CN116904360A - Wettman coagulating strain and application thereof in degradation of industrial production wastewater - Google Patents
Wettman coagulating strain and application thereof in degradation of industrial production wastewater Download PDFInfo
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- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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Abstract
The invention discloses a strain of condensation Wittman's bacterium and application thereof in degrading industrial production wastewater, and belongs to the technical field of biodegradation. The Wittman's bacterium coagulans can adapt to the environment of industrial production wastewater, has good growth and propagation, has good degradation and removal effects on COD components in the wastewater, and the degradation rate can reach 100% at most. Therefore, the strain disclosed by the invention can be suitable for severe environments containing industrial wastewater, can be applied to treatment of industrial wastewater, and is favorable for sustainable development of ecological environments.
Description
Technical Field
The invention relates to a strain of condensation Wittman's bacterium and application thereof in degrading industrial production wastewater, belonging to the technical field of biodegradation.
Background
Industrial wastewater refers to wastewater, sewage and waste liquid generated in the industrial production process. With the rapid development of industry, the variety and quantity of wastewater are rapidly increased, the pollution to water is also increasingly and seriously promoted, and the health and safety of human beings are threatened. Therefore, the treatment of industrial wastewater is more important than the treatment of municipal wastewater for environmental protection. Industrial wastewater often contains a large amount of COD, i.e., chemical oxygen demand in industrial wastewater, which reflects the degree of pollution of the water by reducing substances, and the COD index is also one of the comprehensive indexes of the relative content of organic matters. COD can lead to the incapability of living organisms in general water, the carcasses of the organism silk screen can not be fully oxidized, and the carcasses can be incompletely oxidized by anaerobic bacteria, so that a plurality of organic toxicants are generated, for example, sulfur element in the carcasses can be converted into hydrogen sulfide, nitrogen element can be converted into methylamine and the like, and the substances are substances which are peculiar smell and have strong biological toxicity, enter natural water bodies, destroy the balance of the water bodies, cause death of almost all organisms except microorganisms, not only harm the organisms in the water bodies, such as fish, but also can enter human bodies after being enriched by food chains, and cause chronic poisoning.
At present, the main treatment methods of industrial wastewater in China include an adsorption method, a filtration method, a flocculation precipitation method and a redox method. The adsorption method has lower cost for treating the printing and dyeing wastewater, but the efficiency for treating the wastewater is lower and the problem of adsorbent regeneration is not solved yet. The filtering method can treat printing and dyeing wastewater better but has higher operation difficulty. The flocculation precipitation method requires a large amount of flocculation precipitant to be added, resulting in high cost. The oxidation-reduction method has higher efficiency for treating the printing and dyeing wastewater but can produce secondary pollution.
At present, the related industrial wastewater of microbial degradation comprises the following components: the cationic dye is degraded by the bacillus subtilis composite material through the adsorption mechanism of the bacillus subtilis to the cationic dye, but the current experiment of biodegradation of industrial production wastewater has the defects of low biodegradation efficiency and high cost, and the COD-containing industrial production wastewater is treated by the immobilized bacillus licheniformis in the modified condition of the bacillus subtilis immobilized carrier and the treatment simulated wastewater for 24 hours, so that the COD degradation rate reaches 23.5% at maximum, and the industrial production requirement is difficult to meet. Therefore, the degradation efficiency is further improved, the degradation scheme is optimized, and efficient strains which can adapt to industrial production are screened out.
Disclosure of Invention
The technical problem is that in the field of degrading industrial production wastewater, the biological degradation still has the problems of large equipment occupation area, high cost, low degradation efficiency and incapability of being widely applied to industrial production.
In view of the problems existing at present, the invention extracts a strain of condensation Wittman from sludge of a Changzhou sewage treatment plant, and the strain can be used for degrading COD in industrial production wastewater, thereby providing an effective biological treatment method for treating industrial production wastewater.
The invention provides a strain of condensation Wittman's bacteria, which is preserved in China center for type culture Collection (CGMCC) No.25803 in the year 2022 and has a preservation address of North Chen Xili No. 1 and 3 in the Korean region of Beijing city.
The invention also provides a microbial preparation containing the condensation Wittman.
In one embodiment, the microbial preparation contains a viable count of Weizmann condensation bacteria of at least 2×10 9 CFU/ml。
In one embodiment, the microbial formulation further comprises a lyoprotectant.
The invention also provides a method for preparing the microbial preparation, which comprises the step of fermenting the condensation Wittman in a culture medium.
In one embodiment, the fermentation is at pH7.0-10.0, 100-180r/min,20-40℃for at least 22h.
The invention also provides application of the condensation Wittman or the microbial preparation in degrading industrial wastewater COD.
The invention also provides a method for degrading COD in industrial wastewater, which comprises the step of adding the coagulated Wittman's bacteria or the microbial preparation into the industrial wastewater.
In one embodiment, the COD value of the industrial wastewater is greater than or equal to 205.4mg/L.
In one embodiment, the bacterial liquid is treated with (4X 10) 8 ～2×10 9 ) The final concentration of CFU/mL is added to the wastewater and degraded for at least 6 hours at pH7.0-10.0, 100-180r/min,20-40 ℃.
The invention also provides the application of the Weizhman's bacterium coagulans, or the microbial preparation, or the method for preparing the microbial preparation in preparing sewage treatment agents.
Advantageous effects
The invention separates and obtains a strain of condensation Wittman's bacteria from sludge of a Changzhou sewage treatment plant, the condensation Wittman's bacteria can normally grow under the alkaline condition of pH7.0-10.0, and can degrade COD in industrial production wastewater, and the COD degradation rate of wastewater with 205.4-1002 mg/L COD content reaches more than 99%; for the wastewater with the COD content of 17481.5mg/L, the COD degradation rate reaches 63.82 percent. The degradation process is simple, and the requirements on environment and technology are low.
Preservation of biological materials
The strain of the Welch mannia coagulans, which is named Weizmannia coagulans in taxonomy, is preserved in China general microbiological culture Collection center (CGMCC) No.25803 in day 9 and day 26 of 2022, and has a preservation address of North Chen West Lu No. 1, no. 3 in the Chaoyang area of Beijing city.
Drawings
FIG. 1 is a diagram showing colony morphology and cell morphology of M.Weizhengensis Weizmannia coagulans; a is colony morphology, B is cell morphology.
FIG. 2 is a graph showing the change of the treatment of high-concentration COD wastewater by Wettman coagulans.
Detailed Description
The media involved in some embodiments of the invention are as follows:
LB liquid medium: yeast powder 5 g.L -1 Tryptone 10 g.L -1 Sodium chloride 10 g.L -1 1000mL of distilled water.
LB solid medium: yeast powder 5 g.L -1 Tryptone 10 g.L -1 Sodium chloride 10 g.L -1 1000mL of distilled water and 20g of agar.
pH adjustment of LB liquid Medium and LB solid Medium: the pH of the LB liquid medium or LB solid medium was adjusted to 7, 9, 10, 11, 13, 14 with 5mol/L sodium hydroxide solution and 0.1mol/L hydrochloric acid solution as needed.
In some embodiments of the invention, the COD degradation rate of industrial production wastewater is calculated by:
calculating the degradation rate of COD of the industrial wastewater:
the COD degradation rate of the industrial wastewater (total amount at the COD inlet-total amount at the COD outlet)/total amount at the COD inlet is measured by using a multiparameter water quality measuring instrument.
Technical terms:
sludge: the term "sludge" is taken from a Hemsy sewage treatment plant and has a density of 0.027g.mL -1 The pH was 7.66.
Industrial process wastewater: industrial wastewater from Changzhou sewage treatment plant contains a large amount of organic and inorganic pollutants such as dye, heavy metal, salt, grease and the like, has pH of 8.7, and has the characteristics of complex composition, strong alkalinity, high salinity, large chromaticity, toxic and harmful substances and the like.
The present invention is further described below with reference to examples, but embodiments of the present invention are not limited thereto.
Example 1: screening of strains
(1) Taking a density of 0.027 g.mL from a Changzhou sewage treatment plant -1 10mL of sludge is put into 90mL of LB liquid culture medium, and is cultivated for 4 to 6 days at the temperature of between 30 and 35 ℃ at 140 to 180 rpm;
(2) Inoculating the bacterial liquid in the step (1) into a new 100mL LB liquid culture medium according to the inoculum size of 5-10% of the volume ratio, and culturing for 4-6 d at the temperature of 30-35 ℃ at 140-180 rpm;
(3) Inoculating the bacterial liquid in the step (2) into a new 100mL LB liquid culture medium according to the inoculum size of 5-10% of the volume ratio, and culturing for 4-6 d at the temperature of 30-35 ℃ at 140-180 rpm;
(4) Adding 5 mu L of the bacterial liquid in the step (3) into the sterilized LB liquid culture medium, culturing for 4-6 d in a shaking table at 140-180 rpm and 30-40 ℃, sucking 100 mu L of liquid by using a gun head, and coating on an alkaline LB solid culture medium;
(5) Culturing the alkaline LB solid culture medium coated with the bacterial liquid in an incubator at 37 ℃ for 1-2 d, observing the form of bacterial colonies, picking a small amount of bacterial strains on each bacterial colony, respectively inoculating the bacterial strains into the alkaline LB solid culture medium, culturing for 4-6 d, and coating the bacterial liquid on the alkaline LB solid culture medium;
(6) Repeating the step (5) for a plurality of times until a single strain is obtained in each alkaline LB solid medium;
(7) Selecting single strain from culture medium with single strain, inoculating new 100mL alkaline LB liquid culture medium, culturing at 140-180 rpm and 30-35deg.C for 4-6 d, and measuring OD of bacterial liquid 600 The value reaches 0.8 to obtain the required bacterial liquid.
Example 2: identification of strains
(1) And (3) strain preservation: and (3) slope preservation, namely, preserving the strain in China general microbiological culture Collection center (CGMCC) at the 9 th day of 2022 and the 26 th day of 9 th day, wherein the preservation number is CGMCC No.25803, and the preservation address is North Chen Xiyu No. 1 and No. 3 in the Chaoyang area of Beijing city.
(2) And (3) strain identification: the strain was identified as Weatheroma belvedere Weizmannia coagulans by 16s RNA from Shanghai, inc.
Example 3: strains are cultured at different pH and pH tolerance
(1) The strain obtained in example 1 was inoculated into 100mL of LB liquid medium at a volume ratio of 2%, and cultured at 30℃at 100 r/min;
(2) Continuously shake culturing in LB liquid culture medium with initial pH of 7.0, 9.0, 10.0, 11.0, 13.0 and 14.0 for 48 hr;
(3) OD measurements in different initial pH Medium at intervals of 6h 600 Values.
As shown in Table 3, the Weizmannia coagulans strain enters the log phase earlier at an initial pH of 7, and thus it preferably has an initial pH of 7. And the tolerance of the pH of the Weizmannia coagulans strain is 7.0-10.0.
TABLE 1 OD of Weizmannia coagulans bacteria at different initial pH 600 Value of
Example 4: strains were cultivated at different rotational speeds
(1) The strain obtained in example 1 was inoculated into 100mL of LB liquid medium at a volume ratio of 2%, and cultured at 30℃and pH of 7;
(2) And (3) continuously shake-culturing in LB liquid culture media with shaking table rotating speeds of 0r/min, 100r/min and 140r/min for 48 hours.
(3) OD measurement in different shaking table rotating speed culture media at intervals of 6h 600 Values.
As shown in Table 2, weizmannia coagulans bacteria enter the logarithmic phase of microorganism growth at a rotation speed of 180r/min, so that the rotation speed of the optimal shaking table is 180r/min, and Weizmannia coagulans bacteria can normally grow in the rotation speed range of 100-180 r/min.
TABLE 2 OD of Weizmannia coagulans bacteria at different rotational speeds 600 Value of
Example 5: strain culture at different temperatures and temperature tolerance
(1) The strain obtained in example 1 was inoculated into 100mL of LB liquid medium at an inoculum size of 2% by volume, and cultured at a pH of 7 and a rotation speed of 180 r/min;
(2) Culturing in LB liquid medium at 20deg.C, 30deg.C, 40deg.C, 50deg.C and 60deg.C for 48 hr.
(3) OD measurement in different temperature Medium at intervals of 6h 600 Values.
The Weizmannia coagulans strain entered the log phase early at a temperature of 40℃as shown in Table 3, so that a suitable temperature range is 40℃in comparison. And the temperature tolerance of Weizmannia coagulans bacteria is 20-40 ℃.
TABLE 3 OD of Weizmannia coagulans bacteria at different temperatures 600 Value of
Example 6: application of Welch's bacteria in degradation of high-concentration industrial production wastewater
(1) Strain culture: inoculating Weizmannia coagulans bacteria obtained in example 1 into 100mL LB liquid medium respectively at 2% by volume, shake culturing at 30deg.C, pH7 and rotation speed of 100r/min for 48 hr to reach OD 600 About 1.0, 2×10 9 CFU/ml；
(2) Wastewater treatment: respectively collecting Weizmannia coagulans bacterial solutions obtained after culturing in step (1) for 48 hr, and respectively mixing bacterial solutions by 4×10 8 CFU/mL、8×10 8 CFU/mL、1.2×10 9 CFU/mL、1.6×10 9 CFU/mL and 2X 10 9 The final concentration of CFU/mL was added to 100mL of wastewater.
Treating for 6-72h at 40 ℃ and rotating speed of 180r/min, detecting the residual COD content in the wastewater every 12h, and calculating the COD degradation rate of the strain on the industrial wastewater. The reaction parameters for COD treatment at this time are the optimal growth conditions for the strain. The total COD at the wastewater inlet is 17481.5mg/L.
(3) Results determination: and (3) measuring the change of COD content before and after sewage, calculating degradation rate, and selecting the concentration of the bacterial liquid under the optimal degradation effect.
As is clear from Table 4 and FIG. 2, the amount of the M.Weizmanni coagulans produced in the step (1) was 1.2X10 9 Degrading high concentration industrial production in CFU/mLThe efficiency of the wastewater can reach the maximum, which is 63.82 percent.
Table 4Weizmannia coagulans bacteria treat high concentration COD wastewater for 72h (total amount at wastewater inlet is 17481.5 mg/L)
Example 7: application of Webster coagulans in degradation of low-concentration industrial production wastewater
The specific embodiment is as in example 6, except that the total amount at the industrial wastewater inlet is changed to 205.4mg/L, 321.9mg/L and 1002mg/L, respectively.
As is clear from Table 5, the amount of Weizmannia coagulans bacteria prepared in the step (1) added was 2X 10 9 When CFU/mL is carried out, the efficiency of degrading low-concentration industrial production wastewater (the total amount of wastewater inlet is 205.4 mg/L) can reach the maximum, the degradation is carried out for 12 hours, and the COD degradation rate can reach 100%.
Table 5Weizmannia coagulans bacteria treat COD wastewater of low concentration (total amount at wastewater inlet is 205.4 mg/L)
As is clear from Table 6, the amount of Weizmannia coagulans bacteria prepared in the step (1) added was 2X 10 9 When CFU/mL is carried out, the efficiency of degrading low-concentration industrial production wastewater (the total amount of wastewater inlet is 321.9 mg/L) can reach the maximum, the degradation is carried out for 12 hours, and the COD degradation rate can reach 99.14%.
Table 6Weizmannia coagulans bacteria treat COD wastewater of low concentration (total amount at wastewater inlet: 321.9 mg/L)
As is clear from Table 7, the amount of Weizmannia coagulans bacteria prepared in the step (1) added was 2X 10 9 When CFU/mL, low-concentration industrial wastewater (total wastewater at wastewater inlet) is degradedThe amount is 1002 mg/L), the efficiency can reach the maximum, the degradation is carried out for 12 hours, and the COD degradation rate can reach 100 percent.
Table 7Weizmannia coagulans bacteria treat COD wastewater of low concentration (total amount at wastewater inlet is 1002 mg/L)
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A strain of wegener's condensation (Weizmannia coagulans), wherein the strain of wegener's condensation has been deposited at the China general microbiological culture Collection center (ccm) under the accession number CGMCC No.25803 at month 09 of 2022.
2. A microbial preparation comprising the frozen Weizhman bacterium according to claim 1.
3. The microbial preparation according to claim 2, wherein the viable count of the coagulated Weizmann bacteria in the microbial preparation is not less than 2X 10 9 CFU/ml。
4. A microbial preparation according to claim 3, wherein the microbial preparation further comprises a lyoprotectant.
5. A process for producing the microbial preparation according to any one of claims 2 to 4, characterized in that the M.coagulans according to claim 1 is fermented in a medium.
6. The method of claim 5, wherein the fermentation is performed at a pH of 7.0-10.0, 100-180r/min,20-40℃for at least 22h.
7. Use of the microbial preparation of claim 1 or any one of claims 2 to 4 for degrading industrial wastewater COD.
8. A method for degrading COD in industrial wastewater, characterized in that the condensation of the species weemarginia according to claim 1 or the microbial agent according to any one of claims 2 to 4 is added to industrial wastewater.
9. The method according to claim 8, wherein the cells are treated with (4X 10) 8 ～2×10 9 ) The final concentration of CFU/mL is added into industrial wastewater, and degradation is carried out for at least 6 hours at the pH of 7.0-10.0 and 100-180r/min and the temperature of 20-40 ℃.
10. Use of the microbial preparation of claim 1, or of the microbial preparation of any one of claims 2 to 4, or of the method of claim 5 or 6 for the preparation of a sewage treatment agent.
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| CN117603847A (en) * | 2023-10-30 | 2024-02-27 | 无锡爱科派生物科技有限公司 | Wettman coagulans and application thereof in heavy metal adsorption |
| CN117603847B (en) * | 2023-10-30 | 2024-05-14 | 无锡爱科派生物科技有限公司 | Wettman coagulans and application thereof in heavy metal adsorption |
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