CN109735476A - One plant of Degradation Formaldehyde bacterial strain and its application - Google Patents
One plant of Degradation Formaldehyde bacterial strain and its application Download PDFInfo
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- CN109735476A CN109735476A CN201910215095.1A CN201910215095A CN109735476A CN 109735476 A CN109735476 A CN 109735476A CN 201910215095 A CN201910215095 A CN 201910215095A CN 109735476 A CN109735476 A CN 109735476A
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- methylobacter
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
One plant of Methylobacter of present invention offer (Methylobacterium podarium) bacterium JQ-1, it is CGMCC No.17229 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.The bacterial strain has good tolerance to high-concentration formaldehyde, can be grown on and is with formaldehyde in the culture medium in unique source C, and the formaldehyde of 54.8mg/L all can be degraded in 120h.Invention also provides the culture medium of the bacterial strain and cultural methods.The present invention provides core strain excellent for effective degradation of formaldehyde, reduces the cost of degradation of formaldehyde, has saved resource, and without secondary pollution, and large scale fermentation is suitble to produce and be used for a long time.
Description
Technical field
The present invention relates to microbe fields, specially one plant of Degradation Formaldehyde bacterial strain and its application.
Background technique
Formaldehyde can make protein denaturation, have carcinogenic, teratogenesis, disabling effect, and the exceeded influence human body of formaldehyde in air is strong
Health, formaldehyde in indoor air has no the mode of being effectively treated at present.
Mainstream processing method has 2 kinds at present, the first: adsorption method.Absorption method must have corresponding adsorbent, such as recessed
Convex stick, active carbon etc..Wherein most commonly seen is active carbon adsorption, this method is also the common side of home interior formaldehyde removal
Method.Absorption method is divided into nature absorption again and forces absorption, wherein natural adsorption method refers to furnishes adsorbing medium indoors, passes through
The natural flowing of air is contacted with adsorbing medium, realizes the suction-operated of limited efficacy.This method almost to exceeded formaldehyde without effect,
Or its act on it is small can be ignored, main cause is that air flowing is limited, active carbon (or other adsorbing mediums, the following contents
In equal abbreviation active carbons) it is small to the exposure of air, contact area is small, so that the contact probability very little of formaldehyde and active carbon.Shallowly
Layer contact may be repelled by the zone of saturation of activated carbon surface, not can be carried out sufficient adsorption reaction.Therefore natural absorption method
Formaldehyde removal effect it is very limited.Passive adsorption, which refers to, enters the formaldehyde in air full of work
Property charcoal activated carbon chamber, adsorption reaction occur in activated carbon chamber, air more can completely be connect with active carbon here
Touching, active carbon filters the air, and adsorbs to organic molecules such as formaldehyde in air.The shortcomings that the method is activity
The suction-operated of charcoal can generate saturation, and the active carbon after saturation cannot not only continue formaldehyde adsorption, but also can be to releasing in air
Formaldehyde is put, pollution sources is formed, causes secondary pollution, need to regularly replace active carbon can just carry out, therefore become consumption to active carbon
Material, demand is very big, and active carbon is expensive, forms the very big wasting of resources.
Second: oxidation removal method.This method basic principle is method for oxidation, mainly includes catalysis oxidation, plasma oxygen
The methods of change, high-temp combustion oxidizing process, ozone oxidation.High-temp combustion oxidizing process: using electric energy or burning heat production, by all gas
Component indifference is heated to 800 DEG C or more, makes oxygen and formaldehyde in air that redox reaction occur, generates H2O and CO2.But
This method highly energy-consuming, equipment cost is high, has an impact to indoor air temperature, is not suitable for practical application substantially.Plasma oxidation
Method: using the principle of high voltage gas discharge, carrying out plasma for all constituent of air indifferences, the formaldehyde of plasma with
Redox reaction occurs for oxygen, generates H2O and CO2.This method also needs to expend a large amount of electric energy, there is high-voltage safety hidden danger,
The non-specific removal of PARA FORMALDEHYDE PRILLS(91,95), needs biggish occupied area and bulky equipment volume, is not suitable for practical application；It is catalyzed oxygen
Change removal method such as photocatalyst to aoxidize, ultraviolet light is needed to irradiate TiO2Coating, response area is limited, and oxidation efficiency is low, generates ultraviolet light
Pollution, practical value are little；Ozonation generates the ozone PARA FORMALDEHYDE PRILLS(91,95) gas molecule that generator produces using ozone and carries out
Oxidation, ozone generates generator itself because needing the extreme conditions such as high-voltage electricity or ultraviolet light, thus has some potential safety problems,
Secondly extra pernicious gas O can be generated3, secondary pollution is generated, almost without practical value.Both the above method is for first in air
The removal effect of aldehyde is undesirable.
Summary of the invention
The object of the present invention is to provide a kind of technical solutions overcome the shortage of prior art, that is, provide one plant of Formaldehyde degradation bacterium
Strain and its application.
One plant of Methylobacter bacterium that the object of the invention one is to provideMethylobacterium podarium JQ-1
(hereinafter referred to as JQ-1), deposit number of the JQ-1 bacterial strain in China Committee for Culture Collection of Microorganisms's common micro-organisms center
For CGMCC No. 17229.
JQ-1 is characterized in that the bacterial strain can be with degradation of formaldehyde.JQ-1 is taking formaldehyde as the known Ingredients Inorganic in unique source C
In culture medium, at 30 DEG C, in the case of cell concentration is OD value 0.107, the formaldehyde of 54.8mg/L can all be dropped in 120h
Solution.And the highest tolerable concentration of JQ-1 PARA FORMALDEHYDE PRILLS(91,95) are as follows: 120mg/L.
The purpose of the present invention two is to provide the culture medium and condition of culture of JQ-1.Culture medium: formaldehyde 50mg/L, NH4Cl
0.5g/L、Na2HPO4 3.0g/L、KH2PO4 1.5g/L、MgSO4·7H2O 0.1g/L、1M CaCl20.1g/L；Cultivate item
Part: 3-5% inoculation, 30 DEG C, 120rpm are protected from light culture.
The object of the invention three is to provide JQ-1 bacterial strain and the application in terms of methanal removing, specially provides using JQ-1 as core
The microorganism methanal removing product of heart microbial inoculum.
The present invention compared with the existing technology, has the following advantages that and effect: JQ-1 bacterial strain has high-concentration formaldehyde good
Tolerance, be able to carry out industrial wastewater containing high-concentration formaldehyde, industrial waste gas containing high-concentration formaldehyde processing；JQ-1 strains for degrading first
Aldehyde category biological metabolism method removes formaldehyde, will not generate secondary pollution；The determination of exclusive culture medium and condition of culture ensure that JQ-1 bacterium
Strain excellent proliferative and subculture, can large scale fermentation produce and be used for a long time, reduce costs；JQ-1 bacterial strain does not generate suction
Attached saturation effect, facilitates operation and maintenance.
Preservation explanation:
Strain name: Methylobacter bacterium
Latin name:Methylobacterium podarium
Strain number:Methylobacterium podarium JQ-1
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on January 24th, 2019
Collection is registered on the books number: CGMCC No. 17229
JQ-1 sequencing result explanation: sequence is 16S ribosomal RNA gene, sequence length 1417bp, sequence attachment
Shown in sequence table, ncbi database blast result: Methylobacterium podarium strain LM-Pink 16S
ribosomal RNA gene, partial sequence ident=99.9%。
Detailed description of the invention
Fig. 1 is the bacterium colony photo for the JQ-1 bacterial strain that the separation of embodiment 1 obtains.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material as used in the following examples, reagent etc., are commercially available unless otherwise specified.
Culture medium as used in the following examples is as follows:
Formaldehyde culture medium: formaldehyde 50mg/L, NH4Cl 0.5g/L、Na2HPO4 3.0g/L、KH2PO4 1.5g/L、MgSO4·7H2O
0.1g/L、1M CaCl2 0.1g/L。
The separation identification experiment of 1 JQ-1 of embodiment
Formaldehyde culture medium is prepared, agarose 20g/L, 120 DEG C of sterilizing 30min are added into culture medium, the cooling shape in culture dish
At plate.It is coated on culture plate after the activated sludge of the organic industrial sewage containing formaldehyde is diluted with sterile water, 30 DEG C are protected from light training
It supports.Culture is carried out scribing line to isolate and purify to obtain JQ-1.
The bacterial strain JQ-1 isolated and purified using formaldehyde culture medium culture above-mentioned steps, extracts the total DNA conduct of bacterial strain
Gene magnification template carries out PCR reaction with bacterium 16s rDNA universal primer on PCR amplification instrument.After the reaction was completed, it takes
2ulPCR product carries out 1% agarose gel electrophoresis detection.Confirm PCR amplified fragments.PCR product is solidifying with AxyPrep DNA
Plastic recovery kit recycling, concrete operations are carried out by kit specification.PCR product after taking each purifying agaric, uses survey
Sequence instrument ABI3730-XL carries out DNA sequencing.After carrying out tetraploid rice with the known array in NCBI GenBank, determine
Bacterium is divided into category or kind by bacterial species.
Pcr gene expands to obtain the 16s rDNA genetic fragment about 1.4kb of bacterial strain JQ-1, measure after sequence with NCBI data
Published 16s rDNA sequence carries out online tetraploid rice in library, as the result is shown JQ-1 withMethylobacterium podarium strainHomology highest, reach 99%.
The degradation experiment of 2 JQ-1 PARA FORMALDEHYDE PRILLS(91,95) of embodiment
Formaldehyde culture medium high-temperature sterilization is placed to room temperature, inoculating strain JQ-1, inoculum concentration 3-5%, 30 DEG C of cultivation temperature,
120rpm is protected from light culture.OD600 value and concentration of formaldehyde are measured in 0h, 48h, 120h respectively.The size of OD600 value can be indirect
Cell concentration is represented, is obtained with spectrophotometer method.1 group of blank control group is set simultaneously.The measurement of concentration of formaldehyde is adopted in solution
With People's Republic of China's state environment protecting standard: " the measurement acetylacetone,2,4-pentanedione spectrophotometry HJ 601- of water quality formaldehyde
2011 " method is measured.As a result as follows.
1. growing microorganism situation of table
The variation of 2. concentration of formaldehyde of table
This experiments have shown that: 1.JQ-1 bacterial strain to the formaldehyde in solution have degradation capability；2. cell concentration is OD value at 30 DEG C
In the case of 0.107, JQ-1 can all degrade the formaldehyde of 54.8mg/L in 120h.
The tolerance of 3 JQ-1 PARA FORMALDEHYDE PRILLS(91,95) of embodiment is tested
In order to verify the tolerance degree of bacterial strain PARA FORMALDEHYDE PRILLS(91,95) concentration of the present invention, a series of full nutrition containing formaldehyde for preparing concentration gradients is trained
Base is supported, culture medium prescription is as follows: Glucose 10.0 g/L, NH4Cl 0.5g/L、Na2HPO4 3.0g/L、KH2PO4 1.5g/L、
MgSO4·7H2O 0.1g/L、1M CaCl20.1g/L,Agar 20g/L.Sterilised membrane filter PARA FORMALDEHYDE PRILLS(91,95) reagent is diluted, and is formed
The formaldehyde titer of 1000x.The formaldehyde of various concentration is divided in different triangular flasks.Ultimately form concentration gradient are as follows: 50,
75, the culture medium containing formaldehyde of 100,110,120,130,140,150,160,170,180,190,200,210,220mg/L.120
DEG C 30min sterilizing, is cooled into plate in culture dish, this bacterium of picking on the bacterium colony of pure culture, coating, can be formed
The judgement for being proliferated bacterium colony is the positive.
As a result: JQ-1 can be positive in the culture medium containing formaldehyde of 50-120 mg/L and be proliferated growth.Judge bacterial strain of the present invention
Highest formaldehyde tolerable concentration are as follows: 120mg/L.
Sequence table
<110>Institute of Biology, Gansu Academy of Sciences
<120>one plants of Degradation Formaldehyde bacterial strains and its applications
<130> 1
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1417
<212> DNA
<213> Methylobacterium podarium
<400> 1
ctcagagcga acgctggcgg caggcttaac acatgcaagt cgaacgggct tcttcggaag 60
tcagtggcag acgggtgagt aacacgtggg aacgtgccct tcggttcgga ataactcagg 120
gaaacttgag ctaataccgg atacgccctt acggggaaag gtttaccgcc gaaggatcgg 180
cccgcgtctg attagcttgt tggtggggta acggcctacc aaggcgacga tcagtagctg 240
gtctgagagg atgatcagcc acactgggac tgagacacgg cccagactcc tacgggaggc 300
agcagtgggg aatattggac aatgggcgca agcctgatcc agccatgccg cgtgagtgat 360
gaaggcctta gggttgtaaa gctcttttgt ccgggacgat aatgacggta ccggaagaat 420
aagccccggc taacttcgtg ccagcagccg cggtaatacg aagggggcta gcgttgctcg 480
gaatcactgg gcgtaaaggg cgcgtaggcg gccgattaag tcgggggtga aagcctgtgg 540
ctcaaccaca gaattgcctt cgatactggt tggcttgaga ccggaagagg acagcggaac 600
tgcgagtgta gaggtgaaat tcgtagatat tcgcaagaac accagtggcg aaggcggctg 660
tctggtccgg ttctgacgct gaggcgcgaa agcgtgggga gcaaacagga ttagataccc 720
tggtagtcca cgccgtaaac gatgaatgcc agccgttggt ctgcttgcag gtcagtggcg 780
ccgctaacgc attaagcatt ccgcctgggg agtacggtcg caagattaaa actcaaagga 840
attgacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgcagaa 900
ccttaccatc ccttgacatg gcatgttacc ctgggagacc ggggatcctc ttcggaggcg 960
tgcacacagg tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc 1020
gcaacgagcg caacccacgt ccttagttgc catcattgag ttgggcactc tagggagact 1080
gccggtgata agccgcgagg aaggtgtgga tgacgtcaag tcctcatggc ccttacggga 1140
tgggctacac acgtgctaca atggcggtga cagagggacg cgaaggggcg acctggagca 1200
aatcccgaaa agccgtctca gttcggattg cactctgcaa ctcgggtgca tgaaggcgga 1260
atcgctagta atcgtggatc agcacgccac ggtgaatacg ttcccgggcc ttgtacacac 1320
cgcccgtcac accatgggag ttggtcttac ccgacggcgc tgcgccaacc gcaaggaggc 1380
aggcgaccac ggtagggtca gcgactgggg tgaagtc 1417
Claims (7)
1. one plant of Methylobacter (Methylobacterium podarium) bacterium JQ-1, it is protected in Chinese microorganism strain
The deposit number for hiding administration committee's common micro-organisms center is CGMCC No.17229.
2. one plant of Methylobacter bacterium JQ-1 according to claim 1, it is characterised in that the bacterial strain can be with degradation of formaldehyde.
3. one plant of Methylobacter bacterium JQ-1 according to claim 1, it is characterised in that the highest of the bacterial strain PARA FORMALDEHYDE PRILLS(91,95)
Tolerable concentration are as follows: 120mg/L.
4. one plant of Methylobacter bacterium JQ-1 according to claim 1, it is characterised in that the bacterial strain is at 30 DEG C, thallus
Concentration is in the case of OD value 0.107, the interior formaldehyde by 54.8mg/L of 120h is degradable.
5. a kind of for cultivating the culture medium of bacterial strain described in claim 1, it is characterised in that formula are as follows: formaldehyde 50mg/L,
NH4Cl 0.5g/L、Na2HPO4 3.0g/L、KH2PO4 1.5g/L、MgSO4·7H2O 0.1g/L、1M CaCl2 0.1g/L。
6. a kind of cultural method of bacterial strain described in claim 1, it is characterised in that 3-5% inoculation, 30 DEG C, 120rpm are protected from light training
It supports.
7. a kind of microorganism methanal removing product, it is characterised in that the product carries out methanal removing by core microbial inoculum of JQ-1.
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| CN101993839A (en) * | 2010-07-23 | 2011-03-30 | 浙江工业大学 | Methylobacterium rhodesianum H13 capable of efficiently degrading dichloromethane and application thereof |
| CN102533620A (en) * | 2012-03-01 | 2012-07-04 | 黑龙江省科学院微生物研究所 | Methyl bacterial strain for degrading methane gas |
| CN102586149A (en) * | 2012-03-01 | 2012-07-18 | 黑龙江省科学院微生物研究所 | Methyl bacterium capable of degrading dichloromethane |
| CN102816714A (en) * | 2012-07-09 | 2012-12-12 | 浙江工业大学 | Formaldehyde degrading bacterium and its application |
-
2019
- 2019-03-19 CN CN201910215095.1A patent/CN109735476B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101993839A (en) * | 2010-07-23 | 2011-03-30 | 浙江工业大学 | Methylobacterium rhodesianum H13 capable of efficiently degrading dichloromethane and application thereof |
| CN102533620A (en) * | 2012-03-01 | 2012-07-04 | 黑龙江省科学院微生物研究所 | Methyl bacterial strain for degrading methane gas |
| CN102586149A (en) * | 2012-03-01 | 2012-07-18 | 黑龙江省科学院微生物研究所 | Methyl bacterium capable of degrading dichloromethane |
| CN102816714A (en) * | 2012-07-09 | 2012-12-12 | 浙江工业大学 | Formaldehyde degrading bacterium and its application |
Non-Patent Citations (5)
| Title |
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| HIROSHI YONEMITSU等: "Biodegradation of high concentrations of formaldehyde by lyophilized cells of Methylobacterium sp. FD1", 《BIOSCI BIOTECHNOL BIOCHEM》 * |
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| 金晶: "甲醛降解菌筛选、关键酶基因克隆表达和固定化细胞降解特性的研究", 《中国优秀硕士学位论文全文数据库》 * |
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