CN107446912B - Immobilized saccharomyces cerevisiae engineering bacteria and method for removing heavy metal Pb in wastewater by using same2+In (1) - Google Patents

Immobilized saccharomyces cerevisiae engineering bacteria and method for removing heavy metal Pb in wastewater by using same2+In (1) Download PDF

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CN107446912B
CN107446912B CN201710701087.9A CN201710701087A CN107446912B CN 107446912 B CN107446912 B CN 107446912B CN 201710701087 A CN201710701087 A CN 201710701087A CN 107446912 B CN107446912 B CN 107446912B
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saccharomyces cerevisiae
engineering bacteria
immobilized
heavy metal
cerevisiae engineering
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CN107446912A (en
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包红旭
张欣
杨华
张�浩
苏弘治
张馨月
魏德洲
黄金东
陈杨
王丽涛
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Liaoning University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • C—CHEMISTRY; METALLURGY
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/20—Heavy metals or heavy metal compounds

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Abstract

The invention relates to immobilized saccharomyces cerevisiae engineering bacteria and application thereof in removing heavy metal Pb in wastewater2+Belonging to the technical field of biological treatment of inorganic pollutants in the environment. The strain adopted by the invention is AS (2.1882) purchased from the China general microbiological culture Collection center, and the immobilized saccharomyces cerevisiae engineering bacteria is prepared by using fresh malt juice of a brewery AS a culture medium for activation culture, efficiently adsorbs and strengthens flocculation heavy metal Pb2+. The immobilized saccharomyces cerevisiae engineering bacteria have high-efficiency adsorption capacity, and the good flocculation effect of the immobilized saccharomyces cerevisiae engineering bacteria strengthens heavy metal ions Pb in industrial wastewater2+The removal efficiency of (2) is as high as 95.7%.

Description

Immobilized saccharomyces cerevisiae engineering bacteria and method for removing heavy metal Pb in wastewater by using same2+In (1)
Technical Field
The invention relates to a method for removing heavy metal Pb in industrial wastewater by using immobilized saccharomyces cerevisiae engineering bacteria2+Belonging to the technical field of biological treatment of inorganic pollutants in the environment.
Background
Heavy metal pollution in the environment is increasingly serious, and particularly, heavy metal pollution in sewage in the processes of mining, smelting and chemical production becomes a main aspect of environmental pollution. Heavy metals have high toxicity and can migrate with water after entering the water, thereby seriously threatening the surrounding ecological environment and the living quality of human beings. Heavy metal contaminated wastewater refers to a type of wastewater that is rich in heavy metal ions, which is extremely harmful to the ecological environment. Heavy metals cannot be biodegraded after entering the environment and often participate in food chain circulation and are finally enriched in organisms, damage the normal metabolism of the organisms and finally harm the health of human bodies.
However, the existing treatment method has certain disadvantages and technical obstacles, and more importantly, generates secondary pollution to the environment. The microbial adsorption treatment technology is increasingly concerned by people as an emerging treatment technology which is environment-friendly, efficient and economical.
Disclosure of Invention
Heavy metal Pb is repaired aiming at the current polluted water environment2+The invention provides an immobilized saccharomyces cerevisiae engineering bacterium, overcomes the defects that biological cells are too small and difficult to separate from an aqueous solution, and is easy to cause secondary pollution, maintains the advantages of high efficiency, strong stability, capability of purifying and maintaining efficient strains, and has wide application prospect in the field of wastewater treatment.
The technical scheme adopted by the invention is as follows: an immobilized saccharomyces cerevisiae engineering bacterium comprises the following preparation steps:
1) activating and culturing saccharomyces cerevisiae in a fresh malt juice culture medium;
2) adding the saccharomyces cerevisiae after activated culture into a mixed aqueous solution of polyvinyl alcohol and sodium alginate, and uniformly stirring to obtain a mixed bacterial liquid;
3) and (3) dripping the mixed bacterial liquid into the immobilized solution by using a needle tube, reacting for 10-12h, filtering, and washing obtained solid particles by using normal saline to obtain the immobilized saccharomyces cerevisiae engineering bacteria.
The immobilized saccharomyces cerevisiae engineering bacteria is purchased from China general microbiological culture Collection center and is numbered AS (2.1882).
The immobilized saccharomyces cerevisiae engineering bacteria comprise the following fresh malt extract culture media: the unfermented and hop-free fresh malt juice has sugar degree of 10-15 Bolin and pH of 6.5-7.5, preferably pH of 7.0. The fresh malt extract culture medium is obtained from Shenyang Huarun (snowflake) brewery.
The immobilized saccharomyces cerevisiae engineering bacteria is obtained by the step 1), and the saccharomyces cerevisiae is cultured in a fresh malt juice culture medium for 48 hours at the culture temperature of 34 ℃, the pH value of 6.0 and the oscillation intensity of 170 r/min.
The immobilized saccharomyces cerevisiae engineering bacteria is obtained by step 2), the cultured and activated saccharomyces cerevisiae is added into a mixed aqueous solution of polyvinyl alcohol (PVA) and sodium alginate, so that the final concentrations of the polyvinyl alcohol and the sodium alginate are respectively 7.5-10% and 0.4-1.0% in percentage by weight.
The immobilized saccharomyces cerevisiae engineering bacteria comprise the following steps of 3): in H3BO3Adding 1.0-5.0 wt% of CaCl into the saturated aqueous solution2And adjusting the pH value to 4.0-7.0.
The immobilized saccharomyces cerevisiae engineering bacteria are used for removing heavy metal Pb in water2+The use of (1). The method comprises the following steps: adding the immobilized saccharomyces cerevisiae engineering bacteria into the solution containing heavy metal Pb2+Reacting for 7-8 hours. Preferably, the pH of the industrial wastewater is adjusted to 6 to 7. Preferably, the adding amount of the immobilized saccharomyces cerevisiae engineering bacteria is every time2.5kg of industrial wastewater of cubic meter is added.
The saccharomyces cerevisiae used in the present invention has a short rod shape as shown in fig. 1 and 2. The diameter of the thallus cell is about 4.5-7um, the length is about 9.8-17um, and the two ends of the thallus are relatively round and blunt.
The invention has the beneficial effects that: the immobilized saccharomyces cerevisiae engineering bacteria prepared by the invention have the effect of treating heavy metal Pb in industrial wastewater2+The high-efficiency adsorption capacity of the flocculant and the good flocculation of the flocculant strengthen the heavy metal ions Pb in the industrial wastewater2+The removal efficiency is as high as 95.7%. The invention utilizes the immobilized microorganism technology to fix the screened saccharomyces cerevisiae, thereby forming a high-efficiency, rapid and continuous wastewater treatment system and effectively reducing secondary pollution. The immobilized saccharomyces cerevisiae engineering bacteria have the advantages of high cell density, high reaction speed, strong stability, strong poison resistance, less microorganism loss, easy product separation, less residual sludge and the like. The outstanding advantages make the method attach importance to the wastewater treatment, especially have good effect when treating heavy metal wastewater, show great potential and have wide application prospect in the field of wastewater treatment.
Drawings
FIG. 1 is an infrared spectrum of Saccharomyces cerevisiae.
FIG. 2 is a microscopic photograph of Saccharomyces cerevisiae.
FIG. 3 is a graph showing the relationship between the addition amount of the immobilized Saccharomyces cerevisiae engineering bacteria and the adsorption rate of heavy metal ions.
FIG. 4 is a graph showing the relationship between the pH value of wastewater and the adsorption rate of heavy metal ions.
FIG. 5 is a graph showing the relationship between adsorption time and adsorption rate of heavy metal ions.
FIG. 6 is a diagram showing the effect of treating wastewater containing heavy metal ions in the zinc plant in the cucurbit island;
wherein, 1: raw waste water; 2. treating the waste water; 3. and (4) treating the treated wastewater.
Detailed Description
The invention is further illustrated by the following specific examples, but is not limited thereto. Since the water quality and process flow vary from plant to plant, the key parameters of the present invention can be adjusted appropriately without departing from the spirit of the present invention and the scope of the appended claims.
Example 1
1) The saccharomyces cerevisiae (purchased from China general microbiological culture Collection center, number AS (2.1882)) is inoculated into fresh malt juice culture medium according to the inoculation amount of 2%, and activated and cultured for 48h at 34 ℃, with the pH value of 6.0 and the oscillation intensity of 170 r/min. Then inoculating the strain into a conventional liquid culture medium according to the inoculation amount of 2 percent for multiplication and amplification culture, and obtaining a large amount of saccharomyces cerevisiae thalli after 36 hours.
The fresh malt extract culture medium is as follows: fresh malt juice from Shenyang Huarun (snowflake) brewery, unfermented, and without hops, with sugar degree of 10-15 Bolin and pH of 7.0.
2) Heating and dissolving polyvinyl alcohol (PVA) and sodium alginate in water, cooling, adding the saccharomyces cerevisiae thallus obtained in the step 1) into a mixed aqueous solution of the polyvinyl alcohol and the sodium alginate, and uniformly mixing to ensure that the final concentrations of the PVA and the sodium alginate are 7.5-10% and 0.4-1.0% respectively according to weight percentage, thereby obtaining the mixed bacterial liquid.
3) And (3) dripping the mixed bacterial liquid into the immobilized solution by using a needle tube, reacting for 10-12h, filtering, and washing obtained solid particles by using normal saline to obtain the immobilized saccharomyces cerevisiae engineering bacteria.
The immobilization solution comprises the following components: in H3BO3Adding 1.0-5.0 wt% of CaCl into the saturated aqueous solution2And adjusting the pH value to 4.0-7.0.
Example 2 removal of heavy metal Pb from wastewater by immobilized Saccharomyces cerevisiae engineering bacteria2+In (1)
Firstly, sewage source:
the sewage comes from industrial production wastewater containing heavy metal ions in the zinc plant of the cucurbit island, the pH is 6.57, and Pb is2+The concentration of (B) was 11.30 mg/L.
(II) the addition amount of the immobilized saccharomyces cerevisiae engineering bacteria is used for removing the metal ions Pb2+Influence of (2)
Method: 1000ml of sewage is taken and added with the immobilized saccharomyces cerevisiae engineering bacteria 0.5/1.0/1.5/2.0/2.5/3.0/3.5g prepared in the example 1 respectively to react for 5 hours, and then the Pb in the solution is measured2+The content of (a). The results are shown in FIG. 3.
As can be seen from FIG. 3, the immobilized Saccharomyces cerevisiae engineering bacteria can react with Pb as the addition amount of the immobilized Saccharomyces cerevisiae engineering bacteria increases2+The adsorption rate of (A) is increased. When the addition amount of the immobilized saccharomyces cerevisiae engineering bacteria is 2.5g, the immobilized saccharomyces cerevisiae engineering bacteria have the effect of resisting Pb2+Good adsorption effect on Pb2 +The adsorption rate can reach 95.7%.
(III) influence of pH of wastewater on removal of Metal ions
The method comprises the following steps: taking 1000ml of sewage, respectively using dilute HCl and NaOH to adjust the pH value to 5/6/7/8/9, respectively adding 2.5g of the immobilized saccharomyces cerevisiae engineering bacteria prepared in the example 1, reacting for 7 hours, and then measuring Pb in the solution2+The content of (a). The results are shown in FIG. 4.
As can be seen from FIG. 4, as the pH value of the wastewater increases, the immobilized Saccharomyces cerevisiae engineering bacteria have Pb pairs2+The adsorption rate of (A) is increased. Has better adsorption effect in the pH range of 6-7 and has good adsorption effect on Pb2+The adsorption rate can reach 92.0-95.0%.
(IV) Effect of adsorption time on removal of Metal ions
The method comprises the following steps: taking 1000ml of sewage, adjusting the pH value to 6, adding 2.5g of the immobilized saccharomyces cerevisiae engineering bacteria prepared in the example 1, reacting for 1-8 hours, and then measuring Pb in the solution2+The content of (a). The results are shown in FIG. 5.
As can be seen from FIG. 5, the immobilized engineered Saccharomyces cerevisiae strain has Pb-adsorbing effect with prolonged adsorption time2+The adsorption rate of (A) is increased. When the adsorption is carried out for 7-8 hours, the immobilized saccharomyces cerevisiae engineering bacteria are used for treating Pb2+The adsorption rates of (A) and (B) respectively reach 95.7% of the maximum adsorption rates of the (A) and (B).
Through the screening of conditions, the optimal process conditions are as follows: the immobilized saccharomyces cerevisiae engineering bacteria are uniformly added into the industrial wastewater in a proportion of 2.5kg per cubic meter, have good adsorption effect in the range of pH 6-7 and the adsorption time of 6-7h, and can adsorb Pb2+The adsorption rate can reach 95.0 percentLeft and right.
Meanwhile, the microbial adsorbent shows a relatively ideal adsorption effect and has certain flocculation and decoloration effects on the wastewater containing heavy metal ions in the cucurbit island zinc factory. The effect of the treatment is shown in FIG. 6. Pb in the wastewater after adsorption treatment2+The content of the water-soluble organic acid meets the requirement of national industrial class II emission water quality standard, and shows better industrial application prospect.

Claims (3)

1. Method for removing heavy metal Pb in water by immobilized saccharomyces cerevisiae engineering bacteria2+The application of (1), which is characterized in that: the method comprises the following steps: adding the immobilized saccharomyces cerevisiae engineering bacteria into the solution containing heavy metal Pb2+Adjusting the pH =6-7 of the industrial wastewater, and reacting for 7-8 hours; the preparation method of the immobilized saccharomyces cerevisiae engineering bacteria comprises the following steps:
1) inoculating saccharomyces cerevisiae into a fresh malt juice culture medium, and performing activated culture;
2) adding the saccharomyces cerevisiae after activated culture into a mixed aqueous solution of polyvinyl alcohol and sodium alginate, and uniformly stirring to obtain a mixed bacterial liquid; the final concentration of polyvinyl alcohol and sodium alginate is 7.5-10% and 0.4-1.0% respectively;
3) dripping the mixed bacteria liquid into the immobilized solution by using a needle-shaped tube, reacting for 10-12h, filtering, and washing obtained solid particles by using normal saline to obtain immobilized saccharomyces cerevisiae engineering bacteria; the immobilization solution comprises the following components: in H3BO3Adding 1.0-5.0 wt% of CaCl into the saturated aqueous solution2Adjusting the pH value to 4.0-7.0;
the adding amount of the immobilized saccharomyces cerevisiae engineering bacteria is 2.5kg per cubic meter of industrial wastewater;
the saccharomyces cerevisiae is purchased from China general microbiological culture collection center and is numbered AS (2.1882).
2. The use according to claim 1, wherein the fresh malt extract medium is: the unfermented fresh malt juice without hops has sugar degree of 10-15 Bolin and pH value of 6.5-7.5.
3. The use according to claim 1, wherein in step 1), the saccharomyces cerevisiae is inoculated into a fresh malt extract medium and cultured for 48h at a culture temperature of 34 ℃, a pH =6.0 and a shaking intensity of 170 r/min.
CN201710701087.9A 2017-08-16 2017-08-16 Immobilized saccharomyces cerevisiae engineering bacteria and method for removing heavy metal Pb in wastewater by using same2+In (1) Active CN107446912B (en)

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CN103952393A (en) * 2014-05-15 2014-07-30 郑州大学 Preparation method of microorganism composite immobilized particle for in-situ repair of micro-polluted river
CN105734037A (en) * 2016-02-29 2016-07-06 洪寅 Waste water treatment agent and application thereof
CN105950679A (en) * 2016-05-27 2016-09-21 兄弟科技股份有限公司 Method for preparing D-pantoyl lactone by fermentation

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