CN104673710A - Rhodococcus sp. strain and application thereof - Google Patents
Rhodococcus sp. strain and application thereof Download PDFInfo
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- CN104673710A CN104673710A CN201410851523.7A CN201410851523A CN104673710A CN 104673710 A CN104673710 A CN 104673710A CN 201410851523 A CN201410851523 A CN 201410851523A CN 104673710 A CN104673710 A CN 104673710A
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- 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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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- 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
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
- C02F2101/345—Phenols
Abstract
The invention discloses a rhodococcus sp. strain and application thereof, belonging to the field of environmental microbiological application. The strain is named as (Rhodococcus sp.) CM-HZX1 and is now collected with the serial number of CCTCC (China Center For Type Culture Collection) NO:M 2014329 in CCTCC on July 9th, 2014. The rhodococcus sp.CM-HZX1 has relatively strong phenol degrading capacity, namely that the degradation rate is up to 93.6% after 0.5g/L of phenol is degraded for 24 hours, and the degradation rate is up to more than 90% after 1.5g/L of phenol is degraded for 48 hours. The strain is capable of tolerating the salinity of 4% and strong in adaptability; and the strain is efficient when being used for biological treatment of phenol wastewater so as to be wide in application prospect.
Description
Technical field
The present invention relates to environmental microorganism Application Areas, particularly relate to a kind of Rhodococcus strain and application thereof.
Background technology
Phenolic wastewater is mainly derived from oil refining, coking, coal gas and with in phenol or the phenolic aldehyde production process such as chemical industry, papermaking, smelting, textile printing and dyeing that is raw material, this type of wastewater source is wide, the water yield large, complicated components, toxicity are large, is one of poisonous and harmful waste water being classified as emphasis solution in China's water pollution control.
At present, phenolic wastewater treatment method mainly contains Physical, chemical method and biological process.Physical is common with absorption, extraction process, but poor selectivity, be easy to cause secondary pollution.Chemical method mainly contains chemical oxidization method and chemical precipitation method, but processing cost is high, can not reclaim phenol.Biological process mainly relies on phenols as growth carbon source, utilizes the metabolism of microorganism, the phenolic compound in degradation water, discharges after being translated into inorganics.But this type of is by very large containing influence factors such as phenol amount, waste water ph, salinity and other hazardous contaminants, and directly can not process high density, general phenolic concentration does not adopt biological process higher than the waste water of 100mg/L.
Many scholars are separated to the microbial strains of many degradation of phenol from the with serious pollution environment of phenols, comprise Pseudmonas sp., Bacillus sp., Candida tropicalis, Brucella sp., Bacillus cereus etc., these bacterial classifications can with phenol for single carbon source provides growth, but causes biodegradability to differ greatly due to different microbial strainss and different experiment conditions.
J.S.Lee etc. once screened the yeast (Yarrowia lipolytica Y103) of a strain degradation of phenol, the phenol of the 47mg/L that can degrade; The phenol of R.Metallidurans CH34 strains for degrading 4mmol/L needs 48h; Liu Xing equality obtains a pseudomonas from the activated sludge acclimatization separation Yang Zi ethene group's Waste Water Treatment aeration tank and belongs to bacterial strain, can effectively degrade when this bacterial strain is within the scope of 5 DEG C ~ 35 DEG C and the phenol of mineralising 200mg/L, when the starting point concentration of phenol is more than 1000mg/L, the growth of degradation bacteria is suppressed, can not effective degradation of phenol; Song Jing etc. are separated from Suzhou Creek in Shanghai middle reaches and obtain the general Pseudomonas SZH3 of a strain, and in 60h, degraded 500mg/L reaches 95.4%, and the highest can tolerable concentration be the phenol of 1300mg/L; Wang Qimings etc. isolate the efficient degrading bacteria Rhodococcus sp.D3 bacterial strain of phenol from the mud of the trees deadwood rotted and pollution, can be almost that the phenol of 1000mg/L is degradable in 72h by concentration; Shen Xihui etc. are separated to a strain and can grow, have the bacterial isolates Rhodococcus sp.trainPNAN5 simultaneously degraded with phenol sole carbon source and the energy, are being less than 2mmolL in phenol concentration
-1scope can be degradable.
Publication number is that the Chinese invention of CN 101857846 application discloses that a kind of rhodococcus and microbial inoculum thereof and application, deposit number CGMCC No.3695, rhodococcus (Rhodococcus sp.) phenol content in basal salt media of called after BZ-9 is that after 100mg/L, 3d, degradation rate reaches 100%.Publication number is that the Chinese invention of CN 102399700 application discloses that a kind of phenol degrading fungi and application, deposit number CGMCC No.5190, Cladosporium (Cladosp.orium sp.) phenol content in simulated wastewater of called after PHDE-1 is 1000mg/L, after 10d, degradation rate reaches 96%, phenol content is that after 1400mg/L, 10d, degradation rate reaches 68%.
Comprehensive above-mentioned scientific and technical literature and patent documentation, although the multiple-microorganism filtered out can degradation of phenol to a certain extent, degraded envrionment conditions is harsh, and degradation rate is low, and tolerance phenolic concentration is low, extends to practice and is subject to very large restriction.
Therefore filter out and there is efficient bacterium of removing phenol, for the water body that the industrial phenolic wastewater of process and phenols pollute, there is great utilization and extention and be worth.
Summary of the invention
The invention provides a kind of Rhodococcus strain and application thereof, this Rhodococcus strain can efficient degradation phenols, can be widely used in the biological degradation of phenolic waste water.
A kind of Rhodococcus strain, called after rhodococcus (Rhodococcus sp.) CM-HZX1, deposit number is CCTCC NO:M 2014329, and preservation date is on July 9th, 2014.
Described Rhodococcus strain is deposited in the China typical culture collection center (being called for short CCTCC) of Wuhan University of Wuhan, China city on July 9th, 2014, deposit number is CCTCC NO:M 2014329, called after rhodococcus (Rhodococcus sp.) CM-HZX1.
The 16S rDNA sequence of described rhodococcus (Rhodococcus sp.) CM-HZX1 is as shown in SEQ ID:1.
The biological property of described rhodococcus (Rhodococcus sp.) CM-HZX1 is: bacterium colony is circular, moistening, orange, opaque, be easy to provoke, neat in edge, bacterial strain diameter is 819 ~ 1120nm, and gramstaining reacts and is positive.
The physiological and biochemical property of described rhodococcus (Rhodococcus sp.) CM-HZX1: the experimental result of catalase, hydrogen sulfide production test, nitrate reduction and urease is positive; The experimental result of the acid of glucose fermentation, indoles, V.P, methyl red, gelatin, glycerine product, sorbyl alcohol, N.F,USP MANNITOL, citric acid, salicin, Vitamin C2, phenylalanine is negative.
The invention provides a kind of cultural method of Rhodococcus strain, after being activated by described Rhodococcus strain, be inoculated into that initial pH value is 6 ~ 9, temperature is 25 ~ 35 DEG C, the mass concentration of salt is cultivate in the substratum of 0 ~ 5%.The clearance cultivating the Rhodococcus strain Pyrogentisinic Acid of acquisition under this condition is high.
In bacterial strain shaking table culturing process, the growth of shaking speed on Rhodococcus strain also has impact, is preferably 100 ~ 200r/min, more preferably 150r/min.
The invention provides the application of a kind of described Rhodococcus strain in process phenol waste water.Described waste water not only comprises factory's blowdown water, also comprises the water body polluted by phenols.
The application of described Rhodococcus strain in process phenol waste water, comprising: after described Rhodococcus strain immobilization, then carry out biochemical treatment in waste water.Described Rhodococcus strain immobilization comprises and being immobilized onto in microballoon by bacterial strain, or carries out bio-film colonization and be fixed on filler by bacterial strain; Waste water is through being fixed with microballoon or the filler of bacterial strain, and bacterial strain processes the phenol in waste water.
The invention provides a kind of microbial inoculum comprising described Rhodococcus strain.
Present invention also offers a kind of microbe microsphere comprising described Rhodococcus strain.
In described microbe microsphere, the main component of carrier is the mixture of polyvinyl alcohol and sodium alginate; Mass-transfer performance is good, and the stability of microbe microsphere is strong.
In order to strengthen the stability of microbe microsphere, usually also add appropriate gac, boric acid, calcium chloride etc. in the carrier.
Gac can increase microballoon proportion, weakens Float upward function; Boric acid can increase microballoon specific surface and pore volume, and internal pore structure is conducive to the transmission of reaction substrate and nutritive substance, and microorganism cells is at the apposition growth of carrier inside; Calcium chloride can strengthen the permeability of microbe microsphere, improves the activity of microballoon.
Rhodococcus CM-HZX1 provided by the invention has the ability of stronger degradation of phenol, and after after 0.5g/L phenol degrading 24h, degradation rate reaches 93.6%, 1.5g/L phenol degrading 48h, degradation rate reaches more than 90%; This bacterial strain can tolerate 4% salinity, strong adaptability; The present invention has high efficiency in the biological treatment of phenols wastewater, has broad application prospects.
Accompanying drawing explanation
Fig. 1 is the electronic scanning picture of rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1;
Fig. 2 is the phylogenetic tree that rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1 builds based on 16S rDNA sequence homology;
Fig. 3 is the impact that initial pH value grows rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1;
Fig. 4 is the impact that temperature grows rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1;
Fig. 5 is the impact that rotating speed grows rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1;
Fig. 6 is the impact that salt concn grows rhodococcus of the present invention (Rhodococcus sp.) CM-HZX1.
Embodiment
Below in conjunction with specific embodiment, the present invention is further explained.
The present invention adopts the 4-AA Cerium-group REE of " water and waste water method for monitoring and analyzing (the 4th edition) " to carry out the mensuration of volatile phenol (in phenol).
The separation andpreconcentration of embodiment 1 rhodococcus (Rhodococcus sp.) CM-HZX1
One, the separation of rhodococcus (Rhodococcus sp.) CM-HZX1
Rhodococcus (Rhodococcus sp.) CM-HZX1 is separated to obtain from the active sludge of Huzhou sewage work Aerobic Pond.
(1) substratum
Enrichment medium: yeast powder 5g, peptone 10g, NaCl 10g, adding distil water is settled to 1000mL, pH:7.0 ~ 7.2,121 DEG C of sterilizings 20 minutes.
Minimal medium: (NH
4)
2sO
4: 1g/L, CaCl
2: 0.1g/L, K
2hPO
4: 0.5g/L, KH
2pO
4: 0.5g/L, MgSO
4: 0.5g/L, NaCl:1g/L, a certain amount of phenol, with without phenol distilled water constant volume, PH:7.0 ~ 7.2,121 DEG C of sterilizings 20 minutes.
Solid medium: add the agar of 1.5 ~ 2% in aforesaid liquid medium component.
(2) separation and purification of bacterial strain
Getting 10g active sludge, to be inoculated into phenol concentration be in the 100ml enrichment medium of 100mg/L, 30 DEG C, shaking culture on the shaking table of 150r/min; Get supernatant liquor 10ml after muddiness until solution be transferred in the 100ml enrichment medium that phenol concentration is 200mg/L by clarifying to become, 30 DEG C, shaking culture on the shaking table of 150r/min; Switching like this is until phenol concentration is increased to 500mg/L.Getting the above-mentioned bacterium liquid of 10ml, to join 100ml phenol concentration be in the minimal medium of 500mg/L, until nutrient solution by after clarifying and becoming muddiness, collects supernatant liquor and preserve.
Get the bacterium liquid 1ml after above-mentioned domestication cultivation, be mixed with 10
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6diluent, get 100 ~ 200 μ L flat boards and be coated on solid medium flat board, 30 DEG C of constant temperature dark culturing 1 week (each extent of dilution does three repetitions), observe the growing state of dull and stereotyped upper bacterium colony.Select wherein apparent single bacterium colony separating for several times purifying, obtain the strain of pure bacterial strain 4.
Picking list bacterium is seeded in 50ml minimal medium, 30 DEG C, 150r/min constant-temperature table enrichment culture 7 ~ 10 days, detect the content of phenol in substratum, thus judge the ability of strains for degrading phenol, filter out a strain and take phenol as sole carbon source growth and the bacterial strain with efficient degradation ability, called after CM-HZX1.
Two, the qualification of rhodococcus (Rhodococcus sp.) CM-HZX1
(1) bacterium colony of rhodococcus (Rhodococcus sp.) CM-HZX1, thalli morphology are observed
Thalli morphology is studied: carry out gramstaining to thalline, observe under opticmicroscope 16 × 100 eyepiece.
This bacterial strain main biological property is: at LB cultured on solid medium after 2 days, the bacterium colony of CM-HZX1 bacterial strain present circle, moistening, orange, opaque, be easy to provoke, neat in edge, diameter is about 0.8 ~ 1.12nm, and gramstaining reaction is positive.
The physiological and biochemical property of this bacterial strain: the experimental result of catalase, hydrogen sulfide production test, nitrate reduction and urease is positive; The experimental result of the acid of glucose fermentation, indoles, V.P, methyl red, gelatin, glycerine product, sorbyl alcohol, N.F,USP MANNITOL, citric acid, salicin, Vitamin C2, phenylalanine is negative.
(2) molecular biology identification of rhodococcus (Rhodococcus sp.) CM-HZX1
With the DNA of the enzyme extraction CM-HZX1 bacterial strain of prompt base (Shanghai) trade Co., Ltd in English Weihe River synthesis, the 16SrDNA of this bacterial strain that increases, checks order entrusting Shanghai Li Fei Bioisystech Co., Ltd after the PCR primer purifying of acquisition.
Primer is: 1492R:5 '-GGYTAC CTTGTTACGACTT-3 ' (SEQ ID NO:2),
27F：5’-AGA GTT TGA TCM TGG CTCAG-3’(SEQ ID NO：3)，
(synthesis of prompt base (Shanghai) trade Co., Ltd in the English Weihe River).
PCR amplification system is as shown in table 1.
Table 1 PCR amplification system
| Composition | Content (uL) |
| 2×MightyAmd Buffer Ver.2 | 30 |
| MightyAmd DNA Polymerase | 1.5 |
| DNTP primer 2 7F (2.5mmol/L) | 1.5 |
| DNTP primer 1492R (2.5mmol/L) | 1.5 |
| Template | Trace thalline |
| Ultrapure water | 25.5 |
| Total system | 60 |
| Amount to | 50 |
PCR reaction conditions is:
Acquire through order-checking the 16SrDNA fragment (SEQ ID NO:1) that length is 1381bp, the number of registration in GenBank is KM014567, and base sequence is as shown in SEQ ID NO.1.Found by Blast comparison, CM-HZX1 bacterial strain reaches 96% ~ 99% with the 16SrDNA sequence similarity of rhodococcus kind (Rhodococcus sp.) bacterium reported.With MEGA5.0 software, phylogenetic tree is constructed to these bacteriums, and carry out homology analysis.
As shown in Figure 2, the genetic distance of CM-HZX1 bacterial strain and rhodococcus (Rhodococcus sp.) is nearest, in conjunction with the morphological feature of bacterial strain, tentatively can determine that CM-HZX1 bacterial strain is rhodococcus (Rhodococcus sp.), called after rhodococcus (Rhodococcus sp.) CM-HZX1, and this bacterial strain is delivered to the China typical culture collection center (be called for short CCTCC) being positioned at Wuhan, China university and carry out preservation, deposit number is CCTCC NO:M 2014329, and preservation date is on July 9th, 2014.
The optimal growth condition optimization of embodiment 2 rhodococcus (Rhodococcus sp.) CM-HZX1
Substratum: (NH
4)
2sO
4: 1g/L, CaCl
2: 0.1g/L, K
2hPO
4: 0.5g/L, KH
2pO
4: 0.5g/L, MgSO
4: 0.5g/L, NaCl:1g/L, a certain amount of phenol, with without phenol distilled water constant volume, PH:7.0 ~ 7.2.
(1) initial pH value of medium is on the impact of rhodococcus (Rhodococcus sp.) CM-HZX1 degradation of phenol efficiency
By changing pH value in substratum, investigate initial p H to the impact of rhodococcus (Rhodococcus sp.) CM-HZX1 degradation of phenol efficiency.The phenol substratum 300mL of preparation 500mg/L concentration, switching volume is about 15%, it is in the phenol substratum of 3 ~ 11 that centrifuging and taking bacterium mud adds to pH, simultaneously with do not add rhodococcus (Rhodococcus sp.) CM-HZX1 and be blank, 30 DEG C, under 150r/min shaking table culture condition, separated in time centrifuging and taking supernatant liquor utilizes 4-AA direct spectrophotometry to detect the value of volatile phenol.
As can be seen from Figure 3, bacterial strain of the present invention is when phenol concentration is 500mg/L, and initial p H is 7.0 ~ 7.5, and degradation rate is the highest, illustrates that the degradation effect of this bacterial strain Pyrogentisinic Acid under neutrality alkaline condition on the weak side is best.Illustrate that microorganism tenable environment scope is wide, effective.
(2) bacterial strain optimum growth temperature of the present invention
Prepare the phenol substratum of about 500mg/L concentration, switching volume is about 15%, centrifuging and taking bacterium mud adds in phenol substratum, initial pH is 7, rotating speed is that 150r/min shaking table is cultivated, set temperature is respectively 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, simultaneously with do not add rhodococcus (Rhodococcus sp.) CM-HZX1 and be blank, separated in time centrifuging and taking supernatant liquor utilizes 4-AA direct spectrophotometry to detect the value of volatile phenol, judges optimum growth temperature.
As shown in Figure 4, in the present invention, bacterial strain the suitableeest degraded culture temperature under 500mg/L phenol concentration is 30 DEG C ~ 35 DEG C.
(3) bacterial strain the most suitable growth rotating speed of the present invention
Prepare the phenol substratum of about 500mg/L concentration, switching volume is about 15%, centrifuging and taking bacterium mud adds in phenol substratum, initial pH is 7, temperature is 30 DEG C, arranges the shaking table that rotating speed is respectively 100r/min, 150r/min, 200r/min and cultivates, simultaneously with do not add rhodococcus (Rhodococcus sp.) CM-HZX1 and be blank, separated in time centrifuging and taking supernatant liquor utilizes 4-AA direct spectrophotometry to detect the value of volatile phenol, judges optimum growing condition.
As shown in Figure 5, in the present invention, bacterial strain the suitableeest degraded cultivation rotating speed under 500mg/L phenol concentration is 150r/min.
The salt resistance of the Rhodococcus strain of embodiment 3 degradation of phenol
In the industrial phenolic wastewater of complexity, saltiness is higher, by changing salts contg in substratum, investigates the saline-alkaline tolerance of the Rhodococcus strain of degradation of phenol.Prepare the phenol substratum of about 500mg/L concentration, switching volume is about 15%, centrifuging and taking bacterium mud adds to phenol substratum, initial pH is 7, at 30 DEG C, 150r/min shaking table is cultivated, separated in time section, and centrifuging and taking supernatant liquor utilizes 4-AA direct spectrophotometry to detect the value of volatile phenol.
As shown in Figure 6, bacterial strain of the present invention is when salts contg is 0.5 ~ 4%, and phenol degrading is all about 90%, illustrates that this microorganism belongs to moderate salt-durable microbe.
The degraded of embodiment 4 pairs of Low Concentration Phenols
The phenol minimal medium of preparation 10mg/L, 20mg/L, 50mg/L, 100mg/L concentration, bacterial strain volume of the present invention of transferring is about 15%, and centrifuging and taking bacterium mud adds to phenol minimal medium, and initial pH is 7,30 DEG C, the cultivation of 150r/min shaking table, at separated in time, centrifuging and taking supernatant liquor, utilizes 4-AA direct spectrophotometry to detect residue phenol amount, after 18h, degradation rate is respectively 90.6%, 92.5%, 95.4%, 96.6%.
Embodiment 5 Rhodococcus strain is to the degraded of high concentration phenol
The phenol minimal medium of preparation 1000mg/L, 1500mg/L concentration, bacterial strain volume of the present invention of transferring is about 15%, centrifuging and taking bacterium mud adds to phenol minimal medium, initial pH is 7, and at 30 DEG C, 150r/min shaking table is cultivated, after 2d, centrifuging and taking supernatant liquor, utilize 4-AA direct spectrophotometry to detect residue phenol amount, degradation rate is respectively 95.5%, 97.8%.
Embodiment 6 Rhodococcus strain is to the degraded of different concns gradient phenol
The phenol minimal medium of preparation different concns, be respectively 200mg/L, 300mg/L, 400mg/L, 500mg/L, 600mg/L, 700mg/L, 800mg/L, 900mg/L, bacterial strain volume of the present invention of transferring is about 15%, centrifuging and taking bacterium mud adds to phenol minimal medium, initial pH is 7, 30 DEG C, when 24h cultivated by 150r/min shaking table, centrifuging and taking supernatant liquor, 4-AA direct spectrophotometry is utilized to detect residue phenol amount, degradation rate is respectively 99.1%, 92.0%, 90.6%, 93.6%, 91.2%, 92.4%, 91.6%, 90.3%, during 48h, degradation rate is respectively 99.1%, 99.1%, 98.6%, 91.9%, 91.9%, 90.3%, 90.9%, 92.1%.
The phenol of embodiment 7 Rhodococcus strain to simulated wastewater and the degraded of CODcr
The simulation petrochemical wastewater of preparation 500mg/L concentration phenol, bacterial strain volume of the present invention of transferring is about 15%, centrifuging and taking bacterium mud adds in waste water, 30 DEG C, 150r/min shaking table cultivate 24h time, centrifuging and taking supernatant liquor, utilize 4-AA direct spectrophotometry to detect residue phenol amount, degradation rate reaches more than 90%, utilize dichromate titration to detect chemical oxygen demand (COD), CODcr degraded reaches more than 70%.
Embodiment 8 Rhodococcus strain is applied to the improvement of phenols polluted-water
By bacterial strain of the present invention: with the mixture of polyvinyl alcohol and sodium alginate for carrier, and add gac, boric acid, calcium chloride etc. and carry out embedding and be prepared into microbe microsphere, each material composition and embedding method reference literature: rascal Buddhist nun, Cui Yingde, Yan Zhaoqiang, PVA-Alginate-Active Carbon Copolymer Hydrogel Oxygen Permeability is studied, material Leader, 2008,22 (4).
Sanitary sewage is added to as handling object using the phenolic wastewater of certain petrochemical industry, get its waste water and 30%, 60%, 100% add in microbe microsphere and progressively tame by volume, the domestication time is 3 days, temperature 30 ~ 35 DEG C, make microbe microsphere be suitable for wastewater environment, recover the ability of its dephenolize.
10% be added in reactor constructed in advance by volume by the microbe microsphere being embedded with bacterium of the present invention of having tamed, this pond has the functions such as aeration, water circulation and microballoon retain.
Before administering, water quality is poor, and have peculiar smell, phenol concentration is 100 ~ 150mg/L; After improvement, in water quality, phenol clearance reaches more than 95%.
Embodiment 9 Rhodococcus strain is applied to the improvement of phenolic wastewater
Adopt biological reinforced aerobic fluidized bed technique to carry out phenolic wastewater treatment test, sewage is the coking chemical waste water of certain petro-chemical corporation.
With reference to embodiment 2 by bacterial strain enlarged culturing of the present invention after 3 days, be 10% add in biological fluidized bed by volume by bacterium liquid, meanwhile, add polyurethane sponge filler in pond, the filling ratio of polyurethane sponge filler is 20%; Biofilm 5 ~ 6 days, observes strain growth situation on filler; After biofilm completes, enter coking chemical waste water continuously, detect the phenol content of water inlet and water outlet every day.
Coking chemical waste water phenol removes lab scale operational conditions and processing parameter: the pH regulator of water inlet is 7.0 ~ 8.0, flooding velocity controls at 0.9L/h, hydraulic detention time is 12h, detect influent quality situation, in water body, dissolved oxygen stable is stabilized between 7.0 ~ 8.0 at 4 ~ 6mg/L, pH, and temperature does not need strict control, can change with variation of ambient temperature, detect this pond water outlet phenol content.The phenol concentration of coking chemical waste water water inlet is 400 ~ 600mg/L, and after using bacterial strain process of the present invention, water outlet phenol concentration clearance is more than 93%.
Claims (8)
1. a Rhodococcus strain, is characterized in that, called after rhodococcus (Rhodococcus sp.) CM-HZX1, and deposit number is CCTCC NO:M 2014329, and preservation date is on July 9th, 2014.
2. Rhodococcus strain as claimed in claim 1, it is characterized in that, the 16S rDNA sequence of described rhodococcus (Rhodococcus sp.) CM-HZX1 is as shown in SEQ ID:1.
3. Rhodococcus strain as claimed in claim 1, it is characterized in that, the biological property of described rhodococcus (Rhodococcus sp.) CM-HZX1 is: bacterium colony is circular, moistening, orange, opaque, be easy to provoke, neat in edge, bacterial strain diameter is 819 ~ 1120nm, and gramstaining reaction is positive.
4. a cultural method for Rhodococcus strain, is characterized in that, after the Rhodococcus strain activation described in any one of claims 1 to 3, is inoculated into that initial pH value is 6 ~ 9, temperature is 25 ~ 35 DEG C, the mass concentration of salt is cultivate in the substratum of 0 ~ 5%.
5. the application of Rhodococcus strain as claimed in claim 1 in process phenol waste water.
6. apply as claimed in claim 5, it is characterized in that, comprising: after described Rhodococcus strain immobilization, then carry out biochemical treatment in waste water.
7. one kind comprises the microbial inoculum of Rhodococcus strain according to claim 1.
8. one kind comprises the microbe microsphere of Rhodococcus strain according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN108130288A (en) * | 2017-12-15 | 2018-06-08 | 浙江工业大学 | The application of Rhodococcus ruber and its degradable organic pollutant |
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| CN110564716A (en) * | 2019-08-05 | 2019-12-13 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN111269848A (en) * | 2019-12-30 | 2020-06-12 | 浙江工业大学 | Rhodococcus ruber JJ-3 and application thereof in acrylic acid degradation |
| CN113151093A (en) * | 2021-04-25 | 2021-07-23 | 武汉轻工大学 | Strain capable of degrading 3, 5-dimethylphenol and application thereof |
| CN113215072A (en) * | 2021-06-04 | 2021-08-06 | 湖北大学 | Rhodococcus bifidus with strong phenol reducing capability and domestication method thereof |
| CN115725455A (en) * | 2022-10-07 | 2023-03-03 | 安徽农业大学 | Alpha-naphthol degrading bacterium and application thereof |
| CN115806918A (en) * | 2022-12-23 | 2023-03-17 | 中国科学院青岛生物能源与过程研究所 | Rhodococcus and application thereof |
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| CN105039212A (en) * | 2015-07-09 | 2015-11-11 | 中国海洋石油总公司 | Rhodococcus strain C3, microbial agent containing rhodococcus strain C3, and applications of rhodococcus strain C3 and microbial agent |
| CN105219684A (en) * | 2015-11-06 | 2016-01-06 | 安徽农业大学 | The complex microorganism preparations of degrading phenol endocrine disrupter and preparation method |
| CN105219684B (en) * | 2015-11-06 | 2018-01-23 | 安徽全民环保科技有限公司 | The complex microorganism preparations and preparation method of degrading phenol incretion interferent |
| CN109423458A (en) * | 2017-08-30 | 2019-03-05 | 中国石油化工股份有限公司 | A kind of Rhodococcus sp and its identification method and application |
| CN109423458B (en) * | 2017-08-30 | 2022-08-19 | 中国石油化工股份有限公司 | Rhodococcus and identification method and application thereof |
| CN107828679A (en) * | 2017-10-27 | 2018-03-23 | 中国水产科学研究院南海水产研究所 | The rose-red Rhodococcus strain XHRR1 of ammonia and its application in a kind of purifying aquatic water |
| CN107828679B (en) * | 2017-10-27 | 2021-04-27 | 中国水产科学研究院南海水产研究所 | Rhodococcus roseus strain XHRR1 for purifying ammonia in aquaculture water and application thereof |
| CN108130288B (en) * | 2017-12-15 | 2020-10-09 | 浙江工业大学 | Rhodococcus ruber and application thereof in degrading organic pollutants |
| CN108130288A (en) * | 2017-12-15 | 2018-06-08 | 浙江工业大学 | The application of Rhodococcus ruber and its degradable organic pollutant |
| CN109182205A (en) * | 2018-10-09 | 2019-01-11 | 北京林业大学 | Rhodococcus sp and its application with carbon sequestration capacity |
| CN110564716B (en) * | 2019-08-05 | 2021-07-20 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN110564716A (en) * | 2019-08-05 | 2019-12-13 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN111269848A (en) * | 2019-12-30 | 2020-06-12 | 浙江工业大学 | Rhodococcus ruber JJ-3 and application thereof in acrylic acid degradation |
| CN111269848B (en) * | 2019-12-30 | 2022-03-18 | 浙江工业大学 | Rhodococcus ruber JJ-3 and application thereof in acrylic acid degradation |
| CN113151093A (en) * | 2021-04-25 | 2021-07-23 | 武汉轻工大学 | Strain capable of degrading 3, 5-dimethylphenol and application thereof |
| CN113151093B (en) * | 2021-04-25 | 2022-03-18 | 武汉轻工大学 | Strain capable of degrading 3, 5-dimethylphenol and application thereof |
| CN113215072A (en) * | 2021-06-04 | 2021-08-06 | 湖北大学 | Rhodococcus bifidus with strong phenol reducing capability and domestication method thereof |
| CN115725455A (en) * | 2022-10-07 | 2023-03-03 | 安徽农业大学 | Alpha-naphthol degrading bacterium and application thereof |
| CN115725455B (en) * | 2022-10-07 | 2023-10-13 | 安徽农业大学 | Alpha-naphthol degrading bacteria and application thereof |
| CN115806918A (en) * | 2022-12-23 | 2023-03-17 | 中国科学院青岛生物能源与过程研究所 | Rhodococcus and application thereof |
| CN115806918B (en) * | 2022-12-23 | 2024-02-20 | 中国科学院青岛生物能源与过程研究所 | Rhodococcus and application thereof |
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