CN103482772A - Process technology for removing anthraquinone compounds out of wastewater through phanerochaete chrysosporium mycelium - Google Patents
Process technology for removing anthraquinone compounds out of wastewater through phanerochaete chrysosporium mycelium Download PDFInfo
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Abstract
The invention discloses process technology for removing anthraquinone compounds through phanerochaete chrysosporium mycelium and relates to the field of bioengineering. Phanerochaete chrysosporium is used as a starting strain, phanerochaete chrysosporium mycelium is obtained by steps of liquid shake-flask culture and liquid mycelium expansion culture, and the mycelium is used for removing the anthraquinone compounds out of wastewater. By adopting the process technology, the removal rate of the anthraquinone compounds can reach 70-100%.
Description
Technical field
The present invention relates to technical field of bioengineering, refer in particular to and utilize the Phanerochaete chrysosporium thalline to remove the anthraquinone analog compound in the waste water that contains anthraquinone analog compound.Raw wastewater is concentrated or is diluted to Anthraquinone after 0.05～1 grams per liter, drop into the Phanerochaete chrysosporium thalline, anthraquinone analog compound is removed in the aeration-agitation reaction.The clearance of applying this technique anthraquinone analog compound can reach 60～100%.
Background technology
Anthraquinone compounds is of a great variety, is important industrial chemicals and intermediate, bright-colored, scope is wide, thereby widespread use in the industries such as weaving, printing and dyeing, not only can be used for the printing and dyeing of terylene and BLENDED FABRIC, also can be used for the printing and dyeing of polyamide fibre, acrylic fibers, polypropylene fibre, vinegar fibre, wool, silk.Along with the lifting day by day of environmental requirement, azoic dyestuff forbidding range extension, also make the importance of anthraquinone dye more outstanding, makes its application in printing and dyeing industry more and more extensive simultaneously.But not only the factory effluent water yield is large for anthraquinone analog compound, and this compound colourity is high, and solubleness is low! Fusing point is high! A stable in properties! Be difficult to biological degradation, have again higher lipotropy, in the sewage directly discharged, anthraquinone is by sedimentation! But the effect prolonged stays such as food chain enrichment are in water body! Soil! In settling and air.Therefore explore the effectively technology of degraded anthraquinone analog compound, be of great significance for social harmony and development tool.
At present about the processing patent that contains anthraquinone analog compound waste water, have several, but main method is recycling, microbiological deterioration and the oxidation removal of anthraquinone.As patent " 1; improvement and its recovery method as resource of 4-dihydroxyanthraquinone factory effluent " (application number 00112386.6) discloses and has utilized macroporous resin adsorption, resolve with sodium hydroxide the method for recycling the anthraquinone analog compound in Isosorbide-5-Nitrae-dihydroxyanthraquinone factory effluent again; Patent " recovery method of a kind of Isosorbide-5-Nitrae-dihydroxyanthraquinone processing wastewater " (application number 200410013484.X) discloses the method for utilizing concentrated, cooling, crystallization, separation and the technology recycling anthraquinone such as dry.Patent " Sphingomonas strain and the application in the anthraquinone dye wastewater decolouring thereof " (application number 200410020832.6) discloses Sphingomonas, application in the anthraquinone dye wastewater decolouring, this bacterium decoloring ability is strong, degradation speed is fast, but can only be suitable for lower concentration anthraquinone waste water; Adopt in addition electrochemical oxidation (patent No. 200910029887.6) and photochemical catalytic oxidation (200710011629.6), these two kinds of methods also need further processing.Paper relevant to these methods also has many reports.Other doctor's or master's degree paper has reported that Phanerochaete chrysosporium absorption degradation anthraquinone analog compound mechanism is studied, but does not relate to its Technology.
The inventor is through deep research, find out a kind of utilize Phanerochaete chrysosporium (
phanerochaete chrysosporium) thalline, anthraquinone analog compound technique in ventilation and stirring reaction for some time removal waste water, this process time is short, and clearance is high.
Summary of the invention
The present invention is to provide the biotechnology of anthraquinone analog compound in the Phanerochaete chrysosporium degrading waste water.Waste water is after the Phanerochaete chrysosporium degraded, and its anthraquinone analog compound clearance can arrive 60 ~ 100%.
A kind of Phanerochaete chrysosporium degraded of the present invention anthraquinone analog compound Technology, according to following step, carry out: take Phanerochaete chrysosporium as starting strain, carry out test tube enlarged culturing, liquid shaking bottle cultivation, thalline fluid enlargement culture, centrifugal collection thalline, then remove anthraquinone analog compound in waste water with thalline.
The present invention's bacterial classification used be Phanerochaete chrysosporium (
phanerochaete chrysosporium) any strain bacterial classification, as the bacterial classification of Chinese common micro-organisms culture presevation administrative center (CCGMC), Chinese agriculture microbial strains preservation administrative center (ACCC), Chinese industrial microbial strains preservation administrative center (CICC), Chinese medicine microbial strains preservation administrative center (CMCC), national veterinary microorganism DSMZ (CVCC) and microbiotic bacterial classification preservation pipe reason center etc. preservation.
The applicable waste water of the present invention is any one waste water that contains anthraquinone analog compound, extracts the waste liquid of anthraquinone analog compound as the waste liquid of fermentative Production anthraquinone analog compound, from plant or take the waste liquid of anthraquinone analog compound as synthetic other products of raw material.
Enlarged culturing base in wherein said test tube enlarged culturing is potato sucrose substratum (Zhu Gejian, Wang Zhengxiang chief editor's " industrial microorganism experimental technique handbook ", 1994,367 pages.
During wherein said liquid shaking bottle is cultivated and liquid shaking bottle substratum and liquid thalline substratum in the thalline fluid enlargement culture be: glucose 5～30 grams per liters, peptone 0～5 grams per liter, yeast extract paste 0～5 grams per liter, sal epsom 0.2～1, tween-80 0.5～10, potassium primary phosphate 2～9, pH5～8,120～140 ℃ sterilizing 20～40 minutes.
Wherein said shake-flask culture processing condition are, inoculation 3-5 piece Phanerochaete chrysosporium test tube slant thalline (5 * 5mm) is in the 250mL triangular flask that 40～120 mL substratum are housed, at rotating speed: 150 rev/mins, 25～35 ℃, cultivate 24～72 hours.Wherein said yeast culture technique is, by 1～20%(volume ratio) inoculum size the shake-flask culture seed liquor be inoculated into to liquid thalline enlarged culturing technique be, 25～35 ℃ of temperature, 50～200 rev/mins of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18～72 hours; Nutrient solution centrifugal 5～20 minutes through 3000～6000rpm, obtain the Phanerochaete chrysosporium thalline.
The technique of removing anthraquinone analog compound in waste water with thalline of the present invention is: above-mentioned Phanerochaete chrysosporium thalline with the waste water that contains 0.05～1 grams per liter anthraquinone analog compound (by vacuum concentration or dilution adjusting anthraquinone analog compound concentration) by 1:10～50(mass/volume) mix, 25～60 ℃ of temperature, 50～200 rev/mins of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react 3～15 hours, can remove anthraquinone analog compound.
After using this processing wastewater degraded, the anthraquinone analog compound clearance can reach 70-100%.
According to liquid thalline enlarged culturing technique of the present invention, in conjunction with the ABC of this area, can be according to need of production, increase even level Four thalline tank of secondary, three grades, further expand the scale of production, to carry out suitability for industrialized production.Secondary, the three grades substratum that even the level Four seeding tank adopts are identical with thalline fluid enlargement culture base with the shake-flask culture base, and inoculum size is 5～20%, and culture condition is same as liquid thalline enlarged culturing condition.
Embodiment
According to the anthraquinone analog compound measuring method of this process using, be: accurate absorption 1,8-dihydroxyanthraquinone reference substance solution (0.522 mg/ml) 1,3,5,8,10,15 ml, put respectively in the 25ml volumetric flask, fling to methyl alcohol, residue is with 0.5% magnesium acetate dissolve with ethanol solution and be diluted to scale, shake up, in 425 nm places, make reference with 0.5% magnesium acetate ethanolic soln and measure optical density.With 1,8-dihydroxy-anthracene quinone content (y), optical density (x) is carried out to linear regression analysis, obtain equation of linear regression.The accurate sample solution of drawing repeats aforesaid operations mensuration optical density, utilizes equation of linear regression to calculate the content of general anthraquinone in each sample.
Anthraquinone analog compound clearance Re calculates by following formula (1):
In formula,
r t clearance for t time anthraquinone analog compound;
c 0 (mg/L) be the anthraquinone analog compound concentration of 0 hour;
c t (mg/L) for adsorbing the concentration of t hour anthraquinone analog compound.
In order further to set forth related material and the technique of technical scheme of the present invention, provided following examples, but these embodiment do not limit the scope of the invention in any form.
Embodiment 1
1. the making of test tube slant bacterial classification
Potato sucrose substratum (Zhu Gejian, Wang Zhengxiang chief editor's " industrial microorganism experimental technique handbook " in new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (Chinese common micro-organisms culture presevation administrative center CCGMC5.776), 26 ℃, incubation time 50 hours; 4 ℃ save backup.
2. liquid shaking bottle is cultivated the making of bacterial classification
Accurately take glucose 5 grams, peptone 0 gram, yeast extract paste 5 grams, sal epsom 0.2 gram, tween-80 0.5 gram, potassium primary phosphate 2 grams, water 1000 mL, pH 5, packing 250mL triangular flask, every bottle of 50 grams, totally 20 bottles, 120 ℃ of sterilizings 40 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 * 10 of 4 ℃ of preservations of a ring
6individual spore), at 25 ℃, 150 rev/mins, cultivate 72 hours;
3. thalline fluid enlargement culture
Accurately take glucose 50 grams, peptone 0 gram, yeast extract paste 50 grams, sal epsom 2 grams, tween-80 5 grams, potassium primary phosphate 20 grams, water 10L, be loaded in the seeding tank of 15L, 120 ℃ of sterilizings 20 minutes; After sterilizing is cooling, 25 ℃ of temperature, 50 rev/mins of mixing speed, 0.2: 1 volume/volume of air flow/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 72 hours; Centrifugal 20 minutes of nutrient solution process 3000rpm, obtain the Phanerochaete chrysosporium thalline.
4. anthraquinone analog compound in removal waste water
Take rheum officinale as raw material, adopt decocting method to extract the waste liquid of anthraquinone analog compound through concentrated, the waste liquid 1000L that anthraquinone analog compound concentration is 1 gram/L, thalline with waste liquid by the 1:50(mass/volume) mix, 25 ℃ of temperature, 60 rev/mins of mixing speed, air flow 0.3:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react after 3 hours, the anthraquinone analog compound clearance reaches 71.3%.
Embodiment 2
1. the making of test tube slant bacterial classification
Potato sucrose substratum (Zhu Gejian, Wang Zhengxiang chief editor's " industrial microorganism experimental technique handbook " in new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (Chinese agriculture microbial strains preservation administrative center ACCC 30942), 30 ℃, incubation time 100 hours; 7 ℃ save backup.
2. liquid shaking bottle is cultivated the making of bacterial classification
Accurately take glucose 150 grams, peptone 25 grams, yeast extract paste 25 grams, sal epsom 0.4 gram, tween-80 4 grams, potassium primary phosphate 5 grams, water 10 L, pH 6, packing 250mL triangular flask, every bottle of 100 grams, totally 100 bottles, 130 ℃ of sterilizings 30 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 * 10 of 7 ℃ of preservations of a ring
6individual spore), at 30 ℃, 150 rev/mins, cultivate 50 hours;
3. thalline fluid enlargement culture
Accurately take glucose 2000 grams, peptone 250 grams, yeast extract paste 250 grams, sal epsom 60 grams, tween-80 60 grams, potassium primary phosphate 400 grams, water 100L, be loaded in the seeding tank of 130L, 130 ℃ of sterilizings 30 minutes; After cooling, 30 ℃ of temperature, 125 rev/mins of mixing speed, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 50 hours; Centrifugal 13 minutes of nutrient solution process 4500rpm, obtain the Phanerochaete chrysosporium thalline.
4. anthraquinone analog compound in removal waste water
The Fusarium oxysporum fermentation prepares the waste liquid of anthraquinone analog compound, reach the waste water 1000L of 0.2 gram/L through concentrating the content that makes anthraquinone analog compound, thalline with waste liquid by the 1:30(mass/volume) mix, at temperature 45 C, 120 rev/mins of mixing speed, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), after reacting 7 hours, the anthraquinone analog compound clearance reaches 87.5%.
Embodiment 3
1. the making of test tube slant bacterial classification
Potato sucrose substratum (Zhu Gejian, Wang Zhengxiang chief editor's " industrial microorganism experimental technique handbook " in new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (Chinese industrial microbial strains preservation management center C ICC14067), 35 ℃, incubation time 144 hours; 10 ℃ save backup.
2. liquid shaking bottle is cultivated the making of bacterial classification
Accurately take glucose 300 grams, peptone 50 grams, yeast extract paste 0 gram, sal epsom 10 grams, tween-80 100 grams, potassium primary phosphate 90 grams, water 10L, pH7.5, packing 250mL triangular flask, every bottle of 120 mL, totally 80 bottles, 140 ℃ of sterilizings 20 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 * 10 of 10 ℃ of preservations of a ring
6individual spore), at 35 ℃, 150 rev/mins, cultivate 72 hours;
3. one-level thalline fluid enlargement culture
Accurately take glucose 1500 grams, peptone 250 grams, yeast extract paste 0 gram, sal epsom 50 grams, tween-80 500 grams, potassium primary phosphate 450 grams, water 50L, be loaded in the first class seed pot of 70L, 140 ℃ of sterilizings 40 minutes; After sterilizing is cooling, 35 ℃ of temperature, 200 rev/mins of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 50 hours;
4. secondary thalline fluid enlargement culture
Accurately take glucose 15000 grams, peptone 2500 grams, yeast extract paste 0 gram, sal epsom 500 grams, tween-80 5000 grams, potassium primary phosphate 4500 grams, water 500L, be loaded in the secondary seed tank of 700L, 140 ℃ of sterilizings 40 minutes; After sterilizing is cooling, 35 ℃ of temperature, 200 rev/mins of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18 hours; Centrifugal 5 minutes of nutrient solution process 6000rpm, obtain the Phanerochaete chrysosporium thalline.
5. anthraquinone analog compound in removal waste water
Take anthraquinone analog compound as raw material, under the effect of catalyzer, the waste liquid of synthesizing nitryl anthraquinone, the waste liquid 1000L that to be diluted to Anthraquinone be 0.05 gram/L, thalline with waste liquid by the 1:10(mass/volume) mix, at temperature 60 C, 200 rev/mins of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), after reacting 15 hours, the anthraquinone analog compound clearance reaches 100%.
Claims (8)
1. Phanerochaete chrysosporium degraded anthraquinone analog compound Technology, it is characterized in that carrying out according to following step: take Phanerochaete chrysosporium as starting strain, carry out test tube enlarged culturing, liquid shaking bottle cultivation, thalline fluid enlargement culture, centrifugal collection thalline, then remove anthraquinone analog compound in waste water with thalline.
2. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature is any strain bacterial classification of Phanerochaete chrysosporium at bacterial classification used.
3. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature is any one waste water that contains anthraquinone analog compound at applicable waste water, extracts the waste liquid of anthraquinone analog compound as the waste liquid of fermentative Production anthraquinone analog compound, from plant or take the waste liquid of anthraquinone analog compound as synthetic other products of raw material.
4. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature enlarged culturing base in described test tube enlarged culturing therein is the potato sucrose substratum.
5. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature during described liquid shaking bottle is cultivated therein and liquid shaking bottle substratum and liquid thalline substratum in the thalline fluid enlargement culture be: glucose 5～30 grams per liters, peptone 0～5 grams per liter, yeast extract paste 0～5 grams per liter, sal epsom 0.2～1 grams per liter, tween-80 0.5～10 grams per liter, potassium primary phosphate 2～9 grams per liters, pH5～8,120～140 ℃ sterilizing 20～40 minutes.
6. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature described shake-flask culture processing condition therein is, inoculation 3-5 piece Phanerochaete chrysosporium test tube slant thalline (5 * 5mm) is in the 250mL triangular flask that 40～120 mL substratum are housed, at rotating speed: 150 rev/mins, 25～35 ℃, cultivate 24～72 hours.
7. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature described yeast culture technique therein is, by 1～20%(volume ratio) inoculum size the shake-flask culture seed liquor be inoculated into to liquid thalline enlarged culturing technique be, 25～35 ℃ of temperature, 50～200 rev/mins of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18～72 hours; Nutrient solution centrifugal 5～20 minutes through 3000～6000rpm, obtain the Phanerochaete chrysosporium thalline.
8. a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound Technology according to claim 1, its feature described remove waste water with thalline in the technique of anthraquinone analog compound be: above-mentioned Phanerochaete chrysosporium thalline with the waste water that contains 0.05～1 grams per liter anthraquinone analog compound (by vacuum concentration or dilution adjusting anthraquinone analog compound concentration) by 1:10～50(mass/volume) mix, 25～60 ℃ of temperature, 50～200 rev/mins of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react 3～15 hours, can remove anthraquinone analog compound.
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Non-Patent Citations (2)
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| I. MIELGO等: "A packed-bed fungal bioreactor for the continuous decolourisation of azo-dyes (Orange II)", 《JOURNAL OF BIOTECHNOLOGY》 * |
| 王慧等: "真菌对染料的脱色研究进展", 《应用与环境生物学报》 * |
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