CN109880647A - A kind of method of low-order coal microorganism classification degradation - Google Patents
A kind of method of low-order coal microorganism classification degradation Download PDFInfo
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Abstract
The invention discloses a kind of method of low-order coal microorganism classification degradation, this method comprises: one, coal dust is made in low-order coal;Two, coal dust photooxidation is obtained into photooxidation powdered coal;Three, photooxidation powdered coal is added to progress level-one degradation in the fluid nutrient medium for being inoculated with green spore streptomycete;Four, the level-one degradation coal residue after will be sterilized is added to progress second level degradation in the fluid nutrient medium for be inoculated with pseudomonas putida;Five, the second level degradation coal residue after will be sterilized, which is added in the fluid nutrient medium for be inoculated with Phanerochaete chrysosporium, carries out multi-stage degradation.The present invention successively carries out classification degradation to low-order coal using green spore streptomycete, pseudomonas putida and Phanerochaete chrysosporium, it is utilized respectively the various actives substances such as alkali, enzyme, chelating agent and the surfactant that three kinds of bacterium metabolism generate, to degrading activity point synergistic effect different in low order texture of coal, the degradation rate of low-order coal is improved, is utilized for the further clean and effective of low-order coal and creates basic condition.
Description
Technical field
The invention belongs to the microbiological transformation technology fields of coal, and in particular to a kind of side of low-order coal microorganism classification degradation
Method.
Background technique
It is well known that coal is one of most important energy, Chinese coal reserves account occupies third place in the world, coal production and
Consumption figure ranks always the first in the world.Coal exploitation utilize during, with high-quality high-order coal try to be the first exploitation processing and
It utilizes, low-order coal ratio shared in coal reserves is increasing.Currently, the purposes that China is more than 50% low-order coal is hair
Electricity, followed by coking, gasification, liquefaction, directly burning heating etc..It is had the following problems during the utilization of above-mentioned low-order coal:
(1) generating efficiency is low;Low-order coal has the characteristics that calorific value is low, moisture is high, this results in its thermal efficiency low;
(2) processing conditions is harsh;The processing methods such as low order coal gasification, liquefaction, pyrolysis, coking generally require high temperature height
Pressure is also high to equipment requirement;
(3) Air-pollution From Combustion is big;N, S, heavy metal element and content of ashes are higher in low-order coal, and low-order coal is in combustion process
In in addition to generate carbon dioxide, nytron beyond the region of objective existence, can also generate volume of smoke, nitrogen oxides, oxysulfide, heavy metal chemical combination
The pollutants such as object.
Low-order coal is as fuel directly using the development for being increasingly detrimental to today's society, and therefore, the cleaning of low-order coal is high
Effect processing and utilization is the inevitable approach of coal sustainable development.
The microbial degradation of coal is the state-of-the-art technology after the liquefaction of coal, gasification process technology, is one and is related to micro- life
The frontier of the subjects such as object, biochemistry, zymetology, molecular biology, separation engineering, coal chemistry and mineral processing.Coal
Microbial degradation only need to carry out at room temperature and normal pressure, have reaction condition is mild, consersion unit requires simple, low power consuming,
The advantages that product utilization rate is high.In the clean and effective use aspects of coal, especially low-order coal, development prospect is bright.
Find that microorganism can degrade coal so far from scientist's eighties in last century, by nearly research in 40 years and hair
Exhibition, new technology of the microbial degradation of coal as Coal dressing, although having achieved certain achievement, with the depth of research
Enter, it was found that problems, such as: (1) microorganism is low to the degradation efficiency of coal;(2) bacterium-coal matching grating research is insufficient,
Lack efficient degrading bacteria, and degradation bacteria lacks universality etc.;(3) the Nomenclature Composition and Structure of Complexes complexity of coal causes the microbial degradation of coal to produce
Object is difficult to separate and utilize;(4) mechanism of microbial degradation coal is not still apparent.Problems above is all different degrees of to be restrict
The industrialization of the microbial technology of coal, wherein microorganism low to the degradation rate of coal is most basic and crucial problem.Because
Degradation rate is too low, so that catabolite is less, the purposes and industrialization that will lead to research catabolite are obstructed, and influences subsequent grind
The progress studied carefully;The research of mechanism is emphasis, the apparent microbial degradation process that coal can be instructed from objective law of mechanism, from
And the measure for formulating rationally science improves the microbial degradation rate of coal and the catabolite of design coal is constituted;The separation of catabolite
With efficient using being purpose, the purposes of product is wider, and added value is bigger, will promote people to coal microbial degradation skill in turn
The research of art.So above-mentioned these problems are all badly in need of system in-depth study and breakthrough.
The microbial degradation mechanism of coal is respectively by the sequencing of proposition: enzyme mechanism, the alkali mechanism of action, chelating agent
The mechanism of action, Action of Surfactant mechanism and the ABCDE mechanism of action.The ABCDE mechanism of action indicates the macromolecular structure of coal
It is middle respectively by the structure of functional groups position of enzyme, alkali, chelating agent and Action of Surfactant, which is several mechanism in front
On the basis of propose, it considers that the process of microbial degradation coal is the coefficient result of a variety of mechanism.Often single bacterial strain is not
The active material (enzyme, alkali, chelating agent, surfactant etc.) of many degradation coals of degradation coal can be generated.Currently, researcher is most
Microbial degradation is carried out to coal using single strain, so often degradation rate is lower, even if the matching of individual bacterium and certain coal compared with
Good (the degradation coal activity substance that the bacterium generates is just corresponding with existing degrading activity points a large amount of in certain coal texture), but
The bacterium coal different to other structures functional group, not necessarily degradation rate is high, i.e., bacterial strain lacks universality.So with coal microorganism
Mechanism of degradation is starting point, and the method that research improves coal microbial degradation rate is coal, and especially low-order coal high-efficiency cleaning utilizes
Premise and basis.
Summary of the invention
Technical problem to be solved by the present invention lies in view of the above shortcomings of the prior art, providing, a kind of low-order coal is micro-
The method of biology classification degradation.This method is using green spore streptomycete, pseudomonas putida and Phanerochaete chrysosporium successively to low
Rank coal carries out classification degradation, is utilized respectively the various actives objects such as alkali, enzyme, chelating agent and the surfactant that three kinds of bacterium metabolism generate
Matter acts synergistically to degrading activity point different in low order texture of coal, improves the degradation rate of low-order coal, improve degradations at different levels
The type and quantity of the products such as fine chemicals, liquid fuel in liquid create for the further clean and effective utilization of low-order coal
Basic condition.
In order to solve the above technical problems, technical solution provided by the invention are as follows: a kind of classification degradation of low-order coal microorganism
Method, which is characterized in that method includes the following steps:
Step 1: drying after low-order coal is crushed, then successively through grinding and screening, obtaining granularity is -0.15mm+
The coal dust of 0.075mm;
Step 2: carrying out photooxidation pretreatment to coal dust obtained in step 1 using rotation bed photochemical reactor, obtain
To photooxidation powdered coal;
Green spore streptomycete (Streptomyces is inoculated with Step 3: photooxidation powdered coal obtained in step 2 is added to
Viridosporous in fluid nutrient medium), it is placed in progress level-one degradation in incubator, then level-one catabolite is carried out
Filtering collects level-one degradation solution and level-one degradation coal residue respectively, then level-one degradation coal residue is sterilized;
Step 4: the level-one degradation coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is sterilized;
Step 5: the second level degradation coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid and multi-stage degradation coal residue respectively.
Coal dust, that is, granularity that granularity is -0.15mm+0.075mm in step 1 is more than or equal to 0.075mm and is less than 0.15mm
Coal dust.
The present invention obtains coal dust after crushing and screening low-order coal, then pre-process coal dust progress photooxidation to obtain photooxidation
Coal dust improves the oxygen content in coal dust so that coal dust is easier to be degraded by microorganisms, then successively carries out to photooxidation powdered coal green
The degradation of spore streptomycete (actinomyces) level-one, the degradation of pseudomonas putida (bacterium) second level and Phanerochaete chrysosporium (fungi) three-level
Degradation.Wherein, the mechanism of the green spore streptomycete level-one degradation of actinomyces is alkali effect and Action of Surfactant, level-one degradation process
In weaker alkali act on so that acid stronger group is acted in photooxidation powdered coal, Action of Surfactant makes photooxidation
The hydrophily of powdered coal is enhanced, and going on smoothly for level-one degradation process is conducive to;The secondary degradation of bacterium pseudomonas putida
Mechanism be alkali effect, Action of Surfactant and chelating agent effect, the alkali effect in second level degradation process is stronger, can be further
Acid weaker group in level-one degradation coal residue is set to react, since level-one degradation process consumes acid in photooxidation powdered coal
Property stronger group, to ensure that the alkaline matter generated in second level degradation process is only acted on acid weaker group, is kept away
Exempt to be consumed by acid stronger group, has improved the efficiency of alkali effect degradation, while the surface generated in second level degradation process
The substance of active agent properties further enhances the hydrophily of level-one degradation coal residue, is conducive to generate in second level degradation process
Degradation coal activity substance comes into full contact with degradable active site in level-one degradation coal residue, improves the efficiency of second level degradation,
And chelating agent acts on so that the metal ion for playing " bridge formation " in level-one degradation coal residue is removed, in level-one degradation coal residue
Macromolecular structure generate a degree of collapsing depolymerization, to promote the further exposure in degradable active site, favorably
In subsequent degradation;The mechanism that fungi Phanerochaete chrysosporium is degraded three times is enzyme effect, alkali acts on and Action of Surfactant,
Enzyme effect during multi-stage degradation is stronger, so that second level is degraded, coal residue is under the action of the enzyme generated during multi-stage degradation
It reacts and then degrades, in alkaline matter and second level the degradation coal residue generated during multi-stage degradation after premenstrual two-stage degradation
Left acidic-group further reacts, while Action of Surfactant enhances the hydrophily of second level degradation coal residue, mentions
The high efficiency of multi-stage degradation.Under the action of different microorganisms, respectively containing more in the degradation solutions at different levels that multi-stage degradation obtains
The kind type catabolite different with structure, can be through products such as further isolated fine chemicals, liquid fuels, thus real
The efficient utilization of existing low-order coal.
Microorganism is that microbial technique takes in the clean and effective use aspects of low-order coal to the abundant efficient degradation of low-order coal
The premise and basis that must be broken through.The present invention generates a variety of degradations such as alkali, enzyme, chelating agent and surfactant using three kinds of microorganisms
The active material of coal carries out classification degradation to low-order coal step by step according to different microorganisms to the different role mechanism of low-order coal, after
Grade degradation carries out on the basis of previous stage degradation, and previous stage is degraded to rear class degradation and provides the foundation, so that rear class degradation is more
It easily carries out, while having activated the synergistic effect between each active material, jointly to degrading activity point different in low order texture of coal
Effect, meets the succession of herbaceous of microbe groups in low-order coal weathering degradation process, to improve the degradation of low-order coal
Rate improves the type and quantity of the products such as fine chemicals, liquid fuel in degradation solutions at different levels, is that low-order coal is further clear
Solid foundation has been established in clean efficient utilization.
Rotation bed photochemical reactor in the present invention uses Authorization Notice No. for ZL201621380305.0 utility model
The rotation bed photochemical reactor of patent disclosure.
The method of above-mentioned a kind of low-order coal microorganism classification degradation, which is characterized in that photooxidation described in step 2 is pre-
The condition of processing are as follows: coal amount 20g/L, ultraviolet light in terms of the quality of pc being added in unit volume rotation bed photochemical reactor
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter.It is determined simultaneously by single factor experiment and orthogonal test method of analysis of variance
Obtain the pretreated optimum process condition of above-mentioned photooxidation.Under the pretreated optimum process condition of the photooxidation, revolving bed light
Chemical reactor is best to the photooxidation pretreating effect of coal dust, substantially increases the oxygen content in photooxidation powdered coal, Jin Erti
Microorganism is safer compared to common nitric acid oxidation method to the degradation rate of photooxidation powdered coal in high subsequent hierarchical degradation process
Environmental protection.
A kind of method of above-mentioned low-order coal microorganism classification degradation, which is characterized in that the degradation of level-one described in step 3
Condition are as follows: be inoculated with the additional amount 9.50g/L of photooxidation powdered coal in the fluid nutrient medium of green spore streptomycete, green spore streptomycete
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L in fluid nutrient medium, incubator frequency of oscillation 160r/min, incubation time
10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is not less than 3.0 × 105A/mL.
A kind of method of above-mentioned low-order coal microorganism classification degradation, which is characterized in that the degradation of second level described in step 4
Condition are as follows: be inoculated with it is sterilized in the fluid nutrient medium of pseudomonas putida after level-one degradation coal residue additional amount with one
Photooxidation quality of pc meter 13.00g/L in grade degradation process, the pseudomonas putida in the fluid nutrient medium of pseudomonas putida
Female bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench
Viable bacteria concentration in pseudomonad mother's bacterium solution is not less than 8.0 × 105A/mL.
A kind of method of above-mentioned low-order coal microorganism classification degradation, which is characterized in that multi-stage degradation described in step 5
Condition are as follows: be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation coal residue additional amount with
Photooxidation quality of pc meter 13.00g/L in level-one degradation process, the yellow archespore hair in the fluid nutrient medium of Phanerochaete chrysosporium
Flat lead fungi mother bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;It is described
Spore concentration in Phanerochaete chrysosporium mother's bacterium solution is not less than 2.0 × 105A/mL.
By the growth for cultivate and draw to above-mentioned three kinds of degradation bacterias three kinds of degradation bacterias respectively in conjunction with dry cell weight method
Curve determines lag phase, exponential phase of growth, stationary phase and the decline phase of three kinds of various growth courses of degradation bacteria respectively, then leads to
It crosses single factor experiment and Response Surface Method and the corresponding degradation optimum process conditions at different levels of three kinds of above-mentioned degradation bacterias has been determined respectively.
The degradation that low-order coal is successively carried out under the corresponding degradation optimum process conditions at different levels of above-mentioned three kinds of degradation bacterias, ensure that drops at different levels
Solution preocess all has higher degradation rate to degradation substrate, to improve the total degradation rate of low-order coal, and then improves at different levels
The type and quantity of the products such as fine chemicals, liquid fuel, are conducive to further realize low order on this basis in degradation solution
The efficient utilization of coal.
Structure due to belonging to the coal (mainly lignite) of low-order coal scope has similitude, can use unified coal knot
Structure model is expressed, and after microbial action, and it is little that the structure of coal residue compare variation with corresponding low order coal texture, therefore
Under the premise of degradation technique and identical functional bacteria, object of the low-order coal with corresponding coal residue through containing in obtained degradation solution of degrading
Matter species differences are little, and only the concentration of substance is different.That is, the material concentration in the big degradation solution of degradation rate is big,
The absorbance of the degradation solution of measurement is big, and the material concentration in the small degradation solution of degradation rate is small, and the absorbance of the degradation solution of measurement is small,
Therefore there is determining relationship between low-order coal and the degradation rate and degradation solution absorbance of corresponding coal residue, i.e., it is following three to pass through
The degradation rate and degradation solution absorbance relation equation of the spore streptomycete degradation photooxidation low-order coal of experimental study fitting correspondence establishment,
The degradation rate and degradation solution absorbance relation equation of pseudomonas putida degradation photooxidation low-order coal and Phanerochaete chrysosporium drop
The degradation rate and degradation solution absorbance relation equation for solving photooxidation low-order coal are not only suitable for corresponding microbial degradation photooxidation low order
The degradation solution that coal obtains, and it is suitable for the degradation that the coal residue that corresponding microbial degradation is obtained by other microbial degradations obtains
Liquid.
The method of above-mentioned a kind of low-order coal microorganism classification degradation, which is characterized in that using spectrophotometry to step
Absorbance of the level-one degradation solution at 450nm obtained in three is measured, and it is low then to bring green spore streptomycete degradation photooxidation into
In the level-one degradation rate and level-one degradation solution absorbance relation equation of rank coal, level-one degradation rate is calculated;The green spore strepto-
The level-one degradation rate and level-one degradation solution absorbance relation equation of bacterium degradation photooxidation low-order coal are as follows: η1=0.02466+
0.07453Y1, the goodness of fit determine coefficient be R1 2=0.98392, wherein η1For level-one degradation rate, Y1For the suction of level-one degradation solution
Luminosity.
The method of above-mentioned a kind of low-order coal microorganism classification degradation, which is characterized in that using spectrophotometry to step
Absorbance of the second level degradation solution at 450nm obtained in four is measured, and then brings pseudomonas putida degradation photooxidation into
In the second level degradation rate and second level degradation solution absorbance relation equation of low-order coal, second level degradation rate is calculated;The stench is false
The second level degradation rate and second level degradation solution absorbance relation equation of monad degradation photooxidation low-order coal are as follows: η2=0.02919+
0.06412Y2, the goodness of fit determine coefficient be R2 2=0.99075, wherein η2For second level degradation rate, Y2For the suction of second level degradation solution
Luminosity.
The method of above-mentioned a kind of low-order coal microorganism classification degradation, which is characterized in that using spectrophotometry to step
Absorbance of the multi-stage degradation liquid at 450nm obtained in five is measured, and then brings Phanerochaete chrysosporium degradation photooxidation into
Change in the multi-stage degradation rate of low-order coal and the relation equation of multi-stage degradation liquid absorbance, multi-stage degradation rate is calculated;The Huang
The multi-stage degradation rate of the flat lead fungi degradation photooxidation low-order coal of archespore hair and the relation equation of multi-stage degradation liquid absorbance are as follows: η3=
0.02336+0.08945Y3, the goodness of fit determine coefficient be R3 2=0.97836, wherein η3For multi-stage degradation rate, Y3For three-level
Degradation solution absorbance.
Be measured at 450nm by spectrophotometry, respectively obtain during multi-stage degradation degradation rate at different levels with it is right
Answer the relation equation of degradation solution absorbances at different levels.Degradation rate is used in combination the present invention with absorbance two indices, establishes at different levels
The degradation rate of microbial degradation photooxidation low-order coal and the relation equation to degradation solution absorbance, improve the accurate of degradation rate
Degree, so that degradation rates at different levels can be quickly obtained according to relation equation in the application process of the method for the present invention, convenience and high-efficiency, and can
Degradation processes at different levels are adjusted and are controlled according to degradation rate results at different levels, to be conducive to improve the degradation rate of low-order coal.
A kind of method of above-mentioned low-order coal microorganism classification degradation, which is characterized in that going out in step 3 and step 4
Bacterium condition is equal are as follows: 121 DEG C of sterilizing 20min in high-pressure sterilizing pot.Above-mentioned sterilising conditions can effectively kill level-one degradation coal residue
Remaining pseudomonas putida in the green spore streptomycete of middle remaining and second level degradation coal residue, to avoid level-one degradation coal residue
In influence of the green spore streptomycete to second level degradation process and the pseudomonas putida in second level degradation coal residue three-level is dropped
The influence of solution preocess enhances the degradation effect of second level degradation and multi-stage degradation, further improves the degradation rate of low-order coal.
Compared with the prior art, the present invention has the following advantages:
1, the present invention successively divides low-order coal using green spore streptomycete, pseudomonas putida and Phanerochaete chrysosporium
Grade degradation, according to three kinds of bacterium to the different role mechanism of low-order coal, alkali, enzyme, chelating agent and the table generated using three kinds of bacterium metabolism
The various actives substance such as face activating agent acts synergistically to degrading activity point different in low order texture of coal, improves low-order coal
Degradation rate improves the type and content of the products such as fine chemicals, liquid fuel in degradation solutions at different levels, is that low-order coal is further
Clean and effective using laying a good foundation.
2, the present invention carries out photooxidation processing to the coal dust prepared by low-order coal using rotation bed photochemical reactor, increases
Oxygen content in photooxidation powdered coal improves the microbial degradation activity in photooxidation powdered coal, and then improves photooxidation coal
The degradation rate of powder, method is simple, safety and environmental protection.
3, the present invention carries out classification degradation under the optimum process condition of microbial degradation photooxidation powdered coals at different levels respectively, protects
It has demonstrate,proved degradation processes at different levels and higher degradation rate is all had to degradation substrate, to improve the total degradation rate of low-order coal, in turn
The type and quantity for improving the products such as fine chemicals, liquid fuel in degradation solutions at different levels, are advantageously implemented the height of low-order coal
Effect utilizes.
4, the present invention establishes the relationship of degradation rate and corresponding degradation solutions at different levels absorbance at 450nm in degradation processes at different levels
Equation is concise straight so as to quickly be converted to degradation solution absorbance A 450 with corresponding degradation rate according to the relation equation
It connects, convenience and high-efficiency.
5, contain type chemical combination abundant in the degradation solutions at different levels that low-order coal of the invention is obtained through microorganism classification degradation
Object, can the isolated a variety of valuable compounds of further progress, for cleaning liquid fuel, medicament, raw material of industry field.
Below by drawings and examples, technical scheme of the present invention will be described in further detail.
Detailed description of the invention
Fig. 1 is the degradation rate and degradation solution absorbance relationship Linear Quasi of the green spore streptomycete degradation photooxidation low-order coal of the present invention
Close figure.
Fig. 2 be pseudomonas putida of the present invention degradation photooxidation low-order coal degradation rate and degradation solution absorbance relationship it is linear
Fitted figure.
Fig. 3 is the degradation rate and degradation solution absorbance relation line of Phanerochaete chrysosporium degradation photooxidation low-order coal of the present invention
Property fitted figure.
Specific embodiment
Green spore streptomycete (Streptomyces viridosporous), pseudomonas putida used in the present invention
(Pseudomonas putida) and Phanerochaete chrysosporium (Phanerochaete chrysosporium) are purchased from Chinese common
Microbiological Culture Collection administrative center, number are green spore streptomycete (4.1770), pseudomonas putida (1.1820) and yellow archespore
The flat lead fungi (3.7212) of hair.
The culture medium that green spore streptomycete (Streptomyces viridosporous) of the invention uses is Gause I
Culture medium, consisting of: soluble starch 20g, KNO31g, K2HPO40.5g, MgSO4·7H2O 0.5g, NaCl0.5g,
FeSO4·7H2O 0.01g, (solid medium adds agar 15g), distilled water 1000mL, pH are 7.4~7.6.First by the green of preservation
Spore streptomycete (Streptomyces viridosporous) is brought back to life, and using fluid nutrient medium culture to the third generation, will be cultivated extremely
Third generation 3d and bacteria concentration are not less than 3.0 × 105The green spore streptomycete bacterium solution of a/mL is inoculated with as female bacterium solution, is obtained
It is inoculated with the fluid nutrient medium of green spore streptomycete, is used for level-one degradation and independent degradation low-order coal.
The culture medium that pseudomonas putida (Pseudomonas putida) of the invention uses is LB culture medium, composition
Are as follows: peptone 10g, yeast powder 5g, NaCl 10g, (solid medium adds agar 15g), distilled water 1000mL, pH be 7.4~
7.6.First the pseudomonas putida of preservation (Pseudomonas putida) is brought back to life, using fluid nutrient medium culture to third
In generation, by culture to third generation 2d and bacteria concentration is not less than 8.0 × 105The pseudomonas putida bacterium solution of a/mL is as female bacterium solution
It is inoculated with, obtains the fluid nutrient medium for being inoculated with pseudomonas putida, used for second level degradation and independent degradation low-order coal.
The culture medium that Phanerochaete chrysosporium (Phanerochaete chrysosporium) of the invention uses is improvement
Martin's culture medium, consisting of: peptone 5g, yeast powder 2g, glucose 20g, dipotassium hydrogen phosphate 1g, magnesium sulfate 0.5g, (solid
Culture medium adds agar 15g), distilled water 1000mL, pH are 6.2~6.6.First by the Phanerochaete chrysosporium of preservation
(Phanerochaete chrysosporium) is brought back to life, using fluid nutrient medium culture to the third generation, will culture to the third generation the
2d and spore concentration are not less than 2.0 × 105The Phanerochaete chrysosporium bacterium solution of a/mL is inoculated with as female bacterium solution, is connect
Kind has the fluid nutrient medium of Phanerochaete chrysosporium, uses for multi-stage degradation and independent degradation low-order coal.
In general, using absorbance of the microbial degradation liquid at 450nm or microorganism to the degradation rate of substrate coal as evaluation
The index of coal degradation effect, the degradation rate calculation formula of microbial degradation low-order coal are as follows:
In formula (a), η is degradation rate (%), m0For the quality (g) of low-order coal, m1For obtained coal mass of residue of degrading
(g), A0For the ash content quality (g) in low-order coal, A1For the ash content quality (g) in obtained coal residue of degrading.
Degradation effect is evaluated using absorbance, it is easy to operate, but subsequent calculating is also needed, and degradation effect is evaluated with degradation rate
Fruit, it is more simple and clear but complicated for operation, and microorganism is related to the degradation rate of low-order coal and the granularity of low-order coal, low-order coal
Granularity it is smaller, microorganism is higher to the degradation rate of low-order coal.However when the granularity of low-order coal is larger, degradation can be obtained
Coal residue is sufficiently separated with microbial cells, so that degradation rate accurately be calculated using formula (a);When the granularity mistake of low-order coal
Hour, the coal residue that microbial degradation obtains is separated with microbial cells to be not thorough, and causes each parameter value in formula (a) inaccurate
Really, so that the degradation rate error obtained according to formula (a) is larger.Therefore, above-mentioned two index is used in combination the present invention, first
Degradation rate that the degradation solution and utilization formula (a) obtained after microbial degradation according to the biggish low-order coal of granularity is calculated
Corresponding relationship rule, establishes the degradation rate of microbial degradation photooxidation low-order coal and the relation equation of degradation solution absorbance, then
Applicability analysis is carried out to relation equation.The method for obtaining degradation rate using above-mentioned relation equation is simple and easy, only need to be to degradation
Liquid carries out simple centrifugally operated and obtains degradation solution, measures degradation solution absorbance value, without will degrade obtained coal residue with it is micro-
Biomass is kept completely separate, and reduces operation difficulty, substantially increases the accuracy of degradation rate.
(1) foundation of the degradation rate and degradation solution absorbance relation equation of green spore streptomycete degradation photooxidation low-order coal
Use green spore streptomycete respectively to granularity for -1.7mm+1mm (i.e. granularity be more than or equal to 1mm and be less than 1.7mm), -
1mm+0.7mm (i.e. granularity is more than or equal to 0.7mm and is less than 1mm), (i.e. granularity is more than or equal to 0.5mm and small to -0.7mm+0.5mm
In 0.7mm), -0.5mm+0.25mm (i.e. granularity be more than or equal to 0.25mm and be less than 0.5mm), -0.25mm+0.15m (i.e. granularity
More than or equal to 0.15mm and less than 0.25mm), -0.15mm+0.075mm, (i.e. granularity is more than or equal to 0.075mm and is less than
0.15mm), -0.075mm+0.045mm (i.e. granularity is more than or equal to 0.045mm and is less than 0.075mm) and -0.045mm (i.e. granularity
Less than 0.045mm) photooxidation Inner Mongol Shengli Brown (GSLH) degrade, the photooxidation Inner Mongol Shengli Brown of each granularity
(GSLH) three groups of parallel laboratory tests, the condition of degradation is arranged in degradation are as follows: coal amount is trained with the liquid of the green spore streptomycete of unit volume
Support photooxidation Inner Mongol Shengli Brown (GSLH) quality meter 9.50g/L in base, the green spore chain in the fluid nutrient medium of green spore streptomycete
Mould mother bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;It is described
Viable bacteria concentration in green spore strepto- starter bacterium solution is 3.1 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take the photooxidation Inner Mongol Shengli Brown of each granularity
(GSLH) absorbance values of three groups of parallel laboratory tests are as the A450 value under the granularity;It then is -1.7mm+ for granularity
The photooxidation Inner Mongol Shengli Brown of 1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m
(GSLH) the centrifuged precipitating of catabolite is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, by public affairs
Formula (a) calculates above-mentioned five kinds of granularity photooxidation Inner Mongol Shengli Brown (GSLH) corresponding degradation rate, and takes the photooxidation of each granularity
Change the degradation rate average value of Inner Mongol Shengli Brown (GSLH) three groups of parallel laboratory tests as the degradation rate under the granularity.
According to experimental data, draw green spore streptomycete degrade larger granularity (i.e. -1.7mm+1mm, -1mm+0.7mm, -
0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m) photooxidation Inner Mongol Shengli Brown (GSLH) degradation rate and suction
Photometric relationship Linear Fit Chart, as shown in Figure 1, obtaining degradation rate and the degradation solution suction of green spore streptomycete degradation photooxidation low-order coal
Photometric relationship equation are as follows: η10=0.02466+0.07453Y10, the goodness of fit determine coefficient be R10 2=0.98392, degree of fitting
Preferably, reliability is higher, wherein η10For degradation rate, Y10For degradation solution absorbance.It is -0.15+0.075mm, -0.075 by granularity
A450 value (respectively 2.693,2.587 and of the photooxidation Inner Mongol Shengli Brown (GSLH) of+0.045mm and -0.045mm
2.145) corresponding degradation rate η, is acquired10It is respectively as follows: 22.54%, 21.68% and 18.45%.
(2) applicability point of the degradation rate and degradation solution absorbance relation equation of green spore streptomycete degradation photooxidation low-order coal
Analysis
Using green spore streptomycete respectively to photooxidation Zhaotong County, Yunnan lignite (GZTH), photooxidation Shanxi Huiyuan lignite (GHYH)
It degrades with photooxidation Inner Mongol Yuanbaoshan Power Station lignite (GYBH), the granularity of above-mentioned every kind of photooxidation low-order coal is divided into -1.7mm+
1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m, the photooxidation low order of each granularity
Three groups of parallel laboratory tests, the condition of degradation are as follows: coal amount is with the fluid nutrient medium of the green spore streptomycete of unit volume is arranged in the degradation of coal
Middle photooxidation low order coal quality meter 9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration in the culture medium of green spore strepto- bacteria liquid
180mL/L, incubator frequency of oscillation 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;The green spore strepto- starter bacterium solution
In viable bacteria concentration be 3.1 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take three groups of photooxidation low-order coal of each granularity it is parallel
The absorbance values of experiment are as the A450 value under the granularity;Then for the photooxidation low-order coal catabolite of each granularity
Centrifuged precipitating is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, is calculated by formula (a) above-mentioned
The corresponding degradation rate of five kinds of granularity photooxidation low-order coals, and take the degradation of three groups of parallel laboratory tests of photooxidation low-order coal of each granularity
Rate average value is as the degradation rate under the granularity;Bring the A450 value of the photooxidation low-order coal of each granularity into green spore streptomycete again
The degradation rate and degradation solution absorbance relation equation η for photooxidation low-order coal of degrading10=0.02466+0.07453Y10In acquire pair
The degradation rate answered is as predicted value (ηPrediction), and with the degradation rate (η that is calculated by formula (a)It is practical) research is compared, as a result
As shown in table 1 below.
The degradation rate of the green spore streptomycete of table 1 degradation photooxidation low-order coal and the applicability point of degradation solution absorbance relation equation
Analyse result
As shown in Table 1, according to the degradation rate of green spore streptomycete degradation photooxidation low-order coal and degradation solution absorbance relationship side
The varigrained degradation rate η of three kinds of photooxidation low-order coals that journey obtainsPredictionThe degradation rate η obtained with formula (a)It is practicalBetween it is opposite
Error is smaller, illustrates the degradation rate of green spore streptomycete degradation photooxidation low-order coal from degradation solution absorbance relation equation to different
Photooxidation low-order coal has preferable applicability, can be used under the conditions of level-one degradation technique of the present invention, green spore streptomycete degradation light
Aoxidize the level-one degradation solution absorbance (Y of low-order coal1) and level-one degradation rate (η1) between conversion, that is, be expressed as η1=0.02466
+0.07453Y1, the goodness of fit determine coefficient be R1 2=0.98392.
(3) foundation of the degradation rate and degradation solution absorbance relation equation of pseudomonas putida degradation photooxidation low-order coal
Using pseudomonas putida, to granularity, for -1.7mm+1mm, (i.e. granularity is more than or equal to 1mm and is less than respectively
1.7mm), -1mm+0.7mm (i.e. granularity be more than or equal to 0.7mm and be less than 1mm), (i.e. granularity is more than or equal to -0.7mm+0.5mm
0.5mm and be less than 0.7mm), -0.5mm+0.25mm (i.e. granularity be more than or equal to 0.25mm and be less than 0.5mm), -0.25mm+
0.15m (i.e. granularity is more than or equal to 0.15mm and is less than 0.25mm), (i.e. granularity is more than or equal to 0.075mm to -0.15mm+0.075mm
And less than 0.15mm), -0.075mm+0.045mm (i.e. granularity is more than or equal to 0.045mm and is less than 0.075mm) and -0.045mm
Photooxidation Inner Mongol Shengli Brown (GSLH) of (i.e. granularity is less than 0.045mm) is degraded, the photooxidation Inner Mongol victory of each granularity
Three groups of parallel laboratory tests, the condition of degradation are as follows: coal amount is with unit volume pseudomonas putida is arranged in the degradation of sharp lignite (GSLH)
Fluid nutrient medium in photooxidation quality of pc meter 13.00g/L, the Pseudomonas putida in the culture medium of Pseudomonas putida bacteria liquid
Starter bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The evil
Viable bacteria concentration in smelly pseudomonad mother bacterium solution is 8.2 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take the photooxidation Inner Mongol Shengli Brown of each granularity
(GSLH) absorbance values of three groups of parallel laboratory tests are as the A450 value under the granularity;It then is -1.7mm+ for granularity
The photooxidation Inner Mongol Shengli Brown of 1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m
(GSLH) the centrifuged precipitating of catabolite is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, by public affairs
Formula (a) calculates above-mentioned five kinds of granularity photooxidation Inner Mongol Shengli Brown (GSLH) corresponding degradation rate, and takes the photooxidation of each granularity
Change the degradation rate average value of Inner Mongol Shengli Brown (GSLH) three groups of parallel laboratory tests as the degradation rate under the granularity.
According to experimental data, pseudomonas putida degradation larger granularity (i.e. -1.7mm+1mm, -1mm+ are drawn
0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m) photooxidation Inner Mongol Shengli Brown (GSLH) degradation
Rate and absorbance relationship Linear Fit Chart, as shown in Fig. 2, obtain the degradation rate of pseudomonas putida degradation photooxidation low-order coal with
Degradation solution absorbance relation equation are as follows: η20=0.02919+0.06412Y20, the goodness of fit determine coefficient be R20 2=
0.99075, degree of fitting is preferable, and reliability is higher, wherein η20For degradation rate, Y20For degradation solution absorbance.It is -0.15+ by granularity
The A450 value of the photooxidation Inner Mongol Shengli Brown (GSLH) of 0.075mm, -0.075+0.045mm and -0.045mm is (respectively
5.875,5.321 and 4.827), acquire corresponding degradation rate η20It is respectively as follows: 40.59%, 37.04% and 33.87%.
(4) applicability of the degradation rate and degradation solution absorbance relation equation of pseudomonas putida degradation photooxidation low-order coal
Analysis
Using pseudomonas putida respectively to photooxidation Zhaotong County, Yunnan lignite (GZTH), photooxidation Shanxi Huiyuan lignite
(GHYH) it degrades with photooxidation Inner Mongol Yuanbaoshan Power Station lignite (GYBH), the granularity of above-mentioned every kind of photooxidation low-order coal is divided into-
1.7mm+1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m, the photooxidation of each granularity
Three groups of parallel laboratory tests, the condition of degradation are as follows: coal amount is with the liquid of unit volume pseudomonas putida is arranged in the degradation for changing low-order coal
Photooxidation quality of pc meter 13.00g/L in body culture medium, the Pseudomonas putida starter in the fluid nutrient medium of pseudomonas putida
Bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false
Viable bacteria concentration in unit cell starter bacterium solution is 8.2 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take three groups of photooxidation low-order coal of each granularity it is parallel
The absorbance values of experiment are as the A450 value under the granularity;Then for the photooxidation low-order coal catabolite of each granularity
Centrifuged precipitating is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, is calculated by formula (a) above-mentioned
The corresponding degradation rate of five kinds of granularity photooxidation low-order coals, and take the degradation of three groups of parallel laboratory tests of photooxidation low-order coal of each granularity
Rate average value is as the degradation rate under the granularity;Bring the A450 value of the photooxidation low-order coal of each granularity into Pseudomonas putida again
The degradation rate and degradation solution absorbance relation equation η of bacterium degradation photooxidation low-order coal20=0.02919+0.06412Y20In acquire
Corresponding degradation rate is as predicted value (ηPrediction), and with the degradation rate (η that is calculated by formula (a)It is practical) research is compared, it ties
Fruit is as shown in table 2 below.
The degradation rate of 2 pseudomonas putida of table degradation photooxidation low-order coal and the applicability of degradation solution absorbance relation equation
Analyze result
As shown in Table 2, according to the degradation rate and degradation solution absorbance relationship of pseudomonas putida degradation photooxidation low-order coal
The varigrained degradation rate η of three kinds of photooxidation low-order coals that equation obtainsPredictionThe degradation rate η obtained with formula (a)It is practicalBetween phase
Smaller to error, the degradation rate and degradation solution absorbance relation equation for illustrating pseudomonas putida degradation photooxidation low-order coal be not to
Same photooxidation low-order coal has preferable applicability, can be used under the conditions of second level degradation technique of the present invention, pseudomonas putida
The second level degradation solution absorbance (Y for photooxidation low-order coal of degrading2) and second level degradation rate (η2) between conversion.It is expressed as η2=
0.02919+0.06412Y2, the goodness of fit determine coefficient be R2 2=0.99075.
(5) foundation of the degradation rate and degradation solution absorbance relation equation of Phanerochaete chrysosporium degradation photooxidation low-order coal
Using Phanerochaete chrysosporium, to granularity, for -1.7mm+1mm, (i.e. granularity is more than or equal to 1mm and is less than respectively
1.7mm), -1mm+0.7mm (i.e. granularity be more than or equal to 0.7mm and be less than 1mm), (i.e. granularity is more than or equal to -0.7mm+0.5mm
0.5mm and be less than 0.7mm), -0.5mm+0.25mm (i.e. granularity be more than or equal to 0.25mm and be less than 0.5mm), -0.25mm+
0.15m (i.e. granularity is more than or equal to 0.15mm and is less than 0.25mm), (i.e. granularity is more than or equal to 0.075mm to -0.15mm+0.075mm
And less than 0.15mm), -0.075mm+0.045mm (i.e. granularity is more than or equal to 0.045mm and is less than 0.075mm) and -0.045mm
Photooxidation Inner Mongol Shengli Brown (GSLH) of (i.e. granularity is less than 0.045mm) is degraded, the photooxidation Inner Mongol victory of each granularity
Three groups of parallel laboratory tests, the condition of degradation are as follows: coal amount is with unit volume Huang archespore Mao Pingge is arranged in the degradation of sharp lignite (GSLH)
Photooxidation quality of pc meter 13.00g/L in the fluid nutrient medium of bacterium, the yellow archespore in the fluid nutrient medium of Phanerochaete chrysosporium
The flat lead fungi mother bacterium solution inoculum concentration 90mL/L of hair, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;Institute
Stating the spore concentration in Phanerochaete chrysosporium mother's bacterium solution is 2.3 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take the photooxidation Inner Mongol Shengli Brown of each granularity
(GSLH) absorbance values of three groups of parallel laboratory tests are as the A450 value under the granularity;It then is -1.7mm+ for granularity
The photooxidation Inner Mongol Shengli Brown of 1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m
(GSLH) the centrifuged precipitating of catabolite is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, by public affairs
Formula (a) calculates above-mentioned five kinds of granularity photooxidation Inner Mongol Shengli Brown (GSLH) corresponding degradation rate, and takes the photooxidation of each granularity
Change the degradation rate average value of Inner Mongol Shengli Brown (GSLH) three groups of parallel laboratory tests as the degradation rate under the granularity.
According to experimental data, Phanerochaete chrysosporium degradation larger granularity (i.e. -1.7mm+1mm, -1mm+ are drawn
0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m) photooxidation Inner Mongol Shengli Brown (GSLH) degradation
Rate and absorbance relationship Linear Fit Chart, as shown in figure 3, obtaining the degradation rate of Phanerochaete chrysosporium degradation photooxidation low-order coal
With degradation solution absorbance relation equation are as follows: η30=0.02336+0.08945Y30Its goodness of fit determines that coefficient is R30 2=
0.97836, degree of fitting is preferable, and reliability is higher, wherein η30For degradation rate, Y30For degradation solution absorbance.It is -0.15+ by granularity
The A450 value of the photooxidation Inner Mongol Shengli Brown (GSLH) of 0.075mm, -0.075+0.045mm and -0.045mm is (respectively
5.228,4.409 and 3.511), acquire corresponding degradation rate η30It is respectively as follows: 48.09%, 40.95% and 33.12%.
(6) degradation rate and degradation solution absorbance relation equation of Phanerochaete chrysosporium degradation photooxidation low-order coal is applicable
Property analysis
Using Phanerochaete chrysosporium respectively to photooxidation Zhaotong County, Yunnan lignite (GZTH), photooxidation Shanxi Huiyuan lignite
(GHYH) it degrades with photooxidation Inner Mongol Yuanbaoshan Power Station lignite (GYBH), the granularity of above-mentioned every kind of photooxidation low-order coal is divided into-
1.7mm+1mm, -1mm+0.7mm, -0.7mm+0.5mm, -0.5mm+0.25mm, -0.25mm+0.15m, the photooxidation of each granularity
Three groups of parallel laboratory tests, the condition of degradation are as follows: coal amount is with unit volume Phanerochaete chrysosporium is arranged in the degradation for changing low-order coal
Photooxidation quality of pc meter 13.00g/L in fluid nutrient medium, the yellow archespore hair in the fluid nutrient medium of Phanerochaete chrysosporium are flat
Lead fungi mother bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Huang
Spore concentration in the flat lead fungi mother bacterium solution of archespore hair is 2.3 × 105A/mL.
After degradation, 15min is centrifuged to catabolite under conditions of 10000r/min respectively, obtained supernatant warp
It after filtering, crosses 0.22 μm of filter membrane and carries out secondary filter, obtain secondary filtrate, using deionized water as reference, using TU-1900 type point
Absorbance A 450 of the secondary filtrate of light photometric determination at 450nm, and take three groups of photooxidation low-order coal of each granularity it is parallel
The absorbance values of experiment are as the A450 value under the granularity;Then for the photooxidation low-order coal catabolite of each granularity
Centrifuged precipitating is repeatedly washed, and is removed thallus, is placed at 60 DEG C that drying to constant weight, is calculated by formula (a) above-mentioned
The corresponding degradation rate of five kinds of granularity photooxidation low-order coals, and take the degradation of three groups of parallel laboratory tests of photooxidation low-order coal of each granularity
Rate average value is as the degradation rate under the granularity;The A450 value of the photooxidation low-order coal of each granularity is brought into yellow archespore hair again to put down
The degradation rate and degradation solution absorbance relation equation η of lead fungi degradation photooxidation low-order coal30=0.02336+0.08945Y30In ask
Corresponding degradation rate is obtained as predicted value (ηPrediction), and with the degradation rate (η that is calculated by formula (a)It is practical) research is compared,
As a result as shown in table 3 below.
The degradation rate of 3 Phanerochaete chrysosporium of table degradation photooxidation low-order coal and being applicable in for degradation solution absorbance relation equation
Property analysis result
As shown in Table 3, it is closed according to the degradation rate of Phanerochaete chrysosporium degradation photooxidation low-order coal and degradation solution absorbance
It is the varigrained degradation rate η of three kinds of photooxidation low-order coals that equation obtainsPredictionThe degradation rate η obtained with formula (a)It is practicalBetween
Relative error is smaller, illustrates the degradation rate and degradation solution absorbance relation equation of Phanerochaete chrysosporium degradation photooxidation low-order coal
There is preferable applicability to different photooxidation low-order coals, can be used under multi-stage degradation process conditions of the present invention, yellow archespore hair
The multi-stage degradation liquid absorbance (Y of flat lead fungi degradation photooxidation low-order coal3) and multi-stage degradation rate (η3) between conversion.Indicate
For η3=0.02336+0.08945Y3, the goodness of fit determine coefficient be R3 2=0.97836.
Embodiment 1
The present embodiment the following steps are included:
Step 1: in 100 DEG C of drying 1h after Inner Mongol Huolinhe lignite (HLH) is crushed, then successively through grinding and screening,
Obtain Inner Mongol Huolinhe lignite (HLH) coal dust that granularity is -0.15mm+0.075mm;
Step 2: using rotation bed photochemical reactor to Inner Mongol Huolinhe lignite (HLH) coal dust obtained in step 1
Photooxidation pretreatment is carried out, photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust is obtained;The pretreated condition of photooxidation are as follows:
Coal amount 20g/L in terms of Inner Mongol Huolinhe lignite (HLH) quality of pc being added in unit volume rotation bed photochemical reactor,
Ultraviolet ray intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with rotation before photooxidation pretreatment
Every liter of volumeter of rotated bed photochemical reactor of logical oxygen time 10min;
Green spore is inoculated with Step 3: photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of streptomycete (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, so
Level-one catabolite is filtered afterwards, collects level-one degradation solution and level-one degradation coal residue respectively, then level-one degradation coal is residual
Slag is placed in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to level-one
Absorbance of the degradation solution at 450nm is measured to obtain Y1=1.866, carry it into green spore streptomycete degradation photooxidation low order
The level-one degradation rate and level-one degradation solution absorbance relation equation η of coal1=0.02466+0.07453Y1In, the goodness of fit determines
Coefficient is R1 2=0.98392, obtain η1=0.1637, i.e. η1It is 16.37%;The condition of the level-one degradation are as follows: unit volume
It is inoculated with the additional amount 9.50g/L of photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust in the fluid nutrient medium of green spore streptomycete,
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/ in the fluid nutrient medium of green spore streptomycete
Min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.2 × 105A/mL;
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=2.812, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.2095, i.e. η2It is 20.95%,;The condition of the second level degradation are as follows: unit volume is inoculated with foul smelling
In the fluid nutrient medium of pseudomonad it is sterilized after level-one degradation coal residue additional amount with photooxidation in level-one degradation process
Inner Mongol Huolinhe lignite (GHLH) quality of pc meter 13.00g/L, the Pseudomonas putida in the fluid nutrient medium of pseudomonas putida
Starter bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The evil
Viable bacteria concentration in smelly pseudomonad mother bacterium solution is 8.3 × 105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid respectively and multi-stage degradation coal residue is adopted using deionized water as blank control
Absorbance of the multi-stage degradation liquid at 450nm is measured with spectrophotometry, obtains Y3=3.526, carry it into yellow spore
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of the flat lead fungi degradation photooxidation low-order coal of raw wool3=0.02336+
0.08945Y3In, the goodness of fit determines that coefficient is R3 2=0.97836, obtain η3=0.3388, i.e. η3It is 33.88%;It is described
The condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation
The additional amount of coal residue 13.00g/L in terms of photooxidation Inner Mongol Huolinhe lignite (GHLH) quality of pc in level-one degradation process, it is yellow
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L in the fluid nutrient medium of the flat lead fungi of archespore hair, incubator frequency of oscillation
210r/min, incubation time 14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.1 ×
105A/mL.
Comparative example 1
This comparative example the following steps are included:
Step 1: in 100 DEG C of drying 1h after Inner Mongol Huolinhe lignite (HLH) is crushed, then successively through grinding and screening,
Obtain Inner Mongol Huolinhe lignite (HLH) coal dust that granularity is -0.15mm+0.075mm;
Step 2: using rotation bed photochemical reactor to Inner Mongol Huolinhe lignite (HLH) coal dust obtained in step 1
Photooxidation pretreatment is carried out, photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust is obtained;The pretreated condition of photooxidation are as follows:
Photooxidation Inner Mongol Huolinhe lignite (GHLH) quality of pc of coal amount to be added in unit volume rotation bed photochemical reactor
20g/L, ultraviolet ray intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min are counted, photooxidation is located in advance
The logical oxygen time 10min of every liter of bed photochemical reactor volumeter is rotated before reason;
Green spore is inoculated with Step 3: photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of streptomycete (Streptomyces viridosporous), it is placed in incubator and degrades, it is then right
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution in 450nm
The absorbance at place is measured to obtain Y10=1.866, carry it into the degradation rate of green spore streptomycete degradation photooxidation low-order coal with
Degradation solution absorbance relation equation η10=0.02466+0.07453Y10In, the goodness of fit determines that coefficient is R10 2=
0.98392, obtain η10=0.1637, i.e. η10It is 16.37%;The green spore streptomycete single bacterium degradation photooxidation Inner Mongol Huo Linhe
The condition of lignite are as follows: coal amount is with photooxidation Inner Mongol Huolinhe lignite (GHLH) in the green spore streptomycete fluid nutrient medium of unit volume
Quality of pc meter 9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration 180mL/L in green spore streptomycete fluid nutrient medium, culture
Case frequency of oscillation 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution
It is 3.2 × 105A/mL.
Comparative example 2
This comparative example the following steps are included:
Step 1: in 100 DEG C of drying 1h after Inner Mongol Huolinhe lignite (HLH) is crushed, then successively through grinding and screening,
Obtain Inner Mongol Huolinhe lignite (HLH) coal dust that granularity is -0.15mm+0.075mm;
Step 2: using rotation bed photochemical reactor to Inner Mongol Huolinhe lignite (HLH) coal dust obtained in step 1
Photooxidation pretreatment is carried out, photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust is obtained;The pretreated condition of photooxidation are as follows:
Coal amount 20g/L in terms of Inner Mongol Huolinhe lignite (HLH) quality of pc being added in unit volume rotation bed photochemical reactor,
Ultraviolet ray intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with rotation before photooxidation pretreatment
Every liter of volumeter of rotated bed photochemical reactor of logical oxygen time 10min;
Step 3: photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust obtained in step 2 is added to inoculation foul smelling
It in the fluid nutrient medium of pseudomonad (Pseudomonas putida), is placed in incubator and degrades, then degradation is produced
Object is filtered to obtain degradation solution, the suction using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Luminosity is measured, and obtains Y20=2.991, carry it into degradation rate and the degradation of pseudomonas putida degradation photooxidation low-order coal
Liquid absorbance relation equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, it obtains
To η20=0.2209, i.e. η20It is 22.09%;The pseudomonas putida single bacterium degradation photooxidation Inner Mongol Huolinhe lignite
(GHLH) condition are as follows: coal amount is with photooxidation Inner Mongol Huolinhe lignite in unit volume pseudomonas putida fluid nutrient medium
(GHLH) quality of pc meter 13.00g/L, the Pseudomonas putida starter bacterium solution inoculum concentration in pseudomonas putida fluid nutrient medium
135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The Pseudomonas putida starter bacterium
Viable bacteria concentration in liquid is 8.3 × 105A/mL.
Comparative example 3
Step 1: in 100 DEG C of drying 1h after Inner Mongol Huolinhe lignite (HLH) is crushed, then successively through grinding and screening,
Obtain Inner Mongol Huolinhe lignite (HLH) coal dust that granularity is -0.15mm+0.075mm;
Step 2: using rotation bed photochemical reactor to Inner Mongol Huolinhe lignite (HLH) coal dust obtained in step 1
Photooxidation pretreatment is carried out, photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust is obtained;The pretreated condition of photooxidation are as follows:
Photooxidation Inner Mongol Huolinhe lignite (GHLH) quality of pc of coal amount to be added in unit volume rotation bed photochemical reactor
20g/L, ultraviolet ray intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min are counted, photooxidation is located in advance
The logical oxygen time 10min of every liter of bed photochemical reactor volumeter is rotated before reason;
Yellow archespore is inoculated with Step 3: photooxidation Inner Mongol Huolinhe lignite (GHLH) coal dust will be obtained in step 2 and be added to
In the fluid nutrient medium of the flat lead fungi (Phanerochaete chrysosporium) of hair, it is placed in incubator and degrades, then
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, is existed using spectrophotometry to degradation solution
Absorbance at 450nm is measured, and obtains Y30=2.973, carry it into Phanerochaete chrysosporium degradation photooxidation low-order coal
Degradation rate and degradation solution absorbance relation equation η30=0.02336+0.08945Y30In, the goodness of fit determines that coefficient is
R30 2=0.97836, obtain η30=0.2893, i.e. η30It is 28.93%;In the Phanerochaete chrysosporium single bacterium degradation photooxidation
Cover the condition of Huolinhe lignite (GHLH) are as follows: coal amount is in photooxidation in unit volume Phanerochaete chrysosporium fluid nutrient medium
Cover Huolinhe lignite (GHLH) quality of pc meter 13.00g/L, the yellow archespore Mao Pingge in Phanerochaete chrysosporium fluid nutrient medium
Starter bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Huang spore
Spore concentration in the flat lead fungi mother bacterium solution of raw wool is 2.1 × 105A/mL.
Embodiment 1 and comparative example 1~3 are compared it is found that photooxidation Inner Mongol Huolinhe lignite (GHLH) in embodiment 1
The total degradation rate η of microorganism classification degradationAlways=η1+η2+η3=0.7120, i.e. ηAlways=71.20%, and green spore in comparative example 1~3
Streptomycete, pseudomonas putida and Phanerochaete chrysosporium are respectively to the degradation rate of photooxidation Inner Mongol Huolinhe lignite (GHLH)
η10=16.37%, η20=22.09%, η30=28.93%, i.e., successively using above-mentioned three kinds of bacterium classification degradation photooxidation Inner Mongol Huo Lin
The total degradation rate of river lignite (GHLH) is much larger than the individually degradation to photooxidation Inner Mongol Huolinhe lignite (GHLH) of three kinds of bacterium
Rate illustrates that classification biodegrading process of the invention improves the microbial degradation rate of Inner Mongol Huolinhe lignite (HLH).
Embodiment 2
The present embodiment the following steps are included:
Step 1: then successively being obtained through grinding and screening after Zhaotong County, Yunnan lignite (ZTH) is crushed in 100 DEG C of drying 1h
Zhaotong County, Yunnan lignite (ZTH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to Zhaotong County, Yunnan lignite (ZTH) coal dust obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Zhaotong County, Yunnan lignite (ZTH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Green spore chain is inoculated with Step 3: photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, then
Level-one catabolite is filtered, level-one degradation solution and level-one is collected respectively and degrades coal residue, then level-one is degraded coal residue
It is placed in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, level-one is dropped using spectrophotometry
Absorbance of the liquid at 450nm is solved to be measured to obtain Y1=2.873, carry it into green spore streptomycete degradation photooxidation low-order coal
Level-one degradation rate and level-one degradation solution absorbance relation equation η1=0.02466+0.07453Y1In, the goodness of fit determines system
Number is R1 2=0.98392, obtain η1=0.2388, i.e. η1It is 23.88%;The condition of the level-one degradation are as follows: unit volume connects
Kind has the additional amount 9.50g/L of photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust in the fluid nutrient medium of green spore streptomycete, green spore
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min in the fluid nutrient medium of streptomycete, training
Feeding time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.3 × 105A/mL;
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=3.549, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.2568, i.e. η2It is 25.68%;The condition of the second level degradation are as follows: it is false that unit volume is inoculated with foul smelling
In the fluid nutrient medium of monad it is sterilized after level-one degradation coal residue additional amount with photooxidation cloud in level-one degradation process
Southern Zhaotong Brown Coal (GZTH) quality of pc meter 13.00g/L, the Pseudomonas putida starter in the fluid nutrient medium of pseudomonas putida
Bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false
Viable bacteria concentration in unit cell starter bacterium solution is 8.3 ×.105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid and multi-stage degradation coal residue respectively;Using deionized water as blank control, adopt
Absorbance of the multi-stage degradation liquid at 450nm is measured with spectrophotometry, obtains Y3=3.559, carry it into yellow spore
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of the flat lead fungi degradation photooxidation low-order coal of raw wool3=0.02336+
0.08945Y3In, the goodness of fit determines that coefficient is R3 2=0.97836, obtain η3=0.3417, i.e. η3It is 34.17%;It is described
The condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation
The additional amount of coal residue 13.00g/L, yellow spore in terms of photooxidation Zhaotong County, Yunnan lignite (GZTH) quality of pc in level-one degradation process
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/ in the flat lead fungi fluid nutrient medium of raw wool
Min, incubation time 14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.2 × 105
A/mL.
Comparative example 4
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Zhaotong County, Yunnan lignite (ZTH) is crushed in 100 DEG C of drying 1h
Zhaotong County, Yunnan lignite (ZTH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to Zhaotong County, Yunnan lignite (ZTH) coal dust obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Zhaotong County, Yunnan lignite (ZTH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Zhaotong County, Yunnan lignite (ZTH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Green spore chain is inoculated with Step 3: photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in incubator and degrades, then to drop
Solution product be filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance be measured to obtain Y10=2.873, carry it into the degradation rate and drop of green spore streptomycete degradation photooxidation low-order coal
Solve liquid absorbance relation equation η10=0.02466+0.07453Y1In, the goodness of fit determines that coefficient is R10 2=0.98392, it obtains
To η10=0.2388, i.e. η10It is 23.88%;Green spore streptomycete single bacterium degradation photooxidation Zhaotong County, Yunnan lignite (GZTH)
Condition are as follows: coal amount is with photooxidation Zhaotong County, Yunnan lignite (GZTH) quality of pc in the green spore streptomycete fluid nutrient medium of unit volume
Count 9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration 180mL/L in green spore streptomycete fluid nutrient medium, incubator oscillation frequency
Rate 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.3 ×
105A/mL.
Comparative example 5
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Zhaotong County, Yunnan lignite (ZTH) is crushed in 100 DEG C of drying 1h
Zhaotong County, Yunnan lignite (ZTH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to Zhaotong County, Yunnan lignite (ZTH) coal dust obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Zhaotong County, Yunnan lignite (ZTH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Step 3: it is false that photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust obtained in step 2 is added to inoculation foul smelling
In the fluid nutrient medium of monad (Pseudomonas putida), it is placed in incubator and degrades, then to catabolite
It is filtered to obtain degradation solution, the extinction using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Degree is measured, and obtains Y20=4.420, carry it into the degradation rate and degradation solution of pseudomonas putida degradation photooxidation low-order coal
Absorbance relation equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, it obtains
η20=0.3126, i.e. η20It is 31.26%;The condition of the pseudomonas putida single bacterium degradation are as follows: coal amount is disliked with unit volume
Photooxidation Zhaotong County, Yunnan lignite (GZTH) quality of pc meter 13.00g/L, pseudomonas putida in smelly pseudomonad fluid nutrient medium
Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L in fluid nutrient medium, incubator frequency of oscillation 160r/min, incubation time
12d, 30 DEG C of cultivation temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution is 8.3 × 105A/mL.
Comparative example 6
Step 1: then successively being obtained through grinding and screening after Zhaotong County, Yunnan lignite (ZTH) is crushed in 100 DEG C of drying 1h
Zhaotong County, Yunnan lignite (ZTH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to Zhaotong County, Yunnan lignite (ZTH) coal dust obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Zhaotong County, Yunnan lignite (ZTH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Yellow archespore hair is inoculated with Step 3: photooxidation Zhaotong County, Yunnan lignite (GZTH) coal dust will be obtained in step 2 and be added to
In the fluid nutrient medium of flat lead fungi (Phanerochaete chrysosporium), it is placed in incubator and degrades, it is then right
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution in 450nm
The absorbance at place is measured, and obtains Y30=4.890, carry it into the degradation of Phanerochaete chrysosporium degradation photooxidation low-order coal
Rate and degradation solution absorbance relation equation η30=0.02336+0.08945Y30In, the goodness of fit determines that coefficient is R30 2=
0.97836, obtain η30=0.4608, i.e. η30It is 46.08%;The condition of the Phanerochaete chrysosporium single bacterium degradation are as follows: coal
Amount is in terms of photooxidation Zhaotong County, Yunnan lignite (GZTH) quality of pc in unit volume Phanerochaete chrysosporium fluid nutrient medium
13.00g/L, the Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L in Phanerochaete chrysosporium fluid nutrient medium, incubator
Frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;Spore in the Phanerochaete chrysosporium mother bacterium solution is dense
Degree is 2.2 × 105A/mL.
Embodiment 2 is compared with comparative example 4~6 it is found that photooxidation Zhaotong County, Yunnan lignite (GZTH) is micro- in embodiment 2
The total degradation rate η of biology classification degradationAlways=η1+η2+η3=0.8373, i.e. ηAlways=83.73%, and green spore chain in comparative example 4~6
Mould, pseudomonas putida and Phanerochaete chrysosporium are directly respectively to the degradation rate of photooxidation Zhaotong County, Yunnan lignite (GZTH)
η10=23.88%, η20=31.26%, η30=46.08%, i.e., successively using above-mentioned three kinds of bacterium classification degradation photooxidation Zhaotong County, Yunnan
The total degradation rate of lignite (GZTH), individually to the degradation rate of photooxidation Zhaotong County, Yunnan lignite (GZTH), is said much larger than three kinds of bacterium
Bright classification biodegrading process of the invention improves the degradation rate of Zhaotong County, Yunnan lignite (GZTH).
Embodiment 3
The present embodiment the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shanxi Huiyuan lignite (HYH) is crushed in 100 DEG C of drying 1h
Shanxi Huiyuan lignite (HYH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to lignite (HYH) coal dust in Shanxi Huiyuan obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Shanxi Huiyuan lignite (GHYH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Shanxi Huiyuan lignite (HYH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Green spore chain is inoculated with Step 3: photooxidation Shanxi Huiyuan lignite (GHYH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, then
Level-one catabolite is filtered, level-one degradation solution and level-one is collected respectively and degrades coal residue, then level-one is degraded coal residue
It is placed in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, level-one is dropped using spectrophotometry
Absorbance of the liquid at 450nm is solved to be measured to obtain Y1=1.925, carry it into green spore streptomycete degradation photooxidation low-order coal
Level-one degradation rate and level-one degradation solution absorbance relation equation η1=0.02466+0.07453Y1In, the goodness of fit determines system
Number is R1 2=0.98392, obtain η1=0.1681, i.e. η1It is 16.81%;The condition of the level-one degradation are as follows: unit volume connects
Kind has the additional amount 9.50g/L of photooxidation Shanxi Huiyuan lignite (GHYH) coal dust in the fluid nutrient medium of green spore streptomycete, green spore
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min in the fluid nutrient medium of streptomycete, training
Feeding time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.0 × 105A/mL;
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=2.734, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.2032, i.e. η2It is 20.32%;The condition of the second level degradation are as follows: it is false that unit volume is inoculated with foul smelling
In the fluid nutrient medium of monad it is sterilized after level-one degradation coal residue additional amount with photooxidation mountain in level-one degradation process
Western Huiyuan lignite (GHYH) quality of pc meter 13.00g/L, the Pseudomonas putida starter in the fluid nutrient medium of pseudomonas putida
Bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false
Viable bacteria concentration in unit cell starter bacterium solution is 8.0 × 105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid respectively and multi-stage degradation coal residue is adopted using deionized water as blank control
Absorbance of the multi-stage degradation liquid at 450nm is measured with spectrophotometry, obtains Y3=2.688, carry it into yellow spore
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of the flat lead fungi degradation photooxidation low-order coal of raw wool3=0.02336+
0.08945Y3In, the goodness of fit determines that coefficient is R3 2=0.97836, obtain η3=0.2638, i.e. η3It is 26.38%;It is described
The condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation
The additional amount of coal residue 13.00g/L, yellow spore in terms of photooxidation Shanxi Huiyuan lignite (GHYH) quality of pc in level-one degradation process
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L in the fluid nutrient medium of the flat lead fungi of raw wool, incubator frequency of oscillation
210r/min, incubation time 14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.0 ×
105A/mL.
Comparative example 7
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shanxi Huiyuan lignite (HYH) is crushed in 100 DEG C of drying 1h
Shanxi Huiyuan lignite (HYH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to lignite (HYH) coal dust in Shanxi Huiyuan obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Shanxi Huiyuan lignite (GHYH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Shanxi Huiyuan lignite (HYH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Green spore chain is inoculated with Step 3: photooxidation Shanxi Huiyuan lignite (GHYH) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in incubator and degrades, then to drop
Solution product be filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance be measured to obtain Y10=1.925, carry it into the degradation rate and drop of green spore streptomycete degradation photooxidation low-order coal
Solve liquid absorbance relation equation η10=0.02466+0.07453Y10In, the goodness of fit determines that coefficient is R10 2=0.98392,
Obtain η10=0.1681, i.e. η10It is 16.81%;Green spore streptomycete single bacterium degradation photooxidation Shanxi Huiyuan lignite (GHYH)
Condition are as follows: coal amount is with photooxidation Shanxi Huiyuan lignite (GHYH) coal dust matter in the green spore streptomycete fluid nutrient medium of unit volume
Meter 9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration 180mL/L in green spore streptomycete fluid nutrient medium, incubator oscillation
Frequency 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.0 ×
105A/mL.
Comparative example 8
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shanxi Huiyuan lignite (HYH) is crushed in 100 DEG C of drying 1h
Shanxi Huiyuan lignite (HYH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to lignite (HYH) coal dust in Shanxi Huiyuan obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Shanxi Huiyuan lignite (HYH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Shanxi Huiyuan lignite (HYH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Step 3: it is false that photooxidation Shanxi Huiyuan lignite (HYH) coal dust obtained in step 2 is added to inoculation foul smelling
In the fluid nutrient medium of monad (Pseudomonas putida), it is placed in incubator and degrades, then to catabolite
It is filtered to obtain degradation solution, the extinction using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Degree is measured, and obtains Y20=4.013, carry it into the degradation rate and degradation solution of pseudomonas putida degradation photooxidation low-order coal
Absorbance relation equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, it obtains
η20=0.2865, i.e. η20It is 28.65%;Pseudomonas putida single bacterium degradation photooxidation Shanxi Huiyuan lignite (GHYH)
Condition are as follows: coal amount is with photooxidation Shanxi Huiyuan lignite (HYH) coal dust matter in unit volume pseudomonas putida fluid nutrient medium
Meter 13.00g/L, the Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L in pseudomonas putida fluid nutrient medium, incubator
Frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution
It is 8.0 × 105A/mL.
Comparative example 9
Step 1: then successively being obtained through grinding and screening after Shanxi Huiyuan lignite (HYH) is crushed in 100 DEG C of drying 1h
Shanxi Huiyuan lignite (HYH) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: using rotation bed photochemical reactor to lignite (HYH) coal dust in Shanxi Huiyuan obtained in step 1 into
Row photooxidation pretreatment, obtains photooxidation Shanxi Huiyuan lignite (GHYH) coal dust;The pretreated condition of photooxidation are as follows: coal
Measure the 20g/L in terms of Shanxi Huiyuan lignite (HYH) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light
Intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed light before photooxidation pretreatment
The logical oxygen time 10min of every liter of chemical reactor volumeter;
Yellow archespore hair is inoculated with Step 3: photooxidation Shanxi Huiyuan lignite (GHYH) coal dust will be obtained in step 2 and be added to
In the fluid nutrient medium of flat lead fungi (Phanerochaete chrysosporium), it is placed in incubator and degrades, it is then right
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution in 450nm
The absorbance at place is measured, and obtains Y30=4.890, carry it into the degradation of Phanerochaete chrysosporium degradation photooxidation low-order coal
Rate and degradation solution absorbance relation equation η30=0.02336+0.08945Y30In, the goodness of fit determines that coefficient is R30 2=
0.97836, obtain η30=0.3892, i.e. η30It is 38.92%;Phanerochaete chrysosporium single bacterium degradation photooxidation Shanxi is muddy
The condition of source lignite (GHYH) are as follows: coal amount is with photooxidation Shanxi Huiyuan in unit volume Phanerochaete chrysosporium fluid nutrient medium
Lignite (GHYH) quality of pc meter 13.00g/L, Phanerochaete chrysosporium mother's bacterium solution in Phanerochaete chrysosporium fluid nutrient medium
Inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Huang archespore Mao Pingge
Spore concentration in starter bacterium solution is 2.0 × 105A/mL.
Embodiment 3 is compared with comparative example 7~9 it is found that photooxidation Shanxi Huiyuan lignite (GHYH) is micro- in embodiment 3
The total degradation rate η of biology classification degradationAlways=η1+η2+η3=0.6364, i.e. ηAlways=63.64%, and green spore chain in comparative example 7~9
Mould, pseudomonas putida and Phanerochaete chrysosporium are respectively η to the degradation rate of photooxidation Shanxi Huiyuan lignite (GHYH)10=
23.88%, η20=31.26%, η30=38.92%, i.e., it is successively brown using above-mentioned three kinds of bacterium classification degradation photooxidation Shanxi Huiyuan
The total degradation rate of coal (GHYH), individually to the degradation rate of photooxidation Shanxi Huiyuan lignite (GHYH), illustrates this hair much larger than three kinds of bacterium
Bright classification biodegrading process improves the microbial degradation rate of Shanxi Huiyuan lignite (HYH).
Embodiment 4
The present embodiment the following steps are included:
Step 1: then successively being obtained through grinding and screening after Inner Mongol jet coal (ICY) is crushed in 100 DEG C of drying 1h
Granularity is Inner Mongol jet coal (ICY) coal dust of -0.15mm+0.075mm;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (ICY) coal dust in Inner Mongol obtained in step 1
Photooxidation pretreatment, obtains photooxidation Inner Mongol jet coal (GICY) coal dust;The pretreated condition of photooxidation are as follows: coal amount with
Photooxidation Inner Mongol jet coal (GICY) the quality of pc meter 20g/L being added in unit volume rotation bed photochemical reactor, it is ultraviolet
Luminous intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, with revolving bed before photooxidation pretreatment
The logical oxygen time 10min of every liter of photochemical reactor volumeter;
Green spore strepto- is inoculated with Step 3: photooxidation Inner Mongol jet coal (GICY) coal dust obtained in step 2 is added to
In the fluid nutrient medium of bacterium (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, it is then right
Level-one catabolite is filtered, and collects level-one degradation solution and level-one degradation coal residue respectively, then level-one degradation coal residue is set
In 121 DEG C of sterilizing 20min in high-pressure sterilizing pot, using deionized water as blank control, degraded using spectrophotometry to level-one
Absorbance of the liquid at 450nm is measured to obtain Y1=1.447, carry it into green spore streptomycete degradation photooxidation low-order coal
Level-one degradation rate and level-one degradation solution absorbance relation equation η1=0.02466+0.07453Y1In, the goodness of fit determines coefficient
For R1 2=0.98392, obtain η1=0.1325, i.e. η1It is 13.25%;The condition of the level-one degradation are as follows: unit volume inoculation
There are the additional amount 9.50g/L of photooxidation Inner Mongol jet coal (GICY) coal dust in the fluid nutrient medium of green spore streptomycete, green spore strepto-
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L in the fluid nutrient medium of bacterium, incubator frequency of oscillation 160r/min, when culture
Between 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.2 × 105A/mL.
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=2.173, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.1685, i.e. η2It is 16.85%;The condition of the second level degradation are as follows: it is false that unit volume is inoculated with foul smelling
In the fluid nutrient medium of monad it is sterilized after level-one degradation coal residue additional amount in photooxidation in level-one degradation process
Cover jet coal (GICY) quality of pc meter 13.00g/L, the Pseudomonas putida starter bacterium in the fluid nutrient medium of pseudomonas putida
Liquid inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false single
Viable bacteria concentration in born of the same parents' starter bacterium solution is 8.2 × 105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid respectively and multi-stage degradation coal residue is adopted using deionized water as blank control
Absorbance of the multi-stage degradation liquid at 450nm is measured with spectrophotometry, obtains Y3=2.497, carry it into yellow spore
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of the flat lead fungi degradation photooxidation low-order coal of raw wool3=0.02336+
0.08945Y3In, the goodness of fit determines that coefficient is R3 2=0.97836, obtain η3=0.2467, i.e. η3It is 24.67%;It is described
The condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation
The additional amount of coal residue 13.00g/L, yellow archespore in terms of photooxidation Inner Mongol jet coal (GICY) quality of pc in level-one degradation process
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/ in the fluid nutrient medium of the flat lead fungi of hair
Min, incubation time 14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.4 × 105
A/mL.
Comparative example 10
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Inner Mongol jet coal (ICY) is crushed in 100 DEG C of drying 1h
Granularity is Inner Mongol jet coal (ICY) coal dust of -0.15mm+0.075mm;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (ICY) coal dust in Inner Mongol obtained in step 1
Photooxidation pretreatment, obtains photooxidation Inner Mongol jet coal (GICY) coal dust;The pretreated condition of photooxidation are as follows: coal amount with
Inner Mongol jet coal (ICY) the quality of pc meter 20g/L being added in unit volume rotation bed photochemical reactor, ultraviolet ray intensity
150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, to rotate bed photochemical before photooxidation pretreatment
The logical oxygen time 10min of every liter of reactor volume meter;
Green spore strepto- is inoculated with Step 3: photooxidation Inner Mongol jet coal (GICY) coal dust obtained in step 2 is added to
In the fluid nutrient medium of bacterium (Streptomyces viridosporous), it is placed in incubator and degrades, then to degradation
Product is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance is measured to obtain Y10=1.447, carry it into the degradation rate and degradation solution extinction of green spore streptomycete degradation low-order coal
Spend relation equation η10=0.02466+0.07453Y10In, the goodness of fit determines that coefficient is R10 2=0.98392, obtain η10=
0.1325, i.e. η10It is 13.25%;The condition of green spore streptomycete single bacterium degradation photooxidation Inner Mongol jet coal (GICY) are as follows: add
Coal amount 9.50g/L in terms of photooxidation Inner Mongol jet coal (GICY) quality of pc in the green spore streptomycete fluid nutrient medium of unit volume,
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min in green spore streptomycete fluid nutrient medium,
Incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.2 × 105A/mL.
Comparative example 11
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Inner Mongol jet coal (ICY) is crushed in 100 DEG C of drying 1h
Granularity is Inner Mongol jet coal (ICY) coal dust of -0.15mm+0.075mm;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (ICY) coal dust in Inner Mongol obtained in step 1
Photooxidation pretreatment, obtains photooxidation Inner Mongol jet coal (GICY) coal dust;The pretreated condition of photooxidation are as follows: coal amount with
Inner Mongol jet coal (ICY) the quality of pc meter 20g/L being added in unit volume rotation bed photochemical reactor, ultraviolet ray intensity
150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, to rotate bed photochemical before photooxidation pretreatment
The logical oxygen time 10min of every liter of reactor volume meter;
Step 3: it is false single that photooxidation Inner Mongol jet coal (GICY) coal dust obtained in step 2 is added to inoculation foul smelling
In the fluid nutrient medium of born of the same parents bacterium (Pseudomonas putida), be placed in incubator and degrade, then to catabolite into
Degradation solution is obtained by filtration in row, the absorbance using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
It is measured, obtains Y20=1.998, the degradation rate and degradation solution for carrying it into pseudomonas putida degradation photooxidation low-order coal are inhaled
Photometric relationship equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, obtain η20
=0.1573, i.e. η20It is 15.73%;The condition of pseudomonas putida single bacterium degradation photooxidation Inner Mongol jet coal (GICY)
Are as follows: coal amount is in terms of photooxidation Inner Mongol jet coal (GICY) quality of pc in unit volume pseudomonas putida fluid nutrient medium
13.00g/L, the Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L in pseudomonas putida fluid nutrient medium, incubator oscillation
Frequency 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution is 8.2
×105A/mL.
Comparative example 12
Step 1: then successively being obtained through grinding and screening after Inner Mongol jet coal (ICY) is crushed in 100 DEG C of drying 1h
Granularity is Inner Mongol jet coal (ICY) coal dust of -0.15mm+0.075mm;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (ICY) coal dust in Inner Mongol obtained in step 1
Photooxidation pretreatment, obtains photooxidation Inner Mongol jet coal (GICY) coal dust;The pretreated condition of photooxidation are as follows: coal amount with
Inner Mongol jet coal (ICY) the quality of pc meter 20g/L being added in unit volume rotation bed photochemical reactor, ultraviolet ray intensity
150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, to rotate bed photochemical before photooxidation pretreatment
The logical oxygen time 10min of every liter of reactor volume meter;
Step 3: will obtain photooxidation Inner Mongol jet coal (GICY) coal dust in step 2 is added to that be inoculated with yellow archespore hair flat
In the fluid nutrient medium of lead fungi (Phanerochaete chrysosporium), it is placed in incubator and degrades, then to drop
Solution product be filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance be measured, obtain Y30=2.779, carry it into the degradation rate of Phanerochaete chrysosporium degradation photooxidation low-order coal
With degradation solution absorbance relation equation η30=0.02336+0.08945Y30In, the goodness of fit determines that coefficient is R30 2=
0.97836, obtain η30=0.2719, i.e. η30It is 27.19%;Phanerochaete chrysosporium single bacterium degradation photooxidation Inner Mongol is long
The condition of flame coal (GICY) are as follows: coal amount is with photooxidation Inner Mongol jet coal in unit volume Phanerochaete chrysosporium fluid nutrient medium
(GICY) quality of pc meter 13.00g/L, Phanerochaete chrysosporium mother's bacterium solution inoculation in Phanerochaete chrysosporium fluid nutrient medium
Measure 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Phanerochaete chrysosporium is female
Spore concentration in bacterium solution is 2.4 × 105A/mL.
Embodiment 4 is compared with comparative example 10~12 it is found that photooxidation Inner Mongol jet coal (GICY) is micro- in embodiment 4
The total degradation rate η of biology classification degradationAlways=η1+η2+η3=0.6351, i.e. ηAlways=63.51%, and green spore in comparative example 10~12
Streptomycete, pseudomonas putida and Phanerochaete chrysosporium are respectively η to the degradation rate of photooxidation Inner Mongol jet coal (GICY)10=
13.25%, η20=15.73%, η30=27.19%, i.e., successively using above-mentioned three kinds of bacterium classification degradation photooxidation Inner Mongol jet coal
(GICY) total degradation rate, individually to the degradation rate of photooxidation Inner Mongol jet coal (GICY), illustrates of the invention much larger than three kinds of bacterium
Classification biodegrading process improves the microbial degradation rate of Inner Mongol jet coal (ICY).
Embodiment 5
The present embodiment the following steps are included:
Step 1: then successively being obtained through grinding and screening after refreshing mansion dross coal (SFBN) is crushed in 100 DEG C of drying 1h
Refreshing mansion dross coal (SFBN) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to dross coal (SFBN) coal dust in mind mansion obtained in step 1
Photooxidation pretreatment, obtains photooxidation mind mansion dross coal (GSFBN);The pretreated condition of photooxidation are as follows: coal amount is with list
Refreshing mansion dross coal (SFBN) the quality of pc meter 20g/L being added in the volume rotation bed photochemical reactor of position, ultraviolet ray intensity
150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, to rotate bed photochemical before photooxidation pretreatment
The logical oxygen time 10min of every liter of reactor volume meter;
Green spore chain is inoculated with Step 3: photooxidation mind mansion dross coal (GSFBN) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, then
Level-one catabolite is filtered, level-one degradation solution and level-one is collected respectively and degrades coal residue, then level-one is degraded coal residue
It is placed in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, level-one is dropped using spectrophotometry
Absorbance of the liquid at 450nm is solved to be measured to obtain Y1=1.193, carry it into green spore streptomycete degradation photooxidation low-order coal
Level-one degradation rate and level-one degradation solution absorbance relation equation η1=0.02466+0.07453Y1In, the goodness of fit determines system
Number is R1 2=0.98392, obtain η1=0.1136, i.e. η1It is 11.36%;The condition of the level-one degradation are as follows: unit volume connects
Kind has the additional amount 9.50g/L of photooxidation mind mansion dross coal (GSFBN) coal dust in the fluid nutrient medium of green spore streptomycete, green spore chain
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min in the fluid nutrient medium of mould, culture
Time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.1 × 105A/mL;
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=2.263, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.1743, i.e. η2It is 17.43%;The condition of the second level degradation are as follows: it is false that unit volume is inoculated with foul smelling
In the fluid nutrient medium of monad it is sterilized after level-one degradation coal residue additional amount in level-one degradation process photooxidation mind
Mansion dross coal (GSFBN) quality of pc meter 13.00g/L, the Pseudomonas putida starter in the fluid nutrient medium of pseudomonas putida
Bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false
Viable bacteria concentration in unit cell starter bacterium solution is 8.5 × 105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
It in the fluid nutrient medium of (Phanerochaete chrysosporium), is placed in incubator and carries out multi-stage degradation, then carry out
The supernatant for filtering and collecting multi-stage degradation liquid, using deionized water as blank control, using spectrophotometry to multi-stage degradation liquid
Absorbance at 450nm is measured, and obtains Y3=2.963, carry it into Phanerochaete chrysosporium degradation photooxidation low order
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of coal3=0.02336+0.08945Y3In, the goodness of fit determines
Coefficient is R3 2=0.97836, obtain η3=0.2884, i.e. η3It is 28.84%;The condition of the multi-stage degradation are as follows: unit volume
Be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation coal residue additional amount degraded with level-one
Photooxidation mind mansion dross coal (GSFBN) quality of pc meter 13.00g/L in the process, in the fluid nutrient medium of Phanerochaete chrysosporium
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, culture temperature
30 DEG C of degree;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.3 × 105A/mL.
Comparative example 13
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after refreshing mansion dross coal (SFBN) is crushed in 100 DEG C of drying 1h
The coal dust for being -0.15mm+0.075mm to granularity;
Step 2: carrying out photooxidation pretreatment to coal dust obtained in step 1 using rotation bed photochemical reactor, obtain
To photooxidation mind mansion dross coal (GSFBN) coal dust;The pretreated condition of photooxidation are as follows: coal amount is with unit volume revolving bed
Refreshing mansion dross coal (SFBN) quality of pc the meter 20g/L, ultraviolet ray intensity 150W, revolving speed 120r/ being added in photochemical reactor
Min, oxidization time 42h, oxygen flow 800mL/min are every to rotate bed photochemical reactor volumeter before photooxidation pretreatment
The logical oxygen time 10min risen;
Green spore chain is inoculated with Step 3: photooxidation mind mansion dross coal (GSFBN) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in incubator and degrades, then to drop
Solution product be filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance be measured to obtain Y10=1.193, the degradation rate and degradation solution for carrying it into green spore streptomycete degradation low-order coal are inhaled
Photometric relationship equation η10=0.02466+0.07453Y10In, the goodness of fit determines that coefficient is R10 2=0.98392, obtain η10
=0.1136, i.e. η10It is 11.36%;The condition of green spore streptomycete single bacterium degradation photooxidation mind mansion dross coal (GSFBN) are as follows:
Coal amount 9.50g/ in terms of photooxidation mind mansion dross coal (GSFBN) quality of pc in the green spore streptomycete fluid nutrient medium of unit volume
L, green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/ in green spore streptomycete fluid nutrient medium
Min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.1 × 105A/mL.
Comparative example 14
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after refreshing mansion dross coal (SFBN) is crushed in 100 DEG C of drying 1h
Refreshing mansion dross coal (SFBN) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to dross coal (SFBN) coal dust in mind mansion obtained in step 1
Photooxidation pretreatment, obtains photooxidation mind mansion dross coal (GSFBN) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of refreshing mansion dross coal (SFBN) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Step 3: it is false that photooxidation mind mansion dross coal (GSFBN) coal dust obtained in step 2 is added to inoculation foul smelling
In the fluid nutrient medium of monad (Pseudomonas putida), it is placed in incubator and degrades, then to catabolite
It is filtered to obtain degradation solution, the extinction using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Degree is measured, and obtains Y20=2.603, carry it into the degradation rate and degradation solution of pseudomonas putida degradation photooxidation low-order coal
Absorbance relation equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, it obtains
η20=0.1961, i.e. η20It is 19.61%;The item of pseudomonas putida single bacterium degradation photooxidation mind mansion dross coal (GSFBN)
Part are as follows: coal amount is in terms of photooxidation mind mansion dross coal (SFBN) quality of pc in unit volume pseudomonas putida fluid nutrient medium
13.00g/L, the Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L in pseudomonas putida fluid nutrient medium, incubator oscillation
Frequency 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution is 8.5
×105A/mL.
Comparative example 15
Step 1: then successively being obtained through grinding and screening after refreshing mansion dross coal (SFBN) is crushed in 100 DEG C of drying 1h
Refreshing mansion dross coal (SFBN) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to dross coal (SFBN) coal dust in mind mansion obtained in step 1
Photooxidation pretreatment, obtains photooxidation mind mansion dross coal (GSFBN) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of refreshing mansion dross coal (SFBN) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Yellow archespore hair is inoculated with Step 3: photooxidation mind mansion dross coal (GSFBN) coal dust will be obtained in step 2 and be added to
In the fluid nutrient medium of flat lead fungi (Phanerochaete chrysosporium), it is placed in incubator and degrades, it is then right
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution in 450nm
The absorbance at place is measured, and obtains Y30=3.109, carry it into the degradation of Phanerochaete chrysosporium degradation photooxidation low-order coal
Rate and degradation solution absorbance relation equation η3=0.02336+0.08945Y3In, the goodness of fit determines that coefficient is R30 2=
0.97836, obtain η30=0.3015, i.e. η30It is 30.15%;The Phanerochaete chrysosporium degradation photooxidation mind mansion dross coal
(GSFBN) condition are as follows: coal amount is with photooxidation mind mansion dross coal in unit volume Phanerochaete chrysosporium fluid nutrient medium
(GSFBN) quality of pc meter 13.00g/L, Phanerochaete chrysosporium mother's bacterium solution in Phanerochaete chrysosporium fluid nutrient medium connect
Kind of amount 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Phanerochaete chrysosporium
Spore concentration in female bacterium solution is 2.3 × 105A/mL.
Embodiment 5 and comparative example 13~15 are compared it is found that photooxidation mind mansion dross coal (GSFBN) in embodiment 5
The total degradation rate η of microorganism classification degradationAlways=η1+η2+η3=0.5763, i.e. ηAlways=57.63%, and it is green in comparative example 13~15
Spore streptomycete, pseudomonas putida and Phanerochaete chrysosporium are respectively to the degradation rate of photooxidation mind mansion dross coal (GSFBN)
η10=11.36%, η20=19.16%, η30=30.15%, i.e., photooxidation mind mansion is successively degraded not using the classification of above-mentioned three kinds of bacterium
The total degradation rate of viscous coal (GSFBN), individually to the degradation rate of photooxidation mind mansion dross coal (GSFBN), illustrates this much larger than three kinds of bacterium
The classification biodegrading process of invention improves the microbial degradation rate of refreshing mansion dross coal (SFBN).
Embodiment 6
The present embodiment the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shaanxi jet coal (SXCY) is crushed in 100 DEG C of drying 1h
Shaanxi jet coal (SXCY) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (SXCY) coal dust in Shaanxi obtained in step 1
Photooxidation pretreatment, obtains photooxidation Shaanxi jet coal (GSXCY) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of Shaanxi jet coal (GSXCY) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Green spore chain is inoculated with Step 3: photooxidation Shaanxi jet coal (GSXCY) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in progress level-one degradation in incubator, then
Level-one catabolite is filtered, level-one degradation solution and level-one is collected respectively and degrades coal residue, then level-one is degraded coal residue
It is placed in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, level-one is dropped using spectrophotometry
Absorbance of the liquid at 450nm is solved to be measured to obtain Y1=1.624, carry it into green spore streptomycete degradation photooxidation low-order coal
Level-one degradation rate and level-one degradation solution absorbance relation equation η1=0.02466+0.07453Y1In, the goodness of fit determines system
Number is R1 2=0.98392, obtain η1=0.1457, i.e. η1It is 14.57%;The condition of the level-one degradation are as follows: unit volume connects
Kind has the additional amount 9.50g/L of photooxidation Shaanxi jet coal (GSXCY) coal dust in the fluid nutrient medium of green spore streptomycete, green spore chain
Green spore strepto- starter bacterium solution inoculum concentration 180mL/L, incubator frequency of oscillation 160r/min in the fluid nutrient medium of mould, culture
Time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.4 × 105A/mL;
Step 4: the level-one coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is placed in high pressure sterilization
Pot in 121 DEG C of sterilizing 20min, using deionized water as blank control, using spectrophotometry to second level degradation solution at 450nm
Absorbance be measured, obtain Y2=2.452, carry it into the second level degradation of pseudomonas putida degradation photooxidation low-order coal
Rate and second level degradation solution absorbance relation equation η2=0.02919+0.06412Y2In, the goodness of fit determines that coefficient is R2 2=
0.99075, obtain η2=0.1864, i.e. η2It is 18.64%;The condition of the second level degradation are as follows: it is false that unit volume is inoculated with foul smelling
In the fluid nutrient medium of monad it is sterilized after level-one degradation coal residue additional amount with photooxidation Shan in level-one degradation process
Western jet coal (GSXCY) quality of pc meter 13.00g/L, the Pseudomonas putida starter in the fluid nutrient medium of pseudomonas putida
Bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;The stench is false
Viable bacteria concentration in unit cell starter bacterium solution is 8.2 × 105A/mL;
Step 5: the second level coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid respectively and multi-stage degradation coal residue is adopted using deionized water as blank control
Absorbance of the multi-stage degradation liquid at 450nm is measured with spectrophotometry, obtains Y3=3.177, carry it into yellow spore
The multi-stage degradation rate and multi-stage degradation liquid absorbance relation equation η of the flat lead fungi degradation photooxidation low-order coal of raw wool3=0.02336+
0.08945Y3In, the goodness of fit determines that coefficient is R3 2=0.97836, obtain η3=0.3075, i.e. η3It is 30.75%;It is described
The condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level degradation
The additional amount of coal residue 13.00g/L, yellow spore in terms of photooxidation Shaanxi jet coal (GSXCY) quality of pc in level-one degradation process
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L in the fluid nutrient medium of the flat lead fungi of raw wool, incubator frequency of oscillation
210r/min, incubation time 14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is 2.1 ×
105A/mL.
Comparative example 16
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shaanxi jet coal (SXCY) is crushed in 100 DEG C of drying 1h
Shaanxi jet coal (SXCY) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (SXCY) coal dust in Shaanxi obtained in step 1
Photooxidation pretreatment, obtains photooxidation Shaanxi jet coal (GSXCY) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of Shaanxi jet coal (SXCY) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Green spore chain is inoculated with Step 3: photooxidation Shaanxi jet coal (GSXCY) coal dust obtained in step 2 is added to
In the fluid nutrient medium of mould (Streptomyces viridosporous), it is placed in incubator and degrades, then to drop
Solution product be filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Absorbance be measured to obtain Y10=1.624, carry it into the degradation rate and drop of green spore streptomycete degradation photooxidation low-order coal
Solve liquid absorbance relation equation η10=0.02466+0.07453Y10In, the goodness of fit determines that coefficient is R10 2=0.98392,
Obtain η10=0.1457, i.e. η10It is 14.57%;Green spore streptomycete single bacterium degradation photooxidation Shaanxi jet coal (GSXCY)
Condition are as follows: coal amount is with photooxidation Shaanxi jet coal (GSXCY) quality of pc in the green spore streptomycete fluid nutrient medium of unit volume
Count 9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration 180mL/L in green spore streptomycete fluid nutrient medium, incubator oscillation frequency
Rate 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is 3.4 ×
105A/mL.
Comparative example 17
This comparative example the following steps are included:
Step 1: then successively being obtained through grinding and screening after Shaanxi jet coal (SXCY) is crushed in 100 DEG C of drying 1h
Shaanxi jet coal (SXCY) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (SXCY) coal dust in Shaanxi obtained in step 1
Photooxidation pretreatment, obtains photooxidation Shaanxi jet coal (GSXCY) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of Shaanxi jet coal (GSXCY) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Step 3: it is false that photooxidation Shaanxi jet coal (GSXCY) coal dust obtained in step 2 is added to inoculation foul smelling
In the fluid nutrient medium of monad (Pseudomonas putida), it is placed in incubator and degrades, then to catabolite
It is filtered to obtain degradation solution, the extinction using deionized water as blank control, using spectrophotometry to degradation solution at 450nm
Degree is measured, and obtains Y20=2.263, carry it into the degradation rate and degradation solution absorbance of pseudomonas putida degradation low-order coal
Relation equation η20=0.02919+0.06412Y20In, the goodness of fit determines that coefficient is R20 2=0.99075, obtain η20=
0.1743, i.e. η20It is 17.43%;The condition of pseudomonas putida single bacterium degradation photooxidation Shaanxi jet coal (GSXCY) are as follows:
Coal amount is in terms of photooxidation Shaanxi jet coal (GSXCY) quality of pc in unit volume pseudomonas putida fluid nutrient medium
13.00g/L, the Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L in pseudomonas putida fluid nutrient medium, incubator oscillation
Frequency 160r/min, incubation time 12d, 30 DEG C of cultivation temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution is 8.2
×105A/mL.
Comparative example 18
Step 1: then successively being obtained through grinding and screening after Shaanxi jet coal (SXCY) is crushed in 100 DEG C of drying 1h
Shaanxi jet coal (SXCY) coal dust for being -0.15mm+0.075mm to granularity;
Step 2: being carried out using rotation bed photochemical reactor to jet coal (SXCY) coal dust in Shaanxi obtained in step 1
Photooxidation pretreatment, obtains photooxidation Shaanxi jet coal (GSXCY) coal dust;The pretreated condition of photooxidation are as follows: coal amount
The 20g/L in terms of Shaanxi jet coal (GSXCY) quality of pc being added in unit volume rotation bed photochemical reactor, ultraviolet light intensity
150W, revolving speed 120r/min, oxidization time 42h are spent, oxygen flow 800mL/min is photochemical with revolving bed before photooxidation pretreatment
Learn the logical oxygen time 10min of every liter of reactor volume meter;
Yellow archespore hair is inoculated with Step 3: photooxidation Shaanxi jet coal (GSXCY) coal dust will be obtained in step 2 and be added to
In the fluid nutrient medium of flat lead fungi (Phanerochaete chrysosporium), it is placed in incubator and degrades, it is then right
Catabolite is filtered to obtain degradation solution, using deionized water as blank control, using spectrophotometry to degradation solution in 450nm
The absorbance at place is measured, and obtains Y30=2.974, carry it into the degradation of Phanerochaete chrysosporium degradation photooxidation low-order coal
Rate and degradation solution absorbance relation equation η30=0.02336+0.08945Y30In, the goodness of fit determines that coefficient is R30 2=
0.97836, obtain η30=0.2894, i.e. η30It is 28.94%;Phanerochaete chrysosporium single bacterium photooxidation Shaanxi jet coal
(GSXCY) condition degraded are as follows: coal amount is with photooxidation Shaanxi long flame in unit volume Phanerochaete chrysosporium fluid nutrient medium
Coal (GSXCY) quality of pc meter 13.00g/L, Phanerochaete chrysosporium mother's bacterium solution in Phanerochaete chrysosporium fluid nutrient medium
Inoculum concentration 90mL/L, incubator frequency of oscillation 210r/min, incubation time 14d, 30 DEG C of cultivation temperature;The Huang archespore Mao Pingge
Spore concentration in starter bacterium solution is 2.1 × 105A/mL.
Embodiment 6 and comparative example 16~18 are compared it is found that photooxidation Shaanxi jet coal (GSXCY) in embodiment 6
The total degradation rate η of microorganism classification degradationAlways=η1+η2+η3=0.6396, i.e. ηAlways=63.96%, and it is green in comparative example 16~18
Spore streptomycete, pseudomonas putida and Phanerochaete chrysosporium are directly to the degradation rate of photooxidation Shaanxi jet coal (GSXCY) point
It Wei not η10=14.57%, η20=17.43%, η30=28.94%, i.e., successively using above-mentioned three kinds of bacterium classification degradation photooxidation Shan
The total degradation rate of western jet coal (GSXCY), individually to the degradation rate of photooxidation Shaanxi jet coal (GSXCY), is said much larger than three kinds of bacterium
Bright classification biodegrading process of the invention improves the microbial degradation rate of Shaanxi jet coal (SXCY).
The above is only preferable profile embodiment of the invention, not does any restrictions to the present invention, all
According to inventive technique any simple modification substantially to the above embodiments, change and equivalent structural changes, this is still fallen within
In the protection scope of inventive technique scheme.
Claims (9)
1. a kind of method of low-order coal microorganism classification degradation, which is characterized in that method includes the following steps:
Step 1: drying after low-order coal is crushed, then successively through grinding and screening, obtaining granularity is -0.15mm+0.075mm
Coal dust;
Step 2: carrying out photooxidation pretreatment to coal dust obtained in step 1 using rotation bed photochemical reactor, light is obtained
Aoxidize coal dust;
Green spore streptomycete (Streptomyces is inoculated with Step 3: photooxidation powdered coal obtained in step 2 is added to
Viridosporous in fluid nutrient medium), it is placed in progress level-one degradation in incubator, then level-one catabolite is carried out
Filtering collects level-one degradation solution and level-one degradation coal residue respectively, then level-one degradation coal residue is sterilized;
Step 4: the level-one degradation coal residue after will be sterilized in step 3 is added to and is inoculated with pseudomonas putida
In the fluid nutrient medium of (Pseudomonas putida), it is placed in progress second level degradation in incubator, then degrades and produces to second level
Object is filtered, and collects second level degradation solution and second level degradation coal residue respectively, then second level degradation coal residue is sterilized;
Step 5: the second level degradation coal residue after will be sterilized in step 4 is added to and is inoculated with Phanerochaete chrysosporium
In the fluid nutrient medium of (Phanerochaete chrysosporium), it is placed in incubator and carries out multi-stage degradation, then to three
Grade catabolite is filtered, and collects multi-stage degradation liquid and multi-stage degradation coal residue respectively.
2. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that institute in step 2
State the pretreated condition of photooxidation are as follows: coal amount is to rotate in terms of the quality of pc that unit volume is added in bed photochemical reactor
20g/L, ultraviolet ray intensity 150W, revolving speed 120r/min, oxidization time 42h, oxygen flow 800mL/min, photooxidation pretreatment
The preceding logical oxygen time 10min to rotate every liter of bed photochemical reactor volumeter.
3. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that institute in step 3
State the condition of level-one degradation are as follows: unit volume is inoculated with the additional amount of photooxidation powdered coal in the fluid nutrient medium of green spore streptomycete
9.50g/L, the green spore strepto- starter bacterium solution inoculum concentration 180mL/L in the fluid nutrient medium of green spore streptomycete, incubator oscillation frequency
Rate 160r/min, incubation time 10d, 28 DEG C of cultivation temperature;Viable bacteria concentration in the green spore strepto- starter bacterium solution is not less than 3.0
×105A/mL.
4. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that institute in step 4
State second level degradation condition are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of pseudomonas putida after level-one degradation
The additional amount of coal residue 13.00g/L in terms of photooxidation quality of pc in level-one degradation process, the Liquid Culture of pseudomonas putida
Pseudomonas putida starter bacterium solution inoculum concentration 135mL/L, incubator frequency of oscillation 160r/min, incubation time 12d in base, culture
30 DEG C of temperature;Viable bacteria concentration in the Pseudomonas putida starter bacterium solution is not less than 8.0 × 105A/mL.
5. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that institute in step 5
State the condition of multi-stage degradation are as follows: unit volume be inoculated with it is sterilized in the fluid nutrient medium of Phanerochaete chrysosporium after second level drop
Solve additional amount 13.00g/L in terms of photooxidation quality of pc in level-one degradation process of coal residue, the liquid of Phanerochaete chrysosporium
Phanerochaete chrysosporium mother bacterium solution inoculum concentration 90mL/L in culture medium, incubator frequency of oscillation 210r/min, incubation time
14d, 30 DEG C of cultivation temperature;Spore concentration in the Phanerochaete chrysosporium mother bacterium solution is not less than 2.0 × 105A/mL.
6. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that using light splitting light
Degree method is measured absorbance of the level-one degradation solution at 450nm obtained in step 3, then brings green spore streptomycete drop into
It solves in the level-one degradation rate and level-one degradation solution absorbance relation equation of photooxidation low-order coal, level-one degradation rate is calculated;Institute
State the level-one degradation rate and level-one degradation solution absorbance relation equation of green spore streptomycete degradation photooxidation low-order coal are as follows: η1=
0.02466+0.07453Y1, the goodness of fit determine coefficient be R1 2=0.98392, wherein η1For level-one degradation rate, Y1For level-one
Degradation solution absorbance.
7. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that using light splitting light
Degree method is measured absorbance of the second level degradation solution obtained in step 4 at 450nm, then brings pseudomonas putida into
In the second level degradation rate and second level degradation solution absorbance relation equation of photooxidation low-order coal of degrading, second level degradation rate is calculated;
The second level degradation rate and second level degradation solution absorbance relation equation of the pseudomonas putida degradation photooxidation low-order coal are as follows: η2=
0.02919+0.06412Y2, the goodness of fit determine coefficient be R2 2=0.99075, wherein η2For second level degradation rate, Y2For second level
Degradation solution absorbance.
8. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that using light splitting light
Degree method is measured absorbance of the multi-stage degradation liquid at 450nm obtained in step 5, then brings yellow archespore Mao Pingge into
Bacterium is degraded in the multi-stage degradation rate of photooxidation low-order coal and the relation equation of multi-stage degradation liquid absorbance, and multi-stage degradation is calculated
Rate;The multi-stage degradation rate of the Phanerochaete chrysosporium degradation photooxidation low-order coal and the relation equation of multi-stage degradation liquid absorbance
Are as follows: η3=0.02336+0.08945Y3, the goodness of fit determine coefficient be R3 2=0.97836, wherein η3For multi-stage degradation rate, Y3
For multi-stage degradation liquid absorbance.
9. a kind of method of low-order coal microorganism classification degradation according to claim 1, which is characterized in that step 3 and step
Sterilising conditions in rapid four are equal are as follows: 121 DEG C of sterilizing 20min in high-pressure sterilizing pot.
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