CN111607587A - Method for compositely regulating and controlling filamentous microalgae biomass based on micro-electric field/magnetic field - Google Patents
Method for compositely regulating and controlling filamentous microalgae biomass based on micro-electric field/magnetic field Download PDFInfo
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Abstract
The invention belongs to the technical field of bioengineering or biophysics, and relates to a method for regulating and controlling filamentous microalgae biomass by a micro-electric field/magnetic field composite method; filamentous microalgae has special industrial application advantages in the aspects of harvesting, pest resistance, pollution resistance and the like, and algae cells usually contain high grease, polysaccharide and bioactive substances and can be used as production raw materials in industries such as high value-added health products, animal feeds, biodiesel and the like; the invention utilizes the composite magnetic/electric field to regulate the open growth of the filamentous microalgae, the intensity of the micro electric field in the composite field is within 0-1.2V/cm, the timed stimulation is carried out, and the superimposed magnetic field intensity is applied within the range of 0-60 mT, the composite magnetic/electric field regulation has obvious promotion effect on the growth of the filamentous microalgae, the biomass accumulation and the nutrient removal rate of the filamentous microalgae can be improved, the intervention of the composite magnetic/electric field cannot cause the growth lag of the microalgae, and the method is a novel technology for effectively promoting the biomass growth of the filamentous microalgae.
Description
Technical Field
The invention belongs to the technical field of bioengineering or biophysics, and relates to a method for compositely regulating and controlling filamentous microalgae biomass based on a micro-electric field/magnetic field.
Background
Since microalgae is rich in protein, fat, carbohydrate, vitamins, chlorophyll and the like, has high nutritive value, and is widely applied to food, medicine, cosmetics, fuel, environment purification and the like, a great deal of research is carried out on microalgae growth influencing factors. The filamentous microalgae is easy to harvest, high in oil content, strong in vitality and capable of growing in extreme environments, and low in culture cost, so that the filamentous microalgae is the most potential research direction in the future. The research on the improvement of the culture effect of microorganisms such as microalgae mainly focuses on the aspects of culture condition optimization, traditional mutagenesis screening, molecular biology gene modification and the like. However, the controllability of open environmental conditions is limited, and optimization is difficult to implement; the mutation breeding has poor directionality and huge workload, and the modern genetic engineering means research investment cost is higher. In order to fully develop the potential of the hydrolytic acidification coupled filamentous microalgae open system for wastewater treatment, a new strategy for improving the growth of algae in an economic and efficient manner is needed.
The magnetic field and the electric field are used as safe non-ionizing radiation physical fields, and the device has the advantages of cleanness, no pollution, wide application range, simplicity in operation, easiness in amplification, good effect and the like. In recent years, magnetic/electric field-promoted biochemical processes of microorganisms have been successfully applied. Tu et al reported that static magnetic field acted on microalgae-bacteria symbiosisThe system promotes the accumulation of microalgae biomass and the oxygen production in water; kricelle et al research shows that static magnetic field can increase CO in chlorella2Compared with the control group, the magnetic strength of 60mT acts on chlorella for 1h/d, and CO acts2The biological fixation is increased by 49.7 percent, and CO is increased2The fixation is directly connected with the microalgae harvesting biomass, and the method effectively promotes the growth of the microalgae and the biological fixation of carbon dioxide; the research results of Darcy et al show that: the microalgae treated by 10mT is observed under a transmission electron microscope to have increased chloroplast and reduced thylakoid arrangement, which shows that the photosynthesis is obviously enhanced, and is consistent with the previous research conclusion; han et al studied the use of municipal sewage to culture chlorella, and treated by static magnetic field to reduce the turbidity of the waste water, increase active oxygen, and kill pathogenic bacteria in the water. Qin and other research results show that the pulsed electric field can crack microbial cells, so that the pulsed electric field has the function of inactivating bacteria;
the electric field or the magnetic field and the like can promote the growth of the microalgae to a certain extent, and the physical field has the sterilization effect and reduces the pollution degree, thereby better promoting the growth of the microalgae and improving the nutritive value of the microalgae. However, compared with the separately applied magnetic field and electric field, it is unknown whether the two low-intensity physical fields can play a significant synergistic effect when simultaneously introduced into the system; the electric field and the magnetic field in the physical field act on the microalgae together, and no research report is available; how to combine an electric field and a magnetic field to regulate filamentous microalgae biomass so that microalgae growth metabolism and biomass generate accumulation effect is a problem to be solved urgently.
Disclosure of Invention
In view of the deficiencies in the prior art, the present invention is directed to solving one of the problems set forth above; provides a method for regulating filamentous microalgae biomass by micro-electric field/magnetic field combination.
In order to achieve the purpose, the method for compositely regulating and controlling the biomass of the filamentous microalgae by the micro-electric field/magnetic field comprises the following steps:
(1) culturing filamentous microalgae: firstly, preparing a microalgae culture medium, then inoculating microalgae in a logarithmic growth phase into a culture container containing the culture medium, and then transferring the microalgae into an artificial climate incubator for culture, wherein the illumination condition is set to be 1000-10000 Lux, the temperature is 10-40 ℃, the illumination time is 24 hours, and the batch culture period of filamentous microalgae is 9-15 d;
(2) in the process of culturing the microalgae, providing a magnetic field with certain strength for the microalgae in the culture container through a magnetic field device; wherein the magnetic field device consists of magnets or magnetic induction coils, the magnets are arranged at two sides of the culture container, the adjustment of the intensity of the magnetic field is controlled by increasing or decreasing the magnets at two sides or controlling the intensity of the magnetic field by adjusting the number of turns of the coils or the current intensity; providing an electric field with certain strength for the microalgae in the culture container through an electric field device; wherein the electric field is formed by connecting a conventional electrode with a constant current power supply; the growth of filamentous microalgae biomass is realized through the composite regulation and control of an electric field/a magnetic field.
Preferably, the microalgae culture medium in the step (1) consists of water and culture nutrient salt components; the water is spring water, well water, tap water or agricultural sewage; the nutrient salt for culture comprises the following components: 300mg/L CO (NH)2)2(Urea) 60mg/L KH2PO4、60mg/L MgSO4·7H2O and 1mg/L FeSO4·7H2O。
Preferably, the culture vessel in step (1) is made of transparent glass.
Preferably, the inoculation amount of the filamentous microalgae in the step (1) is 0.1-0.2 g/L, and an air pump or an air compressor is used for continuously ventilating in the culture process to prevent the filamentous microalgae from settling; carbon dioxide with the concentration of 0.5-5% is mixed in the ventilation process, so that the photosynthesis effect is improved.
Preferably, in the step (1), the illumination condition is 5400Lux, the temperature is 25 ℃, and the batch culture period is 9 d.
Preferably, the filamentous microalgae culture vessel in step (1) is sterilized overnight by NaClO before use; the final concentration of the available chlorine is 7.5ppm calculated by the concentration of the available chlorine; the logarithmic growth phase of the filamentous microalgae inoculation is 3-6 days after the microalgae seeds are activated to grow.
Preferably, the action mode of the composite regulation and control of the electric field/magnetic field in the step (2) -the direction of the magnetic induction line of the superposed magnetic field and the direction of the electric field are combined into a certain angle, and the included angle is 0-180 degrees.
Preferably, the adjustment of the magnetic field intensity in the step (2) is controlled by increasing and decreasing magnets on two sides, and the magnetic field intensity is 0-60 mT but cannot be 0; the magnetic field is a static magnetic field, and the time of applying the magnetic field to the microalgae culture process is 1-24 h/d.
Preferably, the electric field intensity in the step (2) is controlled by a power supply, and the electric field intensity is 0-1.2V/cm but cannot be 0; the electric field is applied when the filamentous microalgae grow to the logarithmic growth phase, and the application time is 10-60 min.
The invention has the beneficial effects that:
(1) the method provided by the invention has the advantages of simple required equipment, convenience in operation and low cost; the combined action of the weak electric field and the magnetic field on the growth process of the filamentous microalgae is realized to realize the function of promoting the growth of the microalgae; compared with the effect of only an electric field, the biomass of the microalgae is improved by 10-22% by the composite field; the composite field regulation and control can also play a role in sterilization and disinsection, and solve the problem that the microalgae is polluted by microorganisms such as rotifers and the like in the industrial culture process.
(2) The composite field regulation is not simple accumulation, which cannot achieve good effect; the need to determine when to apply, the intensity of the application, and the time of application; the invention limits the magnetic field intensity to be 30-60 mT, and the application time is 1-24 h/d; the electric field intensity is 0.6-1.2V/cm, the application time is 30-60 min, and based on the conditions, the production cost of the microalgae can be reduced through composite field regulation and control, the biomass accumulation of the microalgae is realized, and the biological compound content and the nutritional value of the microalgae are improved.
Drawings
FIG. 1 shows the biomass concentration of filamentous microalgae of example 1 treated under different conditions.
FIG. 2 shows the percentage of TN removal in the filamentous microalgae of example 1 treated under different conditions.
FIG. 3 shows the biomass concentration of filamentous microalgae of example 2 treated under different conditions.
FIG. 4 shows the percentage of TN removal in the filamentous microalgae of example 2 treated under different conditions.
Detailed Description
It will be understood by those skilled in the art that the following examples are illustrative of the present invention only and should not be construed as limiting in any way.
Example 1:
the influence of the electric/magnetic field on the growth of filamentous microalgae (Tribonema sp.);
(1) firstly, preparing a microalgae culture medium, which comprises the following components: tap water 1L, 300mg of CO (NH)2)260mg KH2PO460mg of MgSO 24·7H2O and 1mg of FeSO4·7H2O; then inoculating microalgae in logarithmic growth phase into a transparent glass culture container containing a culture medium, wherein the culture container is a transparent glass square container with the volume of 10 x 10cm, and the inoculation amount is 0.2 g/L; wherein, the culture container is sterilized by NaClO overnight before use; the final concentration of the available chlorine is 7.5ppm calculated by the concentration of the available chlorine; finally, transferring the culture medium to an artificial climate incubator for culture, and setting the illumination conditions to be 5400Lux, the temperature to be 25 ℃ and the illumination time to be 24 h;
(2) providing a magnetic field for the microalgae in the glass container through a magnetic field device; wherein the magnetic field device consists of magnets which are arranged at two sides of the culture container; providing an electric field for the microalgae in the culture container through an electric field device; wherein the electric field is formed by connecting a conventional electrode with a constant current power supply; the micro-electric field/magnetic field composite regulation is adopted, the magnetic field is a static magnetic field, the average magnetic field intensity is 30mT, and the time of applying the magnetic field to the microalgae in the culture process is 24 h/d; the electric field is applied in the logarithmic growth phase (3d, 4d, 5d, 6d) of the filamentous microalgae, the application intensity is 0.6V/cm, and the application time is 30 min; biomass was harvested after 9d, 9d of culture period.
Meanwhile, a control group is set; group A: blank group (no electric field and magnetic field applied), group B: the intensity was 0.6V/cm with only electric field added, group C: only a magnetic field is added, the intensity is 30 mT; the rest of the operation is the same as the above steps.
From the results in fig. 1, it is seen that the biomass concentration of filamentous algae is increased by 9.52% under the complex field condition, compared with the blank group; 5.23% higher than the electric field alone; an improvement of 7.93% compared to the magnetic field alone; according to the results in FIG. 2, TN removal is particularly remarkable under the condition of a composite field, and is improved by 11.92% compared with a blank group; an improvement of 8.53% compared to the electric field alone; an improvement of 10.42% compared to the magnetic field alone.
Example 2:
the influence of the electric/magnetic field on the growth of filamentous microalgae (Tribonema sp.);
(1) firstly, preparing a microalgae culture medium, which comprises the following components: tap water 1L, 300mg of CO (NH)2)260mg KH2PO460mg of MgSO 24·7H2O and 1mg of FeSO4·7H2O; then inoculating microalgae in logarithmic growth phase into a transparent glass culture container containing a culture medium, wherein the culture container is a transparent glass square container with the volume of 10 x 10cm, and the inoculation amount is 0.2 g/L; wherein, the culture container is sterilized by NaClO overnight before use; the final concentration of the available chlorine is 7.5ppm calculated by the concentration of the available chlorine; finally, transferring the culture medium to an artificial climate incubator for culture, and setting the illumination conditions to be 5400Lux, the temperature to be 25 ℃ and the illumination time to be 24 h;
(2) providing a magnetic field for the microalgae in the glass container through a magnetic field device; wherein the magnetic field device consists of magnets which are arranged at two sides of the culture container; providing an electric field for the microalgae in the culture container through an electric field device; wherein the electric field is formed by connecting a conventional electrode with a constant current power supply; by the composite regulation and control of a micro electric field/a magnetic field, the magnetic field is a static magnetic field, the average magnetic field intensity is 30mT, and the time of applying the magnetic field to the microalgae in the culture process is 1 h/d; the electric field is applied in the logarithmic growth phase (3d, 4d, 5d, 6d) of the filamentous microalgae, the application intensity is 0.6V/cm, and the application time is 60 min; biomass was harvested after 9d, 9d of culture period.
Meanwhile, a control group is set; group A: blank group (no electric field and magnetic field applied), group B: the intensity was 0.6V/cm with only electric field added, group C: only a magnetic field is added, the intensity is 30 mT; the rest of the operation is the same as the above steps.
From the results in FIG. 3, it is seen that the biomass concentration of filamentous algae was increased by 21.55% under the complex field condition, compared with the blank group; an improvement of 16.40% compared to the electric field alone; an increase of 11.11% compared to the magnetic field alone; according to the results in FIG. 4, TN removal is particularly remarkable under the condition of a composite field, and is improved by 12.53% compared with a blank group; 15.2% higher than the electric field alone; an improvement of 4.24% compared to the magnetic field alone.
In conclusion, the micro-electric field/magnetic composite field promotes the accumulation of filamentous biomass within a certain range, improves the growth of filamentous algae, and can be used for improving the permeability and the enzymatic activity of a biological membrane by a micro-electric field, enhancing the action of a nutrient substrate, stimulating the growth and metabolism of microorganisms and improving the redox process of the microorganisms; meanwhile, TN removal is particularly remarkable under the condition of a composite field, and the increase of the percentage content of TN removal indicates that the absorption and utilization of the microalgae on TN are promoted under the action of the composite field, so that the content of N in water is reduced, and the absorption and utilization of nutrient substances in the water are facilitated, thereby playing a role in purifying sewage;
on the other hand, the micro-electric field/magnetic field superposed physical field is utilized to jointly act on the microalgae, the stimulation effect on algae cells is enhanced, the sterilization and disinsection effects are achieved, and the theoretical basis and the reference value are provided for the large-scale commercial culture of the outdoor microalgae.
Description of the drawings: the above embodiments are only used to illustrate the present invention and do not limit the technical solutions described in the present invention; thus, while the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted; all such modifications and variations are intended to be included herein within the scope of this disclosure and the present invention and protected by the following claims.
Claims (9)
1. A method for compositely regulating and controlling filamentous microalgae biomass based on a micro-electric field/magnetic field is characterized by comprising the following specific steps:
(1) culturing filamentous microalgae: firstly, preparing a microalgae culture medium, then inoculating microalgae in a logarithmic growth phase into a culture container containing the culture medium, and then transferring the microalgae into an artificial climate incubator for culture, wherein the illumination condition is set to be 1000-10000 Lux, the temperature is 10-40 ℃, the illumination time is 24 hours, and the batch culture period of filamentous microalgae is 9-15 d;
(2) in the process of culturing the microalgae, providing a magnetic field with certain strength for the microalgae in the culture container through a magnetic field device; wherein the magnetic field device consists of magnets or magnetic induction coils, the magnets are arranged at two sides of the culture container, and the adjustment of the intensity of the magnetic field is controlled by increasing or decreasing the magnets at two sides or controlling the intensity of the magnetic field by regulating the number of turns of the coils and the current intensity; providing an electric field with certain strength for the microalgae in the culture container through an electric field device; wherein the electric field is formed by connecting a conventional electrode with a constant current power supply; the growth of filamentous microalgae biomass is realized through the composite regulation and control of an electric field/a magnetic field.
2. The method for regulating filamentous microalgae biomass based on micro-electric field/magnetic field combination according to claim 1, wherein the microalgae culture medium in step (1) consists of water and culture nutrient salt components; the water is spring water, well water, tap water or agricultural sewage; the nutrient salt for culture comprises the following components: 300mg/L CO (NH)2)2、60mg/L KH2PO4、60mg/LMgSO4·7H2O and 1mg/L FeSO4·7H2O。
3. The method for regulating filamentous microalgae biomass based on micro-electric field/magnetic field combination according to claim 1, wherein the culture container in step (1) is made of transparent glass.
4. The method for compositely regulating and controlling the biomass of filamentous microalgae according to claim 1, wherein the inoculation amount of filamentous microalgae in the step (1) is 0.1-0.2 g/L, and an air pump or an air compressor is used for continuously ventilating in the culture process to prevent the filamentous microalgae from settling; carbon dioxide with the concentration of 0.5-5% is mixed in the ventilation process.
5. The method for compositely regulating and controlling the biomass of filamentous microalgae according to claim 1, wherein the light condition in step (1) is 5400Lux, the temperature is 25 ℃, and the batch culture period is 9 d.
6. The method for regulating and controlling filamentous microalgae biomass based on micro-electric field/magnetic field combination according to claim 1, wherein the filamentous microalgae culture vessel in step (1) is sterilized with NaClO overnight before use; the final concentration of the available chlorine is 7.5ppm calculated by the concentration of the available chlorine; the logarithmic growth phase of the filamentous microalgae inoculation is 3-6 days after the microalgae seeds are activated to grow.
7. The method for compositely regulating and controlling the filamentous microalgae biomass based on the micro-electric field/magnetic field according to claim 1, wherein the composite regulation and control of the electric field/magnetic field in the step (2) is implemented by superposing a magnetic induction line direction of the magnetic field and an electric field direction, and combining the magnetic induction line direction and the electric field direction to form a certain angle, wherein the angle is 0-180 degrees.
8. The method for compositely regulating and controlling the filamentous microalgae biomass based on the micro-electric field/magnetic field according to claim 1, wherein the regulation of the magnetic field intensity in the step (2) is controlled by increasing and decreasing magnets on two sides, and the magnetic field intensity is 0-60 mT but not 0; the magnetic field is a static magnetic field, and the time of applying the magnetic field to the microalgae culture process is 1-24 h/d.
9. The method for compositely regulating and controlling the filamentous microalgae biomass based on the micro-electric field/magnetic field according to claim 1, wherein the electric field intensity in the step (2) is controlled by a power supply, and the electric field intensity is 0-1.2V/cm but cannot be 0; the electric field is applied when the filamentous microalgae grow to the logarithmic growth phase, and the application time is 10-60 min.
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