CN103663721A - Method for degrading gallic acid in wastewater by using Phanerochaete chrysosporium - Google Patents
Method for degrading gallic acid in wastewater by using Phanerochaete chrysosporium Download PDFInfo
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- CN103663721A CN103663721A CN201310407288.XA CN201310407288A CN103663721A CN 103663721 A CN103663721 A CN 103663721A CN 201310407288 A CN201310407288 A CN 201310407288A CN 103663721 A CN103663721 A CN 103663721A
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Abstract
The invention discloses a technique for degrading gallic acid in wastewater by using Phanerochaete chrysosporium, relating to the field of bioengineering. The technique comprises the following steps: by using Phanerochaete chrysosporium as the initial strain, carrying out liquid shake culture and liquid strain amplification culture, transferring the strain solution into the wastewater containing gallic acid, supplementing certain nutrient substances, and carrying out fermentation culture. By using the technique, the removal rate of the gallic acid can reach 60-100%.
Description
Technical field
The present invention relates to technical field of bioengineering, refer in particular to the waste water that utilizes Phanerochaete chrysosporium degraded to contain gallic acid.Raw wastewater is concentrated or is diluted to gallic acid content after 1～50 grams per liter, supplemental medium composition, the gallic acid in access Phanerochaete chrysosporium strain degradation waste water.After Phanerochaete chrysosporium degraded, the clearance of gallic acid can reach 60～100%.
Background technology
(the Dissolved organic matter of dissolved organic matter in natural water body, DOM) main component is natural organic acids (Natural organic acids, NOA), and 15-30% is hydrophilic small molecules organic acid in natural organic acids, gallic acid is as the ubiquitous hydrophilic small molecules organic acid of nature, in drink water purifying process, can bring following problem: the colourity that 1) affects water, mouthfeel and smell: 2) in chlorine disinfectant process, form disinfection byproduct (DBP) (Disinfection byproducts, be called for short DPBs), as trichloromethane, halogen acetic acid class, they can be to organism especially mankind liver in water body and after chloropexia, kidney and central nervous system cause damage, have carcinogenic, teratogenesis, mutagenesis, 3) in the advanced treatment process of follow-up water, can easily cause the pollution of film.Gallic acid is also important source material used in food-processing, daily use chemicals, medicine production, in production process, institute's effluent contains a large amount of gallic acids, it can decompose and produce organic acid and biogas etc. in water body, consumes oxygen in water, causes the hydrobiont anoxics such as fishes and shrimps and death; Cause that water body COD raises etc., bring thus a series of water ecological environment problem.Therefore in efficient degradation water body, gallic acid becomes an importance of water pollution control.
At present about the processing patent that contains gallic acid waste water, there are several, but be mainly the methods such as washing or alkaline extraction, as patent method of recovering and treating gallic acid sludge (application number 200610086067.7), protected waste residue twice washing under 90 ~ 100 degree, the method for twice squeezing reclaims the method for gallic acid; Patent technology for recovering mother solution of gallic acid (patent No. 200910226622.5) has been protected and has been utilized the acid of alkaline extraction gallic acid, then regulates pH8.0-9.0 precipitation to obtain the technique of gallic acid crude product; Patent gallic acid production wastewater, waste residue, useless carbon recycling new technology (number of patent application 201210469098.6) disclose and have produced the waste residue of gallic acid, useless charcoal adopts extrusion process of adverse current secondary washing to reclaim gallic acid technique; A kind of method (application number 2010136553.1) and article " Fenton reaction-neutralization-biological process is processed gallic acid crude wastewater " (magazine " water technology " 2010 of processing the crude wastewater of gallic acid of patent, the 9th phase of 36 volumes, pp106-109), disclose gallic acid crude wastewater through catalyzed oxidation, regulate pH precipitation, charcoal absorption, finally by crossing the method for diluting Air Exposure, these methods all exist to be processed not exclusively, or dilutes with a large amount of water.
The inventor is through deep research, find out a kind of with Phanerochaete chrysosporium (
phanerochaete chrysosporium) as starting strain, the method that degraded contains gallic acid waste water.The method that the method is different from is in the past that used bacterial classification is directly to process the waste liquid (1～50 grams per liter) containing high density gallic acid by the method for fermentation, has saved diluting water and place takies
Summary of the invention
The present invention is to provide the biotechnology of gallic acid in Phanerochaete chrysosporium degrading waste water.Waste water is after Phanerochaete chrysosporium degraded, and its gallic acid clearance can arrive 60 ~ 100%.
A kind of Phanerochaete chrysosporium degraded of the present invention gallic acid 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, by gallic acid in fermentative degradation waste water.
The present invention's bacterial classification used is any bacterial classification of Phanerochaete chrysosporium, 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 gallic acid, as the waste liquid of prodn. of gallic acid by fermentation process, extract the waste liquid of gallic acid or take the waste liquid of gallic acid as synthetic other products of raw material from plant.
Test tube 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).
Liquid shaking bottle substratum and liquid spawn culture medium during wherein said liquid shaking bottle is cultivated and in thalline fluid enlargement culture are: glucose 5～30 grams per liters, 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 liters, pH5～8,120～140 ℃ of sterilizings 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 being equipped with the 250mL triangular flask of 40～120 mL substratum, at rotating speed: 150 revs/min, 25～35 ℃, cultivate 24～72 hours; Wherein said first order seed culture process is, by 1～20%(volume ratio) inoculum size shake-flask culture seed liquor is inoculated in first order seed substratum, 25～35 ℃ of temperature, 50～200 revs/min of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18～72 hours;
In degrading waste water of the present invention, the technique of gallic acid is, the waste water that contains gallic acid makes gallic acid concentration reach 1～50 grams per liter waste water 1000L, 5～30 kilograms of glucose, 0.5～2 kilogram of soybean cake powder through concentrated or dilution; 5～15 grams, copper sulfate, 5～15 grams, ferrous sulfate, 5～20 grams of manganous sulfates, each composition of this substratum can amplify in this ratio; Wherein said degradation technique is: by 2～20%(seed liquor volume/wastewater volume) inoculation level liquid bacterial classification, 25～35 ℃ of temperature, 50～200 revs/min of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 36～168 hours.
Use after this processing wastewater degraded, gallic acid clearance can reach 60-100%.
According to fermentating culturing process of the present invention, in conjunction with the ABC of this area, can be according to need of production, increase even level Four seeding 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 level Four seeding tank adopts are identical with first order seed substratum with 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 gallic acid measuring method of this process using, be: precision takes gallic acid reference substance 10.3 mg, be placed in 100 mL beakers, with hplc grade methanol, dissolve, and be settled in 100mL volumetric flask, be mixed with the standard reserving solution of 10.3mg/mL.Face the used time with 0.45 μ m filtering with microporous membrane, controlling sample size is 2,4,6,8, and 10 μ l utilize high performance liquid chromatograph (HIMADZU (Shimadzu) LC-20AT) to measure gallic acid content.Chromatographic column is Alltima
tMc18 post (4.6 mm * 250 mm, 5 μ m).Condition determination is moving phase: methyl alcohol: 0.1% phosphoric acid solution=10:90; Column temperature: 30 ℃, flow velocity: 0.8 mL/min, detects wavelength: 270 nm.Take sample size as X-coordinate (X), and the corresponding peak area of take carries out linear regression as ordinate zou (Y), tries to achieve the equation of linear regression of gallic acid concentration and peak area.The supernatant liquor of drawing sample liquid (before degraded and in degradation process and after degraded) is a small amount of, with 0.45 μ m filtering with microporous membrane, by above-mentioned chromatographic condition, measure respectively peak area, adopt external standard peak area according to the regression equation calculation gallic acid content of above-mentioned gallic acid concentration of trying to achieve and peak area.
Gallic acid clearance Re calculates by following formula (1):
In formula,
r t clearance for t time gallic acid;
c 0 (mg/L) be the gallic acid concentration of 0 hour;
c t (mg/L) for adsorbing the concentration of t hour gallic acid.
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
Access one ring Phanerochaete chrysosporium bacterial classification in the potato sucrose substratum (Zhu Gejian, Wang Zhengxiang chief editor's " industrial microorganism experimental technique handbook ", 1994,367 pages) of new configuration (
phanerochaete chrysosporium) (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 5 grams of glucose, 0 gram of peptone, 5 grams of yeast extract pastes, 0.2 gram, magnesium sulfate, 0.5 gram of tween 80,2 grams of potassium primary phosphates, water 1000 mL, pH 5, packing 250mL triangular flask, 50 grams every bottle, totally 20 bottles, 120 ℃ of sterilizings 40 minutes; After cooling, 3 Phanerochaete chrysosporium test tube slant thalline of every bottle graft kind (5 * 5mm), at 25 ℃, 150 revs/min, cultivate 72 hours;
3. the making of level liquid bacterial classification
Accurately take 50 grams of glucose, 0 gram of peptone, 50 grams of yeast extract pastes, 2 grams, magnesium sulfate, 5 grams of tween 80s, 20 grams of potassium primary phosphates, water 10L, is loaded in the seeding tank of 15L, 120 ℃ of sterilizings 20 minutes; After sterilizing is cooling, 25 ℃ of temperature, 50 revs/min of mixing speed, 0.2: 1 volume/volume of air flow/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 72 hours;
4. wastewater degradation
Take Turkey-galls as raw material, adopt the waste liquid of High Temperature High Pressure and alkaline extraction gallic acid through concentrated, gallic acid concentration is the waste liquid 1000L of 50 grams/L, 5 kilograms of glucose, 0.5 kilogram of soybean cake powder; 5 grams, copper sulfate, 5 grams, ferrous sulfate, 5 grams of manganous sulfates; Access 2%(seed liquor volume/wastewater volume) inoculation level liquid bacterial classification, 25 ℃ of temperature, 60 revs/min of mixing speed, air flow 0.3:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate after 44 hours, gallic acid clearance reaches 60.2%.
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 150 grams of glucose, 25 grams of peptones, 25 grams of yeast extract pastes,, 0.4 gram, magnesium sulfate, 4 grams of tween 80s, 5 grams of potassium primary phosphates, water 10 L, pH 6, packing 250mL triangular flask, 100 grams every bottle, totally 100 bottles, 130 ℃ of sterilizings 30 minutes; After cooling, 4 Phanerochaete chrysosporium test tube slant thalline of every bottle graft kind (5 * 5mm), at 30 ℃, 150 revs/min, cultivate 50 hours;
3. the making of level liquid bacterial classification
Accurately take 2000 grams of glucose, 250 grams of peptones, 250 grams of yeast extract pastes, 60 grams, magnesium sulfate, 60 grams of tween 80s, 400 grams of potassium primary phosphates, water 100L, is loaded in the seeding tank of 130L, 130 ℃ of sterilizings 30 minutes; After cooling, 30 ℃ of temperature, 125 revs/min of mixing speed, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 50 hours;
4. wastewater degradation
The Turkey-galls of take is prepared the waste liquor of gallic acid as raw material by fermentation of Aspergillus niger, through concentrated be the waste water 1000L that the content of gallic acid reaches 25 grams/L, 20 kilograms of glucose, 1.2 kilograms of soybean cake powders; 10 grams, copper sulfate, 10 grams, ferrous sulfate, 12 grams of manganous sulfates; Press 13%(seed liquor volume/wastewater volume) inoculation level liquid bacterial classification, 30 ℃ of temperature, 120 revs/min of mixing speed, air flow 1:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 100 hours.Now gallic acid clearance reaches 80.3%.
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 300 grams of glucose, 50 grams of peptones, 0 gram of yeast extract paste, 10 grams, magnesium sulfate, 100 grams of tween 80s, 90 grams of potassium primary phosphates, water 10L, pH7.5, packing 250mL triangular flask, every bottle of 120 mL, totally 80 bottles, 140 ℃ of sterilizings 20 minutes; After cooling, 5 Phanerochaete chrysosporium test tube slant thalline of every bottle graft kind (5 * 5mm), at 35 ℃, 150 revs/min, cultivate 72 hours;
3. the making of level liquid bacterial classification
Accurately take 1500 grams of glucose, 250 grams of peptones, 0 gram of yeast extract paste, 50 grams, magnesium sulfate, 500 grams of tween 80s, 450 grams of potassium primary phosphates, water 50L, is loaded in the first class seed pot of 70L, 140 ℃ of sterilizings 40 minutes; After sterilizing is cooling, 35 ℃ of temperature, 200 revs/min of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 50 hours;
4. the making of second-class liquid isolate
Accurately take 15000 grams of glucose, 2500 grams of peptones, 0 gram of yeast extract paste, 500 grams, magnesium sulfate, 5000 grams of tween 80s, 4500 grams of potassium primary phosphates, water 500L, is loaded in the secondary seed tank of 700L, 140 ℃ of sterilizings 40 minutes; After sterilizing is cooling, 35 ℃ of temperature, 200 revs/min of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18 hours;
5. wastewater degradation
Take Microcrystalline Cellulose, gallic acid is raw material, under the effect of catalyzer, adopts the waste liquid of direct esterification synthesizing gallic acid Microcrystalline Cellulose ester, and being diluted to gallic acid content is the waste liquid 1000L of 1 gram/L, 30 kilograms of glucose, 1.8 kilograms of soybean cake powders; 15 grams, copper sulfate, 15 grams, ferrous sulfate, 20 grams of manganous sulfates, press 20%(seed liquor volume/wastewater volume) inoculation level liquid bacterial classification, 34 ℃ of temperature, 190 revs/min of mixing speed, air flow 2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 168 hours.Gallic acid clearance reaches 100%.
Claims (9)
1. Phanerochaete chrysosporium degraded gallic acid waster water process 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, liquid fermenting is removed gallic acid in waste water.
2. a kind of Phanerochaete chrysosporium degraded gallic acid waster water process 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 gallic acid Technology according to claim 1, its feature is any one waste water that contains gallic acid at applicable waste water, as the waste liquid of prodn. of gallic acid by fermentation process, extract the waste liquid of gallic acid or take the waste liquid of gallic acid as synthetic other products of raw material from plant.
4. a kind of Phanerochaete chrysosporium degraded gallic acid Technology according to claim 1, its feature therein enlarged culturing base in described test tube enlarged culturing is potato sucrose substratum.
5. a kind of Phanerochaete chrysosporium degraded gallic acid 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 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, magnesium sulfate 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 ℃ of sterilizings 20～40 minutes.
6. a kind of Phanerochaete chrysosporium degraded gallic acid Technology according to claim 1, its feature therein described shake-flask culture processing condition is, inoculation 3-5 piece Phanerochaete chrysosporium test tube slant thalline (5 * 5mm) is in being equipped with the 250mL triangular flask of 40～120 mL substratum, at rotating speed: 150 revs/min, 25～35 ℃, cultivate 24～72 hours.
7. a kind of Phanerochaete chrysosporium degraded gallic acid Technology according to claim 1, its feature therein described yeast culture technique is, by 1～20%(volume ratio) inoculum size shake-flask culture seed liquor is inoculated in liquid thalline enlarged culturing, 25～35 ℃ of temperature, 50～200 revs/min of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 18～72 hours.
8. a kind of Phanerochaete chrysosporium degraded gallic acid waster water process technology according to claim 1, its feature in the described gallic acid wastewater medium that contains is: the waste water that contains gallic acid makes gallic acid concentration reach 1～50 grams per liter waste water 1000L through concentrated or dilution, 5～30 kilograms of glucose, 0.5～2 kilogram of soybean cake powder; 5～15 grams, copper sulfate, 5～15 grams, ferrous sulfate, 5～20 grams of manganous sulfates, each composition of this substratum can amplify in this ratio.
9. a kind of Phanerochaete chrysosporium degraded gallic acid waster water process technology according to claim 1, its feature therein described degrading waste water technique is: by 2～20%(seed liquor volume/wastewater volume) inoculate level liquid bacterial classification, 25～35 ℃ of temperature, 50～200 revs/min of mixing speed, air flow 0.2～2:1 volume/volume/minute (ventilation gas volume/fermentating liquid volume/minute), cultivate 36～168 hours, i.e. degradable gallic acid.
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| CN105417726A (en) * | 2015-12-15 | 2016-03-23 | 安徽皖维高新材料股份有限公司 | Method for treating polyvinyl alcohol production wastewater through white rot fungi |
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| CN105417726A (en) * | 2015-12-15 | 2016-03-23 | 安徽皖维高新材料股份有限公司 | Method for treating polyvinyl alcohol production wastewater through white rot fungi |
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Application publication date: 20140326 |