CN103849615A - Immobilized algal-bacterial symbiotic system for processing petroleum pollutants and application thereof - Google Patents

Immobilized algal-bacterial symbiotic system for processing petroleum pollutants and application thereof Download PDF

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CN103849615A
CN103849615A CN201310754942.4A CN201310754942A CN103849615A CN 103849615 A CN103849615 A CN 103849615A CN 201310754942 A CN201310754942 A CN 201310754942A CN 103849615 A CN103849615 A CN 103849615A
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algae
cogeneration
immobilized
oil
chlorella
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CN201310754942.4A
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Chinese (zh)
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徐珊
郑斌
陆阿定
杨会成
周宇芳
胡建坤
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浙江省海洋开发研究院
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Publication of CN103849615A publication Critical patent/CN103849615A/en

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Abstract

The invention belongs to the technical field of bioremediation of marine oil pollution, and relates to an immobilized algal-bacterial symbiotic system for processing petroleum pollutants. The immobilized algal-bacterial symbiotic system is prepared by the following steps: (1) selection and axenic cultivation of algae species, firstly carrying out lysozyme and sodium dodecyl sulfate (SDS) sterile pretreatment on chlorella algae fluid, and then inoculating into a BG11 culture solution containing antibiotics to carry out sterile cultivation, so as to obtain sterile chlorella algae fluid for later use; (2) selection and cultivation of petroleum degrading bacteria; (3) preparation of composite carrier bacteria algae gel balls, namely immobilizing the sterile chlorella algae fluid obtained in the step (1) and the petroleum degrading bacteria obtained in the step (2) by adopting sodium alga acid and polyvinyl acetate (PVA) as immobilized carriers, and finally obtaining the immobilized algal-bacterial symbiotic system. The immobilized algal-bacterial symbiotic system can float on the water to carry out degradation of petroleum pollution, and is free of secondary pollution, free of toxic action on microorganisms, rapid in reaction speed, stable and reliable in operation, less in microorganism loss, and free of separation.

Description

A kind of immobilized bacterium algae cogeneration system and application thereof of processing petroleum-type pollutent
Technical field
The invention belongs to petroleum pollution in ocean bioremediation technology field, relate to a kind of immobilized bacterium algae cogeneration system of processing petroleum-type pollutent.
Background technology
Ocean is one of the earth three large ecosystems, is the important barrier that maintains Global Ecological balance, and ocean, in providing ample resources for the mankind, is also being faced with more and more severeer pollution, and its PetroChina Company Limited. is most important pollutent in ocean environment.The main source of petroleum pollution in ocean is oil transportation, oilfield exploitation, seashore oil extraction and the sedimentation of atmosphere petroleum hydrocarbon etc., and according to the statistics of related organization of United Nations, the annual human being's production petroleum pollution in ocean causing of living is about 1 × 10 7t, comprising the pollution that causes of offshore field gas blowout accident and oil tanker accident, its oil spillage is up to 2.2 × 10 6t.Petroleum pollution in ocean has brought serious harm not only to ocean environment and Marine ecosystems, the also direct or indirect existence that affects the mankind and Sustainable development simultaneously, and petroleum pollution in ocean has become one of world's public hazards.Therefore, administer that petroleum pollution in ocean has become that our times various countries scientist is concerned about and one of problem in the urgent need to address.
At present, the intelligence method of petroleum pollution in ocean mainly contains physical mechanical method, chemical method and biological process.Physical mechanical method and the oil removing of chemical method ubiquity thoroughly and easily do not cause the problem of secondary pollution, and biological degradation method is because of advantages such as it possesses that processing costs is low, regulation effect good,, non-secondary pollutions little to environmental influence, embody the superiority that additive method is difficult analogy, its application prospect is very good.Biological process is mainly included in Polluted area microbe inoculation, adds nutrition and the several methods such as electron acceptor(EA) is provided.Wherein, immobilized biotechnology is new a kind of new bio technology of rising, it is that one is utilized physics chemical process by enzyme or biomass cells restriction or is positioned within the scope of a certain particular space, retains its intrinsic catalytic activity, and the technology that can be repeated or use continuously.It is active that biology after immobilization can not only keep for a long time, and the microenvironment of immobilization biological carrier is conducive to shield the harmful competition to organism of indigenous bacterium, phage and toxicant, engulfs and poison, alleviate the shearing force of wave to biological infringement, make it in complex environment, play consistently efficient performance.The ocean water body lower for Pollutant levels, area of space is distributed more widely, immobilized biotechnology has special technical superiority undoubtedly.
At present, immobilized biotechnology most critical be the selection of fixation support material, specific requirement is that the immobilized spherule cost of preparation is low, is easy to that moulding, immobilization time are short, physics and the characteristic such as chemical stability is good, and in immobilization process and after immobilization to biological nontoxic.Immobilization material generally adopts sodium alginate, agar, gelatin, polyvinyl alcohol (PVA) and polyacrylamide gel etc.Agar strength is poor, gelatin internal structure is closely knit, mass-transfer performance is poor, polyacrylamide gel is toxic to biology, sodium alginate and PVA are with respect to other carriers, and gel physical strength and mass-transfer performance are all better, to biological nontoxic, and resistance to Biodegradable is good, it is comparatively suitable fixed cell carrier.And need to meet following special conditions as the immobilization carrier of marine oil degradation bacteria: degradable, floatability, good biocompatibility, little to landscape impact.Therefore raw material sources are extensive, cheap, rigid condition is gentle because having for the immobilization complex carrier that adopts sodium alginate and PVA to make, simple to operate, good biocompatibility, matrix and product are expanded production the advantages such as resistance is little, and become the ideal material of preparing marine oil microorganism immobilization carrier.
Helotism system is to utilize physiological function between algea and bacteria two class biologies act synergistically the ecosystem of purifying water body.The current bacterial classification overwhelming majority of utilizing biological process to process petroleum pollution in ocean is aerobic bacteria, so the height of oxygen level affects its degradation efficiency to oil to a great extent.But by the water body of petroleum pollution because photosynthesis weakens and hydrobiological respiration makes content of oxygen dissolved in water lower, this has just limited the raising of microorganism to oil degradation speed.The photosynthesis of algae provides respiration needed oxygen for bacterium, and lacking in the environment of nitrogen, algae also can be fixed the nitrogen in atmosphere by nitrogen fixation, for bacterium provides nitrogen element and organism, promotes its growth.Due to the restriction of illumination, some hydrocarbon degradation bacterium are suppressed, and form after symbiote with algae, can protect it under high light or drying conditions, to survive.Meanwhile, cogeneration system also provides favourable condition for the growth of algae, the CO that bacterial respiratory produces 2can be utilized by algae, degradation by bacteria oil and the organism that produces also can be utilized by algae.In addition, algae can first be enriched in petroleum hydrocarbon in frustule, bacterium these materials of reentrying, thereby when with algae symbiosis, the growth velocity of degradation bacteria and all will be higher than its single culture to the degradation efficiency of oil.As can be seen here, in algae and ocean, the bacterium of degraded oil forms syntaxial system, for the energy and nutritive substance are provided each other, and mutualism, the degradation efficiency of raising offshore oil.
Summary of the invention
The invention provides a kind of immobilized bacterium algae cogeneration system for the treatment of marine oil pollutant, this immobilized bacterium algae cogeneration system can swim in the degraded of carrying out petroleum pollution on the water surface, can not produce secondary pollution, to microorganism toxicological harmless effect, speed of response is fast, stable and reliable operation, microorganism is run off few, does not need to separate.
The present invention also provides the application of a kind of described immobilized bacterium algae cogeneration system in petroleum pollution degradation.
The technical solution adopted for the present invention to solve the technical problems is:
Process an immobilized bacterium algae cogeneration system for petroleum-type pollutent, it is characterized in that it is to be prepared from by the following method:
(1) selection of algae kind and asepticize are cultivated: chlorella algae liquid first passes through N,O-Diacetylmuramidase and the aseptic pre-treatment of SDS, are then seeded to containing in antibiotic BG11 nutrient solution and carry out asepticize cultivation, obtain aseptic chlorella algae liquid for subsequent use;
The present invention adopts N,O-Diacetylmuramidase/SDS(sodium laurylsulfonate) combination antibiotic carries out axenic purification to chlorella, the formula of BG11 nutrient solution used (conventional medium) is: NaNO31.5g/L, K2HPO40.04g/L, MgSO47H2O0.075g/L, CaCl27H2O0.036g/L, citric acid (Citric acid) 0.006g/L, ferric ammonium citrate (Ammonium ferric citrategreen) 0.006g/L, EDTA0.001g/L, Na2CO30.02g/L, H3BO36.1 × 10-5g/L, MnSO4H2O1.69 × 10-4g/L, ZnSO47H2O2.87 × 10-4g/L, CuSO45H2O2.5 × 10-6g/L, (NH4) 6Mo7O244H2O1.25 × 10-5g/L.
(2) selection of oil degradation bacteria and cultivation: get 6 kinds of oil degradation bacterias, buy in Chinese Sea microbial strains preservation administrative center (MCCC), wherein 3 kinds of Rhodopseudomonass, strain number is 1A00241,1A00269 and 1A00364,2 kinds of bacillus, strain number is 1A0008,1A0009,1 kind of achromobacter, strain number is 1A04524, to above-mentioned 6 kinds of oil degradation bacterias called after Y1, Y2, Y3, Y4, Y5, Y6 successively, and cultivate respectively, incubation time is 3～5 days;
(3) oil degradation bacteria that aseptic chlorella algae liquid step (1) being obtained and step (2) obtain adopts sodium alginate and PVA as being fixed of fixation support, being finally fixed helotism system.The composite carrier bacteria algae gel ball that immobilized bacterium algae cogeneration system mixes.
Micro-algae and bacterium can form helotism system, its synergy can make the biological degradation of pollutent more efficient, but because micro-algae is attaching some bacteriums in process of growth, be in building process at artificial bacterium frond, additional bacterium and micro-algae self the relation with epiphyte very complicated, may mutually promote, suppress, work in coordination with or antagonism, be to build the most efficient oil degradation system, micro-algae carried out to asepticize and cultivate the precondition that is absolutely necessary.By helotism system being processed to consulting of marine oil pollutant pertinent literature, finally select chlorella as algae kind of the present invention, this algae kind is purchased from Wuhan hydrobiont institute of the Chinese Academy of Sciences.
As preferably, step (1) aseptic pretreatment process used is: be taken at the chlorella algae liquid of cultivating 7d in BG11 nutrient solution, the centrifugal supernatant liquor that goes, centrifugal 3～5 times of sterile distilled water repeated washing, adds the Tween80 of 1mL mass concentration 0.005%, the EDTA of 5 0.1mol/L, 30mL sterilized water, shake up and place 10min, then add 1mL0.5mg/ml N,O-Diacetylmuramidase, the SDS of 5 0.25%, shake up, place 10min, centrifugal going after supernatant liquor, then with sterilized water washing 3～5 times.
As preferably, in step (1), containing antibiotic BG11 nutrient solution formula be: NaNO 31.5g/L, K 2hPO 43H 2o0.04g/L, MgSO 47H 2o0.075g/L, CaCl 27H 2o0.036g/L, citric acid (Citric acid) 0.006g/L, ferric ammonium citrate (Ammonium ferric citrategreen) 0.006g/L, EDTA0.001g/L, Na 2cO30.02g/L, H 3bO 36.1 × 10 -5g/L, MnSO 4h 2o1.69 × 10 -4g/L, ZnSO 47H2O2.87 × 10 -4g/L, CuSO 45H 2o2.5 × 10 -6g/L, (NH 4) 6mo 7o 244H 2o1.25 × 10 -5g/L, 1ml/L penicillin and 0.5ml/L kalamycin are as preferably, and step (1) culture condition is: 25 ± 1 ℃ of temperature, and shaking speed 150r/min cultivates, and intensity of illumination is 8000lux, Light To Dark Ratio L/D=14:10.
As preferably, the culture medium prescription of step (2) oil degradation bacteria used is: oil 10g, NH 4nO 33g, KH 2pO 40.5g, K 2hPO 40.5g, liquid microelement 2mL and water 1000mL, pH is 7.5; Trace element fluid component (weight) is: MgSO 40.4%, CuSO 40.1%, MnSO 40.1%, FeSO 4h 2o0.1% and CaCl 20.1%.
As preferably, process for fixation is specially: the mixing solutions that contains algae kind, oil degradation flora and immobilization complex carrier with asepsis injector extraction is to the boric acid solution of mass concentration 4%, pH=6.7, at the crosslinked 20h of 10 ℃ of immobilizations, then bead is taken out, deionized water desalination 2h, and 4 ℃ be stored in aseptic seawater, being fixed helotism system.
As preferably, step (2) chlorella algae liquid, oil degradation bacteria culture fluid are that 1:1:2 mixes with the ratio of fixation support, and the formula (massfraction) of fixation support is 0.3% sodium alginate, 20%PVA, 8%SiO 2and 0.6%CaCO 3.
Oil degradation flora and chlorella are attached to a kind of employing on the floatability degradable biological immobilization carrier that the composite membrane of sodium alginate and PVA makes by the present invention, thereby the immobilized bacterium algae cogeneration system of preparing is expected to the improvement for petroleum pollution in ocean.
The application of described immobilized bacterium algae cogeneration system in petroleum pollution degradation, the composite carrier bacteria algae gel ball of preparation in step (3) is inoculated into the weight containing oil 0.5%(according to 1% mass concentration) 50mL MMC substratum in, 30 ℃, 180rpm shaking table are cultivated 7 days, and nutrient solution is for the mensuration of degradation rate.MMC culture medium prescription used is: NaCl0.024g/L, MgSO 47H 2o0.0007g/L, NH 4nO 30.001g/L, KCl0.0007g/L, KH 2pO 40.002g/L, Na 2hPO 47H 2o0.003g/L, pH is 7.5, adds appropriate trace element and mix after sterilizing.
Compared with prior art, the present invention has the following advantages:
(1) adopt N,O-Diacetylmuramidase/SDS combination antibiotic to carry out degerming purifying to the chlorella of selecting, obtain aseptic chlorella.In process domestication's syntaxial system, this algae kind and high efficient petroleum degrading bacteria mutualism, with natural helotism system and only process compared with offshore oil pollution with mix bacterium agent, not only preserve the advantage of syntaxial system, inherited again mix bacterium agent efficient ability of processing oil under control environment.
(2) adopt helotism system to process offshore oil pollution, utilize on the one hand the advantage degraded oil pollutent of bacterium algae mutualism, algae also grease of enriched in cell in the hydrocarbon compound in effective degraded oil, makes it can be used as biofuel and is used on the other hand.
(3) adopt sodium alginate and the PVA complex carrier as helotism system, on the one hand for helotism system provides a metastable environment, build stable microbial ecosystem, made bacterium algae can continue to bring into play oil polluted environment repair.This complex carrier degradable on the other hand, non-secondary pollution; Floatability, makes degradation bacteria fully contact with petroleum pollution, and degradation efficiency is higher; Toxicological harmless, can not work the mischief to helotism system or ocean other biological.
Embodiment
Below by specific embodiment, technical scheme of the present invention is described in further detail.Should be appreciated that enforcement of the present invention is not limited to the following examples, any pro forma accommodation that the present invention is made and/or change all will fall into protection domain of the present invention.
In the present invention, if not refer in particular to, all part, per-cents are weight unit, and equipment and the raw material etc. adopting all can be buied from market or this area is conventional.Method in following embodiment, if no special instructions, is the ordinary method of this area.
The asepticize of embodiment 1 algae kind is cultivated
Chlorella algae liquid first passes through N,O-Diacetylmuramidase/SDS pre-treatment, is then seeded to containing in the BG11 nutrient solution of different antibiotic combinations and carries out sterile culture.
Above-mentioned pretreatment process is: be taken at the algae liquid of cultivating 7d in BG11 nutrient solution, the centrifugal 10min of 2000r/min, removes supernatant liquor, centrifugal 3～5 times of sterile distilled water repeated washing, adds 1mL Tween80(0.005%), 5 EDTA(0.1mol/L), 30mL sterilized water, shakes up and places 10min.Add 1mL N,O-Diacetylmuramidase, 5 SDS, shake up again, and place 10min.Centrifugal going after supernatant liquor, then with sterilized water washing 3～5 times.
Due to the antibacterial mechanism of action difference of each microbiotic, above-mentioned 6 kinds of microbiotic are combined to (table 1), cultivate for micro-algae asepticize, by dull and stereotyped detection bacteria-eliminating efficacy method be: get the algae liquid of microbiotic culture medium culturing 7d, be diluted to 10 -4g/L, coats BG11 solid plate.Be placed under illumination condition and cultivate again 7d left and right, observe the growing state of dull and stereotyped micro-algae and bacterium.
Table 1 antibiotic combinations
Above-mentioned antibiotic combinations is as shown in table 2 on chlorella growth impact:
Table 2 chlorella is at microbiotic substratum growing state
Antibiotic combinations Growth conditions
A Solution is almost transparent
B Algae biomass is many, and better, solution is compared with deep green in growth
C Algae biomass is medium, and solution is breen
D Algae biomass is medium, and solution is emerald green
By flat band method, algae liquid is carried out to aseptic detection and chlorella growing state discovery on microbiotic substratum, microbiotic A combination has strong rejection to chlorella and bacterium, dull and stereotyped upper algae and bacterium are not all grown, B combination restraining effect minimum, can obtain aseptic algae, on C, D assembled flat, still length has bacterium.
Therefore,, by N,O-Diacetylmuramidase/SDS pre-treatment, combination antibiotic combination (penicillin+gentamicin) method can obtain aseptic chlorella.
Selection and the cultivation of embodiment 2 oil degradation bacterias
The present invention selects 6 kinds of oil degradation bacterias, buy in Chinese Sea microbial strains preservation administrative center (MCCC), wherein 3 kinds of Rhodopseudomonass, strain number is 1A00241,1A00269 and 1A00364,2 kinds of bacillus, strain number is 1A0008,1A0009, a kind of achromobacter, strain number is 1A04524, to above-mentioned 6 kinds of oil degradation bacterias called after Y1, Y2, Y3, Y4, Y5, Y6 respectively.
Above-mentioned steps (2) oil degradation bacteria culture medium prescription used is: oil 10g, NH 4nO 33g, KH 2pO 40.5g, K 2hPO 40.5g, liquid microelement 2mL and water 1000mL, pH is 7.5.
Above-mentioned micro-fluid component is: MgSO 40.4%, CuSO 40.1%, MnSO 40.1%, FeSO 4h 2o0.1% and CaCl 20.1%.
Above-mentioned steps (2) incubation time is 3～5 days.
The preparation of embodiment 3 composite carrier bacteria algae gel balls
The formula (massfraction) of fixation support is 0.3% sodium alginate, 20%PVA, 8%SiO 2and 0.6%CaCO 3.
Get 6 parts of 50mL fixation support solution, in every part, add respectively 25mL to contain bacterium liquid and the 25mL chlorella algae liquid of Y1, Y2, Y3, Y4, Y5, Y6 bacterial strain, mix and splash into boric acid solution (boric acid concentration 4% with syringe afterwards, pH=6.7) in, at the crosslinked 20h of 10 ℃ of immobilizations, then bead is taken out to deionized water desalination 2h, 6 kinds of composite carrier bacteria algae gel balls producing are distinguished to called after B1, B2, B3, B4, B5, B6, and 4 ℃ are stored in aseptic seawater for subsequent use.
Embodiment 4 degradeds of immobilized bacterium algae cogeneration system to petroleum pollution
Above-mentioned 6 kinds of composite carrier bacteria algae gel balls are inoculated into respectively in the 50mL MMC substratum containing oil 0.5% according to 1% concentration, and 30 ℃, 180rpm shaking table are cultivated 7 days, and nutrient solution is for the mensuration of degradation rate.
Above-mentioned MMC culture medium prescription used is: NaCl0.024g/L, MgSO 47H 2o0.0007g/L, NH 4nO 30.001g/L, KCl0.0007g/L, KH 2pO 40.002g/L, Na 2hPO 47H 2o0.003g/L, pH is 7.5, adds appropriate trace element and mix after sterilizing.
The measuring method of petroleum degradation rate is: the separating funnel of above-mentioned 6 kinds of nutrient solutions being poured respectively into 100mL, get after 20mL sherwood oil rinses triangular flask and pour in the lump in separating funnel, add a cover shake well 2min, and note venting, stratification 2min, puts back to lower floor's liquid nutrient medium in shaking flask.Upper solution is collected with 100mL volumetric flask.Pour separating funnel at lower floor's substratum of collecting again, by 20mL sherwood oil flushing shaking flask, repeat to extract 2 times, merge No. 2 extracting solutions in 100mL volumetric flask, use sherwood oil constant volume, at ultraviolet scene photometer 225nm place, measure absorbancy take sherwood oil as reference, according to typical curve, calculate petroleum degradation rate.
Measuring 6 kinds of immobilized bacterium algae cogeneration systems is respectively the petroleum pollution degradation rate of 7 days: the degradation rate of B1 is 34.35%, the degradation rate of B2 is 58.1%, the degradation rate of B3 is 38.97%, the degradation rate of B4 is 43.87%, the degradation rate of B5 is 79.23%, the degradation rate of B6 is 38.77%.
As can be seen here, adopt sodium alginate and PVA, as the helotism system of complex carrier fixed pellet algae and bacillus establishment, petroleum pollution in ocean thing is had to higher degradation rate, promote degradation by bacteria oil aspect marine organisms reparation, thering is good application potential.
Above-described embodiment is preferably scheme of one of the present invention, not the present invention is done to any pro forma restriction, also has other variant and remodeling under the prerequisite that does not exceed the technical scheme that claim records.

Claims (8)

1. process an immobilized bacterium algae cogeneration system for petroleum-type pollutent, it is characterized in that it is to be prepared from by the following method:
(1) selection of algae kind and asepticize are cultivated: chlorella algae liquid first passes through N,O-Diacetylmuramidase and the aseptic pre-treatment of SDS, are then seeded to containing in antibiotic BG11 nutrient solution and carry out asepticize cultivation, obtain aseptic chlorella algae liquid for subsequent use;
(2) selection of oil degradation bacteria and cultivation: get 6 kinds of oil degradation bacterias, buy in Chinese Sea microbial strains preservation administrative center (MCCC), wherein 3 kinds of Rhodopseudomonass, strain number is 1A00241,1A00269 and 1A00364,2 kinds of bacillus, strain number is 1A0008,1A0009,1 kind of achromobacter, strain number is 1A04524, to above-mentioned 6 kinds of oil degradation bacterias called after Y1, Y2, Y3, Y4, Y5, Y6 successively, and cultivate respectively, incubation time is 3 ~ 5 days;
(3) preparation of composite carrier bacteria algae gel ball: the oil degradation bacteria that the aseptic chlorella algae liquid that step (1) is obtained and step (2) obtain adopts sodium alginate and PVA as being fixed of fixation support, being finally fixed helotism system.
2. immobilized bacterium algae cogeneration system according to claim 1, it is characterized in that: step (1) aseptic pretreatment process used is: be taken at the chlorella algae liquid of cultivating 7d in BG11 nutrient solution, the centrifugal supernatant liquor that goes, centrifugal 3 ~ 5 times of sterile distilled water repeated washing, add the Tween80 of 1mL 0.005%, the EDTA of 5 0.1mol/L, 30mL sterilized water, shake up and place 10min, then add 1mL 0.5mg/ml N,O-Diacetylmuramidase, the SDS of 5 0.25%, shake up, place 10min, centrifugal going after supernatant liquor, then with sterilized water washing 3 ~ 5 times.
3. immobilized bacterium algae cogeneration system according to claim 1, is characterized in that: in step (1), containing antibiotic BG11 nutrient solution formula be: NaNO 31.5g/L, K 2hPO 43H 2o 0.04 g/L, MgSO 47H 2o 0.075 g/L, CaCl 27H 2o 0.036 g/L, citric acid 0.006 g/L, ferric ammonium citrate 0.006 g/L, EDTA 0.001 g/L, Na 2cO 30.02 g/L, H 3bO 36.1 × 10-5 g/L, MnSO4 H 2o 1.69 × 10-4 g/L, ZnSO4 7H 2o 2.87 × 10-4 g/L, CuSO4 5H 2o 2.5 × 10-6 g/L, (NH 4) 6mo 7o 244H2O 1.25 × 10-5 g/L, 1ml/L penicillin and 0.5ml/L kalamycin.
4. immobilized bacterium algae cogeneration system according to claim 1, is characterized in that: step (1) culture condition is: 25 ± 1 ℃ of temperature, and shaking speed 150r/min cultivates, and intensity of illumination is 8000lux, Light To Dark Ratio L/D=14:10.
5. immobilized bacterium algae cogeneration system according to claim 1, is characterized in that: the culture medium prescription of step (2) oil degradation bacteria used is: oil 10 g, NH 4nO 33 g, KH 2pO 40.5 g, K 2hPO 40.5 g, liquid microelement 2 mL and water 1000 mL, pH is 7.5; Trace element fluid component (weight) is: MgSO 40.4%, CuSO 40.1%, MnSO 40.1%, FeSO 4h 2o 0.1% and CaCl 20.1%.
6. immobilized bacterium algae cogeneration system according to claim 1, it is characterized in that: process for fixation is specially: the mixing solutions that contains algae kind, oil degradation flora and immobilization complex carrier with asepsis injector extraction is to the boric acid solution of mass concentration 4%, pH=6.7, at the crosslinked 20h of 10 ℃ of immobilizations, then bead is taken out, deionized water desalination 2h, and 4 ℃ be stored in aseptic seawater, being fixed helotism system.
7. immobilized bacterium algae cogeneration system according to claim 1, it is characterized in that: step (2) chlorella algae liquid, oil degradation bacteria culture fluid are that 1:1:2 mixes with the ratio of fixation support, and the formula (massfraction) of fixation support is 0.3% sodium alginate, 20%PVA, 8%SiO 2and 0.6%CaCO 3.
8. an immobilized bacterium algae cogeneration system claimed in claim 1 application in petroleum pollution degradation.
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