CN112522109A - Microalgae immobilization method, immobilized microalgae and application in water pollution treatment - Google Patents

Microalgae immobilization method, immobilized microalgae and application in water pollution treatment Download PDF

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CN112522109A
CN112522109A CN202011533848.2A CN202011533848A CN112522109A CN 112522109 A CN112522109 A CN 112522109A CN 202011533848 A CN202011533848 A CN 202011533848A CN 112522109 A CN112522109 A CN 112522109A
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microalgae
sponge
immobilized
seed liquid
water body
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CN112522109B (en
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刘学阳
向文良
李建
简龙骥
黄元松
梁华忠
郑进
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SICHUAN QINGHE TECHNOLOGY CO LTD
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    • C12N11/08Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/089Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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    • C12N11/10Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate

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Abstract

The invention discloses a microalgae immobilization method and application of microalgae immobilized by the method in slow flow type water body pollution treatment, and relates to the field of water environment treatment. Firstly, carrying out enrichment culture on microalgae, and then centrifugally collecting to prepare a microalgae suspension; mixing the microalgae suspension with an immobilization solution prepared from sodium alginate, and brushing the mixture on a thin sponge sheet to complete the immobilization of the microalgae. In the treatment of water pollution, through the specially designed sponge mounting bracket, lay the sponge that is fixed with little algae in polluting the water according to specific arrangement to utilize the space of sponge to carry out the material exchange, little algae absorb carbon dioxide through photosynthesis and release oxygen, nitrogen and partly phosphorus in the sewage are absorbed by little algae and are used for the increase of biomass, and partly phosphorus is adsorbed by the sponge and so reach the effect of dephosphorization denitrogenation.

Description

Microalgae immobilization method, immobilized microalgae and application in water pollution treatment
Technical Field
The invention relates to the field of water environment treatment, in particular to a microalgae immobilization method, immobilized microalgae and application of immobilized microalgae in water pollution treatment.
Background
Microalgae are autotrophic plants which are widely distributed on land and sea, rich in nutrition and high in photosynthetic efficiency, and polysaccharide, protein, pigment and the like are generated through cell metabolism, so that the microalgae have good development prospects in the fields of food, medicine, genetic engineering, liquid fuel and the like.
Microalgae have their unique role, particularly in the field of environmental monitoring and purification. For example, the growth condition of microalgae can directly reflect the water quality condition, judge toxic gas in the air and break the analysis and detection mode of conventional gas samples. The growth activity of the microalgae in water can remove nitrogen and phosphorus, decompose refractory organic matters, and remove heavy metal ions such as Co, Mn, Hg and the like. The microalgae can also absorb certain concentration of NOx, SOx and H2And S. Therefore, the cultivation of microalgae for environmental protection has been studied in various countries around the world.
However, microalgae have great limitations in practical application of water pollution treatment: if the microalgae is directly thrown into the flowing water body, the microalgae can flow away with the water, and the water quality purification effect can not be realized completely; in addition, if the microalgae in the water body finishes the mission of water quality purification, the problem of the flooding of the microalgae exists if the microalgae is allowed to grow, so that the water quality purification effect cannot be achieved, and the turbidity of the water body algae can be caused; even if the growth of the microalgae is controlled, if the microalgae is not separated from water in time, the microalgae can be released into the water again after death and decomposition. Therefore, to make good use of microalgae for water purification, two problems must be solved: (1) ensuring that the microalgae thrown into the water body cannot float with the water; (2) the microalgae thrown into the water body can be conveniently separated from the water.
In order to solve the above problems, extensive research has been conducted on immobilization of microalgae. Among the different cell immobilization techniques, active entrapment is an immobilization method that is suitable for most algae. Sodium alginate is most widely used in all embedding materials. At present, the most common and mature mode is to use sodium alginate to prepare microalgae into immobilized beads, the immobilized beads have a remarkable effect on sewage purification in a bioreactor, but when the immobilized beads are applied to a natural water body, the problems of easy water drifting and difficult water and algae separation still exist.
Disclosure of Invention
The invention provides a microalgae immobilization method and application of immobilized microalgae in water pollution treatment based on the problems in the prior art, and solves the problems that the microalgae is easy to run off and is difficult to separate from water in the application of pollution treatment of a flowing water body.
A microalgae immobilization method comprises the following steps:
s1: selecting high-quality microalgae seeds, activating, inoculating and culturing to make the concentration of the seed liquid be 8-11 g/L;
s2: inoculating the microalgae seed liquid cultured to the logarithmic phase of growth in the step S1 into a seed tank of 150L for culturing, wherein the culture temperature is 30-35 ℃, the tank pressure is 0.03-0.08MPa, the air quantity is 1.0-2.0vvm, the DO is more than or equal to 20%, and the pH value is 5.5-7.5;
s3: when the seed liquid is cultured for 24-32h in the step S2 and the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 1.5 square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s4: when the seed liquid of the microalgae is cultured for 24-32h in the step S3 and the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 17-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s5: when the seed liquid is cultured for 24-32h in the step S4 and the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 50-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s6: centrifugally collecting and washing the microalgae cultured in the step S5, and preparing a microalgae suspension;
s7, preparing an immobilization solution by using sodium alginate, wherein the mass fraction of the immobilization solution is 0.5-0.8%;
s8: mixing the microalgae suspension with the immobilization solution;
s9: and (4) using sponge as an immobilization carrier, and coating the mixed solution obtained in the step (S8) on the sponge, wherein the sponge is in a sheet shape.
Further, the OD value of the microalgae suspension is between 0.25 and 0.30.
Further, the mixing volume ratio of the microalgae suspension to the immobilization solution is 1: 30-80.
Further, the sponge has the length of 1.0-2.0m, the width of 0.5-1.0m and the thickness of 0.3-0.5 cm.
The application of the immobilized microalgae in slow flow type water body pollution treatment comprises the following steps:
(1) placing the sponge fixed with the microalgae in a polluted water body, performing substance exchange by utilizing gaps of the sponge, absorbing carbon dioxide by the microalgae through photosynthesis to release oxygen, absorbing nitrogen and part of phosphorus in the sewage by the microalgae for increasing biomass, and absorbing part of phosphorus by the sponge to achieve the effects of dephosphorization and denitrification;
(2) taking the sponge fixed with the microalgae out of the water body after 10-15 days, filling new sponge fixed with the microalgae, taking out again after 10-15 days, and circulating for 3-5 times.
Furthermore, the sponge fixed with the microalgae is placed in the polluted water body through a sponge mounting frame; the sponge mounting rack comprises a protective net cover, a rotating rod, an upper transverse rack, a lower transverse rack, a screw and gear shaping; the size of the protective net cover is matched with that of the sponge fixed with microalgae, and the sponge is arranged in the protective net cover in water treatment application; the rotating rods are fixedly connected to the upper end and the lower end of the protection screen, the rotating rods at the upper end of the protection screen penetrate through the upper cross frame, the end heads of the rotating rods at the upper end of the protection screen are provided with gears, and the rotating rods at the lower end of the protection screen are inserted into the lower cross frame; the screw thread of the screw is meshed with a gear of a rotating rod at the upper end of the protective screen, and one end of the screw is provided with a motor for controlling the angle of the protective screen; the other end of the screw rod is provided with a rotating seat; the gear shaping is located the crossbearer bottom down, in water treatment application, inserts the gear shaping bottom of water to fix the sponge mounting bracket in the water.
Furthermore, 5-30 protection screen covers are arranged between the upper transverse frame and the lower transverse frame in a group in parallel.
Furthermore, at least two groups of protection net covers are arranged, and the protection net covers between the two adjacent groups are arranged in a staggered mode.
Furthermore, the protection net cover and the water flow direction have an inclination angle of 30-60 degrees, and the inclination angles of the two adjacent groups are opposite.
Further, the mesh opening of the protective net cover is 10-20 cm; the upper end and the lower end of the protective net cover are provided with connecting parts connected with the rotating rod; the side surface of the sponge is provided with an opening, so that the sponge can be conveniently taken and placed.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention fixes the microalgae on the sponge through the sodium alginate, is used for pollution treatment of the slow-flow type water body, and well solves the problem that the unfixed microalgae or the conventionally immobilized microalgae beads are easy to run off along with water; meanwhile, the microalgae and the water are easy to separate, and the sponge is taken out, so that the biomass of the microalgae is convenient to control, and the nitrogen, phosphorus and other elements absorbed by the microalgae after death and decomposition can be prevented from being released into the water again.
2. The invention takes the flaky sponge (the thickness is 0.3-0.5 cm) as the immobilized carrier of the microalgae, not only has good immobilization effect, but also solves the problem of poor light transmittance of the thicker sponge, is more beneficial to the photosynthesis of the microalgae, and ensures the dephosphorization and denitrification effect of the microalgae.
3. The invention also designs a special sponge mounting rack, the sponge mounting rack can place the sponges fixed with the microalgae in groups in the polluted water body, the sponges and the water flow direction can form a certain inclination angle, and two adjacent groups of sponges are arranged in a staggered mode, and the inclination angles are opposite. Through the arrangement, the contact area of the sponge and water flow can be greatly increased, and the water quality purification effect is enhanced; meanwhile, the sponge can be prevented from being directly impacted by water flow, and the sponge is prevented from being damaged.
4. The sponge is used as the immobilized carrier of the microalgae, so that the microalgae can conveniently play a role in dephosphorization and denitrogenation, and the gaps of the sponge can adsorb pollutants in the water body, thereby playing a role in enhancing the water body purification capacity.
5. The invention utilizes the protective net cover to protect the sponge fixed with microalgae, and improves the shock resistance of the sponge.
Drawings
FIG. 1 is a flow chart of microalgae immobilization according to the present invention. .
Fig. 2 is a schematic structural diagram of a sponge mounting rack according to an embodiment of the present invention.
Fig. 3 is a plan view of a sponge mount according to an embodiment of the present invention.
Fig. 4 is a schematic view of a protective mesh enclosure according to an embodiment of the invention.
The device comprises a protective net cover 1, a rotating rod 2, a gear 3, an upper transverse frame 4, a screw 5, a motor 6, a rotating seat 7, a lower transverse rod 8, a gear shaping 9, a mesh wire 100, an opening 101, a connecting part 102 and a sponge 103.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Culturing and immobilizing microalgae:
s1: selecting high-quality microalgae seeds, activating, inoculating and culturing to make the concentration of the seed liquid be 8-11 g/L;
s2: inoculating the microalgae seed liquid cultured to the logarithmic phase of growth in the step S1 into a seed tank of 150L for culturing, wherein the culture temperature is 30-35 ℃, the tank pressure is 0.03-0.08MPa, the air quantity is 1.0-2.0vvm, the DO is more than or equal to 20%, and the pH value is 5.5-7.5;
s3: after the seed liquid is cultured for 24-32h in the step S2, when the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 1.5 square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s4: after the seed liquid is cultured for 24-32h in the step S3, when the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 17-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s5: when the seed liquid is cultured for 24-32h in the step S4 and the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 50-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s6: centrifugally collecting and washing the microalgae cultured in the step S5, and preparing a microalgae suspension with an OD value of 0.25-0.30;
s7: preparing an immobilization solution by using sodium alginate, wherein the mass fraction of the immobilization solution is 0.5-0.8%;
s8: mixing the microalgae suspension with the immobilized solution at a mixing volume ratio of 1: 30-80;
s9: the mixed solution obtained in step S8 is applied to the sheet-like sponge 103 using the sponge 103 as an immobilization carrier.
The application of the immobilized microalgae in water body pollution treatment comprises the following steps:
in a river channel flowing through an urban area, the narrowest part of the river channel is 10 meters, the widest part of the river channel is 13 meters, the average depth is 1.5 meters, and the flow velocity is less than 1 m/s.
According to the condition of a river channel, a sponge mounting rack with the length of 10 meters is manufactured, and the mounting rack comprises a protective net cover 1, a rotating rod 2, an upper transverse rack 4, a lower transverse rack 8, a screw rod 5 and gear shaping teeth 9; the rotating rod 2 is fixedly connected to the upper end and the lower end of the protective screen 1, the rotating rod 2 at the upper end of the protective screen 1 penetrates through the upper cross frame 4, the end of the rotating rod 2 at the upper end of the protective screen 1 is provided with the gear 3, and the rotating rod 2 at the lower end of the protective screen 1 is inserted into the lower cross frame 8; the thread of the screw rod 5 is meshed with the gear 3 of the rotating rod 2 at the upper end of the protective net cover 1, and one end of the screw rod 5 is provided with a motor 6 for controlling the angle of the protective net cover 1; the other end of the screw 5 is provided with a rotating seat 7; the gear shaping 9 is positioned at the bottom of the lower cross frame 8.
The upper transverse frame 4 and the lower transverse frame 8 are respectively formed by connecting two cross rods, 15 protective net covers 1 are arranged on each cross rod, and each protective net cover 1 is 1m long, 0.5m wide and 2cm thick; and in each guard net 1, a sponge 103 fixed with microalgae is filled in the same size from an opening 101 of a side thereof.
The distance between the protective net covers 1 on the same cross rod is 0.5m, and the protective net covers 1 on the two cross rods are arranged in a staggered mode.
Transversely arrange the sponge mounting bracket in the narrowest place in the river course, gear shaping 9 inserts the riverbed to fix the sponge mounting bracket in the river course.
Starting a motor 6 to drive a screw rod 5 to rotate, driving a rotating rod 2 to rotate by the screw rod 5 through a gear 3 meshed with the screw rod 5, and driving a protective net cover 1 connected with the rotating rod 2 to rotate, so that the group of protective net covers 1 firstly contacting with the upstream water flow is inclined at an inclination angle of 30 degrees with the water flow direction; in the same manner, the other set of protective screen 1 is inclined at an inclination angle of 60 ° to the raw water flow direction.
After 10-15 days, starting the motor 6, rotating the protective net cover 1 to be in the horizontal direction with the water flow, and taking the sponge 103 fixed with the microalgae out of the water body, wherein the opening 101 faces the water incoming direction; then, putting a new sponge 103 fixed with microalgae, adjusting the protective net cover 1 to the original inclination angle again, and taking out the sponge 103 again after 10-15 days; and repeating the process according to the water quality monitoring condition.
The invention solves the problems of microalgae in the application of natural water pollution treatment: (1) the microalgae is easy to run off along with water, and hardly plays any role in treating water pollution after running off; (2) the separation of the microalgae and water is convenient, the biomass of the microalgae in the water can be controlled, the water is prevented from developing into a turbid water body, and elements such as nitrogen and phosphorus absorbed by the microalgae are prevented from being released again after the microalgae die and decomposed; (3) through special sponge mounting bracket, can lay the sponge that is fixed with little algae in polluting the water according to specific arrangement to utilize the space of sponge to carry out the material exchange, little algae absorb carbon dioxide through photosynthesis and release oxygen, nitrogen and partly phosphorus in the sewage are absorbed by little algae and are used for the increase of biomass, and partly phosphorus is adsorbed by the sponge and so reach the effect of dephosphorization denitrogenation.
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 changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (10)

1. A microalgae immobilization method is characterized by comprising the following steps:
s1: selecting high-quality microalgae seeds, activating, inoculating and culturing to make the concentration of the seed liquid be 8-11 g/L;
s2: inoculating the microalgae seed liquid cultured to the logarithmic phase of growth in the step S1 into a seed tank of 150L for culturing, wherein the culture control conditions are that the temperature is 30-35 ℃, the tank pressure is 0.03-0.08MPa, the air quantity is 1.0-2.0vvm, the DO is more than or equal to 20 percent, and the pH value is 5.5-7.5;
s3: after the seed liquid is cultured for 24-32h in the step S2, when the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 1.5 square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s4: after the seed liquid is cultured for 24-32h in the step S3, when the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 17-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s5: when the seed liquid is cultured for 24-32h in the step S4 and the dry weight of the microalgae seed liquid is 20-25g/L, inoculating the microalgae seed liquid into a 50-square tank for heterotrophic culture, wherein the culture control conditions are the same as those in the step S2;
s6: centrifugally collecting and washing the microalgae cultured in the step S5, and preparing a microalgae suspension;
s7: preparing an immobilization solution by using sodium alginate, wherein the mass fraction of the immobilization solution is 0.5-0.8%;
s8: mixing the microalgae suspension with the immobilization solution;
s9: the mixed solution obtained in step S8 is applied to the sponge (103) by using the sponge (103) as an immobilization carrier, and the sponge (103) is in the form of a sheet.
2. The method of claim 1, wherein the microalgae suspension has an OD of 0.25-0.30; the mixing volume ratio of the microalgae suspension to the immobilized solution is 1: 30-80.
3. The method of claim 1, wherein the sponge (103) has a length of 1.0-3.0m, a width of 0.5-1.0m, and a thickness of 0.3-0.5 cm.
4. An immobilized microalgae obtained by the method for immobilizing microalgae according to any one of claims 1 to 3.
5. The application of the immobilized microalgae in slow-flow water body pollution treatment is characterized by comprising the following steps:
s1: placing the immobilized microalgae as claimed in claim 4 in a slow-flow type polluted water body, performing substance exchange by utilizing the gaps of the sponge (103), absorbing carbon dioxide by the microalgae through photosynthesis to release oxygen, absorbing nitrogen and a part of phosphorus in the sewage by the microalgae for increasing biomass, and absorbing a part of phosphorus by the sponge (103), thereby achieving the effects of dephosphorization and denitrification;
s2: after 10-15 days, taking the immobilized microalgae of claim 4 out of the water body, putting in new immobilized microalgae, taking out again after 10-15 days, and circulating for 3-5 times.
6. The use of the immobilized microalgae in the slow-flow type water body pollution treatment according to claim 5, wherein the immobilized microalgae is placed in the polluted water body through a sponge mounting rack; the sponge mounting rack comprises a protective net cover (1), a rotating rod (2), an upper transverse rack (4), a lower transverse rack (8), a screw rod (5) and gear shaping (9); the size of the protective net cover (1) is matched with that of the sponge (103) fixed with microalgae, and the sponge (103) is arranged in the protective net cover (1) in water treatment application; the rotating rod (2) is fixedly connected to the upper end and the lower end of the protective screen (1), the rotating rod (2) at the upper end of the protective screen (1) penetrates through the upper transverse frame (4), a gear (3) is installed at the end of the rotating rod (2) at the upper end of the protective screen (1), and the rotating rod (2) at the lower end of the protective screen (1) is inserted into the lower transverse frame (8); the thread of the screw rod (5) is meshed with a gear (3) of a rotating rod (2) at the upper end of the protective screen (1), and one end of the screw rod (5) is provided with a motor (6) for controlling the angle of the protective screen (1); the other end of the screw rod (5) is provided with a rotating seat (7); the gear shaping (9) is positioned at the bottom of the lower transverse frame (8), and in the water treatment application, the gear shaping (9) is inserted into the bottom of a water body, so that the sponge mounting frame is fixed in the water body.
7. The application of the immobilized microalgae in the slow-flow type water body pollution treatment of claim 6, wherein the protective net covers (1) are arranged between the upper cross frame (4) and the lower cross frame (8) in a group of 5-30.
8. The application of the immobilized microalgae in the slow-flow type water pollution treatment of claim 7, wherein at least two groups of the protection screens (1) are arranged, and the protection screens (1) between two adjacent groups are arranged in a staggered manner.
9. The application of the immobilized microalgae in the slow-flow type water body pollution treatment of claim 8, wherein the angle of inclination of the protective net (1) to the water flow direction is 30-60 degrees, and the angle of inclination of the two adjacent groups is opposite.
10. The application of the immobilized microalgae in the slow-flow type water body pollution treatment according to claim 6, wherein the mesh aperture of the protective net (1) is 10-20 cm; the upper end and the lower end of the protective net cover (1) are provided with connecting parts (102) connected with the rotating rod (2); the side surface of the box body is provided with an opening (101).
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