CN104212728A - High temperature-resistant Candidasp. FD-1 for phenolated water degradation - Google Patents
High temperature-resistant Candidasp. FD-1 for phenolated water degradation Download PDFInfo
- Publication number
- CN104212728A CN104212728A CN201410199771.8A CN201410199771A CN104212728A CN 104212728 A CN104212728 A CN 104212728A CN 201410199771 A CN201410199771 A CN 201410199771A CN 104212728 A CN104212728 A CN 104212728A
- Authority
- CN
- China
- Prior art keywords
- phenol
- degradation
- candidasp
- resistant
- high temperature
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses high temperature-resistant Candidasp. FD-1 for phenolated water degradation. The high temperature-resistant Candidasp. FD-1 is named as Candidasp. FD-1, is preserved in China center for type culture collection which is an international depository authority appointed by the national patent office and is located in Chinese Wuhan university in the Wuhan city, is preserved on May 4, 2014, and has an accession number of CCTCC NO: M2014180. The high temperature-resistant Candidasp. FD-1 for phenolated water degradation has the advantages that 1, the domesticated strain FD-1 has greatly improved phenol degradation efficiency and has the highest adaptable phenol complete-degradation concentration of 1000mg/L, and 2, the Candidasp. FD-1 has the optimal phenol-degradation culture temperature of 25-37 DEG C and the optimal pH of 6.0-7.0, has a wide suitable temperature range and a wide suitable pH range, can resist high-content salt, has the highest salt resistance of 5% considerably higher than the seawater average salinity (3.5%) and is microorganism having a good application prospect.
Description
Technical field
this invention relates toyeast
field.
Background technology
Phenol is the principal pollutant in the trade effluents such as papermaking, coking, oil refining, plastics, weaving, pottery, and phenol and its derivatives belongs to aromatics, is protoplasma poisonous substance, has toxic action to organism
[1], and be difficult to be degraded.Phenolic compound is classified as the one in 129 kinds of priority pollutants Black Lists by American National Environmental Protection Agency
].Toxicity due to phenol relates to hydrobiological Growth and reproduction, and polluted drinking water source, causes severe contamination to water body
[3].Phenolic wastewater is listed in one of harmful waste water of emphasis solution in China's water pollution control.The method removing Phenols In Industrial Liquid Waste class material comprises microbiological deterioration, extraction, charcoal absorption and chemical oxidation etc., wherein utilizes microbiological deterioration to be a kind of method of economical and effective and non-secondary pollution.Chinese scholars has carried out large quantifier elimination to how utilizing microorganism to remove Phenol for Waste Water and derivative thereof, and obtaining some can the bacterial strain of degradation of phenol solution, but substantially seldom has the actual treatment being used in industrial phenolic wastewater.
summary of the invention
Object of the present invention is exactly carried out large quantifier elimination for current Chinese scholars to how utilizing microorganism removing Phenol for Waste Water and derivative thereof, obtaining some can the bacterial strain of degradation of phenol solution, but substantially seldom there is the deficiency of the actual treatment being used in industrial phenolic wastewater, and a kind of resistance to high salt candida tropicalis of phenol water of degrading is provided.
The resistance to high salt candida tropicalis called after of this degraded phenol water: candiyeast FD-1(Candida sp.FD-1), in depositary institution's preservation that State Patent Office specifies, depositary institution's title: China typical culture collection center, preservation date: on May 4th, 2014, preservation place: Wuhan, China Wuhan University, preserving number CCTCCNO:
m2014180
Advantage of the present invention is:
(1) greatly improved by the efficiency of the FD-1 strain degradation phenol of domestication, the highlyest to adapt to and degradable concentration is the phenol of 1000mg/L, when the ratio inoculated and other conditions remain unchanged, the concentration of the bacterium of inoculation is larger, and the time that degraded needs is shorter.
(2) the suitableeest culture temperature of this bacterium degradation of phenol is 25 DEG C ~ 37 DEG C, optimal pH is 6.0 ~ 7.0, the temperature range adapted to and pH scope all wider, and can resistance to high salt, its the highest tolerance of salinity can reach 5%, far exceeding the average salinity (3.5%) of seawater, is a kind of microorganism had a good application prospect.When other conditions are constant, the speed that a small amount of culture medium raw material can increase degraded is added in solution during degraded, but it is comparatively large that excessive interpolation culture medium raw material one is financial loss, and two is that its effect can not proportionally increase, the optimum additional proportion of substratum that this experiment is determined is 0.05%.The suitableeest inoculative proportion is 4%.
figure of description
Accompanying drawing 1 is phenol solution canonical plotting.
Accompanying drawing 2 is the degradation process A graphic representation of FD-1 to different concns phenol solution.
Accompanying drawing 3 is the degradation process B graphic representation of FD-1 to different concns phenol solution.
Accompanying drawing 4 is the degradation process C curve figure of FD-1 to different concns phenol solution.
Accompanying drawing 5 is the degradation process D graphic representation of FD-1 to different concns phenol solution.
Accompanying drawing 6 is the impact element A graphic representation of each factor Degradation of Phenol.
Accompanying drawing 7 is the impact element B graphic representation of each factor Degradation of Phenol.
Accompanying drawing 8 is the impact element C curve figure of each factor Degradation of Phenol.
Accompanying drawing 9 is the impact element D graphic representation of each factor Degradation of Phenol.
Accompanying drawing 10 is the impact element E graphic representation of each factor Degradation of Phenol.
embodiment
Candiyeast FD-1(Candida sp.FD-1) separation of bacterial classification, culture & identification
1.1 substratum
1.1.1 LB substratum
peptone 1%, NaCl 1%, yeast extract paste 0.5%, agar (solid medium) 1.5%.
1.1.2 inorganic salt screening culture medium
k
2hPO
40.5g/L, NaCl 0.2g/L, MnSO
4h
20 0.02g/L, CaCl 0.05g/L, KH
2pO
40.5g/L, MgSO
47H
2o 0.1g/L, Fe
2(SO
4)
3h
2o 0.005g/L, (NH
4)
2sO
41g/L, phenol 1g/L, agar 1.5%(solid medium).
The separation and consentration of 1.2 bacterial classifications
Directly from industrial phenolic wastewater pond, get mud, through diluting 10
1, 10
2, 10
3, 10
4, 10
5, 10
6be applied on LB flat board doubly and cultivate, picking list bacterium colony LB liquid nutrient medium enlarged culturing.
1.3 tamed strain
When the bacterium in LB liquid nutrient medium reaches logarithmic growth after date through cultivating, bacterium liquid being linked into step by step phenol content is 500mg/L, 1000 mg/L, in the LB liquid nutrient medium of 1500 mg/L and 2000 mg/L.It is in the minimal medium of 500 mg/L and 1000 mg/L that the bacterium liquid that then can adapt to peak concentration phenol is inoculated into containing phenol, for subsequent use as fungi preservation using the bacterial strain of the bacterium liquid adapting to maximum concentration.
1.4 identification of strains
1.4.1 gram staining method observes slat chain conveyor colony characteristics, and gramstaining, in its form of basis of microscopic observation.
1.4.2 molecular markers for identification distinguishes 18S rDNA and the ITS sequence of pcr amplification bacterium, then carries out BLAST comparison, then uses MEGA4 software building phylogenetic tree, obtain the Phylogenetic Relationships of FD-1.
1.4.2.1 18S rDNA sequence amplification
Primer: 5 '-CCAACCTGGTTGATCCTGCCAGTA-3 ' (forward); 5 '-CCTTGTTACGACTTCACCTTCCTCT-3 ' (oppositely).
The reaction conditions of pcr amplification 18S rDNA gene is as table 1-1:
The reactions steps of table 1-1 pcr amplification 18S rDNA gene
1.4.2.2 ITS sequence amplification
Primer: 5 '-GTCCACTGGAACCTTTACATTTAG-3 ' (forward); 5 '-TCCTCCGCTTATTGATATGC-3 ' (oppositely).
The reaction conditions of pcr amplification ITS gene is as table 1-2:
The reactions steps of table 1-2 pcr amplification ITS sequence
The mensuration of 1.5 strain growth amounts
by the optical density value of nutrient solution under measurement of ultraviolet-visible spectrophotometer 600nm wavelength
[14].
The mensuration of 1.6 phenol solution standard curve making and phenol content
the light absorption value of phenol under employing 4-peace base quinizine method mensuration 510nm wavelength, the light absorption value recorded by calibration series deducts the light absorption value of zero-dose, and drafting light absorption value for the curve of phenol concentration, then obtains phenol concentration by typical curve Solving Equations.
The optimal conditions of 1.7 FD-1 degradation of phenol solution is inquired into
Inquire into the impact of temperature, pH, LB concentration and NaCl concentration Degradation of Phenol respectively, the gradient test under different condition is set respectively, compares the impact of its Degradation of Phenol efficiency, determine the suitableeest scope under the suitableeest degradation condition or optimal proportions.
Result and discussion
The qualification of 2.1 domestication results and bacterial strain
This Acclimation goes out a bacterial strain, and it can breed and in 24 hours, phenol degrading wherein is complete in the minimal medium of 1000 mg/L, called after FD-1.At LB grow on plates, bacterial plaque subcircular, smooth surface, white is opaque.Through gramstaining, in basis of microscopic observation, its form is similar to yeast, breeds in the mode of sprouting, and gramstaining is positive.Its 18S rDNA and ITS is checked order (sequence number of 18S rDNA and ITS in NCBI is respectively KJ647397 and KJ647398), the display of the result of BLAST comparison, the 18S rDNA of FD-1 and ITS sequence and Candida tropicalis (
candida tropicalis) homology is up to 99%, with the white Cryptococcus bacterium (Cryptococeales closely belonged to Candida tropicalis homology, Cryptococcaceae, Torulopsis) as outgroup, the systematic evolution tree in conjunction with ITS and 18S rDNA sequence shows: FD-1 and Candida tropicalis (
candida tropicalis) be in same large branch, and supporting rate reaches 81, therefore, we can judge, FD-1 belongs to Cryptococcaceae, the sibling species of the Candida tropicalis in Candida.The bacterial strain obtained after domestication is through gram staining method microscopic examination, gramstaining is positive, and breeds in the mode of sprouting, and this bacterium is checked order through 18S rDNA and ITS and identifies, through BLAST comparison also with its Phylogenetic of MEGA software building tree, identify that it is Candida tropicalis.
2.2 phenol solution typical curves and utilize FD-1 to degrade the phenol solution of different concns
Adopt 4-to pacify the light absorption value of base quinizine method mensuration phenol, the results are shown in accompanying drawing 1.
Utilize with phenol be sole carbon source minimal medium cultivate the FD-1 bacterial classification after domestication, after in substratum, phenol is completely degraded, the OD value measuring its bacterium is 0.680.In 50ml reaction system, preparing phenol content is respectively the phenol solution of 200mg/L, 400mg/L, 600mg/L, 800mg/L, account for 6% of cumulative volume by inoculum size and add bacterium liquid, now solution PH is about 5.8, at 35 DEG C, cultivate under 200rpm condition, and detect the change of phenol content and the content of bacterium in phenol solution, see accompanying drawing 2.3.4.5.
In accompanying drawing 2.3.4.5, it is the phenol solution of 200mg/L, 400mg/L, 600mg/L, 800mg/L that A, B, C, D represent FD-1 degradation of phenol content respectively.As can be seen from graphic representation, the bacterium liquid through high concentration phenol domestication taken out from minimal medium, after joining phenol solution, need through certain hour adjustment period, to be allowed for access logarithmic phase, treat that the starting point concentration of degradation of phenol solution is larger, the adaptation time of bacterium is also longer, and the time needed for degraded is also proportionate; The degradation rate of phenol reaches maximum at the logarithmic phase of bacterium; After the growth of bacterium reaches stationary phase, phenol is degraded completely thereupon.Degradation efficiency during FD-1 degraded 400mg/L phenol solution is relatively high, i.e. degradable 16.67mg/L per hour.See accompanying drawing 6.7.8.9.10 (the impact element of each factor Degradation of Phenol).
This Setup Experiments 5 thermogrades study the optimum temperature range of FD-1 degradation of phenol, as shown in Figure 6, are the degradation curve of FD-1 bacterium degraded 400mg/L phenol solution under condition of different temperatures.At 25 DEG C, under 30 DEG C, 35 DEG C, 37 DEG C and 42 DEG C of conditions, through the degraded of 22 hours, the light absorption value of the residual content of phenol was respectively 0.006, and 0,0,0.004,0.078.The temperature that degradation efficiency is the highest is 30 DEG C and 35 DEG C, is 25 DEG C in temperature, under 37 DEG C of conditions, its phenol content remaining after degrading 22 hours is also almost nil, and under 42 DEG C of conditions, phenol only has a small amount of degraded, simultaneously Candida tropicalis also cannot raised growth at such a temperature.Therefore, the optimum temperature range of Candida tropicalis FD-1 is 25 DEG C ~ 37 DEG C.
PH is on the impact of degraded: the phenol solution preparing 400mg/L respectively, by 6 the different initial soln pH regulator arranged, culture condition is still 35 DEG C, 200rpm, its cultivation results as shown in Figure 7, when pH is 4, there is not obvious decline in the phenol light absorption value in phenol solution, and maintains constant after 20 hours, cannot be degraded, illustrate under this pH condition, Candida tropicalis cannot normal growth metabolism, also cannot degradation of phenol.Along with the rising of pH value, when pH is 5, parabolically, under this pH condition is described, Candida tropicalis can eubolism, and by phenol fast degradation, within about 16 hours, the phenol solution of 400mg/L all can be degraded for the light absorption value change curve of phenol; When pH is 6,7, degradation time is used the shortest, is about 15 hours; When pH is increased to 8, Candida tropicalis still can metabolism phenol, but the speed of degraded declines.Therefore, can draw, the optimal pH of candida tropicalis is 6 ~ 7.
Inoculum size on degraded impact by Fig. 8, by the time that Units of Account ratio strain degradation is used, can know that the degradation efficiency of these four kinds of inoculative proportions sorts from high to low is: the degradation efficiency of 4% > 5% > 3% > 6%, 4% inoculum size can reach average degradation of phenol 36.36mg/L per hour.Therefore, can draw, along with the increase of inoculum size, the efficiency of its degraded does not increase thereupon.Although the time accelerates to some extent, for a large amount of consumption of substratum, also directly can cause the increase greatly of Financial cost, in the application of reality, we can consider using most effective inoculum size as actual inoculum size.This tests substratum used is LB substratum, and the cost of material of LB substratum relatively also costly, needs to determine better culture medium prescription further, makes it both efficient, cheap again.
The impact of LB on degraded can be drawn by Fig. 9, and along with the additional proportion of LB medium component increases, the degradation efficiency of phenol is in the trend reduced.Although this illustrates that adding of LB substratum makes the quantity of Candida tropicalis greatly increase, more bacterium does not preferentially utilize phenol as carbon source, but preferentially make use of unnecessary LB composition, which results in the slack-off of phenol degrading speed.Therefore, add the degradation efficiency that a small amount of LB medium component can improve phenol greatly, and for input economically also more satisfactoryization.The suitableeest LB substratum add-on that this laboratory draws is 0.05%.In the wastewater treatment of reality, in line with economic, efficient principle, we can add some nutritive elements in right amount, make the degraded of microorganism reach best effect and top efficiency.
Inorganic salt on the impact of degraded as can be seen from Figure 10, after additional proportion is the sodium-chlor of 0.05% and 0.2%, degradation rate can be made to increase, illustrate that the inorganic salt of suitable proportion are for the growth metabolism of bacterium and the effect organic degraded being played to promotion, when the ratio adding sodium-chlor is 1%, degradation rate reaches maximum value; And the ratio of sodium-chlor is too high, also can affect the metabolism of bacterium, as additional proportion be after the sodium-chlor of 3% and 5% its degradation rate lower than the condition of non-salt adding.The Candida tropicalis FD-1 that this laboratory is separated can grow the organism that also this toxicity of degradation of phenol is higher in the sodium-chlor environment of 5%, this salinity has exceeded the average salinity (35 ‰) of seawater, illustrate that its salt resistance ability is very good, adaptive faculty is also very strong, and what further demonstrate this bacterium can application prospect.
Claims (1)
1. the resistance to high salt Candida tropicalis of a phenol water of degrading, it is characterized in that called after: candiyeast FD-1(Candida sp.FD-1), in depositary institution's preservation that State Patent Office specifies, depositary institution's title: China typical culture collection center, preservation date: on May 4th, 2014, preservation place: Wuhan, China Wuhan University, preserving number CCTCCNO:
m2014180.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410199771.8A CN104212728A (en) | 2014-05-13 | 2014-05-13 | High temperature-resistant Candidasp. FD-1 for phenolated water degradation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410199771.8A CN104212728A (en) | 2014-05-13 | 2014-05-13 | High temperature-resistant Candidasp. FD-1 for phenolated water degradation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104212728A true CN104212728A (en) | 2014-12-17 |
Family
ID=52094638
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410199771.8A Pending CN104212728A (en) | 2014-05-13 | 2014-05-13 | High temperature-resistant Candidasp. FD-1 for phenolated water degradation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104212728A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560746A (en) * | 2014-12-26 | 2015-04-29 | 盐城工学院 | Candida tropicalis capable of treating high-salinity organic chemical wastewater, and application of candida tropicalis |
| CN105884036A (en) * | 2016-04-14 | 2016-08-24 | 湖南大学 | Biological treatment method of high-salinity wastewater containing DCMX (2,4-dichloro-3,5-dimethylphenol) |
| CN107142209A (en) * | 2017-06-29 | 2017-09-08 | 史新义 | A kind of phenol wastewater biodegradable fluid |
| CN108977370A (en) * | 2018-08-08 | 2018-12-11 | 江苏师范大学 | The saccharomycete of one plant of degradation of phenol class compound and its application |
| CN114752501A (en) * | 2021-01-12 | 2022-07-15 | 中国石油大学(华东) | Method for removing phenol and promoting growth of chlorella by co-culture of phycomycetes |
-
2014
- 2014-05-13 CN CN201410199771.8A patent/CN104212728A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560746A (en) * | 2014-12-26 | 2015-04-29 | 盐城工学院 | Candida tropicalis capable of treating high-salinity organic chemical wastewater, and application of candida tropicalis |
| CN104560746B (en) * | 2014-12-26 | 2017-07-28 | 盐城工学院 | One plant can handle high salinity organic chemical waste water candida tropicalis and application thereof |
| CN105884036A (en) * | 2016-04-14 | 2016-08-24 | 湖南大学 | Biological treatment method of high-salinity wastewater containing DCMX (2,4-dichloro-3,5-dimethylphenol) |
| CN105884036B (en) * | 2016-04-14 | 2019-05-03 | 湖南大学 | A method of containing the chloro- 3,5- xylenol high-salt wastewater biological treatment of 2,4- bis- |
| CN107142209A (en) * | 2017-06-29 | 2017-09-08 | 史新义 | A kind of phenol wastewater biodegradable fluid |
| CN107142209B (en) * | 2017-06-29 | 2021-01-08 | 史新义 | Phenol-containing wastewater biological degradation liquid |
| CN108977370A (en) * | 2018-08-08 | 2018-12-11 | 江苏师范大学 | The saccharomycete of one plant of degradation of phenol class compound and its application |
| CN108977370B (en) * | 2018-08-08 | 2021-09-28 | 江苏师范大学 | Yeast for degrading phenol compounds and application thereof |
| CN114752501A (en) * | 2021-01-12 | 2022-07-15 | 中国石油大学(华东) | Method for removing phenol and promoting growth of chlorella by co-culture of phycomycetes |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Oikonomou et al. | Plankton microorganisms coinciding with two consecutive mass fish kills in a newly reconstructed lake | |
| Panwichian et al. | Isolation of purple nonsulfur bacteria for the removal of heavy metals and sodium from contaminated shrimp ponds | |
| CN103013859B (en) | Contaminated soil phenanthrene and application thereof in contaminated soil restoration | |
| Bahig et al. | Isolation, characterization and application of bacterial population from agricultural soil at Sohag Province, Egypt | |
| Arriada et al. | Nannochloropsis oculata growth in produced water: an alternative for massive microalgae biomass production | |
| CN104212728A (en) | High temperature-resistant Candidasp. FD-1 for phenolated water degradation | |
| CN102242071B (en) | Application of Pichia guilliermondii 510-6jm in treatment of petroleum hydrocarbon contaminations | |
| Rai et al. | Potential of cyanobacterial biofilms in phosphate removal and biomass production | |
| CN102586160A (en) | Stenotrophomonas maltophilia DS4 | |
| CN102994428A (en) | Marine surfactant-producing bacterial strain LHOD-1 and application thereof | |
| CN102220264A (en) | Facultatively anaerobic denitrifying bacteria and application thereof in biological denitrification of water body | |
| Rahman et al. | Bacterial diversity of a wooded riparian strip soil specifically designed for enhancing the denitrification process | |
| CN102583780A (en) | Application of Stenotrophomonas maltophilia DS4 for degrading organic pollutants in saponin waste water | |
| CN105695360B (en) | A kind of phenanthrene degradation bacteria Acinetobacter tandoii LJ-5 and its application | |
| Bhatt et al. | Chlorella pyrenoidosa-mediated removal of pathogenic bacteria from municipal wastewater–Multivariate process optimization and application in the real sewage | |
| CN107164277B (en) | Halomonas for degrading phenol | |
| CN104845899A (en) | Application of Rhodococcus sp. 2G in degradation of phthalate | |
| CN103952342A (en) | Pseudomonas geniculata and application thereof | |
| CN104805035A (en) | Rhodococcus sp. 2G used for simultaneous degradation of plurality of phthalic acid esters | |
| Sá et al. | Screening of endoglucanase-producing bacteria in the saline rhizosphere of Rhizophora mangle | |
| CN103387947B (en) | JY-Q bacterial strain of Pseudomonassp and application of bacterial strain in degradation of nicotine in concentrated liquor of reconstituted tobacco | |
| CN116515665A (en) | Pseudomonas capable of degrading microcystin, immobilized microbial agent and application | |
| CN102864086B (en) | Strain capable of degrading polycyclic aromatic hydrocarbons (PAHs) and application thereof in soil remediation | |
| CN106754528B (en) | Pseudomonas flexi and application thereof | |
| CN104745515A (en) | Acinetobacter sp. for degrading polycyclic aromatic hydrocarbon and application of acinetobacter sp. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20141217 |