phanerochaete chrysosporium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.3 multiplied by 10⁵one/mL.

After degradation, respectively centrifuging the degradation products for 15min under the condition of 10000r/min, filtering the obtained supernatant, performing secondary filtration through a filter membrane of 0.22 mu m to obtain secondary filtrate, taking deionized water as reference, measuring the absorbance A450 of the secondary filtrate at 450nm by adopting a TU-1900 type spectrophotometer, and taking the absorbance average value of three parallel experiments of photooxidation low-rank coal of each granularity as the A450 value of the granularity; then for each granularity of lightWashing precipitates obtained by centrifuging degraded products of the oxidized low-rank coal for multiple times, removing thalli, drying at 60 ℃ to constant weight, calculating degradation rates corresponding to the five-granularity photooxidation low-rank coal according to a formula (a), and taking the average value of the degradation rates of three groups of parallel experiments of the photooxidation low-rank coal with each granularity as the degradation rate under the granularity; then substituting the A450 value of the photooxidation low-rank coal with each granularity into a relation equation eta of degradation rate and degradation liquid absorbance of the phanerochaete chrysosporium for degrading the photooxidation low-rank coal₃₀＝0.02336+0.08945Y₃₀The corresponding degradation rate is obtained as a predicted value (eta)_Prediction) And the degradation rate (. eta.) calculated by the formula (a)_{Practice of}) Comparative studies were conducted and the results are shown in table 3 below.

TABLE 3 analysis result of the applicability of the equation of the relationship between the degradation rate of Phanerochaete chrysosporium degrading photooxidation low-rank coal and the absorbance of the degradation liquid

As can be seen from Table 3, the degradation rates eta of the three photo-oxidized low-rank coals with different particle sizes are obtained according to the equation of the relationship between the degradation rate of the phanerochaete chrysosporium for degrading the photo-oxidized low-rank coals and the absorbance of the degradation liquid_PredictionDegradation rate eta obtained by the formula (a)_{Practice of}The relative error between the degradation rate and the degradation liquid absorbance equation is small, which shows that the degradation rate of phanerochaete chrysosporium degrading the photooxidation low-rank coal has good applicability to different photooxidation low-rank coals, and the degradation liquid absorbance (Y) of the phanerochaete chrysosporium degrading the photooxidation low-rank coal can be used under the three-level degradation process condition of the invention₃) And third degree degradation rate (. eta.)₃) The conversion between. I.e. as η₃＝0.02336+0.08945Y₃With a goodness-of-fit determination coefficient of R₃ ²＝0.97836。

Example 1

The embodiment comprises the following steps:

firstly, crushing the Nemontmorillonoid river lignite (HLH), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain Nemontmorillonoid river lignite (HLH) coal powder with the granularity of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the Nemontmorillon Hu Lignite (HLH) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized pulverized coal of the Nemontmorillon Hu lignite (GHLH); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of pulverized coal of the Nemeng Huiyeh Lignite (HLH) added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation coal powder of the Nemenghua lignite (GHLH) obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for primary degradation, then filtering primary degradation products, respectively collecting primary degradation liquid and primary degradation coal residues, placing the primary degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the primary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁1.866, and bringing the degradation product into a relation equation eta of the first-stage degradation rate of streptomyces viridis degrading photooxidation low-order coal and the absorbance of the first-stage degradation liquid₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.1637, i.e. η₁16.37 percent; the first-stage degradation conditions are as follows: 9.50g/L of the adding amount of photooxidation inner Mongolian Huohan lignite (GHLH) coal powder in a unit volume of liquid culture medium inoculated with streptomyces viridis, 180mL/L of the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.2 multiplied by 10⁵Per mL;

step four, adding the sterilized primary coal residue in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for secondary degradation, and then performing secondary degradation on the secondary degradationFiltering the product, respectively collecting the second-stage degradation liquid and the second-stage degradation coal residue, sterilizing the second-stage degradation coal residue in an autoclave at 121 deg.C for 20min, determining the absorbance of the second-stage degradation liquid at 450nm by spectrophotometry with deionized water as blank control to obtain Y₂2.812, the degradation rate of the pseudomonas putida is introduced into a relation equation eta of the secondary degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of a secondary degradation solution₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.2095, i.e. η₂20.95%,; the conditions of the secondary degradation are as follows: the addition amount of the sterilized first-stage degradation coal residue in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L in terms of the mass of the pulverized coal of photooxidation Nenghorghe lignite (GHLH) in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.3 multiplied by 10⁵Per mL;

step five, adding the second-level coal residues sterilized in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium in an incubator for third-level degradation, filtering the third-level degradation products, respectively collecting third-level degradation liquid and third-level degradation coal residues, taking deionized water as a blank control, and measuring the absorbance of the third-level degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃3.526, introducing the degradation product into a relation equation eta of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium degradation photo-oxidation low-order coal₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.3388, i.e. η₃33.88%; the conditions of the third-stage degradation are as follows: the addition amount of the second-stage degradation coal residue after sterilization in the liquid culture medium inoculated with phanerochaete chrysosporium in unit volume is 13.00g/L calculated by the mass of the pulverized coal of the photooxidation inner Mongolian Hughua lignite (GHLH) in the first-stage degradation process, and the addition amount of the second-stage degradation coal residue is 13.00g/L calculated by the mass of the pulverized coal of the phanerochaete chrysosporiumThe inoculation amount of a phanerochaete chrysosporium mother solution in a liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.1 multiplied by 10⁵one/mL.

Comparative example 1

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the Nemontmorillon Hu Lignite (HLH) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized pulverized coal of the Nemontmorillon Hu lignite (GHLH); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of the light oxidation Nemeng Huohu lignite (GHLH) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the light oxidation pretreatment is 10 min;

step three, adding the photo-oxidation coal powder of the Nemeng Huihe lignite (GHLH) obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and measuring the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁₀1.866, and bringing the degradation rate into a relation equation eta of degradation rate of streptomyces viridis degrading the photooxidation low-rank coal and degradation liquid absorbance₁₀＝0.02466+0.07453Y₁₀Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.1637, i.e. η₁₀16.37 percent; the conditions for degrading the photooxidation endomongol hollin river lignite by the streptomyces viridis single bacteria are as follows: the coal adding amount is 9.50g/L calculated by the mass of the light oxidation inner Mongolian Huohui lignite (GHLH) coal powder in a unit volume streptomyces viridis liquid culture medium, and the streptomyces viridis in the streptomyces viridis liquid culture mediumThe inoculation amount of the mother bacteria liquid is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.2 multiplied by 10⁵one/mL.

Comparative example 2

This comparative example comprises the following steps:

step three, adding the photo-oxidation coal powder of the Nemon Hulllin river lignite (GHLH) obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂₀2.991, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degradation photooxidation low-order coal is brought into₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀0.2209, i.e. η₂₀22.09%; the conditions for degrading the photooxidation Nemond Hurrier lignite (GHLH) by the pseudomonas putida are as follows: the coal adding amount is 13.00g/L by the mass of the pulverized coal of photooxidation Nemond Hurrill lignite (GHLH) in a unit volume of pseudomonas putida liquid culture medium, the inoculation amount of pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, and the oscillation frequency of an incubator160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.3 multiplied by 10⁵one/mL.

Comparative example 3

step three, adding the photo-oxidation Nemonthlin river lignite (GHLH) coal powder obtained in the step two into a liquid culture medium inoculated with Phanerochaete chrysosporium, placing the culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀2.973, the degradation rate and degradation liquid absorbance relation equation eta of the phanerochaete chrysosporium degradation photooxidation low-order coal is introduced into₃₀＝0.02336+0.08945Y₃₀Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.2893, i.e. η₃₀28.93%; the conditions for degrading photooxidation inner Mongolian Huohui brown coal (GHLH) by the phanerochaete chrysosporium under the single bacteria are as follows: the coal adding amount is 13.00g/L calculated by the mass of the coal powder of photooxidation inner Mongolian Hughua lignite (GHLH) in a unit volume of phanerochaete chrysosporium liquid culture medium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the Phanerochaete chrysosporium parent strainThe spore concentration in the liquid is 2.1 × 10⁵one/mL.

Comparing example 1 with comparative examples 1 to 3, it can be seen that the total degradation rate η of the microbial graded degradation of photooxidized inner Mongolian Hulllin river lignite (GHLH) in example 1_{General assembly}＝η₁₊η₂+η₃0.7120, i.e. η_{General assembly}71.20%, and the degradation rates of Streptomyces viridogriseus, Pseudomonas putida and Phanerochaete chrysosporium in comparative examples 1-3 to photooxidation Nemonthlin Hoghe lignite (GHLH) were eta_10＝16.37％、η_20＝22.09％、η₃₀The total degradation rate of the photooxidation of the Mongolian Hurrian coal (GHLH) by sequentially adopting the three bacteria is far greater than the degradation rate of the photooxidation of the Mongolian Hurrian coal (GHLH) by respectively and independently adopting the three bacteria, which means that the grading degradation method of the invention improves the microbial degradation rate of the Mongolian Hurrian coal (HLH).

Example 2

The embodiment comprises the following steps:

step one, crushing Yunnan Zhaotong lignite (ZTH), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain Yunnan Zhaotong lignite (ZTH) coal powder with the granularity of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the Yunnan Zhaotong lignite (ZTH) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized Yunnan Zotong lignite (GZTH) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal adding amount is 20g/L (mass/mass) of the pulverized coal of Yunnan Zhaotong lignite (ZTH) added into a unit volume rotating bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter is 10min (volume/volume) of the rotating bed photochemical reactor before the photooxidation pretreatment;

step three, adding the photo-oxidation Yunnan Zhaotong lignite (GZTH) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium into an incubator for primary degradation, then filtering primary degradation products, respectively collecting primary degradation liquid and primary degradation coal residues, and placing the primary degradation coal residues into autoclave for sterilizationSterilizing at 121 deg.C for 20min, taking deionized water as blank control, and measuring absorbance of the first-stage degradation liquid at 450nm by spectrophotometry to obtain Y₁2.873, and the product is brought into a relation equation eta of the first-stage degradation rate and the first-stage degradation liquid absorbance of the streptomyces viridis to degrade the photooxidation low-rank coal₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.2388, i.e. η₁23.88%; the first-stage degradation conditions are as follows: the adding amount of the pulverized coal of photooxidation Yunnan Shoitong lignite (GZTH) in a unit volume of liquid culture medium inoculated with streptomyces viridis is 9.50g/L, the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.3 multiplied by 10⁵Per mL;

step four, adding the sterilized first-stage coal residues in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium in an incubator for secondary degradation, then filtering a secondary degradation product, respectively collecting a secondary degradation liquid and second-stage degradation coal residues, placing the second-stage degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the secondary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂3.549, the degradation rate of the pseudomonas putida is carried into the relation equation eta of the secondary degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of the secondary degradation liquid₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.2568, i.e. η₂25.68 percent; the conditions of the secondary degradation are as follows: the addition amount of the sterilized first-stage degradation coal residue in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L in terms of the mass of pulverized coal of photooxidation Yunnan Zhaotong (GZTH) in the first-stage degradation process, the inoculation amount of pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the malodor is fakeViable bacteria concentration in the monimonas solution was 8.3 ×.10⁵Per mL;

step five, adding the second-level coal residues sterilized in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium into an incubator for third-level degradation, then filtering a third-level degradation product, and respectively collecting a third-level degradation liquid and third-level degradation coal residues; taking deionized water as blank control, and measuring absorbance of the third-stage degradation liquid at 450nm by spectrophotometry to obtain Y₃3.559, introducing the degradation product into a relation equation eta of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium degradation photo-oxidation low-order coal₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.3417, i.e. η₃34.17 percent; the conditions of the third-stage degradation are as follows: the addition amount of the sterilized second-stage degraded coal residue in the liquid culture medium inoculated with the phanerochaete chrysosporium in unit volume is 13.00g/L of the mass of the pulverized coal of photooxidation Yunnan Zhaotong lignite (GZTH) in the first-stage degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.2 multiplied by 10⁵one/mL.

Comparative example 4

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the Yunnan Zhaotong lignite (ZTH) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized Yunnan Zhaotong lignite (ZTH) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal adding amount is 20g/L (mass/mass) of the pulverized coal of Yunnan Zhaotong lignite (ZTH) added into a unit volume rotating bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter is 10min (volume/volume) of the rotating bed photochemical reactor before the photooxidation pretreatment;

step three, adding the photo-oxidation Yunnan Zhaotong lignite (GZTH) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁₀2.873, and the degradation rate and degradation liquid absorbance relation equation eta is introduced into streptomyces viridis for degrading the photooxidation low-rank coal₁₀＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.2388, i.e. η₁₀23.88%; the conditions for degrading the photooxidation Yunnan Zhaotong lignite (GZTH) by the streptomyces viridis are as follows: the coal adding amount is 9.50g/L based on the mass of the pulverized coal in the unit volume of streptomyces viridis liquid culture medium for photooxidation of Yunnan Shogtong lignite (GZTH), the inoculation amount of streptomyces viridis mother liquor in the streptomyces viridis liquid culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.3 multiplied by 10⁵one/mL.

Comparative example 5

This comparative example comprises the following steps:

step three, adding the photo-oxidation Yunnan Zhaotong lignite (GZTH) coal powder obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂₀4.420, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degradation photooxidation low-rank coal is introduced₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀0.3126, i.e. η₂₀31.26%; the conditions for degrading the pseudomonas putida by the single bacterium are as follows: the coal adding amount is 13.00g/L calculated by the mass of pulverized coal of photooxidation Yunnan Zhaotong lignite (GZTH) in a unit volume of pseudomonas putida liquid culture medium, the inoculation amount of pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.3 multiplied by 10⁵one/mL.

Comparative example 6

step three, adding the photo-oxidation Yunnan Zhaotong lignite (GZTH) coal powder obtained in the step two into the inoculated Phanerochaete chrysosporium (Phanerochaete chry)sorrium) in a liquid culture medium, placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀4.890, the degradation rate and degradation liquid absorbance relation equation eta of the phanerochaete chrysosporium degradation photooxidation low-order coal is introduced₃₀＝0.02336+0.08945Y₃₀Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.4608, i.e. eta₃₀46.08%; the conditions for degrading the phanerochaete chrysosporium single strain are as follows: the coal adding amount is 13.00g/L in terms of the mass of the coal powder of photooxidation Yunnan Shoitong brown coal (GZTH) in a unit volume of Phanerochaete chrysosporium liquid culture medium, the inoculation amount of a Phanerochaete chrysosporium mother bacteria liquid in the Phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.2 multiplied by 10⁵one/mL.

As can be seen from comparison of example 2 with comparative examples 4 to 6, the total degradation rate eta of the microbial fractionation of the photooxidized Yunnan Showa lignite (GZTH) in example 2_{General assembly}＝η₁₊η₂+η₃0.8373, i.e. η_{General assembly}83.73%, and the degradation rates of Streptomyces viridis, Pseudomonas putida and Phanerochaete chrysosporium in comparative examples 4-6, which directly degrade the photooxidized Yunnan Showa lignite (GZTH), are eta_10＝23.88％、η_20＝31.26％、η₃₀46.08 percent, namely the total degradation rate of the photooxidation Yunnan Zhaotong lignite (GZTH) degraded by three bacteria in a grading way in sequence is far higher than the degradation rate of photooxidation Yunnan Zhaotong lignite (GZTH) degraded by three bacteria independently, which shows that the grading degradation method of the invention improves the degradation rate of the photooxidation Yunnan Zhaotong lignite (GZTH).

Example 3

The embodiment comprises the following steps:

step one, smashing the muddy source lignite (HYH) in Shanxi province, drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain muddy source lignite (HYH) coal powder with the granularity of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the Shanxi muddy-source lignite (HYH) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized Shanxi muddy-source lignite (GHYH) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal adding amount is 20g/L calculated by the mass of Shanxi muddy source lignite (HYH) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidized muddy brown coal (GHYH) obtained in the step two into a liquid culture medium inoculated with Streptomyces viridosporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for primary degradation, then filtering primary degradation products, respectively collecting primary degradation liquid and primary degradation coal residues, placing the primary degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the primary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁1.925, the degradation rate of the streptomyces viridis is brought into a relation equation eta of the first-stage degradation rate and the first-stage degradation liquid absorbance of the degraded photooxidation low-order coal₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.1681, i.e. η₁16.81%; the first-stage degradation conditions are as follows: 9.50g/L of the adding amount of the photo-oxidation muddy source lignite (GHYH) coal powder in the unit volume of the liquid culture medium inoculated with the streptomyces viridis, 180mL/L of the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.0 multiplied by 10⁵Per mL;

step four, adding the sterilized first-stage coal residue in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for secondary degradation, filtering a secondary degradation product, respectively collecting a secondary degradation liquid and a secondary degradation coal residue, and then mixing the two degradation liquidsSterilizing the second-stage degraded coal residue in autoclave at 121 deg.C for 20min, measuring absorbance of the second-stage degraded solution at 450nm with deionized water as blank control by spectrophotometry to obtain Y₂2.734, the degradation rate of the pseudomonas putida is carried into the relation equation eta of the secondary degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of the secondary degradation liquid₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.2032, i.e.. eta₂20.32%; the conditions of the secondary degradation are as follows: the addition amount of the sterilized first-stage degradation coal residue in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L in terms of the mass of pulverized coal of photooxidation Shanxi muddy source lignite (GHYH) in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.0 multiplied by 10⁵Per mL;

step five, adding the second-level coal residues sterilized in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium in an incubator for third-level degradation, filtering the third-level degradation products, respectively collecting third-level degradation liquid and third-level degradation coal residues, taking deionized water as a blank control, and measuring the absorbance of the third-level degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃2.688, introducing the degradation product into a relation equation eta of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium degradation photo-oxidation low-order coal₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.2638, i.e. η₃26.38%; the conditions of the third-stage degradation are as follows: the addition amount of the second-stage degraded coal residue after the sterilization in the liquid culture medium inoculated with the phanerochaete chrysosporium in unit volume is 13.00g/L in terms of the mass of the pulverized coal of the photo-oxidized Shanxi muddy-source lignite (GHYH) in the first-stage degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, and the culture boxThe oscillation frequency is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.0 multiplied by 10⁵one/mL.

Comparative example 7

This comparative example comprises the following steps:

step three, adding the photo-oxidized muddy brown coal (GHYH) obtained in the step two into a liquid culture medium inoculated with Streptomyces viridosporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and measuring the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁₀1.925, the degradation rate and degradation liquid absorbance relation equation eta of streptomyces viridosporus degradation photooxidation low-order coal is brought into₁₀＝0.02466+0.07453Y₁₀Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.1681, i.e. η₁₀16.81%; the conditions for degrading the photooxidation muddy source lignite (GHYH) by the streptomyces viridis single bacteria are as follows: the coal adding amount is 9.50g/L based on the mass of the light oxidized muddy source lignite (GHYH) coal powder in the streptomyces viridis liquid culture medium per unit volume, the inoculation amount of streptomyces viridis mother liquor in the streptomyces viridis liquid culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the Streptomyces viridocyclorisThe viable bacteria concentration in the mother bacteria liquid is 3.0 × 10⁵one/mL.

Comparative example 8

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the Shanxi muddy-source lignite (HYH) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized Shanxi muddy-source lignite (HYH) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal adding amount is 20g/L calculated by the mass of Shanxi muddy source lignite (HYH) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidized Shanxi muddy source lignite (HYH) coal powder obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂₀4.013, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degradation photooxidation low-rank coal is introduced₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀0.2865, i.e. η₂₀28.65%; the conditions for the pseudomonas putida single bacterium degradation photooxidation of muddy-source lignite (GHYH) in Shanxi are as follows: the coal feeding amount is 13.00g/L calculated by the mass of the photo-oxidation muddy source lignite (HYH) coal powder in the unit volume of the pseudomonas putida liquid culture medium, the inoculation amount of the pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.0 multiplied by 10⁵one/mL.

Comparative example 9

step three, adding the photo-oxidized Shanxi muddy source lignite (GHYH) coal powder obtained in the step two into a liquid culture medium inoculated with Phanerochaete chrysosporium, placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀4.890, the degradation rate and degradation liquid absorbance relation equation eta of the phanerochaete chrysosporium degradation photooxidation low-order coal is introduced₃₀＝0.02336+0.08945Y₃₀Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.3892, i.e. η₃₀38.92 percent; the conditions for degrading the photo-oxidation Shanxi muddy source lignite (GHYH) by the phanerochaete chrysosporium under the single bacteria are as follows: the coal adding amount is 13.00g/L calculated by the mass of light oxidized Shanxi muddy source lignite (GHYH) coal powder in a unit volume of phanerochaete chrysosporium liquid culture medium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.0 multiplied by 10⁵one/mL.

Comparing example 3 with comparative examples 7 to 9, it can be seen that in example 3, the muddy-source brown coal in Shanxi is photooxidizedTotal degradation rate eta of (GHYH) microbial fractionation_{General assembly}＝η₁₊η₂+η₃0.6364, i.e. η_{General assembly}63.64%, and the degradation rates of Streptomyces viridis, Pseudomonas putida and Phanerochaete chrysosporium in comparative examples 7-9 to photooxidation of muddy brown coal (GHYH) in Shanxi were η_10＝23.88％、η_20＝31.26％、η₃₀38.92 percent, namely the total degradation rate of the three bacteria sequentially adopted for graded degradation of the photo-oxidized muddy-source lignite (GHYH) is far higher than the degradation rate of the three bacteria for singly oxidizing the photo-oxidized muddy-source lignite (GHYH), which shows that the graded degradation method of the invention improves the microbial degradation rate of the muddy-source lignite (HYH).

Example 4

The embodiment comprises the following steps:

firstly, crushing the inner flame long-flame coal (ICY), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain coal powder of the inner flame long-flame coal (ICY) with the granularity of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the inner Mongolia long flame coal (ICY) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidation pulverized coal of the inner Mongolia long flame coal (GICY); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of the photo-oxidation inner Mongolia long flame coal (GICY) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation inner Mongolia long flame coal (GICY) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for primary degradation, then filtering a primary degradation product, respectively collecting a primary degradation liquid and a primary degradation coal residue, placing the primary degradation coal residue in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the primary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁1.447, carried into streptomyces viridosporus for degradation and photooxidationEquation eta of relation between first-stage degradation rate of low-rank coal and absorbance of first-stage degradation liquid₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.1325, i.e. η₁13.25%; the first-stage degradation conditions are as follows: 9.50g/L of the adding amount of photooxidation inner Mongolia flameray coal (GICY) coal powder in a unit volume of liquid culture medium inoculated with streptomyces viridis, 180mL/L of the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.2 multiplied by 10⁵one/mL.

Step four, adding the sterilized first-stage coal residues in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium in an incubator for secondary degradation, then filtering a secondary degradation product, respectively collecting a secondary degradation liquid and second-stage degradation coal residues, placing the second-stage degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the secondary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂2.173, the degradation rate of the pseudomonas putida is carried into the relation equation eta of the secondary degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of the secondary degradation liquid₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.1685, i.e. η₂16.85 percent; the conditions of the secondary degradation are as follows: the adding amount of the sterilized first-stage degradation coal residues in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L in terms of the mass of the coal powder of the photooxidation inner Mongolian long flame coal (GICY) in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵Per mL;

step five, adding the second-level coal residue sterilized in the step four into the inoculated phanerochaete chrysosporium (Phanerochaet)e chrysosporium) in a liquid culture medium, placing the liquid culture medium in an incubator for three-stage degradation, then filtering the three-stage degradation product, respectively collecting a three-stage degradation liquid and a three-stage degradation coal residue, taking deionized water as a blank control, and measuring the absorbance of the three-stage degradation liquid at 450nm by adopting a spectrophotometric method to obtain Y₃2.497, introducing the degradation product into a relation equation eta of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium degradation photo-oxidation low-order coal₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.2467, i.e. η₃24.67%; the conditions of the third-stage degradation are as follows: the addition amount of the second-grade degraded coal residue after the sterilization in the liquid culture medium inoculated with the phanerochaete chrysosporium in unit volume is 13.00g/L in terms of the mass of the coal powder of the photooxidation inner Mongolian flame coal (GICY) in the first-grade degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.4 multiplied by 10⁵one/mL.

Comparative example 10

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the inner Mongolia long flame coal (ICY) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidation pulverized coal of the inner Mongolia long flame coal (GICY); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of the coal powder of inner Mongolia long flame coal (ICY) added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation inner Mongolia long flame coal (GICY) coal powder obtained in the step two into the inoculated streptomyces viridisPutting into liquid culture medium of bacteria (Streptomyces viridosporus) in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as blank control, and determining absorbance of the degradation liquid at 450nm by spectrophotometry to obtain Y₁₀The degradation rate of the streptomyces viridis degrading the low-rank coal and the absorbance relation equation eta of the degradation liquid are introduced into 1.447₁₀＝0.02466+0.07453Y₁₀Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.1325, i.e. η₁₀13.25%; the conditions for degrading the photooxidation inner Mongolia long flame coal (GICY) by the streptomyces viridis are as follows: the coal adding amount is 9.50g/L calculated by the mass of the coal powder of photooxidation inner Mongolia flameray (GICY) in a unit volume streptomyces viridis liquid culture medium, the inoculation amount of streptomyces viridis mother liquor in the streptomyces viridis liquid culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.2 multiplied by 10⁵one/mL.

Comparative example 11

This comparative example comprises the following steps:

step three, adding the photo-oxidation inner Mongolia long flame coal (GICY) coal powder obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for degradation, and then filtering degradation products to obtain the productDetermining the absorbance of the degradation liquid at 450nm by spectrophotometry to obtain Y with deionized water as blank control₂₀1.998, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degradation photooxidation low-order coal is brought into₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀0.1573, i.e. η₂₀15.73%; the conditions for degrading the photooxidation inner Mongolia long flame coal (GICY) by the pseudomonas putida are as follows: the coal adding amount is 13.00g/L calculated by the mass of the coal powder in the photo-oxidation inner Mongolia long flame coal (GICY) in the unit volume of the pseudomonas putida liquid culture medium, the inoculation amount of the pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵one/mL.

Comparative example 12

step three, adding the photo-oxidation inner Mongolia long flame coal (GICY) coal powder obtained in the step two into a liquid culture medium inoculated with Phanerochaete chrysosporium, placing the culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀2.779, willThe relation equation eta of degradation rate and degradation liquid absorbance of the phanerochaete chrysosporium carried in to degrade the photo-oxidized low-rank coal₃₀＝0.02336+0.08945Y₃₀Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.2719, i.e. η₃₀27.19%; the conditions for degrading photooxidation inner Mongolia flame coal (GICY) by the Phanerochaete chrysosporium single bacteria are as follows: the coal adding amount is 13.00g/L calculated by the mass of the photo-oxidation inner Mongolia flame coal (GICY) coal powder in a unit volume of phanerochaete chrysosporium liquid culture medium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.4 multiplied by 10⁵one/mL.

As can be seen from comparison of example 4 with comparative examples 10 to 12, in example 4, the total degradation rate eta of the microbiological grade degradation of the photooxidation inner Mongolia long-flame coal (GICY)_{General assembly}＝η₁₊η₂+η₃0.6351, i.e. η_{General assembly}63.51%, and the degradation rates of Streptomyces viridis, Pseudomonas putida and Phanerochaete chrysosporium in comparative examples 10-12 to the photo-oxidation inner Mongolia flame coal (GICY) were η₁₀＝13.25％、η₂₀＝15.73％、η₃₀When the total degradation rate of the photo-oxidation inner flame coal (GICY) degraded by three bacteria in a grading way is 27.19 percent, the total degradation rate of the photo-oxidation inner flame coal (GICY) degraded by the three bacteria in sequence is far greater than the degradation rate of the photo-oxidation inner flame coal (GICY) degraded by the three bacteria alone, and the micro-biological degradation rate of the inner flame coal (ICY) is improved by the grading degradation method.

Example 5

The embodiment comprises the following steps:

step one, grinding Shenfu non-sticky coal (SFBN), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain Shenfu non-sticky coal (SFBN) coal powder with the particle size of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the Shenfu non-sticky coal (SFBN) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidation Shenfu non-sticky coal (GSFBN); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of Shenfu non-sticky coal (SFBN) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for primary degradation, then filtering a primary degradation product, respectively collecting a primary degradation liquid and a primary degradation coal residue, placing the primary degradation coal residue in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the primary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁1.193, and introducing the degradation product into a relation equation eta of the first-stage degradation rate and the first-stage degradation liquid absorbance of the streptomyces viridis to degrade the photooxidation low-rank coal₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.1136, i.e. η₁11.36%; the first-stage degradation conditions are as follows: 9.50g/L of the adding amount of photo-oxidation Shenfu non-caking coal (GSFBN) coal powder in a unit volume of liquid culture medium inoculated with streptomyces viridis, 180mL/L of the inoculation amount of streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.1 multiplied by 10⁵Per mL;

step four, adding the sterilized first-stage coal residues in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium in an incubator for secondary degradation, then filtering a secondary degradation product, respectively collecting a secondary degradation liquid and second-stage degradation coal residues, placing the second-stage degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the secondary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂2.263, which is carried into pseudomonas putida for degrading photo-oxidized low-rank coalEquation eta of relationship between secondary degradation rate and absorbance of secondary degradation liquid₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.1743, i.e. η₂17.43 percent; the conditions of the secondary degradation are as follows: the adding amount of the sterilized first-stage degradation coal residue in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L by the mass of the photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder in the first-stage degradation process, the inoculation amount of the pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.5 multiplied by 10⁵Per mL;

step five, adding the second-level coal residues sterilized in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium in an incubator for third-level degradation, then filtering and collecting the supernatant of the third-level degradation liquid, taking deionized water as a blank control, and measuring the absorbance of the third-level degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃2.963, introducing the degradation product into a relation equation eta of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium degradation photo-oxidation low-order coal₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.2884, i.e. η₃28.84%; the conditions of the third-stage degradation are as follows: the addition amount of the second-stage degraded coal residue after sterilization in a unit volume of the liquid culture medium inoculated with the phanerochaete chrysosporium is 13.00g/L in terms of the mass of the coal powder of photo-oxidation Shenfu non-sticky coal (GSFBN) in the first-stage degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.3 multiplied by 10⁵one/mL.

Comparative example 13

This comparative example comprises the following steps:

step one, crushing Shenfu non-sticky coal (SFBN), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain coal powder with the particle size of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of Shenfu non-sticky coal (SFBN) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium into an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁₀1.193, and introducing the degradation rate into a relation equation eta of the degradation liquid absorbance and the degradation rate of streptomyces viridis degrading low-rank coal₁₀＝0.02466+0.07453Y₁₀Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.1136, i.e. η₁₀11.36%; the conditions for degrading photo-oxidation Shenfu non-sticky coal (GSFBN) by the streptomyces viridis single bacteria are as follows: the coal adding amount is 9.50g/L by the mass of light-oxidation Shenfu non-sticky coal (GSFBN) coal powder in a unit volume streptomyces viridis liquid culture medium, the inoculation amount of streptomyces viridis mother liquor in the streptomyces viridis liquid culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.1 multiplied by 10⁵one/mL.

Comparative example 14

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the Shenfu non-sticky coal (SFBN) coal powder obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder; the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of Shenfu non-sticky coal (SFBN) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for degradation, then filtering a degradation product to obtain a degradation solution, taking deionized water as a blank control, and determining the absorbance of the degradation solution at 450nm by adopting a spectrophotometry method to obtain Y₂₀2.603, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degradation photooxidation low-order coal is introduced into₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀0.1961, i.e. η₂₀19.61%; the pseudomonas putida single bacterium degradation photo-oxidation Shenfu non-caking coal (GSFBN) conditions are as follows: the coal adding amount is 13.00g/L by the mass of light oxidation Shenfu non-caking coal (SFBN) coal powder in a unit volume of pseudomonas putida liquid culture medium, the inoculation amount of pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.5 multiplied by 10⁵one/mL.

Comparative example 15

step three, adding the photo-oxidation Shenfu non-sticky coal (GSFBN) coal powder obtained in the step two into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium into an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀3.109, the degradation rate and degradation liquid absorbance relation equation eta of the phanerochaete chrysosporium degradation photooxidation low-order coal is introduced into₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.3015, i.e. η₃₀30.15 percent; the conditions for degrading photo-oxidation Shenfu non-sticky coal (GSFBN) by the phanerochaete chrysosporium are as follows: the coal adding amount is 13.00g/L calculated by the mass of photooxidation Shenfu non-sticky coal (GSFBN) coal powder in a unit volume of phanerochaete chrysosporium liquid culture medium, the inoculation amount of a phanerochaete chrysosporium mother bacteria liquid in the phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.3 multiplied by 10⁵one/mL.

Comparing example 5 with comparative examples 13 to 15, it can be seen that the total degradation rate η of the microbial degradation of photooxidation Shenfu non-caking coal (GSFBN) in example 5_{General assembly}＝η₁₊η₂+η₃0.5763, i.e. η_{General assembly}57.63%, and in comparative examples 13-15, the degradation rates of Streptomyces viridis, Pseudomonas putida, and Phanerochaete chrysosporium to photo-oxidative Gomphon-non-caking coal (GSFBN) were η_10＝11.36％、η₂₀＝19.16％、η₃₀30.15%, namely, the three bacteria are adopted for grading in sequenceThe total degradation rate of the photo-oxidation Shenfu non-sticky coal (GSFBN) is far greater than that of the photo-oxidation Shenfu non-sticky coal (GSFBN) by three bacteria alone, which shows that the grading degradation method of the invention improves the microbial degradation rate of the Shenfu non-sticky coal (SFBN).

Example 6

The embodiment comprises the following steps:

crushing the Shaanxi flame coal (SXCY), drying for 1h at 100 ℃, and then sequentially grinding and screening to obtain the coal powder of the Shaanxi flame coal (SXCY) with the particle size of-0.15 mm +0.075 mm;

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the long flame coal in Shaanxi (SXCY) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized pulverized coal of the long flame coal in Shaanxi (GSXCY); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of the Shaanxi long flame coal (GSXCY) coal powder added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation Shaanxi long flame coal (GSXCY) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridogrous, placing the liquid culture medium in an incubator for primary degradation, then filtering primary degradation products, respectively collecting primary degradation liquid and primary degradation coal residues, placing the primary degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as blank control, and determining the absorbance of the primary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁1.624, the degradation rate and the absorbance relation equation eta of the first-stage degradation liquid of the streptomyces viridosporus degraded photooxidation low-rank coal are introduced₁＝0.02466+0.07453Y₁Wherein the goodness-of-fit determination coefficient is R₁ ²0.98392 to obtain η₁0.1457, i.e. η₁14.57%; the first-stage degradation conditions are as follows: the adding amount of the light oxidized Shaanxi flame coal (GSXCY) coal powder in the liquid culture medium with the unit volume inoculated with the streptomyces viridisporus is 9.50g/L, and the liquid of the streptomyces viridisporusThe inoculation amount of the streptomyces viridis mother bacteria liquid in the culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.4 multiplied by 10⁵Per mL;

step four, adding the sterilized first-stage coal residues in the step three into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium in an incubator for secondary degradation, then filtering a secondary degradation product, respectively collecting a secondary degradation liquid and second-stage degradation coal residues, placing the second-stage degradation coal residues in an autoclave for sterilization at 121 ℃ for 20min, taking deionized water as a blank control, and measuring the absorbance of the secondary degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂2.452, the pseudomonas putida is introduced into a relation equation eta of the secondary degradation rate of the pseudomonas putida degradation photooxidation low-rank coal and the absorbance of a secondary degradation liquid₂＝0.02919+0.06412Y₂Wherein the goodness-of-fit determination coefficient is R₂ ²0.99075 to obtain η₂0.1864, i.e. η₂18.64%; the conditions of the secondary degradation are as follows: the addition amount of the sterilized first-stage degradation coal residue in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L in terms of the mass of the pulverized coal of photooxidation Changshi coal (GSXCY) in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria liquid in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵Per mL;

step five, adding the second-level coal residues sterilized in the step four into a liquid culture medium inoculated with Phanerochaete chrysosporium (Phanerochaete chrysosporium), placing the liquid culture medium in an incubator for third-level degradation, filtering the third-level degradation products, respectively collecting third-level degradation liquid and third-level degradation coal residues, taking deionized water as a blank control, and measuring the absorbance of the third-level degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃3.177, the three-stage degradation rate and the three-stage degradation liquid absorbance of the phanerochaete chrysosporium for degrading the photo-oxidized low-order coal are broughtEquation of relationship η₃＝0.02336+0.08945Y₃Wherein the goodness-of-fit determination coefficient is R₃ ²0.97836 to obtain η₃0.3075, i.e. η₃30.75 percent; the conditions of the third-stage degradation are as follows: the addition amount of the second-grade degraded coal residue after sterilization in a unit volume of the liquid culture medium inoculated with the phanerochaete chrysosporium is 13.00g/L in terms of the mass of the coal powder of photooxidation Changhua coal (GSXCY) in the first-grade degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.1 multiplied by 10⁵one/mL.

Comparative example 16

This comparative example comprises the following steps:

step two, carrying out photo-oxidation pretreatment on the pulverized coal of the long flame coal in Shaanxi (SXCY) obtained in the step one by adopting a rotary bed photochemical reactor to obtain photo-oxidized pulverized coal of the long flame coal in Shaanxi (GSXCY); the conditions of the photo-oxidation pretreatment are as follows: the coal feeding amount is 20g/L calculated by the mass of the coal powder of the Shaanxi long flame coal (SXCY) added into a unit volume rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min;

step three, adding the photo-oxidation Shaanxi long flame coal (GSXCY) coal powder obtained in the step two into a liquid culture medium inoculated with Streptomyces viridisporus (Streptomyces viridosporus), placing the liquid culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₁₀1.624, the degradation rate and degradation liquid absorbance relation equation eta of the streptomyces viridosporus degradation photooxidation low-rank coal is brought into₁₀＝0.02466+0.07453Y₁₀Wherein the goodness-of-fit determination coefficient is R₁₀ ²0.98392 to obtain η₁₀0.1457, i.e. η₁₀14.57%; the conditions for degrading the light-oxidized Shanxi flame coal (GSXCY) by the single streptomyces viridis are as follows: the coal adding amount is 9.50g/L by the mass of light-oxidized Shanxi flame coal (GSXCY) coal powder in a unit volume of streptomyces viridis liquid culture medium, the inoculation amount of streptomyces viridis mother liquor in the streptomyces viridis liquid culture medium is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is 3.4 multiplied by 10⁵one/mL.

Comparative example 17

This comparative example comprises the following steps:

step three, adding the photo-oxidation coal powder (GSXCY) obtained in the step two into a liquid culture medium inoculated with Pseudomonas putida (Pseudomonas putida), placing the liquid culture medium into an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₂₀2.263, the degradation rate and degradation liquid absorbance relation equation eta of the pseudomonas putida degraded low-order coal is introduced₂₀＝0.02919+0.06412Y₂₀Wherein the goodness-of-fit determination coefficient is R₂₀ ²0.99075 to obtain η₂₀＝0.1743, i.e. η₂₀17.43 percent; the conditions for degrading the Shaanxi long flame coal (GSXCY) by the pseudomonas putida single bacteria are as follows: the coal feeding amount is 13.00g/L calculated by the mass of pulverized coal of photooxidation Changshi flame coal (GSXCY) in unit volume of pseudomonas putida liquid culture medium, the inoculation amount of pseudomonas putida mother bacteria liquid in the pseudomonas putida liquid culture medium is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria solution is 8.2 multiplied by 10⁵one/mL.

Comparative example 18

step three, adding the photo-oxidation Shaanxi Changhover coal (GSXCY) coal powder obtained in the step two into a liquid culture medium inoculated with Phanerochaete chrysosporium, placing the culture medium in an incubator for degradation, then filtering degradation products to obtain degradation liquid, taking deionized water as a blank control, and determining the absorbance of the degradation liquid at 450nm by adopting a spectrophotometry method to obtain Y₃₀2.974, the degradation rate and degradation liquid absorbance relation equation eta of the phanerochaete chrysosporium degradation photooxidation low-order coal is introduced₃₀＝0.02336+0.08945Y₃₀Wherein the goodness-of-fit determination coefficient is R₃₀ ²0.97836 to obtain η₃₀0.2894, i.e. η₃₀28.94%; the degradation condition of the phanerochaete chrysosporium by single-bacterium photooxidation of the Changhua Shaanxi (GSXCY) is as follows: coal charge is in unitThe method comprises the following steps of (1) metering 13.00g/L of photooxidation Shaanxi Changhua coal (GSXCY) coal powder in a voluminous Phanerochaete chrysosporium liquid culture medium, wherein the inoculation amount of a Phanerochaete chrysosporium mother fungus liquid in the Phanerochaete chrysosporium liquid culture medium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration of the phanerochaete chrysosporium mother liquor is 2.1 multiplied by 10⁵one/mL.

Comparing example 6 with comparative examples 16-18, it can be seen that the total degradation rate eta of the microbial graded degradation of photooxidation Shaanxi long flame coal (GSXCY) in example 6_{General assembly}＝η₁₊η₂+η₃0.6396, i.e. η_{General assembly}63.96%, and the degradation rates of Streptomyces viridis, Pseudomonas putida and Phanerochaete chrysosporium in comparative examples 16-18 for directly oxidizing Changhua Shaanxi coal (GSXCY) by light are eta respectively₁₀＝14.57％、η₂₀＝17.43％、η₃₀The total degradation rate of the three bacteria sequentially used for graded degradation of the photooxidation shanxi flame coal (GSXCY) is far greater than the degradation rate of the three bacteria used for singly degrading the photooxidation shanxi flame coal (GSXCY), which means that the graded degradation method of the invention improves the microbial degradation rate of the shanxi flame coal (SXCY).

The above description is only an embodiment of the preferred ingredient range of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

1. The method for degrading the low-rank coal by the microorganism in a grading manner is characterized by comprising the following steps of:

2. The method for microbial graded degradation of low-rank coal according to claim 1, wherein the conditions of the photo-oxidation pretreatment in the second step are as follows: the coal feeding amount is 20g/L calculated by the mass of the coal powder added per unit volume in the rotary bed photochemical reactor, the ultraviolet light intensity is 150W, the rotating speed is 120r/min, the oxidation time is 42h, the oxygen flow is 800mL/min, and the oxygen introducing time per liter calculated by the volume of the rotary bed photochemical reactor before the photo-oxidation pretreatment is 10 min.

3. The method for microbial graded degradation of low-rank coal according to claim 1, wherein the conditions of the primary degradation in the third step are as follows: the adding amount of the photo-oxidative coal powder in the liquid culture medium inoculated with the streptomyces viridis in unit volume is 9.50g/L, the inoculation amount of the streptomyces viridis mother liquor in the liquid culture medium of the streptomyces viridis is 180mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 10d, and the culture temperature is 28 ℃; the viable bacteria concentration in the streptomyces viridis mother bacteria liquid is not less than 3.0 multiplied by 10⁵one/mL.

4. The method of claim 1The method for degrading the low-rank coal in the microbial classification mode is characterized in that the conditions of secondary degradation in the fourth step are as follows: the addition amount of the sterilized first-stage degraded coal residues in the unit volume of the liquid culture medium inoculated with the pseudomonas putida is 13.00g/L of the mass amount of the photo-oxidized coal powder in the first-stage degradation process, the inoculation amount of a pseudomonas putida mother bacteria solution in the liquid culture medium of the pseudomonas putida is 135mL/L, the oscillation frequency of an incubator is 160r/min, the culture time is 12d, and the culture temperature is 30 ℃; the viable bacteria concentration in the pseudomonas putida mother bacteria liquid is not less than 8.0 multiplied by 10⁵one/mL.

5. The method for microbial graded degradation of low-rank coal according to claim 1, wherein the conditions of the third-stage degradation in the fifth step are as follows: the addition amount of the sterilized second-stage degraded coal residue in the liquid culture medium inoculated with the phanerochaete chrysosporium in unit volume is 13.00g/L of the mass of the photo-oxidized coal powder in the first-stage degradation process, the inoculation amount of the phanerochaete chrysosporium mother bacteria liquid in the liquid culture medium of the phanerochaete chrysosporium is 90mL/L, the oscillation frequency of an incubator is 210r/min, the culture time is 14d, and the culture temperature is 30 ℃; the spore concentration in the phanerochaete chrysosporium mother liquor is not less than 2.0 multiplied by 10⁵one/mL.

6. The method for the microbial graded degradation of the low-rank coal according to claim 1, wherein the absorbance of the first-order degradation liquid obtained in the third step at 450nm is measured by a spectrophotometry method, and then the measured absorbance is brought into a relation equation between the first-order degradation rate of streptomyces viridis degrading and photooxidizing the low-rank coal and the absorbance of the first-order degradation liquid, so that the first-order degradation rate is calculated; the relation equation of the first-stage degradation rate of the streptomyces viridocystoris for degrading the photooxidation low-rank coal and the absorbance of the first-stage degradation liquid is as follows: eta₁＝0.02466+0.07453Y₁With a goodness-of-fit determination coefficient of R₁ ²0.98392, where eta₁First order degradation rate, Y₁The absorbance of the first-order degradation liquid is shown.

7. The low-rank coal microorganism according to claim 1The method for the secondary degradation is characterized in that the absorbance of the secondary degradation liquid obtained in the fourth step at 450nm is measured by adopting a spectrophotometry method, and then the absorbance is brought into a relation equation between the secondary degradation rate of the pseudomonas putida for degrading and photo-oxidizing the low-rank coal and the absorbance of the secondary degradation liquid, and the secondary degradation rate is calculated; the relation equation of the secondary degradation rate of the pseudomonas putida degrading and photo-oxidizing low-rank coal and the absorbance of the secondary degradation liquid is as follows: eta₂＝0.02919+0.06412Y₂With a goodness-of-fit determination coefficient of R₂ ²0.99075, where eta₂Is a second level of degradation, Y₂The absorbance of the second-order degradation liquid is shown.

8. The method for the microbial graded degradation of the low-rank coal according to claim 1, wherein the absorbance of the third-level degradation liquid obtained in the fifth step at 450nm is measured by a spectrophotometry method, and then the absorbance is brought into a relational equation between the third-level degradation rate of the phanerochaete chrysosporium degradation and the absorbance of the third-level degradation liquid for the photooxidation of the low-rank coal, so as to calculate the third-level degradation rate; the relation equation of the third-level degradation rate and the third-level degradation liquid absorbance of the phanerochaete chrysosporium for degrading the photo-oxidized low-rank coal is as follows: eta₃＝0.02336+0.08945Y₃With a goodness-of-fit determination coefficient of R₃ ²0.97836, where eta₃Three-stage degradation rate, Y₃The absorbance of the third-order degradation liquid is shown.

9. The method for microbial graded degradation of low-rank coal according to claim 1, wherein the sterilization conditions in the third step and the fourth step are both: sterilizing at 121 deg.C for 20 min.