CN109806846B - Method for removing indigo by co-fermenting sesame straw with tremella and morchella - Google Patents
Method for removing indigo by co-fermenting sesame straw with tremella and morchella Download PDFInfo
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Abstract
The invention relates to a method for removing indigo blue from tremella and morchella co-fermented sesame straw, which comprises the following steps: culturing Tremella strain and Morchella esculenta strain on potato glucose agar plate at 25-28 deg.C and humidity of 70-80%, and perforating to obtain 1.0 cm diameter bacterial sheet; washing, drying and crushing straws, sieving to obtain a dye adsorbent raw material, weighing 10.0-30.0g of the raw material, adding distilled water according to the ratio of the raw material to the water of 1:3-1:5(g/mL), adding an inducer, sterilizing at 121 ℃ for 30min, cooling to room temperature, inoculating strains, inoculating 2 bacterial tablets to each strain, and fermenting at 25-28 ℃ and humidity of 75%; drying and crushing the fermented substrate to obtain a dye adsorbent; adding 20.0g of adsorbent into an aqueous solution with a dye concentration of 200 mg/L, pH 6.0.0 and a NaCl concentration of 0 mol/L according to a solid-to-liquid ratio (m/v, g/mL) of 1:150, and carrying out adsorption filtration at a temperature of 30 ℃ and a rotating speed of 200 r/min; the method for removing the indigo has convenient operation and low cost, and is a green process technology.
Description
Technical Field
The invention belongs to the technical field of industrial wastewater treatment, and particularly relates to a method for removing indigo by utilizing tremella and morchella co-fermented sesame straw to effectively remove dye pollutants in wastewater by utilizing the tremella and morchella co-fermented sesame straw to prepare dye adsorbent to adsorb dye indigo; the dye adsorbent is prepared by co-fermenting the sesame straws with the tremella and the morchella, and the dye adsorbent adsorbs the indigo dye and then is incinerated and returned to the field, so that the accumulation of harmful dye in the environment is eliminated.
Background
The dye wastewater is one of the industrial wastewater which is difficult to treat, and the removal of dye pollutants in the wastewater is a key problem for treating the dye wastewater.
Indigo (Indigo carmine) is one of the oldest dyes and is widely used in plastic printing and dyeing, textile printing and dyeing and the like, such as denim printing and dyeing. However, in the production and use of indigo there is a concomitant production of large amounts of colored waste water. The existing methods for removing indigo in water bodies include adsorption methods, photocatalysis, microbial degradation and the like, wherein the adsorption methods have the advantages of low cost and easiness in operation. Sesame is one of main oil crops in China, sesame straw resources are rich but the treatment technology is not complete, and if the sesame straw is directly discarded or burnt, resource waste and environmental pollution are caused. Sesame straw with rich sources and low cost is selected as an adsorbent, and the indigo in the water body is removed by an adsorption method, so that the method has economic benefits and environmental benefits. However, sesame straw is a lignocellulosic biomass, has the characteristics of stubborn structure and chemical inertness, and has the defect of small dye adsorption (Li comet. microbiological decoloration printing dye and mechanism research thereof [ D ], doctor's paper of Jiangnan university, 2015).
The straw modification can destroy the stubborn structure, modify and/or introduce chemical functional groups, and can obviously improve the adsorption capacity. At present, the main modification method of sesame straw is a chemical method, such as Li comet, etc., which utilizes NaOH to modify sesame straw and improves the adsorption capacity of the sesame straw to dye Congo red (Li H X, Zhang R J, Tang L, et al. conjugate adsorption capacity of lignocellulic biological by sodium hydroxide treatment)]Asian Journal of Chemistry 2015, 27(7): 2373-. Wutao, etc. uses sesame straw as raw material, firstly adopts amylase and proteinase to make enzymolysis, then uses the enzymolysis product of said raw material, and adds KOH and dilute H2SO4And carrying out chemical reaction on the urea to prepare the adsorbing material. (Wutao. sesame straw adsorbing material and its preparation method, CN104028234A [ P ]]. 2014.). However, the chemical method for preparing the adsorbent by modifying the sesame straw needs to use chemical reagents such as acid and alkali to treat the straw, which causes the problems of increased cost, secondary pollution and the like.
The white rot fungi can depolymerize and degrade straws by virtue of the penetration capacity of hyphae and the secretion capacity of extracellular cellulose hydrolase (cellulase, hemicellulase and the like) and lignin degrading enzyme (laccase, manganese peroxidase, lignin peroxidase and the like), so as to achieve the purpose of modifying the straws. White rot fungi are inoculated in straws and fermented under the conditions of certain temperature, humidity and the like, and strains grow and metabolize by taking the straws as substrates, so that chemical functional groups (-OH, -NH, C = O and the like) for adsorbing dyes in the straws are changed, the surfaces of the straws are rougher, and pores are enlarged. In addition, different white rot fungi are selected for combination by utilizing the interaction and substrate difference among the white rot fungi, the metabolic characteristics of the strain can be changed by the white rot fungi co-fermentation technology, and the growth rate and the enzyme secretion capacity of the strain are improved. (Chuangzhen, Zhuliping, Yaoyoueliang, three kinds of white rot fungi and their combination strains lignin degrading enzyme comparative research [ J ]. bacterial science, 2009, 28(4): 577) -583; Elisashvili V, Kachlishvili E. Physiological regulation of laccase and mangase peroxidase production by white-rot basic bacteria [ J ]. Journal of Biotechnology, 2009, 144(1):37-42 ]. The method for preparing the dye adsorbent by co-fermenting the straws with the white rot fungi is a green process technology, has the characteristics of low cost and no secondary pollution, and has a very wide application prospect.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a method for removing indigo by co-fermenting white fungus and morchella esculenta with sesame straw; on the basis of screening white rot fungus strains, tremella and morchella co-fermentation sesame straws are selected to prepare a dye adsorbent, and the dye adsorbent adsorbs dye indigo and then is ashed and returned to the field. The white rot fungi strains screened by the experiment are purchased from China center for industrial microorganism culture preservation management. The bacteria are Lentinus EdodesLentinulasp. (strain number: CICC 14031), black fungusAuricularia auricula(Strain No.: CICC 14008) and Stereum glehniaeGloeostereum incarnatum(Strain No.: CICC 14024) and morchella esculentaMorchellasp. (Strain No.: CICC 14033), TremellaTremellafuciformis(Strain No.: CICC 50179), trametes hirsutusTrametes hirsuta(Strain number: CICC 2671) and Cordyceps militarisCordyceps militaris(Strain No.: CICC 14013).
The purpose of the invention is realized as follows:
a method for removing indigo from white fungus and morchella co-fermented sesame straw comprises the following steps:
step 1), culturing the white rot fungus strain on a potato glucose agar plate for 7-10 days at the temperature of 25-28 ℃ and the humidity of 70-80%, and punching a hole on the agar plate by adopting a puncher with the diameter of 1.0 cm to obtain a fungus piece with the diameter of 1.0 cm;
step 2), washing, drying and crushing the straws, and then sieving the straws with a 20-60-mesh sieve to obtain a raw material for preparing the dye adsorbent;
step 3), weighing 10.0-30.0g of raw materials, adding distilled water according to the ratio of the raw materials to the water of 1:3-1:5(g/mL), adding an inducer, sterilizing at 121 ℃ for 30min, cooling to room temperature, inoculating white rot fungi, inoculating 2 pieces of mycelia of each white rot fungus, and culturing and fermenting for 10-15 days at 25-28 ℃ and humidity of 75%;
step 4), drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to prepare the dye adsorbent;
and 5) after the dye adsorbent is prepared, adsorbing the dye indigo under the following conditions: weighing 20.0g of the prepared adsorbent, adding the adsorbent into an aqueous solution with a dye concentration of 200 mg/L, pH 6.0.0 and a NaCl concentration of 0 mol/L according to a solid-to-liquid ratio (m/v, g/mL) of 1:150, adsorbing for 4 hours at an adsorption temperature of 30 ℃ and at a stirring speed of 200 r/min, and then filtering to calculate the adsorption capacity (mg/g);
and step 6), after adsorbing the dye indigo, the adsorbent is naturally air-dried for 36-48 h, calcined for 4.0-6.0h at the temperature of 500-600 ℃, and naturally cooled to obtain ash and then returned to the field.
The white rot fungi in the step 1) are morchella esculenta and tremella.
The straw in the step 2) is sesame straw.
The addition of inducer in step 3) is 72.5mM acetic acid, Cu per kilogram straw feedstock2+81.8mM and baseLignin 27.9 g, acetic acid 72.5mM/kg, Cu2+81.8mM/kg and alkaline lignin 27.9 g/kg.
In the present invention, the adsorption amount (mg/g) is calculated by the formula (C)0-C) x V/m; in the formula C0Is the initial concentration (mg/L) of the indigo in the solution, C is the concentration (mg/L) of the indigo in the solution after adsorption, V is the volume (L) of the solution, and m is the mass (g) of the adsorbent.
The invention has the following positive effects:
the experimental process and data for verifying the effect of the invention are as follows:
1. culturing white rot fungi: 7 kinds of white rot fungi (mushroom)Lentinulasp. Auricularia auriculaAuricularia auriculaFungus of Demanassia mucilaginosaGloeostereum incarnatumMorchella esculenta (pers.) MoenchMorchellasp. Tremella fuciformis berkTremella fuciformisTrametes hirsuta (Roxb.) KuntzeTrametes hirsutaAnd Cordyceps militarisCordyceps militaris) Respectively culturing the strains on a potato glucose agar plate for 10 days at the temperature of 28 ℃ and the humidity of 75%, and punching the agar plate by adopting a puncher with the diameter of 1.0 cm to obtain bacterial sheets with the diameter of 1.0 cm;
2. pretreatment of sesame straw: washing, drying and crushing the straws, and then sieving the straws with a 40-mesh sieve to obtain a raw material for preparing the dye adsorbent;
3. white rot fungus co-fermentation sesame straw: weighing 20.0g of straw raw material, adding distilled water according to a material-water ratio of 1:5(g/mL), sterilizing at 121 ℃ for 30min, cooling to room temperature, inoculating 2 different white rot fungi, inoculating 2 white rot fungi slices to each white rot fungus, and fermenting at 25 ℃ and humidity of 75% for 14 days.
4. Preparing a dye adsorbent: and drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to obtain the dye adsorbent.
5. Adsorbing the dye: after the dye adsorbent was prepared, the dye indigo was adsorbed under the following conditions: weighing 20.0g of the prepared adsorbent, adding the adsorbent into an aqueous solution with a dye concentration of 200 mg/L, pH 6.0.0 and a NaCl concentration of 0 mol/L according to a solid-to-liquid ratio (m/v, g/mL) of 1:150, adsorbing for 4 hours at an adsorption temperature of 30 ℃ and a stirring speed of 200 r/min, and filtering to calculate the adsorption capacity.
6. Returning to the field after ashing: adsorbing dye indigo blue by the adsorbent, naturally drying for 48 h, calcining at 540 ℃ for 5.0h, naturally cooling to obtain ash, and returning to the field.
In order to show the advancement of preparing the dye adsorbent by co-fermenting the sesame straws with the white rot fungi, the invention adopts 7 kinds of white rot fungi (shiitake mushrooms)Lentinulasp. Auricularia auriculaAuricularia auriculaFungus of Demanassia mucilaginosaGloeostereum incarnatumMorchella esculenta (pers.) MoenchMorchellasp. Tremella fuciformis berkTremella fuciformisTrametes hirsuta (Roxb.) KuntzeTrametes hirsutaAnd Cordyceps militarisCordyceps militaris) Under the conditions, the sesame straw is fermented by white rot fungi independently, namely: weighing 20.0g of straw raw material, adding distilled water according to a material-water ratio of 1:5(g/mL), sterilizing at 121 ℃ for 30min, cooling to room temperature, inoculating 1 white rot fungus, inoculating 4 pieces of white rot fungus, and fermenting at 25 ℃ and humidity of 75% for 14 days. Drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to prepare the dye adsorbent.
The results of comparative analysis of the dye adsorbent prepared from the sesame straw raw material, the dye adsorbent prepared from the sesame straw independently fermented by the white-rot fungi and the dye adsorbent prepared from the sesame straw co-fermented by the white-rot fungi are shown in table 1.
It can be seen that: the adsorption amount of the sesame straw raw material to indigo is 21.90 mg/g, the adsorption amount can be improved by separately fermenting the sesame straw with white rot fungi, wherein white fungusTremella fuciformisThe effect of fermenting the sesame straws is good, the adsorption capacity is 29.07 mg/g, the effect of co-fermenting the sesame straws by white rot fungi is good, wherein the morchella esculentaMorchellasp. and TremellaTremella fuciformisAfter the sesame straw is co-fermented, the adsorption capacity reaches 40.87 mg/g, and compared with the raw material of the sesame straw, the adsorption capacity of the raw material of the sesame straw to indigo is improved by 86.62%.
TABLE 1 adsorption of indigo by different adsorbents
Morchella esculenta (L.) KuntzeMorchellasp. and TremellaTremella fuciformisThe co-fermented sesame straw depolymerizes and degrades the straw by virtue of the penetration capacity of hypha and the secretion capacity of extracellular enzyme in the growth and metabolism process, so that the adsorption quantity of dye indigo is increased.
As the growth and the secretase of the white rot fungi are obviously influenced by the inducer, the influence of the variety and the concentration of the inducer is further examined by adopting a single-factor experiment by taking the adsorption quantity as an index, and the result is shown in a figure 2-a figure 7.
FIGS. 2-7 show the effect of different inducers on the adsorption capacity, and the results show that alkaline lignin, guaiacol, acetic acid, Cu2+And resveratrol, while hydrogen peroxide reduces the adsorption, since hydrogen peroxide is a strong oxidant, inhibiting the growth and metabolism of the strain during fermentation. On the basis of a single-factor experiment result, the alkali lignin, acetic acid and Cu with good effect and low cost are selected2+Three inducers were designed using Box-Behnken protocol (table 2) in response to the surface method to examine inducer interaction and optimize inducer concentration. The Box-Behnken protocol design results (Table 3) were analyzed by Minitab software to obtain: acetic acid concentration (X)1) Acetic acid concentration quadratic term (X)1 2）、Cu2+Second order term of concentration (X)2 2) Interaction of acetic acid concentration and alkaline lignin concentration (X)1×X3) Has very significant influence on the adsorption quantity (p<0.01), acetic acid concentration (X)1) And Cu2+Concentration (X)2) The interaction has a significant influence on the amount of adsorption (p<0.05) (table 4).
The best combination of inducer is acetic acid 72.5mM/kg, Cu2+81.8mM/kg and 27.9 g/kg of alkaline lignin, and the adsorption capacity reaches 74.25mg/g under the optimal combination condition through experimental verification.
TABLE 2 Box-Behnken protocol design Table for optimizing inducer concentration
TABLE 3 adsorption at different inducer concentrations
| Serial number | X1 | X2 | X3 | Adsorption Capacity (mg/g) |
| 1 | -1 | -1 | 0 | 54.36 |
| 2 | -1 | 1 | 0 | 49.90 |
| 3 | 1 | -1 | 0 | 59.06 |
| 4 | 1 | 1 | 0 | 65.84 |
| 5 | 0 | -1 | -1 | 61.89 |
| 6 | 0 | -1 | 1 | 58.32 |
| 7 | 0 | 1 | -1 | 60.30 |
| 8 | 0 | 1 | 1 | 63.33 |
| 9 | -1 | 0 | -1 | 53.02 |
| 10 | 1 | 0 | -1 | 70.34 |
| 11 | -1 | 0 | 1 | 66.32 |
| 12 | 1 | 0 | 1 | 68.05 |
| 13 | 0 | 0 | 0 | 72.30 |
| 14 | 0 | 0 | 0 | 73.42 |
| 15 | 0 | 0 | 0 | 73.01 |
TABLE 4 ANOVA TABLE
| Factors of the fact | Degree of freedom | pValue of |
| |
1 | <0.01* |
| |
1 | 0.33 |
| |
1 | 0.11 |
| X1* |
1 | <0.01** |
| X2* |
1 | <0.01** |
| X3* |
1 | 0.06 |
| X1* |
1 | <0.05* |
| X1* |
1 | <0.01** |
| X2* |
1 | 0.14 |
| Lack of |
3 | 0.06 |
*p<0.05 means significance;. Thep<0.01 means very significant
Therefore, the method for removing indigo by co-fermenting the white fungus and the morchella esculenta with the sesame straw provided by the invention comprises the following main steps: pretreating sesame straws, co-fermenting the sesame straws with tremella and morchella to prepare a dye adsorbent, and adding 72.5mM/kg of inducer acetic acid and Cu in the fermentation process2+81.8mM/kg and 27.9 g/kg of alkaline lignin, and ashing the dye indigo to the field after the dye indigo is adsorbed by the adsorbent. The dye adsorbent prepared by co-fermenting the sesame straws with the white rot fungi has the characteristics of low cost and no secondary pollution, and is incinerated and returned to the field after adsorbing the dye indigo, so that the accumulation of the dye in the environment is thoroughly eliminated; the method for removing indigo by using white fungus and morchella co-fermented sesame straw is convenient to operate, low in cost and a green process technology.
Drawings
FIG. 1 is a process step diagram of the present invention.
FIG. 2 is a graph showing the effect of an alkaline lignin inducer on adsorption capacity.
FIG. 3 is a graph showing the effect of guaiacol inducer on adsorption capacity.
FIG. 4 is a graph showing the effect of an acetic acid inducer on adsorption capacity.
FIG. 5 is Cu2+Graph of the influence of inducer on adsorption.
FIG. 6 is a graph of the effect of an inducer of resveratrol on the amount of adsorption.
FIG. 7 is a graph showing the effect of hydrogen peroxide inducer on adsorption capacity.
Detailed Description
Example 1: as shown in figure 1, the method for removing indigo from tremella and morchella co-fermented sesame straw provided by the invention comprises the following steps:
step 1), white rot fungus culture: 2 kinds of white rot fungi (morchella esculenta)Morchellasp. Tremella fuciformis berkTremella fuciformis) Respectively culturing the strains on a potato glucose agar plate for 8 days at the temperature of 25 ℃ and the humidity of 80%, and punching holes on the agar plate by adopting a puncher with the diameter of 1.0 cm to obtain bacterial sheets with the diameter of 1.0 cm;
step 2), pretreatment of sesame straw: washing, drying and crushing the straws, and then sieving the straws with a 20-mesh sieve to obtain a raw material for preparing the dye adsorbent;
step 3), co-fermenting the sesame straws by white rot fungi: weighing 10.0g straw raw material, adding distilled water according to the material-water ratio of 1:3 (g/ml), adding inducer acetic acid 72.5mM/kg and Cu2+81.8mM/kg and alkaline lignin 27.9 g/kg, sterilized at 121 ℃ for 30min and then cooled to room temperature. Inoculating 2 different white rot fungi, inoculating 2 pieces of mycelia of each white rot fungus, and fermenting at 28 deg.C and humidity of 75% for 10 days;
step 4), preparing a dye adsorbent: drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to obtain a dye adsorbent;
step 5), dye adsorption: after the dye adsorbent was prepared, indigo was adsorbed under the following conditions: weighing 20.0g of prepared adsorbent, adding the adsorbent into an aqueous solution with a dye concentration of 200 mg/L, pH 6.0.0 and a NaCl concentration of 0 mol/L according to a solid-to-liquid ratio (m/v, g/mL) of 1:150, adsorbing for 4 hours at an adsorption temperature of 30 ℃ and a stirring speed of 200 r/min, and then filtering, wherein the adsorption capacity is calculated to be 74.04 mg/g;
step 6), returning to the field after ashing: the adsorbent adsorbs dye indigo, then is naturally air-dried for 24 h, is calcined for 6.0h at 500 ℃, and is naturally cooled to obtain ash and then is returned to the field.
Example 2: as shown in figure 1, the method for removing indigo from tremella and morchella co-fermented sesame straw provided by the invention comprises the following steps:
step 1), white rot fungus culture: 2 kinds of white rot fungi (morchella esculenta)Morchellasp. Tremella fuciformis berkTremella fuciformis) Respectively culturing the strains on a potato glucose agar plate for 7 days at the temperature of 28 ℃ and the humidity of 70%, and punching holes on the agar plate by adopting a puncher with the diameter of 1.0 cm to obtain bacterial sheets with the diameter of 1.0 cm; step 2), pretreatmentSesame straw: washing, drying and crushing the straws, and then sieving the straws with a 60-mesh sieve to obtain a raw material for preparing the dye adsorbent;
step 3), co-fermenting the sesame straws by white rot fungi: weighing 30.0g straw raw material, adding distilled water according to the material-water ratio of 1:4 (g/mL), adding inducer acetic acid 72.5mM/kg and Cu2+81.8mM/kg and alkaline lignin 27.9 g/kg, sterilized at 121 ℃ for 30min and then cooled to room temperature. Respectively inoculating 2 different white rot fungi, inoculating 2 pieces of mycelia of each white rot fungus, and fermenting at 27 deg.C and humidity of 75% for 15 days;
step 4), preparing a dye adsorbent: drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to obtain a dye adsorbent;
step 5), dye adsorption: after the dye adsorbent was prepared, indigo was adsorbed under the following conditions: weighing 20.0g of prepared adsorbent, adding the adsorbent into an aqueous solution with a dye concentration of 200 mg/L, pH 6.0.0 and a NaCl concentration of 0 mol/L according to a solid-to-liquid ratio (m/v, g/mL) of 1:150, adsorbing for 4 hours at an adsorption temperature of 30 ℃ and a stirring speed of 200 r/min, and then filtering, wherein the adsorption capacity is calculated to be 73.67 mg/g;
step 6), returning to the field after ashing: adsorbing dye indigo blue by the adsorbent, naturally drying for 36 h, calcining for 4.0 h at 600 ℃, naturally cooling to obtain ash, and returning to the field.
Claims (1)
1. A method for removing indigo from white fungus and morchella co-fermented sesame straw is characterized by comprising the following steps:
step 1), culturing the white rot fungus strain on a potato glucose agar plate for 7-10 days at the temperature of 25-28 ℃ and the humidity of 70-80%, and punching a hole on the agar plate by adopting a puncher with the diameter of 1.0 cm to obtain a fungus piece with the diameter of 1.0 cm; the white rot fungi are morchella esculenta and tremella;
step 2), washing, drying and crushing the straws, and then sieving the straws with a 20-60-mesh sieve to obtain a raw material for preparing the dye adsorbent; the straw is sesame straw;
step 3), weighing 10.0-30.0g of raw materials prepared in the step 2), wherein the ratio of the raw materials to water in grams per milliliter is 1:3-1:5 adding distilled water, adding inducer, sterilizing at 121 deg.C for 30min, cooling to room temperature, inoculating white rot fungi, inoculating 2 pieces of mycelia of each white rot fungi, and fermenting at 25-28 deg.C and humidity of 75% for 10-15 days; the inducer is added by adding 72.5mM acetic acid and Cu per kilogram straw raw material2+81.8mM and alkaline lignin 27.9 g;
step 4), drying and crushing the fermented substrate, and sieving the substrate with a 40-mesh sieve to prepare the dye adsorbent;
and 5) adsorbing the indigo under the following conditions after preparing the dye adsorbent: weighing 20.0g of prepared adsorbent, adding the adsorbent into an aqueous solution with the dye concentration of 200 mg/L, pH 6.0.0 and the NaCl concentration of 0 mol/L according to the solid-to-liquid ratio of 1:150 g/ml, adsorbing for 4 hours at the adsorption temperature of 30 ℃ and the stirring speed of 200 r/min, and then filtering to calculate the adsorption capacity;
and step 6), after adsorbing the dye indigo, the adsorbent is naturally air-dried for 36-48 h, calcined for 4.0-6.0h at the temperature of 500-600 ℃, and naturally cooled to obtain ash and then returned to the field.
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| CN109007790A (en) * | 2018-09-07 | 2018-12-18 | 南阳理工学院 | A kind of preparation method of Chinese yam mycoplasma powder and its Chinese yam mycoplasma powder of preparation |
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