CN105018392B - One plant of long-chain alkane degrading bacterium and its application - Google Patents
One plant of long-chain alkane degrading bacterium and its application Download PDFInfo
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Abstract
The present invention relates to one plant of long-chain alkane degrading bacterium and its application.Its technical solution is：Bacterial strain provided by the present invention is acinetobacter calcoaceticus (Acinetobacter sp.) C3, and by China typical culture collection center preservation, preserving number is：CCTCC NO：M2015394, preservation date are on June 19th, 2015；The GenBank accession number of the 16S rDNA of acinetobacter calcoaceticus (Acinetobacter sp.) C3 is KR072554；Strains A cinetobacter sp.C3 are one plant of Gram-negative bacteria, and bacterium colony is regular circular, and neat in edge, surface is smooth, and colony colour is white；By being negative under the microscope after Gram's staining, thalline is in rod-short.The degradation bacteria generates a kind of phosphatide type biological surfactant in fermentative degradation process, can promote long chain alkane solubilising, emulsification, wetting and osmosis, long chain alkane that can efficiently in treatment of Coking Wastewater and petrochemical wastewater（C8~C40）, realize to the processing of industrial wastewater biological reinforced deep regenerative.
Description
Technical field
The invention belongs to environment pollutant biological treatment technical field.More particularly to one plant of long-chain alkane degrading bacterium and its answer
With.
Background technology
It is fine with cooling, emulsification, anti-corrosion, wear-resistant and clean long chain alkane class in metallurgical production with expanding economy
The demand of chemicals is increasing, and extensive use result in Metallurgical Waste Water there are long chain alkane material, this kind of material due to
Chemical property is stable, hydrophobicity is strong, makes bioavailability poor, it is difficult to which natural degradation, is that severe persistent destroys ecological environment
One of organic pollution.Such as, coking wastewater also contains substantial amounts of long chain alkane after biochemical treatment, and causes water quality not reach work
The standard of industry recycled water.Therefore, research effectively removes the control method of long chain alkane to keeping benign circulation of the eco-environment, promoting
Economic and society sustainable development tool is of great significance.
Long chain alkane pollutant Treatment process in industrial wastewater mainly has Physical, chemical method and bioanalysis.Although
Physical and chemical method, which administer long chain alkane pollutant, can obtain preferable effect, but cost it is high and easily cause secondary pollution and
Limit practical application.Bioanalysis administer long chain alkane pollutant have high treating effect, cost it is low, it is easy to operate, green,
Non-secondary pollution and get more and more people's extensive concerning the advantages that have a wide range of application.
At present, the bacterium and fungi for the petroleum hydrocarbon degrading microorganism reported have 100 more belong to, bacterium is more, fungi compared with
It is few.Energy decomposing petroleum hydrocarbon mainly has in bacterium：Pseudomonas, achromobacter, alcaligenes, bacillus, section
Bacillus, Micrococcus, Flavobacterium and acinetobacter etc..These bacteriums are to the degradation efficiency of petroleum alkane up for carrying
Height, i.e., cannot meet the practical application of alkane degradation, particularly very few to the strain report of acinetobacter.Such as a kind of " viscous crude
Degradation bacteria and its application "（CN 101921722 A）The characteristics of patented technology, the technology is to obtain the not lever for the viscous crude that can degrade
Bacterium, but exist and adapt to environment the shortcomings such as not strong, degradation efficiency is low；" high-efficiency grease degradation bacteria and its application "（CN
103525720 A）The characteristics of patented technology, the technology is to obtain the acinetobacter calcoaceticus for the grease that can degrade, mainly for commercial grease
And the degraded of food garbage pollutant, there is also poor operability, degradation efficiency it is low and the deficiencies of the scope of application of bacterial strain is limited.
The content of the invention
It is contemplated that overcome the deficiencies in the prior art, it is therefore an objective to provide one plant of long-chain alkane degrading bacterium, which can be high
Long chain alkane component in effect ground degraded industrial wastewater, is realized to the processing of industrial wastewater biological reinforced deep regenerative.
To achieve the above object, the technical solution adopted by the present invention is：
One plant of long-chain alkane degrading bacterium, from the contaminated soil of wuhan petrochemical industry factory, through manually enrichment, screening and separation
Obtained by purifying.The bacterial strain is acinetobacter calcoaceticus (Acinetobacter sp.) C3, (military by China typical culture collection center
Chinese university) preservation, preserving number is：CCTCC NO：M 2015394, preservation date are on June 19th, 2015；The acinetobacter calcoaceticus
The GenBank accession number of the 16S rDNA of (Acinetobacter sp.) C3 is KR072554；Strains A cinetobacter
Sp.C3 is one plant of Gram-negative bacteria, and bacterium colony is regular circular, and neat in edge, surface is smooth, and colony colour is white；It is logical
It is negative under the microscope after crossing Gram's staining, thalline is in rod-short.
Strains A cinetobacter sp.C3 provided by the invention are applied to the C8 in treatment of Coking Wastewater and petrochemical wastewater
~ C40 components, C8 ~ C40 components have obtained effective removal；A kind of phospholipid biological surface is generated in fermentative degradation process to live
Property agent.Since it is with stronger emulsification, solubilization, wetting and osmosis, the dissolubility of long chain alkane is added
And transmission speed, so as to improve degradation efficiencies of the strains A cinetobacter sp.C3 to long chain alkane.
Compared with prior art, the present invention has the following advantages：
1st, strains A cinetobacter sp.C3 provided by the invention generate a kind of phospholipid life in fermentative degradation process
Thing surfactant, yield are 3 ~ 10g/L；The surfactant can be dropped the surface tension of solution by initial 80.03mN/m
Low to arrive 20.79mN/m, 72h internal emulsifications ability is up to 60 ~ 100%.
2nd, strains A cinetobacter sp.C3 provided by the invention have efficient degraded long chain alkane ability, are applicable in
In effective degraded of the long chain alkane to varying environment, various concentrations：Drop to concentration for the hexadecane of 500 ~ 5000mg/L
Solution rate is 85 ~ 100%；Degradation rate to the bavin wax mixture that concentration is 500 ~ 10000mg/L is 78 ~ 92%；To metallurgy industry（Solely
Vertical coke-oven plant, integrated iron and steel works and New Coal Chemical enterprise etc.）Middle COD is water outlet water after the wastewater treatment of 80 ~ 400mg/L
The COD of matter is 30 ~ 60mg/L, reaches the standard of industrial regeneration water（COD≤60mg/L）；To independent coke-oven plant, steel joint enterprise
The removal rate of long chain alkane is respectively 52 ~ 73%, 87 ~ 93% and 88.5 ~ 95.2% in the waste water of industry and New Coal Chemical enterprise.
Therefore, strains A cinetobacter sp.C3 provided by the invention are applied to controlling for coking wastewater and petrochemical wastewater
Reason, there is good degradation effect, realizes to the processing of industrial wastewater biological reinforced deep regenerative.
Brief description of the drawings
Fig. 1 is the Gram's staining figure of the strains A cinetobacter sp.C3 of the present invention；
Fig. 2 is the infrared absorption spectroscopy for the biosurfactant that strains A cinetobacter sp.C3 shown in Fig. 1 is produced
Figure；
Fig. 3 is the biosurfactant critical micelle concentration that strains A cinetobacter sp.C3 shown in Fig. 1 is produced
Measurement chart；
Fig. 4 is the measure for the biosurfactant emulsifiability that strains A cinetobacter sp.C3 shown in Fig. 1 is produced
Figure；
Fig. 5 is degradation curve figures of the strains A cinetobacter sp.C3 shown in Fig. 1 to petroleum hydrocarbon；
Fig. 6 is strains A cinetobacter sp.C3 shown in Fig. 1 to the gas chromatography mass spectrometry figure before coking wastewater degradation；
Fig. 7 is strains A cinetobacter sp.C3 shown in Fig. 1 to the gas chromatography mass spectrometry figure after coking wastewater degradation.
Embodiment
The present invention is further described with reference to the accompanying drawings and detailed description, not to its protection domain
Limitation.
Embodiment 1
The acquisition and identification of strains A cinetobacter sp.C3.The acquisition of strains A cinetobacter sp.C3 and mirror
Fixed step is：
Step 1: sample collection and enrichment culture
Pedotheque is gathered from wuhan petrochemical industry factory director's phase polluted soil, the pedotheque of collection is added distilled water
In, shake up, centrifuge 10min in 1000 × g, supernatant is taken, by inoculum concentration 4%（V ︰ v）It is 500mg/L to be inoculated in hexadecane concentration
LB fluid nutrient mediums in, cultivate 24h in the constant-temperature table that rotating speed is 125r/min and temperature is 35 DEG C, obtain just cultivate
Liquid.
Take first nutrient solution 4mL to transfer in the minimal medium that hexadecane concentration is 500mg/L, be in rotating speed
36h is cultivated in 125r/min and the constant-temperature table that temperature is 35 DEG C, obtains nutrient solution.
Nutrient solution 4mL switchings are taken, in the minimal medium of sole carbon source, to transfer 6 times, to transfer in containing hexadecane again
During gradual increase hexadecane concentration, concentration is followed successively by：1000mg/L、1500mg/L、2000mg/L、3000mg/L、
4000mg/L and 5000mg/L；Then the training culture 36h in the constant-temperature table that rotating speed is 125r/min and temperature is 35 DEG C, obtains
Whole nutrient solution.
Step 2: isolate and purify
Whole 1 mL of nutrient solution is taken, gradient dilution is 10 successively by volume by it with sterile water-1, 10-2, 10-3, 10-4, 10-5, 10-6, 10-7With 10-8Bacteria suspension, then take the bacteria suspension for each concentration gradient that 200 μ L have diluted equably to be coated on just
Hexadecane concentration is on the solid LB tablets of 1000mg/L（Each concentration gradient does three repetitions）, in 35 DEG C of constant incubator
Middle culture；After bacterium colony is grown, picking colony is on the solid inorganic salt culture medium flat plate that hexadecane concentration is 1000mg/L
Line, is upside down in 1 ~ 2d of constant temperature incubation in 35 DEG C of biochemical cultivation cases, picking single bacterium colony, is existed bacterium colony using plate streaking partition method
The flat lining out purifying of inorganic salts, repeats line 4 ~ 5 times, obtains single bacterium colony after purification.All single bacterium colonies of picking are inoculated in
Hexadecane concentration is in the minimal medium of 1000mg/L, is trained in the shaking table that rotating speed is 125r/min and temperature is 35 DEG C
48h is supported, using the residual concentration of hexadecane in minimal medium after gas chromatography detection degraded, determines degradation rate highest
Bacterial strain, numbering C3.
Step 3: the identification of bacterial strain
Bacterial strain C3 is inoculated on LB solid mediums, the constant temperature incubation 48h in 35 DEG C of biochemical cultivation cases, bacterium colony is rule
Circle, neat in edge, surface is smooth, and colony colour is white；By being negative under the microscope after Gram's staining, such as scheme
Thalline shown in 1 is in rod-short；The 16S rDNA gene orders length of bacterial strain C3 is 1414bp, as shown in sequence table, is submitted to
After GenBank, the accession number of sequence is KR072554, through NCBI Blast retrieve, bacterial strain C3 of the invention with
The similitude of Acinetobacter sp. is up to 99%.Therefore according to the morphological feature and physiological and biochemical property and its 16S of bacterial strain C3
RDNA gene sequence characteristics, determine that bacterial strain C3 belongs to acinetobacter（Acinetobacter sp.）, it is named as
Acinetobacter sp.C3。
The LB culture medium prescriptions are：Yeast extract 5g, peptone 10g, sodium chloride 10g, distilled water 1L.The richness
The pH value for collecting culture medium is 6.8 ~ 7.2, spare in 121.3 DEG C of 20 min of moist heat sterilization.
The minimal medium formula is：Ammonium nitrate 1g, disodium hydrogen phosphate 1.42g, potassium dihydrogen phosphate 1.36g, seven
Water magnesium sulfate 0.432g, calcium chloride 0.006g, micro- 1mL, distilled water 1L.The pH value of culture medium is 6.8~7.2,
121.3 DEG C moist heat sterilization 20min.Wherein trace element formula：Manganese sulfate monohydrate 1.69g/L, CoCL2 6H2O 0.24g/L, boron
Sour 1.16g/L, Sodium Molybdate Dihydrate 0.024g/L, ferrous sulfate heptahydrate 2.78g/L, white vitriol 1.15g/L, cupric sulfate pentahydrate
0.38g/L。
The hexadecane is with the product after 0.2 μm of filtering with microporous membrane.
The LB solid culture based formulas：2% is added on LB culture mediums（w/v）Agar, high-temperature heat sterilization.
The inorganic salts solid culture based formulas：2% is added on minimal medium formula（w/v）Agar, high temperature
Moist heat sterilization.
Embodiment 2
Influence of the temperature to strains A cinetobacter sp.C3 degraded hexadecanes.By strains A cinetobacter
Sp.C3 is seeded in the minimal medium that hexadecane concentration is 1000mg/L, is 150r/min and cultivation temperature in rotating speed
48h is cultivated in respectively 25 DEG C, 30 DEG C, 35 DEG C and 40 DEG C of constant-temperature table, using gas chromatography detection temperature to bacterial strain
The influence of Acinetobacter sp.C3 degraded hexadecanes, the results are shown in Table 1.
Influence of 1 temperature of table to strains A cinetobacter sp.C3 degraded hexadecanes
| Temperature（℃） | 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 have necessarily hexadecane in the range of 25 ~ 40 DEG C
Degradation effect, at 35 DEG C, degradation effect is optimal.Strains A cinetobacter sp.C3 at 40 DEG C of higher temperature to also having
Preferable degradation rate, up to 86.3%.
This example demonstrates that degradabilities of the strains A cinetobacter sp C3 to long chain alkane in different temperature environments
Can be preferably.
Embodiment 3
Influence of the pH value to strains A cinetobacter sp.C3 degraded hexadecanes.By strains A cinetobacter
Sp.C3 is seeded in the minimal medium that hexadecane concentration is 1000mg/L, and it is respectively 6.5,6.8,7.0,7.2 to adjust pH value
With 7.5,48h is cultivated in the constant-temperature table that rotating speed is 150r/min and temperature is 35 DEG C, pH pairs is detected using gas chromatography
The influence of strains A cinetobacter sp.C3 degraded hexadecanes, the results are shown in Table 2.
Influence of 2 pH value of table to strains A cinetobacter sp.C3 degraded hexadecanes
| pH | Degradation rate（%） |
| 6.5 | 79.7 |
| 6.8 | 90.2 |
| 7.0 | 99.1 |
| 7.2 | 97.5 |
| 7.5 | 83.2 |
From table 2 it can be seen that when pH value is 7.0, strains A cinetobacter spC3 are high to the degradation rate of hexadecane
Up to 99.1%, pH value has preferable degradation rate 6.8 ~ 7.2（More than 90%）.
This example demonstrates that strains A cinetobacter sp.C3 are to long in neutral and wider meta-alkalescence pH value range
The degradation property of alkane is preferable.
Embodiment 4
Influence of the hexadecane initial concentration to strains A cinetobacter sp.C3 degradation properties.By bacterial strain
Acinetobacter sp.C3 are seeded to the inorganic salts training that hexadecane initial concentration is 500 ~ 5000mg/L and pH value is 7.0
Support in base, 48h is cultivated in the constant-temperature table that rotating speed is 150r/min and temperature is 35 DEG C, positive ten are detected using gas chromatography
The residual concentration of six alkane, the results are shown in Table 3.
Influence of the 3 hexadecane initial concentration of table to strains A cinetobacter sp.C3 degradation properties
| Hexadecane concentration（mg/L） | Degradation rate（%） |
| 500 | 99.4 |
| 1000 | 100 |
| 2000 | 98.4 |
| 4000 | 96.3 |
| 5000 | 85 |
As can be seen from Table 3 strains A cinetobacter sp C3 to the degradation rate of hexadecane up to 85 ~ 100%.
This example demonstrates that degradation properties of the strains A cinetobacter sp C3 to long chain alkane in wider concentration range
Preferably, guarantee is provided for its application in the actual environment.
Embodiment 5
The extraction for the biosurfactant that strains A cinetobacter sp.C3 are produced and performance study.By 1 ~ 2%（v/
v）Strains A cinetobacter sp C3 bacterial suspension inoculation to hexadecane concentration be 500 ~ 5000mg/L inorganic salts train
Support in base, cultivate 48h in the constant-temperature table that rotating speed is 150r/min and temperature is 35 DEG C, obtain zymotic fluid；Then will fermentation
Liquid centrifuges 15 ~ 20min under the conditions of 8000 ~ 12000r/min and 4 ~ 8 DEG C, collects supernatant；Supernatant pH value is adjusted to 2 ~ 3；Again
By chloroform/methanol solution（2/1, v/v）︰ pH are that the volume ratio of 2 ~ 3 supernatant is that 1 ︰ 0.5 ~ 1 carries out extractive distillation；Will extraction
After organic phase afterwards is evaporated under reduced pressure under 20 ~ 30 DEG C and -0.1MPa, yellow surfaces activating agent is obtained, yield is 3 ~ 10g/L.
Using infrared analysis and thin layer chromatography (TLC) to biological caused by strains A cinetobacter sp.C3
Surfactant carries out category authentication, and CMC value, oil extraction circle value and the emulsifiability to biosurfactant are surveyed
It is fixed.
The results are shown in Figure 2 for biosurfactant IR spectrum scanning, 3436cm-1Locate to absorb for-OH, 1649cm-1Place
For NH+Absorb；1543cm-1And 1389cm-1For the absorption of C=O；1079cm-1Place is probably the absorption of PO-R；950cm-1For P=0
Absorption；668cm-1Locate the absorption for P=S, 511cm-1For the absorption of P-S.The biosurfactant is initially identified as phosphatide
Class activating agent.
The biosurfactant of extraction is put into chromatography cylinder and carries out thin-layer chromatography with capillary point sample in silica gel thin sheet.
Solvent is chloroform/methanol/water=65/15/2 (v/v/v).It is color developing agent with perchloric acid, the characteristic color reaction of solution is aobvious blue
Color, it is phospholipid surfactant to illustrate this surfactant.It is consistent with IR spectrum scanning qualification result, therefore, it is possible to identify bacterium
The surfactant that strain Acinetobacter sp.C3 are produced is phospholipid surfactant.
The measurement result of CMC value is as shown in figure 3, when surfactant concentration is less than 35mg/L, with surfactant
Concentration increase, the surface tension of water accordingly reduces, and when surfactant concentration is higher than 35mg/L, increases surfactant concentration
It cannot make the surface tension of water further reduce, show the biosurfactant that strains A cinetobacter sp.C3 are produced
Critical micelle concentration be about 35mg/L, the surface tension of water can be reduced to 20mN/m.
Oil extraction circle value：Culture dish is taken, adds water, adds 0.1ml hexadecanes on the water surface, form oil film.Added at oil film center
The biosurfactant of extraction, center oil film are pressed against surrounding and form a circle, measure a diameter of 80mm of circle.
The results are shown in Figure 4 for the emulsifiability for the biosurfactant that strains A cinetobacter sp.C3 are produced,
72h internal emulsifications ability is 60 ~ 100%.Illustrate the surfactant emulsifies better performances that bacterial strain C3 is produced, there is solubilising well
Effect, emulsifying capacity and wetting penetrating power.
Embodiment 6
Degradeds of the strains A cinetobacter sp.C3 to certain independent Coking Plant Wastewater.It is biochemical to gather certain independent coke-oven plant
Waste water afterwards, contains a certain amount of C8 ~ C16 alkane in waste water, COD is 200 ~ 300mg/L.By strains A cinetobacter
Sp.C3 is seeded in the flask equipped with waste water after independent coke-oven plant's biochemistry, and in rotating speed be 150r/min and temperature is 35 DEG C
48h is cultivated in constant-temperature table, which 50 ~ 58.5mg/L can be reduced to, reach the mark of industrial regeneration water
It is accurate（COD≤60mg/L）；GC-MS analyses C8 ~ C16 in the coking wastewaters of strains A cinetobacter sp.C3 before and after the processing
The removal rate of alkane is 52 ~ 73%, and most of alkane is removed.Strains A cinetobacter sp.C3 to except long chain alkane with
Outer organic matter also has preferable degradation capability.
Embodiment 7
Degradeds of the strains A cinetobacter sp.C3 to certain New Coal Chemical enterprise wastewater.Gather certain New Coal Chemical
Waste water after enterprise's biochemistry, the long chain alkane of substantial amounts of C16 ~ C24 is contained in waste water, COD is 300 ~ 400mg/L.By bacterial strain
Acinetobacter sp.C3 are seeded in the flask equipped with the New Coal Chemical enterprise wastewater, rotating speed for 150r/min and
Temperature is to cultivate 48h in 35 DEG C of constant-temperature table, New Coal Chemical enterprise wastewater COD can be reduced to 40 ~ 60mg/L, reached
The standard of industrial regeneration water（COD≤60mg/L）；The Jiao of strains A cinetobacter sp.C3 before and after the processing is used in GC-MS analyses
The removal rate for changing C16 ~ C24 alkane in waste water is 87 ~ 93%.Strains A cinetobacter sp.C3 centering long chain alkanes have very
Good degradation effect.
Embodiment 8
Degradeds of the strains A cinetobacter sp.C3 to petroleum hydrocarbon.Strains A cinetobacter sp.C3 are seeded to
Containing 1%（V ︰ v）Bavin wax（Chai You ︰ paraffin=1: 1, v ︰ v）Minimal medium in（Long chain alkane carbon number for C20 ~
C30）, cultivate in the constant-temperature table that rotating speed is 150r/min and temperature is 35 DEG C, trained every 12h using Their Determination by Spectrophotometry
The degradation rate of nutrient solution petrochina hydrocarbon, experimental result are as shown in Figure 5.As shown in Figure 5, after 96 ~ 156h, the bavin of 500 ~ 10000mg/L
The degradation rate of wax mixed solution is up to 78 ~ 92%.
Embodiment 9
Degradeds of the strains A cinetobacter sp.C3 to certain joint Coking Plant Wastewater.Gather certain joint coke-oven plant biochemistry
Processed waste water, contains substantial amounts of C30 ~ C40 long chain alkanes after advanced treating in waste water, COD is 80 ~ 200mg/L, as a result
As shown in fig. 6, fail to reach industrial regeneration water water standard.Strains A cinetobacter sp.C3 are seeded to new equipped with this
In the flask of moulded coal chemical industry enterprise wastewater, 48h is cultivated in the constant-temperature table that rotating speed is 150r/min and temperature is 35 DEG C, can be incited somebody to action
Water outlet COD is reduced to 30 ~ 60mg/L, and the results are shown in Figure 7, reaches the standard of industrial regeneration water（COD≤60mg/L）；GC-MS
Analysis is with the removal rate of C30 ~ C40 long chain alkanes in strains A cinetobacter sp.C3 coking wastewaters before and after the processing
88.5~95.2%。
Present embodiment has the following advantages：
1st, the strains A cinetobacter sp.C3 that present embodiment provides generate one kind in fermentative degradation process
Phosphatide type biological surfactant, yield are 3 ~ 10g/L；The surfactant can be by the surface tension of solution by initial
80.03mN/m is reduced to 20.79mN/m, and 72h internal emulsifications ability is up to 60 ~ 100%.
2nd, the strains A cinetobacter sp.C3 that present embodiment provides have efficient degraded long chain alkane energy
Power, suitable for effective degraded of the long chain alkane to varying environment, various concentrations：To concentration be 500 ~ 5000mg/L positive 16
The degradation rate of alkane is 85 ~ 100%；Degradation rate to the bavin wax mixture that concentration is 500 ~ 10000mg/L is 78 ~ 92%；To metallurgy
Industry（Independent coke-oven plant, integrated iron and steel works and New Coal Chemical enterprise etc.）Middle COD is the wastewater treatment of 80 ~ 400mg/L
Afterwards, the COD of effluent quality is 30 ~ 60mg/L, reaches the standard of industrial regeneration water（COD≤60mg/L）；To independent coke-oven plant, steel
In the waste water of iron integrated complex and New Coal Chemical enterprise the removal rate of long chain alkane be respectively 52 ~ 73%, 87 ~ 93% and 88.5 ~
95.2%。
Therefore, the strains A cinetobacter sp.C3 that present embodiment provides are applied to coking wastewater and petrochemical industry
The improvement of waste water, there is good degradation effect, realizes to the processing of industrial wastewater biological reinforced deep regenerative.
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 (1)
1. a kind of application of long-chain alkane degrading bacterium, it is characterised in that the degradation bacteria is applied to treatment of Coking Wastewater and petrochemical industry is given up
C8~C40 components in water；The degradation bacteria is acinetobacter calcoaceticus (Acinetobacter sp.) C3, by Chinese Typical Representative culture
Collection preservation, preserving number are：CCTCCNO：M 2015394, preservation date are on June 19th, 2015；The acinetobacter calcoaceticus
The GenBank accession number of the 16S rDNA of (Acinetobacter sp.) C3 is KR072554；
Strains A cinetobacter sp.C3 are one plant of Gram-negative bacteria, and bacterium colony is regular circular, neat in edge, surface
Smooth, colony colour is white；By being negative under the microscope after Gram's staining, thalline is in rod-short；
The degradation bacteria generates a kind of phosphatide type biological surfactant in fermentative degradation process.
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| CN106834189B (en) * | 2017-03-09 | 2020-02-14 | 武汉科技大学 | Biosurfactant producing bacterium and application thereof |
| CN111394271B (en) * | 2019-09-12 | 2021-06-15 | 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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| CN102399719A (en) * | 2011-10-21 | 2012-04-04 | 中国人民解放军海军医学研究所 | Bacterium DW3 capable of degrading marine diesel oil pollutants |
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