CN104928207B - Huo Shi enterobacteria CL2013 and the method for preparing Cr VI remediation microbial inoculum - Google Patents
Huo Shi enterobacteria CL2013 and the method for preparing Cr VI remediation microbial inoculum Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
Abstract
For one plant of Huo Shi enterobacteria (Enterobacter hormaechei) CL.2013 bacterial strain that separation screening comes out from chromium slag storage yard soil in preservation on January 27 in 2015, deposit number was CGMCC No.10451.The bacterial strain is cultivated on LB culture medium, and cell shape is rod-shaped, Gram-negative；Culture 3 days, colony colour be it is milky white, colonial morphology be it is round, smooth, determine that the bacterial strain is Huo Shi enterobacteria (Enterobacter hormaechei) by biochemical reactions and molecular biology identification.Huo Shi enterobacteria CL.2013 bacterial strain has powerful reduction to Cr VI.The invention also discloses the methods for preparing Cr VI remediation microbial inoculum with Huo Shi enterobacteria CL.2013 bacterial strain and marigold processing-waste and waste residue, and method is simple, low in cost, easy to use without secondary pollution, have a vast market foreground.
Description
Technical field
The present invention relates to microorganism and application fields, have the Huo Shi intestines of reduction more particularly to a kind of pair of Cr VI
It is prepared by the preparation of bacillus and the Huo Shi enterobacteria fermentation liquid and the utilization Huo Shi enterobacteria and marigold processing-waste and waste residue
The method of Cr VI remediation microbial inoculum.
Background technique
Chromium (VI) be it is second largest in underground water, soil and bed mud endanger heavy metal, it is single in U.S. EPA priority pollutant name
Name the 6th, is 117 kinds of one of priority pollutants done great harm to human body, has acute, subacute, chronic toxicity to human body, have
Carcinogenesis, aberration inducing, mutagenesis.Due to chromate ion (CrO4 2-) and sulfate ion (SO4 2-) structure it is similar
Property, chromate ion are easy to enter biomembrane by sulfate ion channel, and further with intracytoplasmic Cucumber
Redox reaction occurs, the active oxygen generated therewith is very big to the injury of cell.Due to high water solubility, the poison of Cr VI
Property is significantly larger than trivalent chromium, about the 100 of trivalent chromium toxicity times.Cr VI into human body cell can also cause DNA break, carefully
Born of the same parents' chromosome is distorted.Chromium (VI) in chromium slag is unstable, enters environment after being soluble in acid or water, pollutes.According to
" China Environmental State Bulletin in 2011 " of Chinese Ministry of Environmental Protection's in June, 2012 publication, by the end of the year 2011, the accumulative disposition chromium slag in the whole nation is super
Cross 4,000,000 tons, but this does not imply that left over by history chromium slag has been administered and has been finished, and under slag muck soil pollutant in rainwash
It is migrated with being also occurred that under Biogeochemistry, causes greater area of soil/contaminated sites.With these harmful substances
A large amount of discharges, mineral products and Soil Surrounding environment are threatened agricultural production, grain security and people by different degrees of combined pollution
Body health.
Chromium mainly exists in the form of Cr (III) and Cr (VI) in the environment, and wherein Cr (III) solubility is small, mobility
It is weak, small toxicity, and Cr (VI) is soluble easily in water, oxidisability is strong, and toxicity is big, is approximately 100 times of Cr (III).Therefore, Cr (VI) is turned
Become Cr (III), be not only a kind of effective removing toxic substances mode, and is also the committed step for removing chromium from environment eventually.
There are immobilization/stabilisation, electronation, chemical leaching, electricity for hexavalent chromium polluted traditional administering method at present
Dynamic reparation and phytoremediation etc., but big multipair site requirements is higher, and limitation is more, and biologic treating technique and conventional processing technique
Compare, have the advantages that quickly, safety, expense it is low etc., therefore be known as and a kind of emerging environmental-friendly replace technology.From sexavalence
Separate microorganism in pollution of chromium medium, and be targetedly applied to during the repairing and treating of contaminated soil and water body,
It is a kind of ideal restorative procedure.
Marigold is rich in natural carotenol, is the current primary raw material for extracting natural carotenol.Natural carotenol is as one
The important natural pigment of kind, the increasingly favor by the domestic enterprises such as food, medicine outward, but bring environment is dirty therewith
Dye problem also becomes the restraining factors of marigold industry development, and predominant emissions are fresh marigold flowers by micro- in production process
Acid waste liquid after biofermentation and the discarded colored slag for having extracted lutein.Wherein contain a large amount of lactic acid, amino acid, humic acid
Equal organic principles, also containing ingredients such as a large amount of lignin, cellulose, microelements, if arbitrarily discharge is not only polluted by it
Environment, and waste valuable resource.As the cultivation of marigold and production scale expand, sewage resource is squeezed to marigold
Rationally processing has caused the concern of society gradually.Chinese patent 201410283919.6 discloses a kind of useless using fresh marigold flower
The method that water prepares organic liquid fertilizer.This method mainly passes through potassium hydroxide and is adjusted to neutrality acid waste liquid, after neutralization
Neutralizer be stored in fermentation vat, under the conditions of 4 DEG C to 37 DEG C, fermentation time is no less than 1 month temperature, be made organic liquor
Body fertilizer.Chinese patent 201410225056.7 discloses a kind of integrated treatment side of the squeezing sewage of marigold extraction lutein
Method.However, marigold squeezing sewage acidity is high, the content of organic matter is big, there is that energy consumption is high, difficult up to standard etc. asks during processing
Topic.Therefore, the exploitation of the application technology as the second resource of marigold processing waste water and waste residue is very urgent.
Summary of the invention
The purpose of the present invention is specific aim the shortcomings of the prior art, provide a kind of using marigold processing waste water and useless
The method of slag production Cr VI microorganism remediation microbial inoculum.
Technical scheme is as follows:
One plant of Huo Shi enterobacteria (Enterobacter hormaechei) CL.2013 bacterial strain was protected on January 27th, 2015
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.10451, address: north
No. 3 Institute of Microorganism, Academia Sinica of institute of the Chaoyang District Jing Shi North Star West Road 1.
The Huo Shi enterobacteria CL.2013 is that separation screening comes out from chromium slag storage yard soil.The bacterial strain is trained in LB
It supports and is cultivated on base, cell shape is rod-shaped, Gram-negative；Culture 3 days, colony colour be it is milky white, colonial morphology be circle
Shape, smooth (see figure 1).It is identified through Physiology and biochemistry, feature is shown in Table 1；
Using bacterial 16 S rDNA universal primer, Huo Shi enterobacteria CL.2013 bacterial strain bacterial strain 16S is expanded with the method for PCR
RDNA Partial Fragment, the sequencing of PCR product obtain the fragment sequence, the 16S rDNA (length of sequencing gained CL.2013 bacterial strain
1405bp) result is as follows:
CCGAAGGTTAAGCTACCTACTTCTTTTGCAACCCACTCCCATGGTGTGACGGGCGGTGTGTACAAGGCC
CGGGAACGTATTCACCGTGGCATTCTGATCCACGATTACTAGCGATTCCGACTTCATGGAGTCGAGTTGCAGACTCC
AATCCGGACTACGACGCACTTTATGAGGTCCGCTTGCTCTCGCGAGGTCGCTTCTCTTTGTATGCGCCATTGTAGCA
CGTGTGTAGCCCTACTCGTAAGGGCCATGATGACTTGACGTCATCCCCACCTTCCTCCAGTTTATCACTGGCAGTCT
CCTTTGAGTTCCCGGCCTAACCGCTGGCAACAAAGGATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATTTCA
CAACACGAGCTGACGACAGCCATGCAGCACCTGTCTCAGAGTTCCCGAAGGCACCAAAGCATCTCTGCTAAGTTCTC
TGGATGTCAAGAGTAGGTAAGGTTCTTCGCGTTGCATCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCG
TCAATTCATTTGAGTTTTAACCTTGCGGCCGTACTCCCCAGGCGGTCGACTTAACGCGTTAGCTCCGGAAGCCACGC
CTCAAGGGCACAACCTCCAAGTCGACATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACG
CTTTCGCACCTGAGCGTCAGTCTTTGTCCAGGGGGCCGCCTTCGCCACCGGTATTCCTCCAGATCTCTACGCATTTC
ACCGCTACACCTGGAATTCTACCCCCCTCTACAAGACTCTAGCCTGCCAGTTTCGAATGCAGTTCCCAGGTTGAGCC
CGGGGATTTCACATCCGACTTGACAGACCGCCTGCGTGCGCTTTACGCCCAGTAATTCCGATTAACGCTTGCACCCT
CCGTATTACCGCGGCTGCTGGCACGGAGTTAGCCGGTGCTTCTTCTGCGGGTAACGTCAATCGACAAGGTTATTAAC
CTTATCGCCTTCCTCCCCGCTGAAAGTACTTTACAACCCGAAGGCCTTCTTCATACACGCGGCATGGCTGCATCAGG
CTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTCTCAGTTCCAGTGTGGCTG
GTCATCCTCTCAGACCAGCTAGGGATCGTCGCCTAGGTGAGCCGTTACCCCACCTACTAGCTAATCCCATCTGGGCA
CATCCGATGGCAAGAGGCCCGAAGGTCCCCCTCTTTGGTCTTGCGACGTTATGCGGTATTAGCTACCGTTTCCAGTA
GTTATCCCCCTCCATCAGGCAGTTTCCCAGACATTACTCACCCGTCCGCCACTCGTCAGCAAAGCAGCAAGCTGCTT
GCCTGTTACCGTTCGTGCTGCATGGT
Knot frame application BLAST tool is sequenced in above-mentioned gained and carries out genetic homology and similarity analysis, it is homologous through Blast
Property retrieval, it is known that the 16S rDNA sequence of CL.2013 bacterial strain is same with Huo Shi enterobacteria (Enterobacter hormaechei)
Property highest in source has reached 99%-100% (being shown in Table 2).
It is analyzed according to BLAST as a result, the affiliated biological bacterium taxonomic information of CL.2013 bacterial strain is as follows:
domain Bacteria (0/20/659718)
phylum “Pmteobacteria” (0/20/183305)
class “Gammapwteobacteria” (0/20/83510)
order “Enterobacteriales” (0/20/18387)
family “Enterobacteriaceae”(0/20/18387)
genus “Enterobacter hormaechei”(0/20/1678)
Therefore, which is accredited as Huo Shi enterobacteria (Enterobacter hormaechei), and is named as Huo Shi
Enterobacteria (Enterobacter hormaechei) CL.2013.
Microorganism will receive the limitation of many environmental factors during removing chromium (VI).This is the life of microorganism itself
What long metabolic process was determined.Therefore, microorganisms reduction chromium (VI) process when, should emphatically investigate various environment because
Son removes the influence of chromium (VI) effect to it, is practical application of the microorganism in future in terms of except chromium (VI) and the technique except chromium (VI)
Certain basis is established in design.Studies have shown that Huo Shi enterobacteria (Enterobacter hormaechei) CL.2013 difference pH
Value, temperature, Cr (VI) initial concentration all have a certain impact to the ability of micro-reduction Cr (VI).Experiments have shown that bacterial strain
The optimal pH of CL.2013 is 8,30 DEG C of optimum temperature.800 mgL are up in Cr (VI) initial concentration-1When, CL.2013 bacterial strain
Also there is certain reduction to Cr (VI), compared with existing Cr (VI) reducing bacteria having found both at home and abroad, reducing power
Powerful is more.The research plays certain facilitation to the practical application of bioanalysis removal Cr (VI).This is to administer Cr from now on
(VI) high pollution soil provides a kind of efficient strain.
The method that present invention marigold processing-waste and waste residue prepare Cr VI remediation microbial inoculum, includes the following steps:
(1) preparation strain activation and culture base adds glucose 1g/L in LB culture medium, and sterilize 15-30min, for use；
(2) the Huo Shi enterobacteria (Enterobacter of the invention that activated seed will be deposited in slant tube is prepared
Hormaechei) in the culture medium that CL.2013 bacterial strain access (1) prepares, in 28-35 DEG C, 120-180r/min shaken cultivation
12-16h obtains activated seed；
(3) it prepares liquid seed culture fluid sodium hydrate aqueous solution and adjusts the processing waste water that marigold extracts lutein
PH to 6.5-7.5, in 115-121 DEG C of sterilizing 15-30min；
(4) activated seed of inoculation (2) preparation in the culture solution that liquid seeds take (3) to prepare, inoculum concentration 8%- are prepared
10%, in 30-37 DEG C, 120-200r/min shaken cultivation 24-48 h, obtain liquid seeds；
(5) preparing zymocyte liquid and being inoculated with volume ratio in culture solution fermentor prepared by (3) is 2%-15%(4) preparation
Liquid seeds, in 25-35 DEG C, lead to filtrated air culture 12-48 h, when viable count reaches 5 × 108When CFU/ ml, liquid is obtained
Zymocyte liquid；
(6) it prepares after marigold flower slag charcoal dries the marigold flower slag for having extracted lutein and is crushed to 20-100
Mesh is put into pipe reaction furnace, and before heating, it is 250cm that 20-30min gas velocity is led in furnace3The nitrogen of/min, makes reacting furnace
Interior is oxygen-free environment, and 300-600 DEG C is heated under conditions of 5 DEG C/min of heating rate, 3-6h is kept, is then cooled to room
Temperature obtains black bio carbon；
(7) it prepares Cr VI and repairs solid-state microbial inoculum using the biological carbon prepared as carrier, according to zymocyte liquid: biological carbon
For the mass ratio of 1 ︰ 1.2-5, (5) are prepared into zymocyte liquid and are added in the biological carbon of (7) preparation, is sufficiently mixed uniformly ventilation
It is dry to microbial inoculum water content 30% hereinafter, packaging, is stored in shady and cool dry place, obtain Cr VI reparation solid-state microbial inoculum.
Cr VI of the invention repairs the application method of solid-state microbial inoculum: it is different by hexavalent chromium polluted degree, it takes a certain amount of
Cr VI repair solid-state microbial inoculum mix, stir evenly with contaminated soil, holding soil moisture content 10 days between 50%-60%
It is above.
The test of the repairing hexavalent chromium polluted soil of solid-state microbial inoculum is repaired with Cr VI of the invention
1, test method
Uncontaminated soil is taken, is air-dried, grinds, cross 10 meshes.Potassium bichromate solution is added in soil after sieving, soil is made
The contaminated soil of earth hexavalent chromium polluted horizontal respectively 250mg/kg, 500mg/kg, 1000mg/kg.By each level of pollution
Soil weighs the soil of 3 100g respectively, repairs solid-state microbial inoculum with Cr VI prepared by the present invention and handles in each level of pollution
It is middle to take mass parts for 10%, 20%, 30% percentage respectively, renovation agent is mixed with soil, is stirred, soil moisture content control
It between 50%-60%, is placed in 30 DEG C of incubator and cultivates, after keeping 10d, measure the concentration of Cr VI in soil.
2, test result
Test result is shown in Table 3.
Experiments have shown that the additive amount of renovation agent is the 10% of contaminated soil when hexavalent chromium polluted level is 250mg/kg
When, content of 6-valence Cr ions can be down to 98.78%, and the additive amount effect for being further added by renovation agent is also not significantly different；Pollution of chromium water
When putting down as 500mg/kg, the optimum addition of renovation agent is the 30% of contaminated soil, and it is reachable that content of 6-valence Cr ions reduces amplitude maximum
74.20%；It is extremely significant to be higher than other two processing；When pollution of chromium level is 1000mg/kg, renovation agent optimum addition is
The 30% of contaminated soil, content of 6-valence Cr ions reduction amplitude are extremely significant to be higher than other two processing up to 58.78%.
Beneficial effects of the present invention
The fermentation liquid of Huo Shi enterobacteria (Enterobacter hormaechei) CL.2013 bacterial strain of the invention have compared with
The effect of strong reduction of hexavalent chromium, under aerobic, medium temperature and alkaline condition, which has stronger reduction to Cr (VI),
Relative to other chromium reducing bacterias for needing better nutritivity condition and adaptive capacity to environment difference, which will more save the cost and environment
It is adaptable.800 mgL are up in Cr (VI) initial concentration-1When, CL.2013 bacterial strain also has certain go back to Cr (VI)
Original work are used, with it is existing both at home and abroad it has been found that Cr (VI) reducing bacteria compared with, powerful more of reducing power.The research is to bioanalysis
The practical application of removal Cr (VI) plays certain facilitation.
The present invention utilizes the fermentation liquid of marigold processing-waste culture Huo Shi enterobacteria CL.2013 bacterial strain, and utilizes for ten thousand longevity
Chrysanthemum processing waste residue is prepared into marigold flower slag charcoal, then by the fermentation liquid and charcoal system of Huo Shi enterobacteria CL.2013 bacterial strain
Standby to repair solid-state microbial inoculum at Cr VI, preparing raw material is trade waste, and process does not have secondary pollution, belongs to environment friend
Good type technique.Cr VI of the invention repairs that solid-state microbial inoculum preparation process is simple, raw material use marigold processing waste liquid and
Waste residue, therefore cost is very cheap.Contain a large amount of carbons and some other nutrient in solid-state microbial inoculum of the invention, can be micro-
Biology and soil plant growth provide carbon source abundant, are conducive to Microbial biomass C L.2013 strain growth, and have infertile field effect.By
Experiments have shown that Cr VI of the invention, which repairs solid-state microbial inoculum, to be had repair time short (10 days), modification effect good (slight pollution is repaired
Again in 98% or more, serious pollution reparation 58% or more), as bioremediation agents, this has reached extraordinary effect.Therefore,
The present invention has a vast market foreground in repairing hexavalent chromium soil pollution.
Detailed description of the invention
Fig. 1 is Huo Shi enterobacteria CL.2013 colonial morphology of the present invention.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated.
Embodiment 1
One plant of Huo Shi enterobacteria (Enterobacter hormaechei) CL.2013 bacterial strain was protected on January 27th, 2015
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.10451, address: north
No. 3 Institute of Microorganism, Academia Sinica of institute of the Chaoyang District Jing Shi North Star West Road 1.
The Huo Shi enterobacteria CL.2013 is that separation screening comes out from chromium slag storage yard soil.The bacterial strain is trained in LB
It supports and is cultivated on base, cell shape is rod-shaped, Gram-negative；Culture 3 days, colony colour be it is milky white, colonial morphology be circle
It is shape, smooth.Through Physiology and biochemistry identification, 16S rDNA Phylogenetic Analysis, which is identified and is named as Huo Shi enterobacteria
(Enterobacter hormaechei) CL.2013.
Embodiment 2
Cr VI, which is prepared, using Huo Shi enterobacteria CL.2013 and marigold waste water, waste residue of the invention repairs solid-state microbial inoculum,
Its method includes:
(1) preparation strain activation and culture base adds glucose 1g/L in LB culture medium, and sterilize 15-30min, for use；
(2) the Huo Shi enterobacteria (Enterobacter of the invention that activated seed will be deposited in slant tube is prepared
Hormaechei) in the culture medium that CL.2013 bacterial strain access (1) prepares, in 28-35 DEG C, 120-180r/min shaken cultivation
12-16h obtains activated seed；
(3) the processing waste water sodium hydrate aqueous solution tune that marigold is extracted lutein by liquid seed culture fluid is prepared
PH to the 6.5-7.5 for saving waste water, in 115-121 DEG C of sterilizing 15-30min；
(4) activated seed of inoculation (2) preparation in the culture solution that liquid seeds take (3) to prepare, inoculum concentration 8%- are prepared
10%, in 30-37 DEG C, 120-200r/min shaken cultivation 24-48 h, obtain liquid seeds；
(5) preparing zymocyte liquid and being inoculated with volume ratio in culture solution fermentor prepared by (3) is 2%-15%(4) preparation
Liquid seeds, in 25-35 DEG C, lead to filtrated air culture 12-48 h, when viable count reaches 5 × 108When CFU/ ml, liquid is obtained
Zymocyte liquid；
(6) it prepares after marigold flower slag charcoal dries the marigold flower slag for having extracted lutein and is crushed to 20-100
Mesh is put into pipe reaction furnace, and before heating, it is 250cm that 20-30min gas velocity is led in furnace3The nitrogen of/min, makes reacting furnace
Interior is oxygen-free environment, and 300-600 DEG C is heated under conditions of 5 DEG C/min of heating rate, 3-6h is kept, is then cooled to room
Temperature obtains black bio carbon；
(7) it prepares Cr VI and repairs solid-state microbial inoculum using the biological carbon prepared as carrier, according to zymocyte liquid: biological carbon
For the mass ratio of 1 ︰ 1.2-5, (5) are prepared into zymocyte liquid and are added in the biological carbon of (7) preparation, is sufficiently mixed uniformly ventilation
It is dry to microbial inoculum water content 30% hereinafter, packaging, is stored in shady and cool dry place, obtain Cr VI reparation solid-state microbial inoculum.
Cr VI of the invention repairs the application method of solid-state microbial inoculum: it is different by hexavalent chromium polluted degree, it takes a certain amount of
Cr VI repair solid-state microbial inoculum and mix, stir evenly with contaminated soil, holding soil moisture content in 50%-60%, 10 days with
It is upper.
Claims (2)
1. a kind of method for preparing Cr VI remediation microbial inoculum with Huo Shi enterobacteria CL.2013 bacterial strain, the Huo Shi enterobacteria
(Enterobacter hormaechei) CL.2013 bacterial strain is preserved in Chinese microorganism strain preservation on January 27th, 2015
Administration committee's common micro-organisms center, deposit number are CGMCC No.10451, and the preparation method includes the following steps:
(1) preparation strain activation and culture base adds glucose 1g/L in LB culture medium, and sterilize 15-30min, for use；
(2) activated seed is prepared by the Huo Shi enterobacteria being deposited in slant tube (Enterobacter hormaechei)
In the culture medium that CL.2013 bacterial strain access (1) prepares, in 28-35 DEG C, 120-180r/min shaken cultivation 12-16h, obtain
Activated seed；
(3) it prepares liquid seed culture fluid sodium hydrate aqueous solution and adjusts the pH that marigold extracts the processing waste water of lutein
To 6.5-7.5, in 115-121 DEG C of sterilizing 15-30min；
(4) it prepares and is inoculated with activated seed prepared by (2) in the culture solution that liquid seeds take (3) to prepare, inoculum concentration 8%-10%,
In 30-37 DEG C, 120-200r/min shaken cultivation 24-48 h, liquid seeds are obtained；
(5) it prepares zymocyte liquid and is inoculated with volume ratio in culture solution prepared by (3) as the liquid strain of 2%-15%(4) preparation
Son leads to filtrated air culture 12-48 h, when viable count reaches 5 × 10 in 25-35 DEG C8When CFU/ ml, liquid fermentation bacterium is obtained
Liquid；
(6) it prepares after marigold flower slag charcoal dries the marigold flower slag for having extracted lutein and is crushed to 20-100 mesh,
It is put into pipe reaction furnace, before heating, it is 250cm that 20-30min gas velocity is led in furnace3The nitrogen of/min, make be in reacting furnace
Oxygen-free environment is heated to 300-600 DEG C under conditions of 5 DEG C/min of heating rate, keeps 3-6h, is then cooled to room temperature,
Obtain black bio carbon；
(7) prepare Cr VI and repair solid-state microbial inoculum using the biological carbon prepared as carrier, according to zymocyte liquid: biological carbon is 1 ︰
(5) are prepared zymocyte liquid and are added in the biological carbon of (7) preparation by the mass ratio of 1.2-5, are sufficiently mixed uniformly aeration-drying
Cr VI reparation solid-state microbial inoculum is obtained hereinafter, packaging, is stored in shady and cool dry place 30% to microbial inoculum water content.
2. a kind of application method of the Cr VI remediation microbial inoculum using the preparation of claim 1 the method, the method is by six
The degree of valence pollution of chromium is different, takes a certain amount of Cr VI to repair solid-state microbial inoculum and mixes, stirs evenly with contaminated soil, keeps
Soil moisture content was in 50%-60%, 10 days or more.
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| CN107629976A (en) * | 2016-07-18 | 2018-01-26 | 中国石油天然气集团公司 | A kind of microorganism and its microbial inoculum and application |
| CN109593667A (en) * | 2017-09-30 | 2019-04-09 | 天津科技大学 | The identification of Cr (VI) reducing bacteria and its cultural method |
| CN110331110B (en) * | 2019-07-26 | 2021-07-13 | 中南民族大学 | Method for efficiently reducing hexavalent chromium in water and recovering chromium by utilizing waste molasses and bacteria |
| CN113583896B (en) * | 2021-07-02 | 2024-04-02 | 王学芳 | Enterobacter cholerae and application thereof |
| CN114231520A (en) * | 2021-12-08 | 2022-03-25 | 上海康恒环境修复有限公司 | Carbon-based biological agent for efficiently converting heavy metal chromium in soil |
| CN115404179A (en) * | 2022-03-29 | 2022-11-29 | 湖南省蔬菜研究所 | Complex microbial inoculant containing achromobacter and enterobacter and application thereof |
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