CN103482772B - 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 Phanerochaete chrysosporium thalline to remove containing the anthraquinone analog compound in the waste water of anthraquinone analog compound.Concentrated by raw wastewater or be diluted to Anthraquinone after 0.05 ~ 1 grams per liter, dropping into Phanerochaete chrysosporium thalline, anthraquinone analog compound is removed in aeration-agitation reaction.The clearance 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, thus 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.Simultaneously along with the lifting day by day of environmental requirement, azoic dyestuff forbidding range extension, also makes the importance of anthraquinone dye more outstanding, makes its application in printing and dyeing industry more and more extensive.But the anthraquinone analog compound not only factory effluent water yield is large, 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 of directly discharge, anthraquinone passes through sedimentation! Food chain enrichment etc. effect can prolonged stay in water body! Soil! In settling and air.Therefore explore the technology of effectively degraded anthraquinone analog compound, the harmony for society is of great significance with development tool.
Have several about the process patent containing anthraquinone analog compound waste water at present, but main method is the recycling of anthraquinone, microbiological deterioration and oxidation removal.As patent " 1; improvement of 4-dihydroxyanthraquinone factory effluent and its recovery method as resource " (application number 00112386.6) discloses and utilizes macroporous resin adsorption, the method of the anthraquinone analog compound recycled in Isosorbide-5-Nitrae-dihydroxyanthraquinone factory effluent is resolved again with sodium hydroxide; Patent " recovery method of a kind of Isosorbide-5-Nitrae-dihydroxyanthraquinone processing wastewater " (application number 200410013484.X) discloses the method utilizing the technology recycling anthraquinones such as concentrated, cooling, crystallization, separation and drying.Patent " Sphingomonas strain and the application in anthraquinone dye wastewater decolouring thereof " (application number 200410020832.6) discloses Sphingomonas, application in 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 electrochemical oxidation (patent No. 200910029887.6) and photochemical catalytic oxidation (200710011629.6) in addition, these two kinds of methods also need further process.Also many reports are had with these method correlative theses.Other doctor's or master's degree paper reports Phanerochaete chrysosporium absorption degradation anthraquinone analog compound mechanism and is studied, but does not relate to its Technology.
The present inventor through deep research, find out one utilize Phanerochaete chrysosporium (
phanerochaete chrysosporium) thalline, remove anthraquinone analog compound technique in waste water in ventilation and stirring reaction for some time, 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 Phanerochaete chrysosporium degrading waste water.Waste water is after 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, carry out according to following step: take Phanerochaete chrysosporium as starting strain, carry out test tube enlarged culturing, liquid submerged culture, thalline fluid enlargement culture, collected by centrifugation thalline, then removes 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 China General Microbiological culture presevation administrative center (CCGMC), Chinese agriculture Microbiological Culture Collection administrative center (ACCC), Chinese industrial Microbiological Culture Collection administrative center (CICC), Chinese medicine Microbiological Culture Collection administrative center (CMCC), National Veterinary Culture Collection (CVCC) and microbiotic bacterial classification preservation pipe reason center etc. preservation.
The present invention the waste water that is suitable for be that any one contains the waste water of anthraquinone analog compound, as the waste liquid of fermentative Production anthraquinone analog compound, extract from plant anthraquinone analog compound waste liquid or take anthraquinone analog compound as the waste liquid of other products of Material synthesis.
Enlarged culturing base in wherein said test tube enlarged culturing is potato sucrose substratum (" the industrial microorganism experimental technique handbook " of Zhu Gejian, Wang Zhengxiang chief editor, 1994,367 pages.
Liquid submerged culture base in wherein said liquid submerged culture and in thalline fluid enlargement culture and liquid thalline substratum are: glucose 5 ~ 30 grams per liter, peptone 0 ~ 5 grams per liter, yeast extract paste 0 ~ 5 grams per liter, magnesium sulfate 0.2 ~ 1, tween-80 0.5 ~ 10, potassium primary phosphate 2 ~ 9, pH5 ~ 8,120 ~ 140 DEG C of sterilizings 20 ~ 40 minutes.
Wherein said shake-flask culture processing condition are, inoculation 3-5 block Phanerochaete chrysosporium test tube slant thalline (5 × 5mm), in the 250mL triangular flask that 40 ~ 120 mL substratum are housed, at rotating speed: 150 revs/min, 25 ~ 35 DEG C, is cultivated 24 ~ 72 hours.Wherein said yeast culture technique is, by 1 ~ 20%(volume ratio) inoculum size shake-flask culture seed liquor is inoculated into liquid thalline enlarged culturing technique and is, temperature 25 ~ 35 DEG C, mixing speed 50 ~ 200 revs/min, 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, obtains Phanerochaete chrysosporium thalline.
The technique that thalline of the present invention removes anthraquinone analog compound in waste water is: above-mentioned Phanerochaete chrysosporium thalline with contain 0.05 ~ 1 grams per liter anthraquinone analog compound waste water (by vacuum concentration or dilute regulate anthraquinone analog compound concentration) by 1:10 ~ 50(mass/volume) mix, temperature 25 ~ 60 DEG C, mixing speed 50 ~ 200 revs/min, air flow 0.2 ~ 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react 3 ~ 15 hours, can anthraquinone analog compound be removed.
After using this processing wastewater to degrade, 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 according to need of production, increase secondary, three grades of even level Four thalline tanks, expand the scale of production further, to carry out suitability for industrialized production.Secondary, three grades even level Four seeding tank adopt substratum identical with thalline fluid enlargement culture base with Shake flask medium, 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 that this technique adopts be: accurate absorption 1,8-dihydroxyanthraquinone reference substance solution (0.522 mg/ml) 1,3,5,8,10,15 ml, put in 25ml volumetric flask respectively, 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), (x) linear regression analysis is carried out to optical density, obtain equation of linear regression.Accurate pipette samples solution repeats aforesaid operations and measures 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):
(1)
In formula,
r t for the clearance of t time anthraquinone analog compound;
c 0 (mg/L) be the anthraquinone analog compound concentration of 0 hour;
c t (mg/L) be the concentration of absorption t hour anthraquinone analog compound.
In order to set forth the material and progress involved by technical scheme of the present invention further, give following examples, but these embodiments do not limit the scope of the invention in any form.
Embodiment 1
1. the making of test tube slant bacterial classification
At potato sucrose substratum (" the industrial microorganism experimental technique handbook " of Zhu Gejian, Wang Zhengxiang chief editor of new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (China General Microbiological culture presevation administrative center CCGMC5.776), 26 DEG C, incubation time 50 hours; 4 DEG C save backup.
2. the making of liquid submerged culture bacterial classification
Accurately take glucose 5 grams, peptone 0 gram, yeast extract paste 5 grams, 0.2 gram, magnesium sulfate, tween-80 0.5 gram, potassium primary phosphate 2 grams, water 1000 mL, pH 5, packing 250mL triangular flask, 50 grams every bottle, totally 20 bottles, 120 DEG C of sterilizings 40 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 × 10 that 4 DEG C, a ring is preserved
6individual spore), at 25 DEG C, 150 revs/min, cultivate 72 hours;
3. thalline fluid enlargement culture
Accurately take glucose 50 grams, peptone 0 gram, yeast extract paste 50 grams, 2 grams, magnesium sulfate, tween-80 5 grams, potassium primary phosphate 20 grams, water 10L, is loaded in the seeding tank of 15L, 120 DEG C of sterilizings 20 minutes; After sterilizing cooling, temperature 25 DEG C, mixing speed 50 revs/min, air flow 0.2: 1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 72 hours; Nutrient solution centrifugal 20 minutes through 3000rpm, obtains Phanerochaete chrysosporium thalline.
4. remove anthraquinone analog compound in waste water
Take rheum officinale as raw material, decocting method is adopted to extract the waste liquid of anthraquinone analog compound through concentrated, anthraquinone analog compound concentration is the waste liquid 1000L of 1 gram/L, thalline and waste liquid are by 1:50(mass/volume) mix, temperature 25 DEG C, mixing speed 60 revs/min, air flow 0.3:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react after 3 hours, anthraquinone analog compound clearance reaches 71.3%.
Embodiment 2
1. the making of test tube slant bacterial classification
At potato sucrose substratum (" the industrial microorganism experimental technique handbook " of Zhu Gejian, Wang Zhengxiang chief editor of new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (Chinese agriculture Microbiological Culture Collection administrative center ACCC 30942), 30 DEG C, incubation time 100 hours; 7 DEG C save backup.
2. the making of liquid submerged culture bacterial classification
Accurately take glucose 150 grams, peptone 25 grams, yeast extract paste 25 grams, 0.4 gram, magnesium sulfate, tween-80 4 grams, potassium primary phosphate 5 grams, water 10 L, pH 6, packing 250mL triangular flask, 100 grams every bottle, totally 100 bottles, 130 DEG C of sterilizings 30 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 × 10 that 7 DEG C, a ring is preserved
6individual spore), at 30 DEG C, 150 revs/min, cultivate 50 hours;
3. thalline fluid enlargement culture
Accurately take glucose 2000 grams, peptone 250 grams, yeast extract paste 250 grams, 60 grams, magnesium sulfate, tween-80 60 grams, potassium primary phosphate 400 grams, water 100L, is loaded in the seeding tank of 130L, 130 DEG C of sterilizings 30 minutes; After cooling, temperature 30 DEG C, mixing speed 125 revs/min, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 50 hours; Nutrient solution centrifugal 13 minutes through 4500rpm, obtains Phanerochaete chrysosporium thalline.
4. remove anthraquinone analog compound in waste water
Fusarium oxysporum fermentation is for the waste liquid of anthraquinone analog compound, the content of anthraquinone analog compound is made to reach the waste water 1000L of 0.2 gram/L through concentrated, thalline and waste liquid are by 1:30(mass/volume) mix, at temperature 45 C, mixing speed 120 revs/min, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react after 7 hours, anthraquinone analog compound clearance reaches 87.5%.
Embodiment 3
1. the making of test tube slant bacterial classification
At potato sucrose substratum (" the industrial microorganism experimental technique handbook " of Zhu Gejian, Wang Zhengxiang chief editor of new configuration, 1994,367 pages) middle access one ring Phanerochaete chrysosporium bacterial classification (Chinese industrial Microbiological Culture Collection administrative center CICC14067), 35 DEG C, incubation time 144 hours; 10 DEG C save backup.
2. the making of liquid submerged culture bacterial classification
Accurately take glucose 300 grams, peptone 50 grams, yeast extract paste 0 gram, 10 grams, magnesium sulfate, 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 DEG C of sterilizings 20 minutes; After cooling, every bottle graft enters the Phanerochaete chrysosporium bacterial classification (1 × 10 that 10 DEG C, a ring is preserved
6individual spore), at 35 DEG C, 150 revs/min, cultivate 72 hours;
3. one-level thalline fluid enlargement culture
Accurately take glucose 1500 grams, peptone 250 grams, yeast extract paste 0 gram, 50 grams, magnesium sulfate, tween-80 500 grams, potassium primary phosphate 450 grams, water 50L, is loaded in the first class seed pot of 70L, 140 DEG C of sterilizings 40 minutes; After sterilizing cooling, temperature 35 DEG C, mixing speed 200 revs/min, 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, 500 grams, magnesium sulfate, tween-80 5000 grams, potassium primary phosphate 4500 grams, water 500L, be loaded in the secondary seed tank of 700L, 140 DEG C of sterilizings 40 minutes; After sterilizing cooling, temperature 35 DEG C, mixing speed 200 revs/min, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18 hours; Nutrient solution centrifugal 5 minutes through 6000rpm, obtains Phanerochaete chrysosporium thalline.
5. remove anthraquinone analog compound in waste water
Take anthraquinone analog compound as raw material, under the effect of catalyzer, the waste liquid of synthesizing nitryl anthraquinone, be diluted to the waste liquid 1000L that Anthraquinone is 0.05 gram/L, thalline and waste liquid are by 1:10(mass/volume) mix, at temperature 60 C, mixing speed 200 revs/min, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), react after 15 hours, anthraquinone analog compound clearance reaches 100%.
Claims (1)
1. a Phanerochaete chrysosporium degraded anthraquinone analog compound technique, it is characterized in that carrying out according to following step: take Phanerochaete chrysosporium as starting strain, carry out test tube enlarged culturing, liquid submerged culture, thalline fluid enlargement culture, collected by centrifugation thalline, then removes anthraquinone analog compound in waste water with thalline; Anthraquinone analog compound clearance reaches 70 ~ 100%;
Bacterial classification used is any strain bacterial classification of Phanerochaete chrysosporium;
The waste water that is suitable for be fermentative Production anthraquinone analog compound waste liquid, extract from plant anthraquinone analog compound waste liquid or take anthraquinone analog compound as the waste liquid of other products of Material synthesis;
Enlarged culturing base in wherein said test tube enlarged culturing is potato sucrose substratum;
Liquid submerged culture base in wherein said liquid submerged culture and in thalline fluid enlargement culture and thalline liquid nutrient medium are: glucose 5 ~ 30 grams per liter, peptone 0 ~ 5 grams per liter, yeast extract paste 0 ~ 5 grams per liter, magnesium sulfate 0.2 ~ 1 grams per liter, tween-80 0.5 ~ 10 grams per liter, potassium primary phosphate 2 ~ 9 grams per liter, pH5 ~ 8,120 ~ 140 DEG C of sterilizings 20 ~ 40 minutes;
Wherein said shake-flask culture processing condition are, inoculation 3-5 block, size be 5 × 5mm Phanerochaete chrysosporium test tube slant thalline in the 250mL triangular flask that 40 ~ 120 mL substratum are housed, at rotating speed: 150 revs/min, 25 ~ 35 DEG C, cultivate 24 ~ 72 hours;
Wherein said thalline fluid enlargement culture technique is, be by volume 1 ~ 20% inoculum size shake-flask culture seed liquor is inoculated into thalline fluid enlargement culture, temperature 25 ~ 35 DEG C, mixing speed 50 ~ 200 revs/min, air flow 0.2 ~ 2:1 ventilation gas volume/fermentating liquid volume/minute, cultivate 18 ~ 72 hours; Nutrient solution centrifugal 5 ~ 20 minutes through 3000 ~ 6000rpm, obtains Phanerochaete chrysosporium thalline;
Described a kind of Phanerochaete chrysosporium degraded anthraquinone analog compound technique is: above-mentioned Phanerochaete chrysosporium thalline with contain that to regulate anthraquinone analog compound concentration to be the waste water of 0.05 ~ 1 grams per liter anthraquinone analog compound by vacuum concentration or dilute be that 1:10 ~ 50 mix by quality and volume ratio, temperature 25 ~ 60 DEG C, mixing speed 50 ~ 200 revs/min, air flow 0.2 ~ 2:1 ventilation gas volume/fermentating liquid volume/minute, react 3 ~ 15 hours, can anthraquinone analog compound be removed.
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Effective date of registration: 20210126 Address after: No. 159, Chengjiang Middle Road, Jiangyin City, Wuxi City, Jiangsu Province Patentee after: Jiangyin Intellectual Property Operation Co., Ltd Address before: Zhenjiang City, Jiangsu Province, 212013 Jingkou District Road No. 301 Patentee before: JIANGSU University |
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