CN114437940A - Fungus flocculant, preparation method and application of fungus flocculant - Google Patents
Fungus flocculant, preparation method and application of fungus flocculant Download PDFInfo
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Abstract
The invention relates to the technical field of water treatment, in particular to a fungal flocculant, a preparation method and application of the fungal flocculant, and provides the following scheme, wherein the fungal flocculant comprises Aspergillus oryzae (Aspergillus oryzae) CGMCC 3.16041. The preparation method comprises the following steps: inoculating aspergillus oryzae spores into a liquid culture medium, performing shake culture on a shaking table until mycelium pellets are formed, discarding supernatant of the liquid culture medium, separating the mycelium pellets from the liquid culture medium, washing the mycelium pellets with deionized water for three to five times, and performing vacuum filtration to obtain mycelium serving as a fungal flocculant. The invention screens the aspergillus oryzae fungi, utilizes hypha and extracellular secretion of the aspergillus oryzae fungi to have good flocculation effect on fresh water microalgae, is used for treating and repairing micro-polluted algae-containing water, discusses the action mechanism of the aspergillus oryzae fungi on the fresh water microalgae by researching the preparation method, the composition structure and the chemical components of the flocculant, and provides technical and theoretical support for further engineering application of the technology and bioremediation of the micro-polluted algae-containing water.
Description
Technical Field
The invention relates to the technical field of water treatment, in particular to a fungal flocculant, a preparation method and application thereof.
Background
In recent years, under the influence of human activities and climate change, the global water environment is constantly changed, the eutrophication degree of water bodies such as lakes and reservoirs is increasingly intensified, the quantity and the area of the water bodies which can be used as drinking water sources are sharply reduced due to the outbreak of algae, and the drinking water safety is seriously influenced;
the existing methods for removing microalgae in water comprise methods such as fishing, filtering, flotation, oxidation and the like and combined processes of the methods, and although the efficiency is high, the technologies can cause chemical pollution and high energy consumption; the flocculation sedimentation method is used as an important technology in water treatment, is widely applied due to simple process, convenient operation, economy and high efficiency, the type and the quality of the flocculating agent are key factors influencing the flocculation effect, the traditional flocculating agent, such as polyaluminium chloride (PAC), polyferric chloride (PFC) and Polyacrylamide (PAM), has lower use cost, but has the problems of low efficiency, high residue and the like, such as Alzheimer disease possibly caused by aluminum (Al) residues, strong carcinogen and neurotoxin to human beings caused by Acrylamide Monomer (AM), secondary pollution caused by ferric ion easily remained by ferric iron flocculating agent and easy corrosion to equipment;
therefore, the invention provides a fungal flocculant, a preparation method and application of the fungal flocculant.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a fungal flocculant, a preparation method and application of the fungal flocculant.
Bioflocculation is an effective strategy for removing microalgae, and particularly, filamentous fungus balls are used as a pre-enrichment step, a flocculant does not need to be separated and purified, so that energy consumption can be effectively reduced, cost can be reduced, and most microalgae biomass can be harvested through simple filtration; meanwhile, the microalgae is fixed in the mycelial pellets and is easy to separate by gravity sedimentation, and the structure of algae cells is complete in the flocculation process, so that the release of algal toxin is avoided, and a new idea is provided for large-scale harvesting of the microalgae;
not all microbial flocculants are suitable for removing microalgae in drinking water source water, and a great deal of experimental research shows that the filamentous fungus Aspergillus oryzae microbial flocculant can achieve a good flocculation effect. The strain is Aspergillus oryzae which is separated, identified and cultured by the applicant, and is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC 3.16041;
the invention provides a fungus strain Aspergillus oryzae (A.oryzae) with high flocculation effect on microalgae, namely microcystis aeruginosa. Systematic analysis and identification are carried out on the bacterial strain, the flocculation effect and the flocculation mode of the bacterial strain A.oryzae on microcystis aeruginosa are researched, and the influence of flocculation conditions such as different temperatures, rotating speeds, flocculation time, pH and the like on the flocculation effect of flocculation substances and the flocculation efficiency of the flocculation substances are researched.
In order to achieve the purpose, the invention adopts the following technical scheme:
a fungal flocculant comprises Aspergillus oryzae (CGMCC 3.16041).
The application of the fungal flocculant in removing microalgae in water.
A preparation method of a fungal flocculant comprises the following steps:
s1: inoculating Aspergillus oryzae spores into a liquid culture medium, and performing shake culture on a shaking table until mycelium pellets are formed;
s2: and discarding the supernatant of the liquid culture medium, separating mycelium pellets from the liquid culture medium, washing the mycelium pellets three to five times by using deionized water, and performing vacuum filtration to obtain the mycelium serving as the fungal flocculant.
Further, the temperature of the liquid culture medium in S1 is 20-25 deg.C, and shaking culture is carried out at 150rpm for 5-7 days until the diameter of mycelium pellet is 0.5-0.9 cm.
Further, the preparation method of the liquid culture medium comprises the following steps: mixing yeast extract 10 parts, peptone 20 parts, glucose 20 parts, adenine sulfate 0.03 parts and 1000 parts distilled water, and autoclaving at 121 deg.C for 20min to obtain liquid culture medium.
Further, the liquid culture medium comprises the following components in percentage by weight: 10 parts of yeast extract, 20 parts of peptone, 20 parts of glucose, 0.03 part of adenine sulfate and 1000 parts of distilled water.
The fungus flocculating agent prepared by the preparation method.
The application of the fungal flocculant in removing microalgae in water.
Further, the method comprises the following steps: adding Aspergillus oryzae pellet with concentration of 6.5-14.0mg/mL into microalgae with cell concentration of 1.5mg/mL, adjusting pH of the culture solution to 4.0-9.0, and flocculating at 15-30 deg.C and 50-200 rpm for 0.5-6 hr.
Further, the microalgae comprise the freshwater microalgae microcystis aeruginosa.
The invention has the beneficial effects that:
1. the invention aims at the problems that flocculant residue exists commonly in chemical flocculation and algae cell structures are damaged possibly, and provides a method for removing microalgae by utilizing filamentous fungi flocculation;
2. according to the invention, the filamentous fungus Aspergillus oryzae is used as the flocculant, and the fermentation product and extracellular secretion substances do not need to be further purified, so that the preparation cost of the flocculant is low, the extracellular secretion substances are protein, polysaccharide, lipid nucleic acid and the like, and the flocculant is safe and non-toxic; filamentous fungus Aspergillus oryzae cultured for 6 days in a shaking table is spherical particles with the diameter of 6-9mm, and can be separated by a simple filtering device after effectively adsorbing microalgae; compared with other methods for removing the microalgae, the method has the advantages of higher effect, shorter time and larger application potential, and provides effective basis and new selection for removing the microalgae in the water body.
Drawings
FIG. 1 is a diagram showing a developmental tree of the filamentous fungus Aspergillus oryzae obtained in example 1;
FIG. 2 is a graph showing the analysis of the removal rate of algal cells from the algae-containing water by the flocculant in example 1, wherein a is a graph showing the change in flocculation rate with the amount of addition of the filamentous fungus flocculant, b is a graph showing the change in flocculation rate with the flocculation time, c is a graph showing the change in flocculation rate with the flocculation temperature, and d is a graph showing the change in flocculation rate with pH;
FIG. 3 is a graph of a fit of adsorption isotherms of the filamentous fungal flocculant obtained in example 1 on algae-containing water, and b is a graph of a fit of the adsorption kinetics of the filamentous fungal flocculant obtained in example 1 on algae-containing water;
FIG. 4 is an SEM photograph of the filamentous fungal flocculant obtained in example 1 before and after flocculation;
FIG. 5 shows X-ray photoelectron spectra of fungus-microalgae obtained in examples 1-2, wherein (a) shows XPS spectra of fungus-microalgae, and (b-e) shows high resolution 1s XPS spectra of C, O, N and P derived from fungus-microalgae.
FIG. 6 is a chart of the infrared analysis spectra of the filamentous fungus flocculants, microalgae and flocculated flocs obtained in examples 1-2.
The fungus of the invention is Aspergillus oryzae (Aspergillus oryzae), with a preservation date: 04/09/2021; is preserved in China general microbiological culture Collection center (CGMCC), and the preservation number is CGMCC 3.16041.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
The aspergillus oryzae has the excellent characteristics of clear physiological, biochemical and genetic backgrounds, nonpathogenicity, strong protein secretion capacity, easy separation and culture and the like, so the aspergillus oryzae is used as a microorganism with food safety level; the extracellular secretion of Aspergillus oryzae comprises protein, extracellular polysaccharide, fat and nucleic acid.
A preparation method of a fungus flocculating agent comprises the following specific steps:
step S1: inoculating Aspergillus oryzae spores into a liquid culture medium, and performing shake culture on a shaking table until mycelium pellets are formed, wherein the formula of the liquid culture medium is as follows: 10 parts of yeast extract, 20 parts of peptone, 20 parts of glucose, 0.03 part of adenine sulfate and 1000 parts of distilled water. And (3) carrying out autoclaving at 121 ℃ for 20min to obtain the liquid fermentation medium.
The specific process is as follows: adding Aspergillus oryzae spore into flocculation culture medium, shake culturing at 20-25 deg.C and 150rpm for 5-7d until the diameter of most mycelium pellet is 0.5-0.9 cm;
step S2: taking a liquid fermentation culture medium, removing supernatant, washing the separated mycelium pellets three to five times by using deionized water, and obtaining the mycelium as a microbial flocculant by using a vacuum filtration device.
The application of the microbial flocculant in efficiently collecting microalgae biomass is characterized by comprising the following specific processes: adding Aspergillus oryzae pellet with concentration of 6.5-14.0mg/mL (dry weight) into microalgae culture solution with cell concentration (dry weight) of 1.5mg/mL, adjusting pH of the culture solution to 4.0-9.0, flocculating at 15-30 deg.C and 50-200 rpm for 0.5-6 hr, flocculating to adsorb microalgae, and collecting the pellet by fishing or filtering to flocculate and settle microalgae in the culture solution.
The concentration of the microbial flocculant is preferably 8.0-14.0 mg/mL. The invention collects the microalgae preferably under the conditions of stirring speed of 50-150 r/min, temperature of 20-30 ℃ and time of 0.5-6 hours.
The microalgae of the invention are fresh water algae, microcystis aeruginosa and are available on the market; the culture medium of the microalgae culture solution is BG11 culture medium.
Based on the preparation method of the fungus flocculant, the invention provides the fungus flocculant prepared by the preparation method and application of the fungus flocculant in removing microalgae in water.
Also, the present invention provides a fungal flocculant, comprising Aspergillus oryzae (Aspergillus oryzae) CGMCC 3.16041, in some embodiments, the fungal flocculant comprises a mixture of a compound secreted by Aspergillus oryzae and an extracellular active substance, e.g., protein, extracellular polysaccharide, fat, nucleic acid; and other related reagents can be added as required by the person skilled in the related art, but the main effective component of the reagent is aspergillus oryzae.
Implementation 1: preparation of microbial flocculant
Inoculating Aspergillus oryzae in YPDA culture medium (YPDA liquid culture medium: yeast extract 10g, peptone 20g, glucose 20g, adenine sulfate 0.03g, deionized water 1L, 120 deg.C, 20min autoclaving), adding Aspergillus oryzae spore into flocculation culture medium, shake culturing at 25 deg.C and 150rpm for 6d until most mycelium pellet diameter is 0.5-0.9cm, discarding supernatant, washing mycelium with deionized water three to five times, and vacuum filtering to obtain mycelium as microbial flocculant.
Implementation 2: microalgae culture
At 1.6X 106Inoculating Microcystis aeruginosa in deionized water containing BG11 culture medium, shaking, and culturing at 25 deg.C under 4000lx illumination for 14 days with cell density of 1.6 × 109one/mL, this culture was used in the following examples. BG11 culture medium: NaNO3 1.5g/L,Na2CO3 0.02g/L,CaCl2·2H2O 0.036g/L,MgSO4·7H2O 0.075g/L,K2HPO40.04g/L, 0.006g/L ferric ammonium citrate, 0.006g/L citric acid, Na2·EDTA 0.001g/L。
Example 3:
molecular characterization of Aspergillus oryzae
1) Inoculating spores of Aspergillus oryzae in flocculation medium, and shake culturing at 25 deg.C and 150rpm for 3 d. Extracting hypha genome DNA, sending to a company for genome sequencing, screening single-copy direct homologous genome comparison clusters of coding proteins in the genome, selecting seeds which are close to the genetic relationship with a target strain, and constructing a phylogenetic tree (figure 1).
Example 4: research on removal effect of mycelium pellets on microalgae
1) Adding mycelium pellets with the diameter of about 0.7cm into the culture solution of the microcystis aeruginosa according to the addition of 11mg/mL, shaking the microcystis aeruginosa culture solution for 1 to 6 hours in a shaking table at 150rpm, and sampling at regular intervals.
2) After flocculation of the algae cells, the cultures of the treated and control groups were aliquoted and sampled from the bottom two thirds of the height, and the absorbance value of the samples at 686nm wavelength was determined by uv spectrophotometer to characterize the microalgae content in the suspension (fig. 2).
3) And (3) placing part of the mycelium pellets on a glass slide, and observing the distribution characteristics of the microalgae in the mycelium pellets under an optical microscope.
The mycelium pellet has a certain removing effect on microalgae, and a large number of microalgae cells with the diameter of about 8 mu m are found in the mycelium pellet, which shows that the mature mycelium pellet has a certain flocculation effect on the microalgae cells. FIG. 2a shows the change curve of flocculation rate of filamentous fungi obtained in different culture time at different dosages, and the flocculation rate is above 90% when the culture time is 5-7 days and the dosage of the filamentous fungi exceeds 11mg/mL, which indicates that the filamentous fungi can effectively flocculate microalgae. FIG. 2b shows the flocculation rate as a function of flocculation time at different rotation speeds, and the flocculation rate gradually increases with the increase of the flocculation time, indicating that the effect of the fungi on the microalgae is influenced by the flocculation time. The flocculation efficiency increased from 57% to 93% at a flocculation time of 5h, increasing the stirring speed from 50rpm to 100 rpm. However, further increasing the stirring speed to 200rpm resulted in a decrease in flocculation efficiency (5%). Experimental data indicate that excessive shear force may cause microalgae cells to be detached from fungal hyphae. Figure 2c shows the effect of different flocculation temperatures on flocculation efficiency, with the flocculation rate showing a tendency to increase and then decrease as the flocculation temperature increases. The increase in temperature may favor chemisorption between the fungal hyphae and the microalgae cells, while the temperature effect may affect the growth of the fungus, i.e. the total surface area of fungal hyphae in contact with the microalgae, thereby affecting the flocculation efficiency. FIG. 2d is a graph showing the change of Zeta potential of the supernatant fluid under different pH conditions, wherein the Zeta potential gradually increases with the increase of pH, and the potential after flocculation is higher than the potential before flocculation, indicating that the microalgae surface has negative charges, and the part of positive charges on the fungal surface neutralizes the negative charges on the microalgae to increase the potential after flocculation. Higher flocculation rates were achieved at different pH conditions, indicating that chemisorption might also play an important role in flocculation in addition to electrical neutralization.
The research determines the optimal flocculation condition according to a single-factor experiment, 50mL of the Microcystis aeruginosa culture solution is taken, 11.0mg/mL of the prepared microbial flocculant is added into the Microcystis aeruginosa culture solution, the temperature is 25 ℃, the speed is 100r/min, and the flocculation rate is 99.2 percent after 5 hours of flocculation (figure 2).
Example 5: experimental study on adsorption of mycelium pellets on microalgae
On the basis of the preliminary experiment, 3 influencing factors of pH value, flocculation time and flocculation rotating speed are selected for carrying out adsorption experiment research.
1) Adsorption isotherms were determined by analyzing the experimental data of different initial algal cell mass concentrations on the adsorption of microalgae cells by filamentous fungi at different temperatures (15, 20, 25 ℃), by using Freundlich adsorption isotherm equations and Langmuir adsorption isotherm equations for adsorption isotherm analysis, and by studying what effects the adsorption behavior of filamentous fungi on algal cells is dominated by, Langmuir and Freundlich equations fit to20.9970, 0.9974, 0.9876 and 0.9916, 0.9685 and 0.9866 respectively, the fitting property of the two is high, and the single-layer adsorption and the multi-layer adsorption are simultaneously carried out in the process of adsorbing the algae cells by the filamentous fungi (figure 3 a).
2) Adsorption kinetics 50mL of 1.152g/L algae solution was adsorbed at 100r/min, and samples were taken at 0, 10, 20, 30, 60, 90, 120, 150, 180, 210, 240, 270 and 300min to determine the amount of adsorbed microalgae.
For a simulated water sample experiment, the flocculation rate is over 90 percent when the reaction time is 30min, the adsorption balance is reached when the reaction time is 90min, and the adsorption quantity of the microalgae is 93.3 mg/ml.
In order to research the reaction rate change and the adsorption capacity change of the fungus hypha adsorbing the algae cells, a quasi-first-stage and quasi-second-stage kinetic model is utilized to analyze experimental data so as to better understand the potential mechanism quasi-first-stage and quasi-second-stage kinetic equation of the fungus hypha in the process of adsorbing the algae cells, which is shown in an equation (1) and an equation (2):
the experimental results show that the flocculation time of the water sample after the pH is adjustedCan be greatly reduced, which shows that the adsorption effect of the filamentous fungi on the microalgae is better (figure 3 b). R after fitting quasi-first order kinetic equation2At 0.995, R after fitting the quasi-second order kinetics equation2Is 0.999. K1＝0.54，Qe＝94.7796，K2The adsorption kinetics process is more consistent with a simulated second-order kinetics model when the adsorption kinetics process is 0.008 and Qe is 95.75, and the fitted Qe is also very close to the experimentally obtained Qe. Adsorption is primarily based on chemisorption (associated with chemical bonds and functional groups), including physisorption (e.g., electrostatic interactions).
Example 6: sample characterization before and after flocculation
In FIG. 4, (a), (b), and (c) correspond to scanning electron micrographs of filamentous fungi at magnifications of 5000 ×, 10000 ×, 20000 ×, respectively, and they are shown in the following figures: the Aspergillus oryzae is a spatial reticular structure formed by cross-linking, intertwining and supporting filamentous hyphae, and the surface of the spatial reticular structure is wrinkled. The morphology of the flocculated fungi and microalgae is shown in FIG. 4 (d, e, f). Compared with separately cultured hypha particles, with the extension of flocculation time, some spherical blue algae cells can be clearly observed to be adsorbed on the fungal hyphae, and some spherical blue algae cells are wrapped in the hypha cross-linked reticular interior. The mycosphaerella gradually turns green, and the color of the culture solution tends to be clear and transparent, which indicates that the microalgae are captured and fixed by the fungal mycelium. The adsorption and fixation efficiency of the hypha on the microalgae reaches 99%. SEM showed that fungal mycelia cross-linked with the microalgae particles while the algal cell structure was intact (b in fig. 3), and the overall results demonstrate that filamentous fungi efficiently flocculated microalgae without disrupting algal cell structure.
FIG. 5 is a plot of X-ray photoelectron spectra of flocculated fungi-microalgae, with peaks associated with C- (C, H) bonds at a binding energy of 284.8eV, possibly from lipid or amino acid side chains. The peak at 286.3eV is associated with C-O, N in ethers, alcohols, amines and amides. The peak at 287.9eV is a C ═ O bond or O — C — O bond, including amides, carbonyls, carboxylates, esters, acetals, and hemiacetals. The peak at 292.8eV may be due to an unsaturated conjugated system (pi-pi transition), such as carbonyl C ═ O and aromatic rings, there is an interaction between pi electron systems of fungal-microalgal aromatic rings, while pi electron systems of aromatic rings may be involved in microbial flocculant-on-microalgalAdsorption behavior. The O peaks (O1s, and O1sA) can be broken down into two bonds: the peak at 530.8eV is due to carboxylate, carbonyl, ester or amide O-C bonds and the peak at 532.3eV is due to C ═ O from alcohol, hemiacetal or acetal groups. The N peaks (N1s and N1sA) are due to two different bonds: the peak at 399.9eV is attributed to-NH from the aprotic nitrogen amide or amine2And C-NH2Group correlation, another peak at 402.1eV is due to protonated amines, commonly found in amino acids and amino sugars. The content of P element in the microalgae is low, the content of P element in the fungi is high, and the content of P element in the flocculated fungi-microalgae floc is between that of the fungi and the microalgae, which indicates that the fungi flocculate the microalgae successfully. And (3) the relative content of O-C after the fungi adsorb the microalgae is found to be changed in a semi-quantitative way by combining related parameters of an X-ray photoelectron spectrogram, so that the O-C is supposed to participate in the reaction process of adsorbing the microalgae by the fungi. Due to the interaction between microalgae and fungi, the intensities of the C-C, C-O, C-N and pi-bond absorption peaks after flocculation are reduced to different degrees, presumably during the adsorption of microalgae by fungi, CH-, COOH-and pi-on the fungal surface take part in the reaction.
FIG. 6 is the infrared spectra of microalgae, fungi and flocculated fungi-microalgae, and it can be seen from FIG. 6 that the structures of the thallus functional groups before and after flocculation have not changed significantly, wherein the molecular functional groups at 3420cm-1The wider absorption band is the stretching vibration of the associated OH-functional group, and the strength of the flocculated fungus-microalgae is slightly reduced; at 2926cm-1The characteristic peak is aliphatic chain-CH2And (4) stretching the groups. 1648cm-1C ═ O and 1075cm-1The intensity of the characteristic peak of the C-O single bond is weakened, which indicates that the active site in the flocculant is occupied by the microalgae. 3281cm-1And 1541cm-1NH-sum at 1648cm-1The amide group, hydroxyl group and carboxyl group of C ═ O play an important role in flocculation. The XPS result is consistent with FTIR analysis, which proves that the flocculating agent contains abundant hydroxyl, amino and carboxyl, and plays a key role in the flocculation process.
The above embodiments show that the filamentous fungus bioflocculant provided by the invention not only has high-efficiency algae removal and turbidity removal capability, but also is simple to prepare and convenient to add.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A fungal flocculant, characterized in that: comprises Aspergillus oryzae (CGMCC 3.16041).
2. Use of the fungal flocculant of claim 1 for removing microalgae from water.
3. The preparation method of the fungal flocculant is characterized by comprising the following steps of:
s1: inoculating Aspergillus oryzae spores into a liquid culture medium, and performing shake culture on a shaking table until mycelium pellets are formed;
s2: and discarding the supernatant of the liquid culture medium, separating the mycelium pellets from the liquid culture medium, washing the mycelium pellets three to five times by using deionized water, and performing vacuum filtration to obtain the mycelium serving as the fungal flocculant.
4. The method for preparing the fungal flocculant according to claim 3, wherein the temperature of the liquid medium in S1 is 20-25 ℃, and the liquid medium is shake-cultured at 150rpm for 5-7 days until the diameter of the mycelium pellet is 0.5-0.9 cm.
5. The method according to claim 4, wherein the method for preparing the liquid culture medium comprises: mixing yeast extract 10 parts, peptone 20 parts, glucose 20 parts, adenine sulfate 0.03 parts and 1000 parts distilled water, and autoclaving at 121 deg.C for 20min to obtain liquid culture medium.
6. The method for preparing the fungal flocculant according to claim 5, wherein the liquid culture medium comprises the following components in percentage by weight: 10 parts of yeast extract, 20 parts of peptone, 20 parts of glucose, 0.03 part of adenine sulfate and 1000 parts of distilled water.
7. A fungal flocculant produced by the production process according to any one of claims 3 to 6.
8. Use of the fungal flocculant of claim 7 for removing microalgae from water.
9. The use of the fungal flocculant of claim 8 for removing microalgae from water, comprising the steps of: adding Aspergillus oryzae pellet with concentration of 6.5-14.0mg/mL into microalgae with cell concentration of 1.5mg/mL, adjusting pH of the culture solution to 4.0-9.0, and flocculating at 15-30 deg.C and 50-200 rpm for 0.5-6 hr.
10. The use of a fungal flocculant of claim 8 for the removal of microalgae in water, wherein the microalgae comprise the freshwater microalgae microcystis aeruginosa.
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| CN113562936A (en) * | 2021-08-20 | 2021-10-29 | 河南功能高分子膜材料创新中心有限公司 | Water-based ink wastewater treatment process |
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