CN106916765A - A kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc - Google Patents
A kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc Download PDFInfo
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
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Abstract
A kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc, belongs to the applied technical field of microorganism fungus kind.The penicillium janthinellum (Penicillium janthinellum) BC109 2, China Committee for Culture Collection of Microorganisms's common micro-organisms center, abbreviation CGMCC have been preserved in, deposit number is CGMCC No.13561.Step of the present invention is：First, screen to separate and purify from soil using dilution spread flat band method and obtain heavy metal zinc resistant strain；2nd, will screen in heavy metal zinc resistant strain fluid nutrient medium of the addition containing heavy metal zinc for obtaining, carry out adsorption conditionses optimization；3rd, under optimal adsorption conditionses, the bacterial strain is added in the waste water containing heavy metal zinc, that is, is completed；The heavy metal zinc of penicillium janthinellum BC109 2 has height endurability.It is an advantage of the invention that advantages of good adsorption effect, it is simple to operate, with low cost, to environment do not exist secondary pollution.
Description
Technical field
The present invention relates to a kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc, belong to microorganism fungus kind
Applied technical field.More specifically, the present invention relates to it is a kind of for adsorbing zinc in waste water penicillium janthinellum (Penicillium janthinellum) BC109-2。
Background technology
Since the thirties in 20th century, as industrial or agricultural is developed rapidly, the application of heavy metal is also more and more extensive, therewith
The pollution problem for coming also is highlighted all the more.Heavy metal pollution is mostly derived from the industrial wastewater and biochemistry of the industries such as mining, smelting, plating
The random discharge of sewage.At present, because exploitation of mineral resources, sewage irrigation etc. cause China to be ploughed by heavy metal pollutions such as Zn, Cd
Nearly 20,000,000 hm2, account for the 1/5 of total area under cultivation.Zinc is a kind of important essential trace element in Animal nutrition, but it is dense
High risks, the World Health Organization can be caused to biology in the case of spending height（WHO）Acceptable maximum in suggestion drinking water
Zn2+Concentration is 5 mg/L.Zinc can not be biodegradable, and the mutual conversion between various forms can only occur, and can be by food
Thing chain is enriched with, and triggers human health risk.In order to the water body of heavy metal pollution is repaired, the restorative procedure bag commonly used at present
Include physics and chemical method.And the physical method such as traditional filtering, ion exchange is relatively costly, there is significant limitation, because
The methods such as this thing followed chemical method, electrolysis are used widely, but chemical method has secondary pollution, electrolysis
Then it is only applicable to the recovery of precious metal.With the development of modern biotechnology, it is biological prosthetic as a kind of environmental protection, low cost,
Efficient biological technique method turns into study hotspot in recent years.
Microorganism can be applied to the reparation of heavy metal-polluted water, and microbe absorption counterweight is pointed in existing numerous researchs
The repair of metal, by screening the microorganism of preventing from heavy metal, is reached using the potential suction-operated of their heavy metals
To the purpose of repairing heavy metal pollution.Report in recent years to Zn2+The bacterium for having suction-operated includes pseudomonad
（Pseudomonas）Adsorbance be 88.23 mg/g, streptomyces (Streptomyces) adsorbance be 54 mg/g, gemma bar
Bacterium (Geobacillus thermodenitrificans) adsorbance is 48.26 mg/g etc.；Fungi includes Penicillium notatum
(Penicillium sp.) adsorbance be 2.22 mg/g and sickle-like bacteria (Fusarium sp.) adsorbance is 3.79 mg/g etc.
Deng.Shaken cultivation in fluid nutrient medium is inoculated into by by strain, can by just turning into after the processes such as filtering, washing, drying
Adsorbent, the cumbersome and harsh condition of culture of operating procedure makes it be dfficult to apply to industrialized production.
The content of the invention
The problems such as the object of the invention is directed to cumbersome current biological adsorption agent preparation process and high cost, and for providing
Kind utilization penicillium janthinellum (Penicillium janthinellum) absorption heavy metal in waste water（Zn）Method, the method is green
Colour circle is protected, and is had broad application prospects.
Technical scheme：A kind of penicillium janthinellum (Penicillium janthinellum) BC109-2,
China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, abbreviation CGMCC, deposit number is CGMCC
No.13561。
A kind of method that heavy metal in waste water zinc is adsorbed using the penicillium janthinellum BC109-2, by 0.1mL after purification
BC109-2 bacteria suspensions, control 108Cfu/mL is added in the PDB fluid nutrient mediums of 100mL, natural pH conditions, and cultivation temperature is
30 DEG C, shake flask culture determines growth curve, after culture 10-16h, the culture bacterium solution in growth logarithmic phase is obtained, by this
Bacterium solution is inoculated into containing carrying out adsorption conditionses optimization in various concentrations heavy metal zinc, the PDB fluid nutrient mediums of difference pH.
The heavy metal for being adsorbed is zinc, and penicillium janthinellum BC109-2 heavy metal zinc has height endurability.
The microassembly robot bacterial strain BC109-2 forms mycelium pellet in the fluid nutrient mediums of PDB containing zinc, and hyphal surface is in fold
Shape.
It is 8 containing Zn in 30 DEG C, pH2+PDB fluid nutrient mediums in adsorbance be 217.47 mg/g.
The present invention filters out a kind of microorganism penicillium janthinellum using dilution spread flat band method from soil
(Penicillium janthinellum), its heavy metal zinc has height endurability, and has preferable adsorption effect to zinc.
The microassembly robot bacterial strain BC109-2 zinc initial concentration be 3 mmol/L, pH be 8 PDB fluid nutrient mediums in expand
Exponential phase is reached during big culture 10-12 h, collects thalline is then centrifuged for.
The thalline that will be collected by centrifugation is added in the waste water containing heavy metal zinc, is stirred under conditions of being 10-35 DEG C in temperature,
Adsorption time reaches saturation for 10 h, that is, complete.The wherein mass volume ratio g/mL of thalline weight in wet base and the waste water containing heavy metal zinc
For（2~4）︰ 1000.
In the PDB fluid nutrient mediums that 30 DEG C, pH are 8, the h of 150r/min shaking tables shaken cultivation 24, the microassembly robot bacterial strain
BC109-2 is to Zn2+Adsorbance with ZnSO4·7H2The increase of O initial concentrations and increase, and increasing degree gradually tends to flat
It is slow, in Zn2+When initial concentration is 3 mmol/L, adsorbance reaches 217.47 mg/g（Accompanying drawing 4）.
Beneficial effects of the present invention：Microassembly robot bacterial strain BC109-2 cultures are easy, it is not necessary to by filtering, washing, dry
The complicated processes such as do, grind, sieving and being made biological adsorption agent, the effect of Adsorption of Heavy Metals zinc can reach 217.47 mg/g, and
Thalline is easily separated from waste water, and environmental protection does not result in secondary pollution.
Biological material specimens preservation：The bacterial strain be penicillium janthinellum (Penicillium janthinellum) BC109-
2, full name isPenicillium janthinellumBC109-2, sequence has been uploaded in Genbank databases, strain sequence
Number be KX891188.Penicillium janthinellum BC109-2 has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center, abbreviation CGMCC, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number is CGMCC No.13561, is protected
The Tibetan date is on January 16th, 2017.
Brief description of the drawings
Fig. 1 bacterial strains BC109-2 resistance to zinc in the medium.
Fig. 2 is containing Zn2+PDB fluid nutrient mediums in grow microassembly robot bacterial strain BC109-2 formed mycelium pellet.
Fig. 3 shows BC109-2 hyphal surface folds under a scanning electron microscope.
Fig. 4 contains Zn2+Waste strength is with BC109-2 to Zn2+The relation of adsorbance.
Specific embodiment
Below technical scheme is described in detail with instantiation.
Screening and culture penicillium janthinellum (Penicillium janthinellum) culture medium used by BC109-2 is such as
Under：
1）LB culture mediums（1000mL）：Tryptone 10.0g；Yeast extract 5.0g；NaCl 10.0g；Agar powder 10.0 ~
15.0g（Fluid nutrient medium is not added with agar）, pH 7.0.
2）PDA culture medium：The g of potato 200, the g of glucose 20, agar 15-20 g, ultra-pure water 1000 mL, natural pH, 1
×10520 min of Pa sterilizings.
3）PDB culture mediums：It is not added with the PDA of agar.
Embodiment 1：The screening of microassembly robot bacterial strain and identification
1）The screening of microassembly robot bacterial strain BC109-2
Soil sample is gathered near discarded Tong Xin Electroplate Factories, sampling depth is 15 ~ 20cm.Weigh pedotheque 5g, be added to containing
In the conical flask of 45mL sterilized waters, then shaking table room temperature concussion 10min takes 1mL sludge suspensions and is added to the examination containing 9mL sterilized waters
Guan Zhong, fully mixes, stepwise dilution to 10-7Sludge suspension.10 are taken respectively-2、10-3、10-4、10-5、10-6、10-7Dilution
1mL coats Zn on superclean bench2+Concentration is on the LB solid mediums of 30mg/L, 30 DEG C in constant incubator
Under the conditions of cultivate 48h.After bacterium colony grows, the difference visibly different bacterial strain of picking colony feature, using trilinear method line transposing
To containing Zn2+LB flat boards on continue to cultivate, then transposing, on PDA plate, is purified fungi after obtaining bacterial strain by separating for several times,
One plant is finally given in the Zn containing 2100mg/L2+PDA plate on normal growth bacterial strain, numbering is BC109-2.
2）The identification of BC109-2
（a）Extracting genome DNA
According to the Ezup pillar fungal genomic DNA extraction agent boxes of Shanghai Sheng Gong companies（SK8259）Operation.
（b）PCR is expanded
Primer 1：5 ' one TCCGTAGGTG AACCTGCGG mono- 3 ',
Primer 2：5 ' one TCCTCCGCTT ATTGATATGC mono- 3 ',
Reaction system：
PCR cycle condition：94 DEG C of first stage, 4min；Second stage（30 circulations）94 DEG C, 45s, 55 DEG C, 45s, 72 DEG C,
1min；72 DEG C of phase III, 10min.
DNA purpose bands, PCR primer PCR primer direct Sequencing needed for the cutting of PCR primer electrophoretic band.DNA sequencing is entrusted
Support Shanghai Sheng Gong companies are carried out.
ITS sequence total length 552bp, sequence such as SEQ ID NO that sequencing is obtained：Shown in 1.
Blast Homology search analyses are carried out in Genbank databases, it is found that the bacterium reaches with microassembly robot homology
100%, combining form identification, the bacterial strain be penicillium janthinellum (Penicillium janthinellum), full name isPenicillium janthinellumBC109-2, sequence has been uploaded in Genbank databases, and strain sequence number is
KX891188。
Embodiment 2：The influence that zinc components grow to penicillium janthinellum BC109-2
In fresh PDA liquid medium, 30 DEG C, 150 r/min shaken cultivations treat bacterium solution OD to picking bacterial strain respectively600Reach
When 0.2 ~ 0.4, each 2mL of bacterium solution is taken respectively and is placed in 250mL triangular flasks, plus PDA liquid medium is to 50mL.In 30 DEG C, 150r/
Min continues shaken cultivation, treats OD600Value reaches 0.2 or so, and addition final concentration is respectively the ZnSO of 1,2,5 mmol/L4·7H2O
Solution, while doing control experiment, each concentration sets three parallel repetitions.Continue will above 12 triangular flasks in 30 DEG C, 150
2d is cultivated in the shaken cultivation case of r/min, at regular intervals（0、6、12、18、24、30、36、42、48 h）Take l mL bacterium solutions
Determine OD600Value.Three repetition summations of each sample are averaged as the OD of the sample600Value.
Thalline is free of Zn in the medium2+When growing state preferably, and reach highest in culture 24h or so biomass.With
Zn in culture medium2+The increase of concentration, thalli growth substantially slows down, biomass reduction, illustrates Zn2+Have one to the growth of bacterial strain
Fixed inhibitory action, but the bacterial strain is in Zn2+Concentration be 3 mmol/L culture medium in remain to growth, show that bacterial strain BC109-2 has
Certain resistance to zinc, specific visible Figure of description 1.
30 DEG C, pH be 8 containing Zn2+PDB fluid nutrient mediums in grow microassembly robot bacterial strain BC109-2 formed mycelium pellet
（Fig. 2）, and hyphal surface fold is shown under a scanning electron microscope（Fig. 3）.
Embodiment 3：Bacterial strain is to Zn2+Absorption
100 mL PDA liquid mediums are added in 250 mL triangular flasks, bacterial strain BC109-2 is inoculated into and is contained 0,1,2 respectively
And the ZnSO of 3mmol/L4·7H2In the culture medium of O, the pH value of culture medium is adjusted to 1 mol/L HCl and 1 mol/L NaOH
8, it is subsequently placed in 30 DEG C, cultivates in 150 r/min shaking tables.Three groups of repetitions of Setup Experiments.After cultivating 24 h, 8000 g centrifugations 15
Washed three times with the 0.9% NaCl aqueous solution after min.Weighed being dried under 80 DEG C of constant temperature of cell, then use nitric acid and perchloric acid
Zn is determined after (7 ︰ 3, V/V) micro-wave digestion2+Content.
It is 8 in pH value of solution, and under conditions of reaching saturated adsorption time, thalline is to Zn2+Adsorbance with ZnSO4·
7H2The increase of O initial concentrations and increase, and increasing degree gradually tends towards stability, in Zn2+When initial concentration is 3 mmol/L, inhale
Attached amount reaches 217.47 mg/g（Fig. 4）.
Sequence table
<210> SEQ ID NO：1
<211> 552
<212> DNA
<213>Penicillium janthinellum BC109-2
<400>1
gcttaagttc agcgggtatc cctacctgat ccgaggtcaa cctgagaaag attgaggggg 60
gtcgccggcg ggcgccggcc gggcctacag agcgggtgac gaagccccat acgctcgagg 120
accggacgcg gtgccgccgc tgcctttcgg gcccgccccc cgggagccgg ggggcggggg 180
cccaacacac aagccgtgct tgagggcagc aatgacgctc ggacaggcat gccccccgga 240
ataccagggg gcgcaatgtg cgttcaaaga ctcgatgatt cactgaattc tgcaattcac 300
attacttatc gcatttcgct gcgttcttca tcgatgccgg aaccaagaga tccgttgttg 360
aaagttttaa ctgatttagc taatcgctca gacagcaatc ttcagacagc gttcaggggg 420
ggcttcggcg ggcgcgggcc cgggggcggg tgccccccgg cggccatgac ggcgggcccg 480
ccgaagcaac taggtatgat aaacacgggt gggaggttgg acccagaggg ccctcactcg 540
gtaatgatcc tt 552
Claims (5)
1. a kind of penicillium janthinellum (Penicillium janthinellum) BC109-2, it has been preserved in Chinese microorganism strain
Preservation administration committee common micro-organisms center, abbreviation CGMCC, deposit number is CGMCC No.13561.
2. a kind of method that utilization penicillium janthinellum BC109-2 adsorbs heavy metal in waste water zinc, it is characterised in that purify 0.1mL
BC109-2 bacteria suspensions afterwards, control 108Cfu/mL is added in the PDB fluid nutrient mediums of 100mL, natural pH conditions, culture temperature
It is 30 DEG C to spend, shake flask culture, determines growth curve, after culture 10-16h, obtains the culture bacterium solution in growth logarithmic phase,
The bacterium solution is inoculated into containing carrying out adsorption conditionses optimization in various concentrations heavy metal zinc, the PDB fluid nutrient mediums of difference pH.
3. the method that utilization penicillium janthinellum BC109-2 according to claim 2 adsorbs heavy metal in waste water zinc, its feature
It is that penicillium janthinellum BC109-2 heavy metal zinc has height endurability.
4. the method that utilization penicillium janthinellum BC109-2 according to claim 2 adsorbs heavy metal in waste water zinc, its feature
It is that the microassembly robot bacterial strain BC109-2 forms mycelium pellet in the fluid nutrient mediums of PDB containing zinc, hyphal surface is in accordion.
5. the method that utilization penicillium janthinellum BC109-2 according to claim 2 adsorbs heavy metal in waste water zinc, its feature
It in 30 DEG C, pH is 8 containing Zn to be2+PDB fluid nutrient mediums in adsorbance be 217.47 mg/g.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108097711A (en) * | 2017-12-18 | 2018-06-01 | 江南大学 | A kind of method for repairing Grown In Zinc Contaminated Soil using penicillium janthinellum joint Indian mustard |
| CN109652346A (en) * | 2019-02-19 | 2019-04-19 | 北京本农科技发展有限公司 | Zinc adsorbs the application of bacterial strain and the cultural method of zinc absorption bacterial strain |
| CN109967040A (en) * | 2017-12-28 | 2019-07-05 | 营口富里泥炭科技有限公司 | A kind of preparation method of novel sludge compound adsorbent |
| CN111849787A (en) * | 2020-08-04 | 2020-10-30 | 安徽工业大学 | Endophytic fungus and application thereof in dye pollutant treatment |
| CN115011488A (en) * | 2022-05-31 | 2022-09-06 | 贵州省生物研究所 | Penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings |
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| CN1704470A (en) * | 2004-06-02 | 2005-12-07 | 广西大学 | High metal-salt-resistant micro penicillium purpurogerum |
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| CN1704470A (en) * | 2004-06-02 | 2005-12-07 | 广西大学 | High metal-salt-resistant micro penicillium purpurogerum |
Non-Patent Citations (2)
| Title |
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| KX891188: "Penicillium janthinellum strain BC109-2 internal transcribed spacer 1", 《GENBANK》 * |
| 韦明肯等: "青霉菌Penicillium janthinellum菌株GXCR的高抗重金属盐及其对Cu2+和Zn2+离子吸收的特性", 《菌物学报》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108097711A (en) * | 2017-12-18 | 2018-06-01 | 江南大学 | A kind of method for repairing Grown In Zinc Contaminated Soil using penicillium janthinellum joint Indian mustard |
| CN109967040A (en) * | 2017-12-28 | 2019-07-05 | 营口富里泥炭科技有限公司 | A kind of preparation method of novel sludge compound adsorbent |
| CN109652346A (en) * | 2019-02-19 | 2019-04-19 | 北京本农科技发展有限公司 | Zinc adsorbs the application of bacterial strain and the cultural method of zinc absorption bacterial strain |
| CN109652346B (en) * | 2019-02-19 | 2020-11-10 | 北京本农科技发展有限公司 | Application of zinc adsorption strain and culture method of zinc adsorption strain |
| CN111849787A (en) * | 2020-08-04 | 2020-10-30 | 安徽工业大学 | Endophytic fungus and application thereof in dye pollutant treatment |
| CN111849787B (en) * | 2020-08-04 | 2021-09-07 | 安徽工业大学 | Endophytic fungus and application thereof in dye pollutant treatment |
| CN115011488A (en) * | 2022-05-31 | 2022-09-06 | 贵州省生物研究所 | Penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings |
| CN115011488B (en) * | 2022-05-31 | 2023-03-24 | 贵州省生物研究所 | Penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings |
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