CN105018392A - Long-chain alkane degrading bacterium and application thereof - Google Patents
Long-chain alkane degrading bacterium and application thereof Download PDFInfo
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Abstract
The invention relates to a long-chain alkane degrading bacterium and application thereof. According to the technical scheme, the bacterium is Acinetobacter sp. C3 and collected by China center for type culture collection (CCTCC), wherein CCTCC NO:M2015394, and the collection date is June 19, 2015. The GenBank accession number of 16S rDNA of the Acinetobacter sp. C3 is KR072554. The Acinetobacter sp. C3 is a Gram-negative bacterium, bacterial colonies are regularly round, the edges are neat, surfaces are smooth, and the color of the bacterial colonies is white; through Gram staining, the bacterium is negative under a microscope, and bacterium bodies are in a short rod shape. In the fermentative degradation process, the degrading bacterium generates phosphatide-type biological surfactant, can promote long-chain alkane solubilization, emulsifying, moisturizing and permeating and efficiently degrade long-chain alkane (C8-C40) in coke wastewater and petrochemical wastewater, and achieves biological enhancement depth regeneration treatment of industrial wastewater.
Description
Technical field
The invention belongs to environment pollutant biological treatment technical field.Be specifically related to strain long-chain alkane degrading bacterium and an application thereof.
Background technology
Along with expanding economy, increasing by the demand of cooling, emulsification, anticorrosion, wear-resistant and clean long chain alkane class fine chemicals in metallurgical production, use to result in Metallurgical Waste Water widely and there is long chain alkane material, this kind of material due to stable chemical nature, hydrophobicity strong, make bioavailability poor, being difficult to natural degradation, is that severe persistent destroys one of organic pollutant of ecotope.As, coking chemical waste water also contains a large amount of long chain alkanes after biochemical treatment, and causes water quality not reach the standard of industrial regeneration water.Therefore, the research control method of effectively removing long chain alkane to keeping benign circulation of the eco-environment, the Sustainable development tool that facilitates economic and social is of great significance.
Long chain alkane pollutant Treatment process in trade effluent mainly contains Physical, chemical method and biological process.Although Physical and chemical method administer long chain alkane pollutant can obtain good effect, cost is high and easily cause secondary pollution and limit practical application.Advantages such as biological process is administered long chain alkane pollutant and had that treatment effect is good, cost is low, simple to operate, green, non-secondary pollution and applied range and getting more and more people's extensive concerning.
At present, the bacterium of the petroleum hydrocarbon degrading microorganism reported and fungi have genus more than 100, and bacterium is more, and fungi is less.Can the mainly containing of decomposing petroleum hydrocarbon in bacterium: Rhodopseudomonas, achromobacter, Alkaligenes, bacillus, genus arthrobacter, micrococcus sp, Flavobacterium and acinetobacter etc.The degradation efficiency of these bacteriums to petroleum alkane awaits improving, and namely can not meet the practical application of alkane degradation, particularly very few to the bacterial classification report of acinetobacter.As " a kind of thick oil-degrading bacteria and application thereof " (CN 101921722 A) patented technology, the feature of this technology is the acinetobacter calcoaceticus obtaining energy degrade viscous crude oil, but there is the shortcomings such as strong to environmental adaptation, degradation efficiency is low; " high-efficiency grease degradation bacteria and application thereof " (CN 103525720 A) patented technology, the feature of this technology obtains degrading the acinetobacter calcoaceticus of grease, mainly for the degraded of commercial grease and food garbage pollutent, also there is the deficiencies such as poor operability, the degradation efficiency scope of application that is low and bacterial strain is limited.
Summary of the invention
The present invention is intended to overcome the deficiencies in the prior art, and object is to provide a strain long-chain alkane degrading bacterium, and this bacterial classification can be degraded the long chain alkane component in trade effluent efficiently, realizes the biological reinforced deep regenerative process of trade effluent.
For achieving the above object, the technical solution used in the present invention is:
One strain long-chain alkane degrading bacterium, derives from the contaminated soil of wuhan petrochemical industry factory, obtained through artificial enrichment, screening and separation and purification.This bacterial strain is acinetobacter calcoaceticus (Acinetobacter sp.) C3, and by China typical culture collection center (Wuhan University) preservation, preserving number is: CCTCC NO:M 2015394, and preservation date is on June 19th, 2015; The GenBank accession number of the 16S rDNA of described acinetobacter calcoaceticus (Acinetobacter sp.) C3 is KR072554; Strains A cinetobacter sp.C3 is a strain Gram-negative bacteria, and bacterium colony is the circle of rule, neat in edge, smooth surface, and colony colour is in white; By being negative under the microscope after gramstaining, thalline is rod-short.
Strains A cinetobacter sp.C3 provided by the invention is applied to the C8 ~ C40 component in treatment of Coking Wastewater and petrochemical wastewater, and C8 ~ C40 component obtains effective removal; A kind of phospholipid bio-surfactant is generated in fermentative degradation process.Because it has stronger emulsifying effect, solublization, wetting and osmosis, add solvability and the transmission speed of long chain alkane, thus improve the degradation efficiency of strains A cinetobacter sp.C3 to long chain alkane.
The present invention compared with prior art, has the following advantages:
1, strains A cinetobacter sp.C3 provided by the invention generates a kind of phospholipid bio-surfactant in fermentative degradation process, and productive rate is 3 ~ 10g/L; The surface tension of solution can be reduced to 20.79mN/m by initial 80.03mN/m by this tensio-active agent, and 72h internal emulsification ability reaches 60 ~ 100%.
2, strains A cinetobacter sp.C3 provided by the invention has efficiently degraded long chain alkane ability, is applicable to effective degraded of the long chain alkane to varying environment, different concns: be the degradation rate of the n-hexadecane of 500 ~ 5000mg/L to concentration be 85 ~ 100%; Be the degradation rate of the bavin wax mixture of 500 ~ 10000mg/L to concentration be 78 ~ 92%; After being the wastewater treatment of 80 ~ 400mg/L to COD in metallurgy industry (independent coke-oven plant, Steel Complex and New Coal Chemical enterprise etc.), the COD of effluent quality is 30 ~ 60mg/L, reaches the standard (COD≤60mg/L) of industrial regeneration water; 52 ~ 73%, 87 ~ 93% and 88.5 ~ 95.2% are respectively to the clearance of long chain alkane in the waste water of independent coke-oven plant, Steel Complex and New Coal Chemical enterprise.
Therefore, strains A cinetobacter sp.C3 provided by the invention is applied to the improvement of coking chemical waste water and petrochemical wastewater, has good degradation effect, achieves the biological reinforced deep regenerative process of trade effluent.
Accompanying drawing explanation
Fig. 1 is the gramstaining figure of strains A cinetobacter sp.C3 of the present invention;
The infrared absorpting light spectra of the bio-surfactant that Fig. 2 produces for the sp.C3 of strains A cinetobacter shown in Fig. 1;
The mensuration figure of the bio-surfactant micelle-forming concentration that Fig. 3 produces for the sp.C3 of strains A cinetobacter shown in Fig. 1;
The mensuration figure of the bio-surfactant emulsifying property that Fig. 4 produces for the sp.C3 of strains A cinetobacter shown in Fig. 1;
Fig. 5 is for the sp.C3 of strains A cinetobacter shown in Fig. 1 is to the degradation curve figure of petroleum hydrocarbon;
Fig. 6 is for the sp.C3 of strains A cinetobacter shown in Fig. 1 is to the gas chromatography mass spectrometry figure before coking wastewater degradation;
Fig. 7 is for the sp.C3 of strains A cinetobacter shown in Fig. 1 is to the gas chromatography mass spectrometry figure after coking wastewater degradation.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is conducted further description, the restriction not to its protection domain.
embodiment 1
The acquisition of strains A cinetobacter sp.C3 and qualification.The acquisition of strains A cinetobacter sp.C3 and the step of qualification are:
Step one, sample collecting and enrichment culture
Pedotheque is gathered from wuhan petrochemical industry factory director phase polluted soil, the pedotheque gathered is added in distilled water, shake up, in the centrifugal 10min of 1000 × g, get supernatant, v) being inoculated in n-hexadecane concentration by inoculum size 4%(v ︰ is in the LB liquid nutrient medium of 500mg/L, be 125r/min and temperature is cultivate 24h in the constant-temperature table of 35 DEG C at rotating speed, obtains just nutrient solution.
Getting just nutrient solution 4mL, to transfer in n-hexadecane concentration be in the minimal medium of 500mg/L, be 125r/min and temperature is cultivate 36h in the constant-temperature table of 35 DEG C at rotating speed, obtains nutrient solution.
Get again nutrient solution 4mL transfer in containing n-hexadecane be in the minimal medium of sole carbon source, transfer 6 times, increase the concentration of n-hexadecane in switching process gradually, concentration is followed successively by: 1000mg/L, 1500mg/L, 2000mg/L, 3000mg/L, 4000mg/L and 5000mg/L; Then at rotating speed, to be 125r/min and temperature be in the constant-temperature table of 35 DEG C that training cultivates 36h, obtains whole nutrient solution.
Step 2, separation and purification
Get whole nutrient solution 1 mL, with sterilized water by its by volume successively gradient dilution be 10
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6, 10
-7with 10
-8bacteria suspension, the bacteria suspension then getting each concentration gradient that 200 μ L have diluted is coated on solid LB flat board that n-hexadecane concentration is 1000mg/L (each concentration gradient does three repetitions) equably, cultivates in the constant incubator of 35 DEG C; After growing bacterium colony, picking colony is that the solid inorganic salt culture medium flat plate of 1000mg/L is rule in n-hexadecane concentration, be upside down in constant temperature culture 1 ~ 2d in 35 DEG C of biochemical cultivation cases, picking list bacterium colony, adopt plate streaking partition method by bacterium colony at the flat lining out purifying of inorganic salt, repeat line 4 ~ 5 times, obtain the single bacterium colony after purifying.The all single colony inoculation of picking is in the minimal medium of 1000mg/L in n-hexadecane concentration, be 125r/min and temperature in rotating speed be cultivate 48h in the shaking table of 35 DEG C, vapor-phase chromatography is adopted to detect the residual concentration of n-hexadecane in the rear minimal medium of degraded, determine the bacterial strain that degradation rate is the highest, be numbered C3.
The qualification of step 3, bacterial strain
Be inoculated in by bacterial strain C3 on LB solid medium, constant temperature culture 48h in 35 DEG C of biochemical cultivation cases, bacterium colony is the circle of rule, neat in edge, smooth surface, and colony colour is in white; By being negative under the microscope after gramstaining, thalline is rod-short as shown in Figure 1; The 16S rDNA gene order length of bacterial strain C3 is 1414bp, and as shown in sequence table, after being submitted to GenBank, the accession number of sequence is KR072554, retrieves through NCBI Blast, and the similarity of bacterial strain C3 of the present invention and Acinetobacter sp. reaches 99%.Therefore, according to the morphological specificity of bacterial strain C3 and physiological and biochemical property and 16S rDNA gene sequence characteristic thereof, determine that bacterial strain C3 belongs to acinetobacter (Acinetobacter sp.), called after Acinetobacter sp.C3.
Described LB culture medium prescription is: yeast extractive substance 5g, peptone 10g, sodium-chlor 10g, distilled water 1L.The pH value of described enrichment medium is 6.8 ~ 7.2, for subsequent use at 121.3 DEG C of moist heat sterilization 20 min.
Described minimal medium formula is: ammonium nitrate 1g, Sodium phosphate dibasic 1.42g, potassium primary phosphate 1.36g, magnesium sulfate heptahydrate 0.432g, calcium chloride 0.006g, micro-1mL, distilled water 1L.The pH value of substratum is 6.8 ~ 7.2, at 121.3 DEG C of moist heat sterilization 20min.Wherein trace element formula: manganese sulfate monohydrate 1.69g/L, CoCL2 6H2O 0.24g/L, boric acid 1.16g/L, Sodium Molybdate Dihydrate 0.024g/L, iron vitriol 2.78g/L, Zinc Sulphate Heptahydrate 1.15g/L, cupric sulfate pentahydrate 0.38g/L.
Described n-hexadecane be with 0.2 μm of filtering with microporous membrane after product.
Described LB solid culture based formulas: add 2%(w/v on LB substratum) agar, high-temperature heat sterilization.
Described inorganic salt solid culture based formulas: add 2%(w/v on minimal medium formula) agar, high-temperature heat sterilization.
embodiment 2
Temperature is on the impact of strains A cinetobacter sp.C3 degraded n-hexadecane.Strains A cinetobacter sp.C3 being seeded to n-hexadecane concentration is in the minimal medium of 1000mg/L, be that 150r/min and culture temperature are respectively in the constant-temperature table of 25 DEG C, 30 DEG C, 35 DEG C and 40 DEG C and cultivate 48h at rotating speed, adopt vapor-phase chromatography detected temperatures on the impact of strains A cinetobacter sp.C3 degraded n-hexadecane, the results are shown in Table 1.
Table 1 temperature is on the impact of strains A cinetobacter sp.C3 degraded n-hexadecane
| Temperature (DEG C) | Degradation rate (%) |
| 25 | 82.8 |
| 30 | 96.4 |
| 35 | 98.6 |
| 40 | 86.3 |
As can be seen from Table 1, strains A cinetobacter sp C3 has certain degradation effect to n-hexadecane within the scope of 25 ~ 40 DEG C, and 35 DEG C time, degradation effect is best.Strains A cinetobacter sp.C3 also has good degradation rate at comparatively high temps 40 DEG C, reaches 86.3%.
The present embodiment illustrates that strains A cinetobacter sp C3 is better to the degradation property of long chain alkane in different temperature environments.
embodiment 3
PH value is on the impact of strains A cinetobacter sp.C3 degraded n-hexadecane.Strains A cinetobacter sp.C3 being seeded to n-hexadecane concentration is in the minimal medium of 1000mg/L, adjust pH is respectively 6.5,6.8,7.0,7.2 and 7.5, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, adopt vapor-phase chromatography to detect the impact of pH on strains A cinetobacter sp.C3 degraded n-hexadecane, the results are shown in Table 2.
Table 2 pH value is on the impact of strains A cinetobacter sp.C3 degraded n-hexadecane
| pH | Degradation rate (%) |
| 6.5 | 79.7 |
| 6.8 | 90.2 |
| 7.0 | 99.1 |
| 7.2 | 97.5 |
| 7.5 | 83.2 |
As can be seen from Table 2, when pH value is 7.0, strains A cinetobacter spC3 is to the degradation rate of n-hexadecane up to 99.1%, and pH value has good degradation rate (being greater than 90%) 6.8 ~ 7.2.
The present embodiment illustrates that the degradation property of strains A cinetobacter sp.C3 to long chain alkane in neutral and that meta-alkalescence is wider pH value range is better.
embodiment 4
N-hexadecane starting point concentration is on the impact of strains A cinetobacter sp.C3 degradation property.Strains A cinetobacter sp.C3 to be seeded to n-hexadecane starting point concentration be 500 ~ 5000mg/L and pH value is in the minimal medium of 7.0, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, adopt vapor-phase chromatography to detect the residual concentration of n-hexadecane, the results are shown in Table 3.
Table 3 n-hexadecane starting point concentration is on the impact of strains A cinetobacter sp.C3 degradation property
| N-hexadecane concentration (mg/L) | Degradation rate (%) |
| 500 | 99.4 |
| 1000 | 100 |
| 2000 | 98.4 |
| 4000 | 96.3 |
| 5000 | 85 |
The degradation rate of strains A cinetobacter sp C3 to n-hexadecane reaches 85 ~ 100% as can be seen from Table 3.
The present embodiment illustrates that strains A cinetobacter sp C3 is better to the degradation property of long chain alkane in wider concentration range, for its application in actual environment provides guarantee.
embodiment 5
The extraction of the bio-surfactant that strains A cinetobacter sp.C3 produces and performance study.By 1 ~ 2%(v/v) the bacterial suspension inoculation of strains A cinetobacter sp C3 be in the minimal medium of 500 ~ 5000mg/L to n-hexadecane concentration, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, obtain fermented liquid; Then by fermented liquid centrifugal 15 ~ 20min under 8000 ~ 12000r/min and 4 ~ 8 DEG C condition, supernatant liquor is collected; Adjust supernatant liquor pH value to 2 ~ 3; By chloroform/methanol solution, (2/1, v/v) ︰ pH is the volume ratio of the supernatant liquor of 2 ~ 3 is that 1 ︰ 0.5 ~ 1 carries out extractive distillation again; By the organic phase after extraction after underpressure distillation under 20 ~ 30 DEG C and-0.1MPa, obtain yellow surfaces promoting agent, productive rate is 3 ~ 10g/L.
Adopt infrared analysis and thin layer chromatography (TLC) to carry out category authentication to the bio-surfactant that strains A cinetobacter sp.C3 produces, and the CMC value of bio-surfactant, oil extraction circle value and emulsifying property are measured.
Bio-surfactant Infrared spectrum scanning result as shown in Figure 2,3436cm
-1place is-OH absorption, 1649cm
-1place is NH
+absorb; 1543cm
-1and 1389cm
-1for the absorption of C=O; 1079cm
-1place may be the absorption of PO-R; 950cm
-1for the absorption of P=0; 668cm
-1place is the absorption of P=S, 511cm
-1for the absorption of P-S.This bio-surfactant is initially identified as phospholipid promoting agent.
By extract bio-surfactant with kapillary point sample in silica gel thin sheet, put into chromatography cylinder and carry out thin-layer chromatography.Developping agent is chloroform/methanol/water=65/15/2 (v/v/v).Be developer with perchloric acid, the characteristic color reaction of solution is aobvious blue, illustrates that this tensio-active agent is phospholipid surfactant.Consistent with Infrared spectrum scanning qualification result, therefore, can identify that the tensio-active agent that strains A cinetobacter sp.C3 produces is phospholipid surfactant.
The measuring result of CMC value as shown in Figure 3, when surfactant concentration is lower than 35mg/L, along with surfactant concentration increases, the corresponding reduction of surface tension of water, when surfactant concentration is higher than 35mg/L, increasing surfactant concentration can not make the surface tension of water reduce further, shows that the micelle-forming concentration of the bio-surfactant that strains A cinetobacter sp.C3 produces is about 35mg/L, the surface tension of water can be reduced to 20mN/m.
Oil extraction circle value: get culture dish, add water, the water surface adds 0.1ml n-hexadecane, forms oil film.Add the bio-surfactant of extraction at oil film center, center oil film is pressed against surrounding and forms a circle, and the diameter measuring circle is 80mm.
As shown in Figure 4,72h internal emulsification ability is 60 ~ 100% to the emulsifying property result of the bio-surfactant that strains A cinetobacter sp.C3 produces.Illustrate to there is the surfactant emulsifies better performances that bacterial strain C3 produces good solubilizing effect, emulsifying capacity and wetting penetrating power.
embodiment 6
Strains A cinetobacter sp.C3 is to the degraded of certain independent Coking Plant Wastewater.Gather waste water after certain independent coke-oven plant biochemistry, containing a certain amount of C8 ~ C16 alkane in waste water, COD is 200 ~ 300mg/L.Strains A cinetobacter sp.C3 is seeded in the flask that waste water after this independent coke-oven plant biochemistry is housed, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, this independent Coking Plant Wastewater COD can be reduced to 50 ~ 58.5mg/L, reach the standard (COD≤60mg/L) of industrial regeneration water; In coking chemical waste water before and after GC-MS analysis strains A cinetobacter sp.C3 process, the clearance of C8 ~ C16 alkane is 52 ~ 73%, and most of alkane is removed.Strains A cinetobacter sp.C3 also has good degradation capability to the organism except long chain alkane.
embodiment 7
Strains A cinetobacter sp.C3 is to the degraded of certain New Coal Chemical enterprise wastewater.Gather waste water after certain New Coal Chemical enterprise biochemistry, the long chain alkane containing a large amount of C16 ~ C24 in waste water, COD is 300 ~ 400mg/L.Strains A cinetobacter sp.C3 is seeded in the flask that this New Coal Chemical enterprise wastewater is housed, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, this New Coal Chemical enterprise wastewater COD can be reduced to 40 ~ 60mg/L, reach the standard (COD≤60mg/L) of industrial regeneration water; In coking chemical waste water before and after GC-MS analysis strains A cinetobacter sp.C3 process, the clearance of C16 ~ C24 alkane is 87 ~ 93%.Strains A cinetobacter sp.C3 centering long chain alkane has good degradation effect.
embodiment 8
Strains A cinetobacter sp.C3 is to the degraded of petroleum hydrocarbon.Strains A cinetobacter sp.C3 is seeded to containing 1%(v ︰ v) bavin wax (Chai You ︰ paraffin=1: 1, in v ︰ minimal medium v) (long chain alkane carbonatoms is C20 ~ C30), be 150r/min and temperature at rotating speed be cultivate in the constant-temperature table of 35 DEG C, utilize the degradation rate of Their Determination by Spectrophotometry nutrient solution Petroleum Hydrocarbon every 12h, experimental result as shown in Figure 5.As shown in Figure 5, after 96 ~ 156h, the degradation rate of the bavin wax mixing solutions of 500 ~ 10000mg/L reaches 78 ~ 92%.
embodiment 9
Strains A cinetobacter sp.C3 is to the degraded of certain associating Coking Plant Wastewater.Gather waste water after certain associating coke-oven plant biochemical treatment, after advanced treatment in waste water containing a large amount of C30 ~ C40 long chain alkanes, COD is 80 ~ 200mg/L, and result as shown in Figure 6, fails to reach industrial regeneration water water quality standard.Strains A cinetobacter sp.C3 is seeded in the flask that this New Coal Chemical enterprise wastewater is housed, be 150r/min and temperature at rotating speed be cultivate 48h in the constant-temperature table of 35 DEG C, water outlet COD can be reduced to 30 ~ 60mg/L, result as shown in Figure 7, reaches the standard (COD≤60mg/L) of industrial regeneration water; In coking chemical waste water before and after GC-MS analysis strains A cinetobacter sp.C3 process, the clearance of C30 ~ C40 long chain alkane is 88.5 ~ 95.2%.
This embodiment has the following advantages:
1, the strains A cinetobacter sp.C3 that this embodiment provides generates a kind of phospholipid bio-surfactant in fermentative degradation process, and productive rate is 3 ~ 10g/L; The surface tension of solution can be reduced to 20.79mN/m by initial 80.03mN/m by this tensio-active agent, and 72h internal emulsification ability reaches 60 ~ 100%.
2, the strains A cinetobacter sp.C3 that this embodiment provides has efficiently degraded long chain alkane ability, is applicable to effective degraded of the long chain alkane to varying environment, different concns: be the degradation rate of the n-hexadecane of 500 ~ 5000mg/L to concentration be 85 ~ 100%; Be the degradation rate of the bavin wax mixture of 500 ~ 10000mg/L to concentration be 78 ~ 92%; After being the wastewater treatment of 80 ~ 400mg/L to COD in metallurgy industry (independent coke-oven plant, Steel Complex and New Coal Chemical enterprise etc.), the COD of effluent quality is 30 ~ 60mg/L, reaches the standard (COD≤60mg/L) of industrial regeneration water; 52 ~ 73%, 87 ~ 93% and 88.5 ~ 95.2% are respectively to the clearance of long chain alkane in the waste water of independent coke-oven plant, Steel Complex and New Coal Chemical enterprise.
Therefore, the strains A cinetobacter sp.C3 that this embodiment provides is applied to the improvement of coking chemical waste water and petrochemical wastewater, has good degradation effect, achieves the biological reinforced deep regenerative process of trade effluent.
Sequence table
<1> 1414
<2> DNA
<3> acinetobacter calcoaceticus Acinetobacter sp.
<4>
1 CCATGCAGTC GAGCGGAGTG ATGGTGCTTG CACTATCACT TAGCGGCGGA
51 CGGGTGAGTA ATGCTTAGGA ATCTGCCTAT TAGTGGGGGA CAACATCTCG
101 AAAGGGATGC TAATACCGCA TACGTCCTAC GGGAGAAAGC AGGGGATCAC
151 TTGTGACCTT GCGCTAATAG ATGAGCCTAA GTCGGATTAG CTAGTTGGTG
201 GGGTAAAGGC CTACCAAGGC GACGATCTGT AGCGGGTCTG AGAGGATGAT
251 CCGCCACACT GGGACTGAGA CACGGCCCAG ACTCCTACGG GAGGCAGCAG
301 TGGGGAATAT TGGACAATGG GGGGAACCCT GATCCAGCCA TGCCGCGTGT
351 GTGAAGAAGG CCTTATGGTT GTAAAGCACT TTAAGCGAGG AGGAGGCTCT
401 TTTGGTTAAT ACCCAAGATG AGTGGACGTT ACTCGCAGAA TAAGCACCGG
451 CTAACTCTGT GCCAGCAGCC GCGGTAATAC AGAGGGTGCA AGCGTTAATC
501 GGATTTACTG GGCGTAAAGC GCGCGTAGGC GGCCAATTAA GTCAAATGTG
551 AAATCCCCGA GCTTAACTTG GGAATTGCAT TCGATACTGG TTGGCTAGAG
601 TGTGGGAGAG GATGGTAGAA TTCCAGGTGT AGCGGTGAAA TGCGTAGAGA
651 TCTGGAGGAA TACCGATGGC GAAGGCAGCC ATCTGGCCTA ACACTGACGC
701 TGAGGTGCGA AAGCATGGGG AGCAAACAGG ATTAGATACC CTGGTAGTCC
751 ATGCCGTAAA CGATGTCTAC TAGCCGTTGG GGCCTTTGAG GCTTTAGTGG
801 CGCAGCTAAC GCGATAAGTA GACCGCCTGG GGAGTACGGT CGCAAGACTA
851 AAACTCAAAT GAATTGACGG GGGCCCGCAC AAGCGGTGGA GCATGTGGTT
901 TAATTCGATG CAACGCGAAG AACCTTACCT GGCCTTGACA TAGTAGAAAC
951 TTTCCAGAGA TGGATTGGTG CCTTCGGGAA TCTACATACA GGTGCTGCAT
1001 GGCTGTCGTC AGCTCGTGTC GTGAGATGTT GGGTTAAGTC CCGCAACGAG
1051 CGCAACCCTT TTCCTTACTT GCCAGCATTT CGGATGGGAA CTTTAAGGAT
1101 ACTGCCAGTG ACAAACTGGA GGAAGGCGGG GACGACGTCA AGTCATCATG
1151 GCCCTTACGG CCAGGGCTAC ACACGTGCTA CAATGGTCGG TACAAAGGGT
1201 TGCTACCTAG CGATAGGATG CTAATCTCAA AAAGCCGATC GTAGTCCGGA
1251 TTGGAGTCTG CAACTCGACT CCATGAAGTC GGAATCGCTA GTAATCGCGG
1301 ATCAGAATGC CGCGGTGAAT ACGTTCCCGG GCCTTGTACA CACCGCCCGT
1351 CACACCATGG GAGTTTGTTG CACCAGAAGT AGCTAGCCTA ACTGCAAAGA
1401 GGGCGGTACC A
Claims (3)
1. a long-chain alkane degrading bacterium, it is characterized in that described degradation bacteria is acinetobacter calcoaceticus (Acinetobacter sp.) C3, by China typical culture collection center preservation, preserving number is: CCTCC NO:M 2015394, and preservation date is on June 19th, 2015; The GenBank accession number of the 16S rDNA of described acinetobacter calcoaceticus (Acinetobacter sp.) C3 is KR072554; Strains A cinetobacter sp.C3 is a strain Gram-negative bacteria, and bacterium colony is the circle of rule, neat in edge, smooth surface, and colony colour is in white; By being negative under the microscope after gramstaining, thalline is rod-short.
2. an application for long-chain alkane degrading bacterium, is characterized in that described degradation bacteria is applied to the C8 ~ C40 component in treatment of Coking Wastewater and petrochemical wastewater.
3. the application of long-chain alkane degrading bacterium according to claim 2, is characterized in that described degradation bacteria generates a kind of phospholipid bio-surfactant in fermentative degradation process.
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| CN105733976A (en) * | 2015-12-29 | 2016-07-06 | 中国科学院烟台海岸带研究所 | Composite inoculant for degrading petroleum, preparation method and applications thereof |
| CN106834189A (en) * | 2017-03-09 | 2017-06-13 | 武汉科技大学 | One plant produces biosurfactant bacterium and its application |
| CN110846257A (en) * | 2019-12-11 | 2020-02-28 | 中冶华天工程技术有限公司 | Microbial bacterium for degrading long-chain alkane and application thereof |
| CN111394271A (en) * | 2019-09-12 | 2020-07-10 | 浙江大学 | Acinetobacter for producing surfactant and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105733976A (en) * | 2015-12-29 | 2016-07-06 | 中国科学院烟台海岸带研究所 | Composite inoculant for degrading petroleum, preparation method and applications thereof |
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| CN106834189A (en) * | 2017-03-09 | 2017-06-13 | 武汉科技大学 | One plant produces biosurfactant bacterium and its application |
| CN106834189B (en) * | 2017-03-09 | 2020-02-14 | 武汉科技大学 | Biosurfactant producing bacterium and application thereof |
| CN111394271A (en) * | 2019-09-12 | 2020-07-10 | 浙江大学 | Acinetobacter for producing surfactant and application thereof |
| CN110846257A (en) * | 2019-12-11 | 2020-02-28 | 中冶华天工程技术有限公司 | Microbial bacterium for degrading long-chain alkane and application thereof |
| CN113684153A (en) * | 2021-09-05 | 2021-11-23 | 东北石油大学 | Long-chain alkane degrading bacterium and application thereof |
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| CN105018392B (en) | 2018-05-15 |
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