CN102071146A - Culture method and applications of Ganoderma weberianum mycelial pellets - Google Patents
Culture method and applications of Ganoderma weberianum mycelial pellets Download PDFInfo
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- CN102071146A CN102071146A CN 201010524508 CN201010524508A CN102071146A CN 102071146 A CN102071146 A CN 102071146A CN 201010524508 CN201010524508 CN 201010524508 CN 201010524508 A CN201010524508 A CN 201010524508A CN 102071146 A CN102071146 A CN 102071146A
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- lucidum pompon
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- 239000008188 pellet Substances 0.000 title abstract description 41
- 238000012136 culture method Methods 0.000 title abstract 3
- 241001480598 Ganoderma weberianum Species 0.000 title abstract 2
- 229940097275 indigo Drugs 0.000 claims abstract description 83
- 235000000177 Indigofera tinctoria Nutrition 0.000 claims abstract description 82
- COHYTHOBJLSHDF-UHFFFAOYSA-N indigo powder Natural products N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 claims abstract description 82
- 238000000034 method Methods 0.000 claims abstract description 70
- 230000015556 catabolic process Effects 0.000 claims abstract description 57
- 238000006731 degradation reaction Methods 0.000 claims abstract description 57
- 239000002351 wastewater Substances 0.000 claims abstract description 51
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 241001388119 Anisotremus surinamensis Species 0.000 claims description 73
- 238000004043 dyeing Methods 0.000 claims description 56
- 238000004042 decolorization Methods 0.000 claims description 55
- 239000000243 solution Substances 0.000 claims description 54
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 27
- 229910052739 hydrogen Inorganic materials 0.000 claims description 27
- 239000001257 hydrogen Substances 0.000 claims description 27
- 239000002609 medium Substances 0.000 claims description 11
- 239000001963 growth medium Substances 0.000 claims description 10
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 239000011259 mixed solution Substances 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 244000061456 Solanum tuberosum Species 0.000 claims description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
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- 235000013619 trace mineral Nutrition 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 235000019156 vitamin B Nutrition 0.000 claims description 3
- 239000011720 vitamin B Substances 0.000 claims description 3
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Abstract
The invention provides a culture method and applications of Ganoderma weberianum mycelial pellets, belonging to the biological technical field. The culture method comprises the following steps: preculturing on a shelving medium slant to activate strains, transferring the activated strains to plating medium to culture and obtain plating strains; and transferring the plating strains to liquid medium to perform shaking culture under controlled conditions and obtain the mycelial pellets. The mycelial pellet culture prepared by the method is mixed with 0.5-10 times of indigo solution or denim printing and dying wastewater by volume to perform shaking culture for decoloration and degradation under the conditions that the pH value is 3-7, the temperature is 13-38 DEG C and the rotational speed is 160rpm or the conditions that the pH value is 2-6, the temperature is 23-43 and the rotational speed is 160rpm. The invention has the following advantages: the cell immobilization cost is saved, industrial production is easy to realize; decoloration and degradation can be performed without using toxic and expensive medium, thus the method is green and environmentally friendly; and the service life of the mycelial pellets is long, the mycelial pellets can be used repeatedly; and the method is convenient and extensive to operate and the cost is low.
Description
Technical field
The present invention relates to the cultural method and the application thereof of a kind of weber of Ganderma lucidum pompon, belong to biological technical field, specifically at first make the aggegation automatically under the situation of not adding entrapment media of weber glossy ganoderma cell, coagulate wadding formation mycelium pellet, utilize the mycelium pellet that is obtained that indigo and dyeing waste water are carried out decolored degradation again and handle.
Background technology
2006,536.8 hundred million tons of China's wastewater emission amounts, wherein trade effluent is 240.2 hundred million tons, 19.8 hundred million tons of dyeing waste waters, quantity discharged occupies the 5th of each industrial sector total emission volumn of the whole nation, and gross contamination emission then occupies the 6th of each industrial sector.Therefore, dyeing industry is that one of industry is polluted in China's priority industry.At present, the total dyestuff that surpasses 10,000 kinds is widely used in production processes such as yarn fabric printing and dyeing, produces a large amount of colorful wastewaters.Because dyestuff just is designed to waterproof, anti-illumination, oxidation resistant difficult degradation compound when synthetic, dyeing waste water is the trade effluent that a class is difficult to degrade.Dissimilar dye structures differ greatly, and its decolored degradation mechanism is different, therefore, should develop integrated different Technology according to the characteristic of printing and dyeing, improve the processing efficiency of waste water from dyestuff.
For many years, the fashionable whole world of jeans, it is flourish to drive " cowboy " industry, and there is thousands of family denim factory in the whole nation.Therefore, denim dyeing waste water quantity discharged is very big.Denim after making clothes, also will be made fade treatment by physics (as polishing), chemistry (as oxidizer treatment) or biological methods such as (washing as enzyme) mainly with the indigo mazarine of dying.Factory effluent is because of containing the indigo faint blue that is of higher concentration.So the key of handling the denim dyeing waste water is its main dyestuff---indigo decolouring.
Treatment method of printing and dying wastewater roughly is divided three classes, i.e. physics method, chemical method and biological process, each tool relative merits.Physics and method of chemical treatment are of limited application, and running cost is higher, and also there is secondary pollution problem in chemical method.Advantages such as it is low that biological process has running cost, and treatment effect is stable, but also have the defective of the not too high and long reaction time of colourity decreasing ratio.But biological process is extremely praised highly because of environmental protection, mainly carries out the dyestuff degraded with immobilized enzyme, bacterium or fungi.Yet, the cost height of immobilized enzyme, and active low not long with the life-span; The many structures to substrate of dyestuff bacterium for degrading have the specificity of height, are unsuitable for the various treatment of dyeing wastewater of structure.Compare with bacterium, fungi has that degraded spectrum is wide, degraded concentration is low, degraded is fast and can degrade advantages such as the pollutent that is difficult to degrade and growth vigor and anti-murder by poisoning performance are good, the self-flocculation of particularly outstanding is many fungies can pass through cell and immobilization, thereby aspect environmental improvement, demonstrate wide application prospect.
Screened the fungi of some energy degradation of dye or dyeing waste water, as Phanerochaete chrysosporium, variegated rainbow conk, variable color bolt bacterium, hair bolt bacterium, pycnoporus samguineus etc.But these fungies all also are not very high to the degradation efficiency of dyestuff especially dyeing waste water.Phanerochaete chrysosporium be find the earliest with the dye decolored bacterium of studying at most, in the biological decolouring of various wastewater, utilize, the decolorization of dyeing rate is up to 96 hours 93%.
At present used denim treatment method of printing and dying wastewater mainly is the method that traditional physics, chemistry and biology (mainly utilizing bacterium) or wherein two or three combine, and has problems such as the not high and long reaction time of colourity removal efficiency.The present invention at first utilizes the weber ganoderma strain capable that early stage, screening obtained to produce the mycelium pellet culture, carries out the denim dyestuff again---and biological decolouring indigo or the denim dyeing waste water is degraded.Through condition optimizing, obtained to handle making indigo in 3.8 and 13.2 hours respectively or denim decolorization of dyeing rate reaches 99.0% effect.
Summary of the invention
The object of the present invention is to provide the cultural method of a kind of weber of Ganderma lucidum pompon.
Another object of the present invention is that the weber Ganderma lucidum pompon that utilizes the present invention to obtain carries out indigo, denim decolorization of dyeing degraded.
The cultural method of weber Ganderma lucidum pompon provided by the present invention is included under the aseptic condition, the weber Ganderma lucidum strain is inoculated in the slant medium cultivates in advance; Then, under aseptic condition, from slant strains, take mycelia with transfering loop and forward on the plate culture medium, put into 25～32 ℃ of incubators and cultivate and obtained activated spawn in 2～5 days; Under aseptic condition, take mycelia from the activated spawn plate culture medium again, be uniformly dispersed, add and be equipped with in the liquid nutrient medium of weber glossy ganoderma, shaking culture is 3～6 days under 25～32 ℃, 120～200 rev/mins condition, obtains weber Ganderma lucidum pompon.The top condition that weber Ganderma lucidum pompon is cultivated is that slant strains is inoculated on the plate culture medium, cultivates 4 days in 28 ℃ incubator, is inoculated in the liquid nutrient medium 28 ℃, 160 rev/mins shaking culture 5 days then.
Contain potato 200g, glucose 20g, KH in the cultural method of a kind of weber of Ganderma lucidum pompon, every liter of described plate culture medium
2PO4 3g, MgSO
47H
2O 1.5g, VITMAIN B1 1mg, agar 15g, pH nature.
Contain potato 200g, glucose 20g, KH in the cultural method of a kind of weber of Ganderma lucidum pompon, every liter of described liquid nutrient medium
2PO
43g, MgSO
47H
2O 1.5g, vitamins B
12mg, yeast extract paste 5g and 70mL trace element solution, surplus is a water, pH4.8;
The cultural method of a kind of weber of Ganderma lucidum pompon, the trace element solution of described liquid nutrient medium are meant and contain MnSO
4H
2O 0.5g/L, NaCl 1g/L, FeSO
47H
2O 0.1g/L, CoCl
20.1g/L, ZnSO
47H
2O 0.1g/L, CuSO
45H
2O 0.1g/L, KA1 (SO
4)
212H
2O 0.01g/L, H
3BO
30.01g/L, Na
2MoO
42H
2The aqueous solution of O 0.01g/L.
Another object of the present invention is that the weber Ganderma lucidum pompon that utilizes aforesaid method to obtain carries out indigo, denim decolorization of dyeing degraded, realizes by the following method:
1) the weber Ganderma lucidum pompon that aforesaid method obtained is in the application of indigo decolored degradation, realize by the following method: the weber Ganderma lucidum pompon that will utilize aforesaid method to obtain mixes with the indigo solution of 0.5～10 times of volume, the potential of hydrogen of mixed solution is adjusted into pH3～7, shaking culture under 13～38 ℃, 160 rev/mins condition promotes indigo decolored degradation.Best decolored degradation condition is under the top condition that above-mentioned described weber Ganderma lucidum pompon is cultivated, the weber Ganderma lucidum pompon that obtains is mixed with isopyknic indigo solution, the pH value of adjusting mixed solution is 5, handled 1 hour, make the percent of decolourization of the indigo solution of 20mg/L reach 92%, can make 99.0% dye decoloredly in promptly 3.8 hours, its decolored degradation ability is 0.67mg/h/gFW, mycelium pellet used 30 days repeatedly, can also keep per hour 37% percent of decolourization.
2) the weber Ganderma lucidum pompon that aforesaid method obtained is in the application of denim decolorizing printing and dyeing waste water degraded, realize by the following method: the weber Ganderma lucidum pompon culture that will utilize aforesaid method to obtain mixes with the denim dyeing waste water of 0.5～10 times of volume, the potential of hydrogen of mixed solution is adjusted into pH2～6, shaking culture under 23～43 ℃, 160 rev/mins condition promotes the degraded of denim decolorization of dyeing.Best decolored degradation condition is under the top condition that above-mentioned described weber Ganderma lucidum pompon is cultivated, and the weber Ganderma lucidum pompon that obtains is mixed with isopyknic denim dyeing waste water, and the pH value of adjusting mixed solution is 3, handles 4 hours, can make isopyknic OD
650=0.52 denim decolorization of dyeing rate reaches 79%, 13.2 hour can make 99.0% denim decolorizing printing and dyeing waste water, and its decolored degradation ability is 7ml/h/gFW, and mycelium pellet used 30 days repeatedly, can also keep per 4 hours 29% percent of decolourization.
The invention has the advantages that:
1) weber Ganderma lucidum pompon cultural method does not produce the fringe cost of cell fixation process; Used all is conventional method and apparatus, amplifies through the fermentor tank Chinese style, realizes large scale culturing easily;
2) handle indigo or denim decolorization of dyeing degradation efficiency height with weber Ganderma lucidum pompon, under top condition, 6 day age, mycelium pellet can make 99.0% indigo or denim dyeing waste water generation decolored degradation at 3.8 and 13.2 hours respectively;
3) carry out indigo with a weber Ganderma lucidum pompon or the degraded of denim decolorizing printing and dyeing waste water need not to use poisonous and expensive amboceptor, green, environmental protection, low consumption;
4) mycelium pellet long service life and using repeatedly, mycelium pellet were used 30 days continuously in the laboratory, still can keep higher decolored degradation activity;
5) easy to operate extensive, with low cost, except that the acquisition of bacterial classification activation with mycelium pellet needs the aseptic technique, other especially dyestuff handle and can carry out under field conditions (factors), indigo or the denim dyeing waste water of prolonged and repeated interpolation is not seen any pollution, and to produce the material with strong anti-microbial activity relevant with glossy ganoderma fermentation for this.
Description of drawings
Fig. 1 cultivates schema for weber Ganderma lucidum pompon of the present invention;
Fig. 2 is weber glossy ganoderma profile variation figure in the culturing process of the present invention;
The synoptic diagram of the mycelium pellet that the difference that Fig. 3 obtains for the present invention is big or small;
Fig. 4 shakes the graphic representation of the indigo decolored degradation of bottle vibration influence for the present invention;
Fig. 5 is the graphic representation of the indigo decolored degradation of pH value influence of the present invention;
Fig. 6 is the graphic representation of the indigo decolored degradation of temperature effect of the present invention;
Fig. 7 is the graphic representation of mycelium pellet culture under the optimal conditions of the present invention to indigo decolored degradation;
Fig. 8 for same bottle mycelium pellet culture under the optimal conditions of the present invention to the time changing curve figure of indigo decolored degradation speed;
Fig. 9 influences the graphic representation of denim decolorizing printing and dyeing waste water degraded for pH value of the present invention;
Figure 10 is the graphic representation of temperature effect denim decolorizing printing and dyeing waste water degraded of the present invention;
Figure 11 is the graphic representation of mycelium pellet culture under the optimal conditions of the present invention to the degraded of denim decolorizing printing and dyeing waste water;
Figure 12 is the time changing curve figure of mycelium pellet culture under the optimal conditions of the present invention to denim decolorization of dyeing degradation rate.
Embodiment
Below be the measuring method of indigo and denim decolorizing printing and dyeing waste water rate among the embodiment:
Add dyestuff or waste water in the mycelium pellet nutrient solution, the OD value at sampling and measuring 650nm place immediately behind the mixing is counted A
0As required, the OD value of sampling and measuring after reaction for some time counted A
1Calculate percent of decolourization as follows:
The test triplicate is averaged.
One, the preparation of weber Ganderma lucidum pompon
Under aseptic condition, from slant strains, take mycelia with transfering loop and forward on the plate culture medium, put into 25～32 ℃ of incubators and cultivate and can obtain activated spawn as depicted in figs. 1 and 2 in 2～5 days.Under aseptic condition, take mycelia from the activated spawn flat board again, be uniformly dispersed, add and be equipped with in weber liquid nutrient medium of Ganderma lucidum pompon, shaking culture is 3～6 days under 25～32 ℃, 120～200 rev/mins condition, can obtain a weber Ganderma lucidum pompon.The top condition of actication of culture is to cultivate 4 days in 28 ℃ of incubators; A top condition that obtains weber Ganderma lucidum pompon is 28 ℃, 160 rev/mins shaking culture 5 days.Figure 3 shows that the mycelium pellet that utilizes all size that present method obtains.
Used culture medium prescription is in the aforesaid method: contain potato 200g, glucose 20g, KH in every liter of the plate culture medium
2PO
43g, MgSO
47H
2O 1.5g, VITMAIN B1 1mg, agar 15g, pH nature.
Contain potato 200g, glucose 20g, KH in every liter of the liquid nutrient medium
2PO
43g, MgSO
47H
2O 1.5g, vitamins B
12mg, yeast extract paste 5g and 70mL trace element solution, surplus is a water, pH4.8; Wherein said trace element solution is meant and contains MgSO
47H
2O 3g/L, MnSO
4H
2O 0.5g/L, NaCl 1g/L, FeSO
47H
2O 0.1g/L, CoCl
20.1g/L, ZnSO
47H
2O 0.1g/L, CuSO
45H
2O 0.1g/L, KA1 (SO
4)
212H
2O 0.01g/L, H
3BO
30.01g/L, Na
2MoO
42H
2The aqueous solution of O 0.01g/L.
Two, weber Ganderma lucidum pompon is to indigo decolored degradation
The above-mentioned weber Ganderma lucidum pompon culture that utilizes method to obtain is mixed with indigo solution, carry out the optimization of decolored degradation condition.Carrying out indigo degradation condition when optimizing, theory in line with the green degraded, without hazardous and noxious substances (as amboceptor), only bigger to dye decolored degradation influence factor (as shaking bottle vibration, pH value and temperature) is probed into, and follow-up optimization is all based on the optimum result of front.
At first, determine to shake the effect of bottle vibration.Shaking bottle vibration and leaving standstill under the cultivation, a weber Ganderma lucidum pompon culture all can be degraded indigo, still shakes decolored degradation speed that bottle oscillation energy quickens dyestuff greatly as shown in Figure 4.Therefore, follow-up optimization all will be carried out shaking under bottle condition.
Secondly, determine best pH.Culture in the same culturing bottle evenly is divided in 5 different triangular flasks, regulates the pH value to needed value, add respectively indigo in right amount, observe shake-flask culture after 2 hours colour-change and measure percent of decolourization.Fig. 5 shows that weber Ganderma lucidum pompon culture all can be degraded indigo in the scope of pH3-7, but decolored degradation is the fastest when pH5.So, follow-up optimization all will pH5 and shake the bottle condition under carry out.
And then, determine optimum temps.Under 160 rev/mins the condition of shaking bottle, pH5.0, observe the influence of differing temps to decolored degradation.As shown in Figure 6, in 13-38 ℃ scope, weber Ganderma lucidum pompon culture all can be degraded indigo, but decolored degradation is the fastest in the time of 28 and 33 ℃, almost completely decolouring in 1.5 hours.So follow-up probe into all will and be shaken under the condition of bottle and carry out at 28-33 ℃, pH5.
At last, determine decolored degradation ability and work-ing life under the optimal conditions.In cultivating 6 days 100 ml shake flask mycelium pellet cultures, add indigo solution, be adjusted to pH5,33 ℃, 160 rev/mins condition shaking culture 1 hour, per 10 minutes sampling and measuring and calculate percent of decolourization.After finishing the mensuration decolorization experiment, filter immediately and collect mycelium pellet, fully blot with thieving paper, claim fresh weight, the amount of Units of Account time, unit fresh weight mycelium pellet catalysis dye decolored degradation draws the decolored degradation ability.Fig. 7 shows that along with the prolongation of time, indigo decolored degradation increases, but the decolorization rate maintenance is constant.Through conversion, handled 1 hour under the top condition, can make the percent of decolourization of the indigo solution of 20mg/L reach 92%, can make 99.0% indigo decolouring in promptly 3.8 hours, its decolored degradation ability is 0.67mg/h/gFW.
Give every day in the same bottle mycelium pellet culture and add indigo solution, reaction is 1 hour under our optimized conditions, measures decolored degradation speed, to estimate work-ing life.As seen from Figure 8, mycelium pellet long service life and using repeatedly, mycelium pellet used 30 days continuously in the laboratory, still can keep higher decolored degradation activity.
Add indigo solution in utilizing the weber Ganderma lucidum pompon culture that method of the present invention obtains, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH4, leaves standstill to cultivate 2 hours under 28 ℃ of conditions, and indigo percent of decolourization is 41% as shown in Figure 4.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH4, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 88% as shown in Figure 4.
Comparing embodiment 1 and 2 as can be known, shaking culture help the decolouring, following examples all adopt shaking culture.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH3, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 35% as shown in Figure 5.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH4, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 92% as shown in Figure 5.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 96% as shown in Figure 5.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH6, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 16% as shown in Figure 5.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH7, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 15% as shown in Figure 5.
Comparing embodiment 3,4,5,6 and 7 as can be known, under 160 rev/mins of shaking culture conditions, pH5 to the decolouring the most favourable, following examples all adopt the culture condition of 160 rev/mins of vibrations and pH5 to carry out indigo decolored degradation.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 13 ℃, 160 rev/mins condition, and indigo percent of decolourization is 39% as shown in Figure 6.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 18 ℃, 160 rev/mins condition, and indigo percent of decolourization is 73% as shown in Figure 6.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 23 ℃, 160 rev/mins condition, and indigo percent of decolourization is 81% as shown in Figure 6.
Embodiment 11
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 94% as shown in Figure 6.
Embodiment 12
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 95% as shown in Figure 6.
Embodiment 13
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add indigo solution, making its concentration is 20mg/L, and the potential of hydrogen of solution is adjusted into pH5, and shaking culture is 2 hours under 38 ℃, 160 rev/mins condition, and indigo percent of decolourization is 82% as shown in Figure 6.
Comparing embodiment 8,9,10,11,12 and 13 as can be known, under the culture condition of 160 rev/mins of vibrations, pH5,28 ℃ are the most favourable to decolouring, so adopt 160 rev/mins of vibrations, pH5 and 28 ℃ as the indigo decolored degradation conditions of optimizing.Handled with this understanding 1 hour, mycelium pellet can make the percent of decolourization of the indigo solution of 20mg/L reach 92% as shown in Figure 7, can make 99.0% dye decoloredly in promptly 3.8 hours, and its decolored degradation ability is 0.67mg/h/gFW.
Under similarity condition, add indigo solution every day in the mycelium pellet culture of same bottle 100mL, and 1 hour decolored degradation speed of assaying reaction is to estimate work-ing life.The result as shown in Figure 8, the the 1st, 5,10,20 and 30 day the percent of decolourization that uses is respectively 92%, 88%, 78%, 47% and 37%, promptly in the time of being followed the trail of, mycelium pellet can be degraded indigo all the time, therefore have long work-ing life, just its degradation efficiency has the trend that reduces gradually with prolonging duration of service.If wish to reach 99.0% percent of decolourization, then need 3.8,4.6,6.2,15.0 and 20.0 hours respectively with above-mentioned the 1st, 5,10,20 and 30 day mycelium pellet mixture process.
Three, weber Ganderma lucidum pompon is degraded to the denim decolorization of dyeing
The weber Ganderma lucidum pompon culture that utilizes the present invention to obtain is mixed with indigo dyeing waste water, carry out the decolored degradation condition optimizing.
At first, determine best pH.Add 150 milliliters of ox cloth dyeing waste waters (OD595=0.52) at 150 milliliters of mycelium pellet cultures, evenly divide behind the mixing and install in 5 different triangular flasks, regulate the pH value to needed value, further each bottle is distributed into 3 bottles again, observation is shaken bottle (160 rev/mins) and is cultivated the colour-change after 4 hours and measure percent of decolourization.As seen from Figure 9, under the shaking culture condition, Ganoderma lucidum mycelium all can degrade by catalysis denim decolorizing printing and dyeing waste water, but decolored degradation is the fastest when pH3 in the scope of pH3-6.So follow-up probing into will and be shaken under the condition of bottle and carry out at pH3.
Secondly, determine optimum temps.Observing differing temps under 160 rev/mins the condition of shaking bottle, pH3 influences decolored degradation.As shown in Figure 10, at pH3, shake under bottle oscillating condition cultivates, Ganoderma lucidum mycelium all can the degraded of catalysis denim decolorizing printing and dyeing waste water in 23-43 ℃ scope, but decolored degradation is the fastest in the time of 28 and 33 ℃, react 4 hours, almost completely decolours.
At last, determine decolored degradation ability and work-ing life under the optimal conditions.Add isopyknic waste water (OD=0.52) in bottle mycelium pellet culturing mixt cultivating the 6th day shake, shaking culture is 4 hours under pH3,33 ℃, 160 rev/mins condition, and sampling in per 30 minutes is measured and calculated percent of decolourization, and the result as shown in figure 11.After finishing the mensuration decolorization experiment, filter immediately and collect mycelium pellet, fully blot with thieving paper, claim fresh weight, the amount of Units of Account time, unit fresh weight mycelium pellet catalysis dye decolored degradation draws corresponding decolored degradation ability.Figure 12 shows that along with the prolongation of time, waste water decolours gradually, and it is constant that its percent of decolourization keeps, and 4 hours is 79%.Calculate that by this desiring to reach 99.0% percent of decolourization then needs 13.2 hours.Through conversion, per hour be equivalent to, to make 7 milliliters of OD be 0.52 waste water decoloring to every gram fresh weight mycelium pellet.
Embodiment 14
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH2, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 43% as shown in Figure 9.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 79% as shown in Figure 9.
Embodiment 16
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH4, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 40% as shown in Figure 9.
Embodiment 17
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH5, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 29% as shown in Figure 9.
Embodiment 18
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH6, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 18% as shown in Figure 9.
Comparing embodiment 14,15,16,17 and 18 as can be known, under 160 rev/mins of shaking culture conditions, pH3 is the most favourable to waste water decoloring, following examples all adopt the culture condition of 160 rev/mins of vibrations and pH3 to carry out the degraded of young cloth decolorizing printing and dyeing waste water.
Embodiment 19
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 23 ℃, 160 rev/mins condition, and indigo percent of decolourization is 52% as shown in figure 10.
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 28 ℃, 160 rev/mins condition, and indigo percent of decolourization is 79% as shown in figure 10.
Embodiment 21
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 33 ℃, 160 rev/mins condition, and indigo percent of decolourization is 79% as shown in figure 10.
Embodiment 22
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 38 ℃, 160 rev/mins condition, and indigo percent of decolourization is 70% as shown in figure 10.
Embodiment 23
In the weber Ganderma lucidum pompon culture that utilizes method of the present invention to obtain, add isopyknic OD
650=0.52 denim dyeing waste water, and the potential of hydrogen of solution is adjusted into pH3, shaking culture is 4 hours under 43 ℃, 160 rev/mins condition, and indigo percent of decolourization is 60% as shown in figure 10.
Comparing embodiment 19,20,21,22 and 23 as can be known, under the culture condition of 160 rev/mins of vibrations, pH3,28 ℃ and 33 ℃ are the most favourable to decolouring, so adopt 160 rev/mins of vibrations, pH5 and 28 ℃ as the denim decolorization of dyeing degradation conditions of optimizing.Handled with this understanding 4 hours, mycelium pellet can make isopyknic OD
650=0.52 denim decolorization of dyeing rate reaches 79% (Figure 11), can make 99.0% decoloring dye waste water in promptly 13.2 hours, and its decolored degradation ability is 7ml/h/gFW.
Under similarity condition, the per 5 days isopyknic OD of adding in same mycelium pellet culture
650=0.52 denim dyeing waste water, 4 hours decolored degradation speed of assaying reaction is to estimate work-ing life.The result as shown in figure 12, the the 1st, 5,10,20 and 30 day the degradation rate that uses is respectively 76%, 73%, 62%, 37% and 29%, promptly in the time of being followed the trail of, cultivate the mycelium pellet denim dyeing waste water of degrading all the time, therefore have long work-ing life, just its degradation efficiency has the trend that reduces gradually with prolonging duration of service.If wish to reach 99.0% percent of decolourization, then need 13.2,14.8,19.4,35.9 and 53.9 hours respectively with above-mentioned the 1st, 5,10,20 and 30 days mycelium pellet mixture process.
Claims (9)
1. the cultural method of a weber Ganderma lucidum pompon is characterized in that described method is included under the aseptic condition, the weber Ganderma lucidum strain is inoculated in the slant medium cultivates in advance; Then under aseptic condition, from slant strains, take mycelia with transfering loop and forward on the plate culture medium, put into 25～32 ℃ of incubators and cultivate and obtained activated spawn in 2～5 days; Under aseptic condition, take mycelia from the activated spawn flat board again, be uniformly dispersed, add and be equipped with in weber liquid nutrient medium of Ganderma lucidum pompon, shaking culture is 3～6 days under 25～32 ℃, 120～200 rev/mins condition, obtains weber Ganderma lucidum pompon.
2. the cultural method of weber Ganderma lucidum pompon according to claim 1, it is characterized in that the top condition that described weber Ganderma lucidum pompon is cultivated is that slant strains is inoculated on the plate culture medium, in 28 ℃ incubator, cultivated 4 days, be inoculated in the liquid nutrient medium 28 ℃, 160 rev/mins shaking culture 5 days then.
3. the cultural method of weber according to claim 1 Ganderma lucidum pompon is characterized in that containing potato 200g, glucose 20g, KH in every liter of the described plate culture medium
2PO
43g, MgSO
47H
2O 1.5g, VITMAIN B1 1mg, agar 15g, pH nature.
4. according to the cultural method of claim 1 weber Ganderma lucidum pompon, it is characterized in that containing potato 200g, glucose 20g, KH in every liter of the described liquid nutrient medium
2PO
43g, MgSO
47H
2O1.5g, vitamins B
12mg, yeast extract paste 5g and 70mL trace element solution, surplus is a water, pH4.8.
5. according to the cultural method of claim 3 weber Ganderma lucidum pompon, it is characterized in that described trace element solution is meant contains MnSO
4H
2O 0.5g/L, NaCl 1g/L, FeSO
47H
2O 0.1g/L, CoCl
20.1g/L, ZnSO
47H
2O 0.1g/L, CuSO
45H
2O 0.1g/L, KA1 (SO
4)
212H
2O0.01g/L, H
3BO
30.01g/L, Na
2MoO
42H
2The aqueous solution of O 0.01g/L.
6. the described method of claim 1 is cultivated the weber Ganderma lucidum pompon culture of acquisition to indigo decolored degradation application, it is characterized in that realizing by the following method: the weber Ganderma lucidum pompon culture that will utilize aforesaid method to obtain mixes with the indigo solution of 0.5～10 times of volume, the potential of hydrogen of mixed solution is adjusted into pH3～7, shaking culture under 13～38 ℃, 160 rev/mins condition is carried out indigo decolored degradation.
7. weber Ganderma lucidum pompon culture according to claim 6 is used indigo decolored degradation, and the optimal ph that it is characterized in that weber Ganderma lucidum pompon culture and indigo solution mixed solution is 5.
8. a weber Ganderma lucidum pompon culture that is obtained by the described method cultivation of claim 1 is to denim decolorization of dyeing degraded application, it is characterized in that realizing by the following method: the weber Ganderma lucidum pompon culture that will utilize aforesaid method to obtain mixes with the denim dyeing waste water of 0.5～10 times of volume, the potential of hydrogen of mixed solution is adjusted into pH2～6, shaking culture under 23～43 ℃, 160 rev/mins condition promotes the degraded of denim decolorization of dyeing.
9. weber Ganderma lucidum pompon culture according to claim 8 is characterized in that to denim decolorization of dyeing degraded application the optimal ph of weber Ganderma lucidum pompon culture and denim dyeing waste water mixed solution is 3.
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| CN102523921A (en) * | 2012-01-11 | 2012-07-04 | 福建农林大学 | Preparation method and products of needle mushroom mycelium pellets |
| CN102523920A (en) * | 2012-01-11 | 2012-07-04 | 福建农林大学 | Preparation method of Grifola frondosa mycelial pellets and products thereof |
| CN108497153A (en) * | 2018-03-01 | 2018-09-07 | 广州大学 | A kind of Ganoderma Lucidum pompon chewing gum and preparation method thereof |
| CN108587924A (en) * | 2018-04-20 | 2018-09-28 | 湖南农业大学 | A kind of Ganoderma Lucidum pompon product |
| CN108902615A (en) * | 2018-08-31 | 2018-11-30 | 广州大学 | A kind of guava ganoderma lucidum biological drink and preparation method thereof |
| CN109258289A (en) * | 2018-08-31 | 2019-01-25 | 广州大学 | A kind of mung bean ganoderma lucidum biological drink and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102523921A (en) * | 2012-01-11 | 2012-07-04 | 福建农林大学 | Preparation method and products of needle mushroom mycelium pellets |
| CN102523920A (en) * | 2012-01-11 | 2012-07-04 | 福建农林大学 | Preparation method of Grifola frondosa mycelial pellets and products thereof |
| CN102523921B (en) * | 2012-01-11 | 2013-12-04 | 福建农林大学 | Preparation method and products of needle mushroom mycelium pellets |
| CN102523920B (en) * | 2012-01-11 | 2013-12-04 | 福建农林大学 | Preparation method of Grifola frondosa mycelial pellets and products thereof |
| CN108497153A (en) * | 2018-03-01 | 2018-09-07 | 广州大学 | A kind of Ganoderma Lucidum pompon chewing gum and preparation method thereof |
| CN108587924A (en) * | 2018-04-20 | 2018-09-28 | 湖南农业大学 | A kind of Ganoderma Lucidum pompon product |
| CN108902615A (en) * | 2018-08-31 | 2018-11-30 | 广州大学 | A kind of guava ganoderma lucidum biological drink and preparation method thereof |
| CN109258289A (en) * | 2018-08-31 | 2019-01-25 | 广州大学 | A kind of mung bean ganoderma lucidum biological drink and preparation method thereof |
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