CN107937382A - A kind of preparation method of immobilized microalgae - Google Patents
A kind of preparation method of immobilized microalgae Download PDFInfo
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- CN107937382A CN107937382A CN201711428923.7A CN201711428923A CN107937382A CN 107937382 A CN107937382 A CN 107937382A CN 201711428923 A CN201711428923 A CN 201711428923A CN 107937382 A CN107937382 A CN 107937382A
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Abstract
The invention discloses a kind of preparation method of immobilized microalgae, comprise the following steps：Under aseptic technique, chlorella is inoculated into culture medium, when light dark cycle alternate culture 60 75 is small, obtains micro algae culturing liquid；The pH value of micro algae culturing liquid is adjusted to 4.5 6, adds sodium citrate, is stirred 20 30 minutes, adds calcium chloride, stirs 40 50 minutes, is centrifugally separating to obtain microalgae liquid；Prepare the carrier solution containing sodium alginate and magnesium chloride, microalgae liquid is added in carrier solution, instill mass fraction with syringe as in 2.6% 3.2% calcium chloride solution after mixing, 8 10 DEG C fix 2 2.5 it is small when after take out, with deionized water desalination 12 it is small when.The immobilized microalgae prepared using the method for the present invention is had the advantages that treatment effect is stable, can effectively prevent secondary pollution.
Description
Technical field
The present invention relates to microalgae sewage-treating agent preparation field, more particularly to a kind of preparation method of immobilized microalgae.
Background technology
With the large-scale development of China's livestock and poultry breeding industry, the livestock and poultry cultivation waste of nearly 1,000,000,000 tons of generation per annual meeting, to having
The water resource of limit causes serious threat.
At present, the processing mode of aquaculture wastewater mainly has two kinds：First, being handled using methane-generating pit anaerobic fermentation, but produce
The nutritive salt such as the nitrogen phosphorus in the biogas slurry of raw enormous amount still containing high concentration, with the propulsion of the Course of Urbanization, disappear
Receive biogas slurry arable land it is increasingly insufficient, the biogas slurry of generation is directly discharged in water body, it will causes natural water serious eutrophication；
Second, bioanalysis removes, biological treatment is to be degraded using microorganisms such as bacterium, fungies to the pollutant in sewage and/or together
Change and absorb, realize the purification of sewage, but due to aquaculture wastewater after Anaerobic Treatment general available carbon source it is too low, can not
Meet the needs of heterotrophism function bacterium, therefore to nitrate and phosphatic poor processing effect, it is necessary to physical filtering and chemisorbed
It is used in combination Deng various ways, this not only adds processing cost, also increases potential secondary pollution risk.
Microalgae facture is sewage treatmentmethod emerging at present, the abundant nitrogen contained in livestock and poultry cultivation sewage, phosphorus etc.
Nutriment can provide nutrition for microalgae, can be joined at the same time with the large-scale production phase lotus root of microalgae using microalgae treated sewage, realize
Resource recycles.
But since microalgae cell is generally less than 30 μm, negatively charged, density causes frustule to exist close to water, these characteristics
Often microalgae cell is caused largely to be lost in processing water, this not only can secondary pollution in stable suspended state in water
Water is handled, and causes microalgae cell biomass in treated sewage to be difficult to largely maintain（It is typically only 0.2-0.6g/L）.Low
Culture density causes removal efficiency low so that treatment effect stability is poor.
The content of the invention
Based on technical problem existing for background technology, the present invention proposes a kind of preparation method of immobilized microalgae, uses
Immobilized microalgae prepared by the method for the present invention has the advantages that treatment effect is stable, can effectively prevent secondary pollution.
The present invention provides a kind of preparation method of immobilized microalgae, comprise the following steps：
S1, under aseptic technique, by chlorella with 1 × 106A/mL-1 × 107The inoculum concentration of a/mL is inoculated into culture medium
In, it is 25-27 DEG C in temperature, under the conditions of rotating speed is 110-130 revs/min, when light dark cycle alternate culture 60-75 is small,
Obtain micro algae culturing liquid；
The pH value of the micro algae culturing liquid obtained in S1, is adjusted to 4.5-6 by S2, adds sodium citrate, is stirred 20-30 minutes, is added
Enter calcium chloride, stir 40-50 minutes, centrifuged with 2000-2200 revs/min of revolution, obtain microalgae liquid；
S3, prepare the carrier solution containing sodium alginate and magnesium chloride, and the microalgae liquid obtained in S2 is added in carrier solution, mixes
Instilled after closing uniformly with syringe in the calcium chloride solution that mass fraction is 2.6%-3.2%, when 8-10 DEG C of fixed 2-2.5 is small after
Take out, with deionized water desalination 1-2 it is small when.
Preferably, the raw material of culture medium includes according to parts by weight in S1：1-2 parts of glucose, 4-6 parts of starch sugars, 0.01-
0.03 part of zinc sulfate, 0.1-0.3 parts of magnesium sulfate, 0.15-0.3 parts of ammonium sulfate, 0.2-0.4 parts of potassium dihydrogen phosphates, 0.1-0.3 parts of nitre
Sour sodium, 9-12 part sodium chloride and 240-260 parts of deionized waters.
Preferably, in S1 light dark cycles alternate culture, when the time cultivated under light environment is 8-10 small, illumination
Intensity is 3500-3800Lux, and when the time cultivated under dark surrounds is 11-13 small, light environment, dark surrounds circulation are handed over
For progress.
Preferably, the pH value of micro algae culturing liquid is adjusted to 5.1-5.4 in S2.
Preferably, the amount that sodium citrate is added in S2 is the 1.0%-2.5% of micro algae culturing liquid weight.
Preferably, the amount that sodium citrate is added in S2 is the 1.5%-1.7% of micro algae culturing liquid weight.
Preferably, the amount that calcium chloride is added in S2 is the 3%-5% of micro algae culturing liquid weight.
Preferably, the amount that calcium chloride is added in S2 is the 3.8%-4.3% of micro algae culturing liquid weight.
Preferably, the sodium alginate and mass fraction for being 2%-3% containing mass fraction in carrier solution in S3 are 0.6-
0.8% magnesium chloride.
Preferably, the amount of the carrier solution used in S3 is 2.3-2.7 times of microalgae liquid weight.
The preparation method of immobilized microalgae provided by the invention mainly includes culture, harvesting and the microalgae immobilization of chlorella
Three parts.The culture of chlorella uses light dark cycle alternate culture method, and preferred light environment, dark surrounds culture
Time, when light dark cycle alternate culture 60-75 is small after, terminate culture.Invention technician is had found in culture 60-75
Terminate chlorella that culture obtains in hour period after livestock and poultry cultivation sewage is entered, the speed of its growth is better than at other
Between terminate the chlorella that culture obtains in section.The harvesting of chlorella adds sodium citrate, chlorine by adjusting micro algae culturing liquid pH value
Changing the mode of calcium makes chlorella flocculate and then is separated from liquid, and this method is easy to operate, and energy consumption is low, is adapted to industry should
With.The immobilization of chlorella prepares method of the diameter in 4-6 millimeters of microalgae gel ball using alginate carrier, and in seaweed
Magnesium chloride is added in sour sodium carrier solution, there is broken, deformation the effect for preventing microalgae gel ball.
Using immobilized microalgae prepared by the method for the present invention using microalgae in growth course to organic matter in aquaculture wastewater,
The absorption of nutrient ingredients such as nitrogen, phosphorus, synthesize own cells skeleton to purify livestock and poultry cultivation sewage, and can remove removing heavy metals and difficulty
Degradation of organic substances.Microalgae improves absorption and enrichment to materials such as nitrogen, phosphorus and heavy metals after the method for the present invention immobilization,
Have the advantages that reaction speed is fast, resistance to murder by poisoning ability is strong, stable and reliable operation, microalgae cell are lost in less, product is easily separated, can have
Effect prevents processing water secondary pollution.
Embodiment
In the following, technical scheme is described in detail by specific embodiment.
Embodiment 1
The present invention proposes a kind of preparation method of immobilized microalgae, comprises the following steps：
S1, under aseptic technique, by chlorella vulgaris with 5.4 × 106The inoculum concentration of a/mL is inoculated into culture medium,
Temperature is 26 DEG C, under the conditions of rotating speed is 122 revs/min, carries out light dark cycle alternate culture, 9 are cultivated under light environment
Hour, intensity of illumination 3700Lux, when the time cultivated under dark surrounds is 11 small, light environment, dark surrounds circulation are handed over
For culture 68 it is small when after, obtain micro algae culturing liquid；
The pH value of the micro algae culturing liquid obtained in S1, is adjusted to 5.2 by S2, adds 1.58% lemon of micro algae culturing liquid weight
Sour sodium, is stirred 25 minutes, adds 4.1% calcium chloride of micro algae culturing liquid weight, is stirred 44 minutes, with 2100 revs/min turn
Number centrifuges, and obtains microalgae liquid；
The carrier solution for the magnesium chloride that the sodium alginate and mass fraction that S3, preparation are 2.6% containing mass fraction are 0.68%, by S2
In obtained microalgae liquid be added in carrier solution, it is molten to instill the calcium chloride that mass fraction is 2.9% with syringe after mixing
In liquid, 9 DEG C fix 2.3 it is small when after take out, with deionized water desalination 1.2 it is small when, being fixed microalgae gel ball.Wherein,
The amount of carrier solution is 2.6 times of microalgae liquid weight.
The raw material of culture medium includes according to parts by weight in S1：1.6 parts of glucose, 5.2 portions of starch sugars, 0.02 part of zinc sulfate,
0.19 part of magnesium sulfate, 0.23 part of ammonium sulfate, 0.28 part of potassium dihydrogen phosphate, 0.21 part of sodium nitrate, 11 parts of sodium chloride and 250 parts go from
Sub- water.
Embodiment 2
The present invention proposes a kind of preparation method of immobilized microalgae, comprises the following steps：
S1, under aseptic technique, by chlorella vulgaris with 1.6 × 106The inoculum concentration of a/mL is inoculated into culture medium,
Temperature is 25 DEG C, under the conditions of rotating speed is 128 revs/min, carries out light dark cycle alternate culture, 10 are cultivated under light environment
Hour, intensity of illumination 3600Lux, when the time cultivated under dark surrounds is 12 small, light environment, dark surrounds circulation are handed over
For culture 73 it is small when after, obtain micro algae culturing liquid；
The pH value of the micro algae culturing liquid obtained in S1, is adjusted to 5.3 by S2, adds 1.67% lemon of micro algae culturing liquid weight
Sour sodium, is stirred 22 minutes, adds 3.9% calcium chloride of micro algae culturing liquid weight, is stirred 48 minutes, with 2200 revs/min turn
Number centrifuges, and obtains microalgae liquid；
The carrier solution for the magnesium chloride that the sodium alginate and mass fraction that S3, preparation are 2.8% containing mass fraction are 0.62%, by S2
In obtained microalgae liquid be added in carrier solution, it is molten to instill the calcium chloride that mass fraction is 2.7% with syringe after mixing
In liquid, 10 DEG C fix 2.4 it is small when after take out, with deionized water desalination 1.4 it is small when, being fixed microalgae gel ball.Wherein,
The amount of carrier solution is 2.4 times of microalgae liquid weight.
Wherein, the raw material of culture medium includes according to parts by weight in S1：1.9 parts of glucose, 4.3 portions of starch sugars, 0.01 part of sulphur
Sour zinc, 0.12 part of magnesium sulfate, 0.17 part of ammonium sulfate, 0.38 part of potassium dihydrogen phosphate, 0.27 part of sodium nitrate, 10 parts of sodium chloride and 256
Part deionized water.
Embodiment 3
The present invention proposes a kind of preparation method of immobilized microalgae, comprises the following steps：
S1, under aseptic technique, by chlorella vulgaris with 8.9 × 106The inoculum concentration of a/mL is inoculated into culture medium,
Temperature is 27 DEG C, under the conditions of rotating speed is 113 revs/min, carries out light dark cycle alternate culture, 8 are cultivated under light environment
Hour, intensity of illumination 3800Lux, when the time cultivated under dark surrounds is 13 small, light environment, dark surrounds circulation are handed over
For culture 62 it is small when after, obtain micro algae culturing liquid；
The pH value of the micro algae culturing liquid obtained in S1, is adjusted to 5.1 by S2, adds 1.51% lemon of micro algae culturing liquid weight
Sour sodium, is stirred 28 minutes, adds 4.2% calcium chloride of micro algae culturing liquid weight, is stirred 41 minutes, with 2000 revs/min turn
Number centrifuges, and obtains microalgae liquid；
The carrier solution for the magnesium chloride that the sodium alginate and mass fraction that S3, preparation are 2.3% containing mass fraction are 0.77%, by S2
In obtained microalgae liquid be added in carrier solution, it is molten to instill the calcium chloride that mass fraction is 3.1% with syringe after mixing
In liquid, 8 DEG C fix 2.1 it is small when after take out, with deionized water desalination 1.8 it is small when, being fixed microalgae gel ball.Wherein,
The amount of carrier solution is 2.5 times of microalgae liquid weight.
Wherein, the raw material of culture medium includes according to parts by weight in S1：1.2 parts of glucose, 5.8 portions of starch sugars, 0.03 part of sulphur
Sour zinc, 0.27 part of magnesium sulfate, 0.28 part of ammonium sulfate, 0.23 part of potassium dihydrogen phosphate, 0.14 part of sodium nitrate, 9 parts of sodium chloride and 241 parts
Deionized water.
Embodiment 4
The pH value of micro algae culturing liquid can also be adjusted in the preparation method of immobilized microalgae in embodiment 1, wherein S2
4.8。
Embodiment 5
The pH value of micro algae culturing liquid can also be adjusted in the preparation method of immobilized microalgae in embodiment 1, wherein S2
5.9。
Embodiment 6
The pH value of micro algae culturing liquid can also be adjusted in the preparation method of immobilized microalgae in embodiment 1, wherein S2
5.6。
Embodiment 7
The addition of sodium citrate can also be microdisk electrode in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 1.2% of liquid weight.
Embodiment 8
The addition of sodium citrate can also be microdisk electrode in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 2.1% of liquid weight.
Embodiment 9
The addition of calcium chloride can also be micro algae culturing liquid in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 3.3% of weight.
Embodiment 10
The addition of calcium chloride can also be micro algae culturing liquid in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 4.8% of weight.
Embodiment 11
The addition of sodium citrate can also be microdisk electrode in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 2.4% of liquid weight, the addition of calcium chloride can also be the 3.6% of micro algae culturing liquid weight.
Embodiment 12
The addition of sodium citrate can also be microdisk electrode in the preparation method of immobilized microalgae in embodiment 1, wherein S2
The 1.3% of liquid weight, the addition of calcium chloride can also be the 4.6% of micro algae culturing liquid weight.
Chlorella involved in S1 of the present invention is preferably chlorella vulgaris（Chlorella vulgaris）, it is inoculated into culture
Chlorella in base is preferably the chlorella of activated mistake, and activation side well known by persons skilled in the art can be selected in activation method
Method, preferably slant activation method.The adjusting of micro algae culturing liquid pH can preferably be added by the way of hydrochloric acid is added in S2 of the present invention
Mass fraction is the hydrochloric acid of 30%-40%.
The methods of inoculation, stirring, centrifugation involved in the present invention, configuration of culture medium be this area it is known often
Know, details are not described herein.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of immobilized microalgae, it is characterised in that comprise the following steps：
S1, under aseptic technique, by chlorella with 1 × 106A/mL-1 × 107The inoculum concentration of a/mL is inoculated into culture medium
In, it is 25-27 DEG C in temperature, under the conditions of rotating speed is 110-130 revs/min, when light dark cycle alternate culture 60-75 is small,
Obtain micro algae culturing liquid；
The pH value of the micro algae culturing liquid obtained in S1, is adjusted to 4.5-6 by S2, adds sodium citrate, is stirred 20-30 minutes, is added
Enter calcium chloride, stir 40-50 minutes, centrifuged with 2000-2200 revs/min of revolution, obtain microalgae liquid；
S3, prepare the carrier solution containing sodium alginate and magnesium chloride, and the microalgae liquid obtained in S2 is added in carrier solution, mixes
Instilled after closing uniformly with syringe in the calcium chloride solution that mass fraction is 2.6%-3.2%, when 8-10 DEG C of fixed 2-2.5 is small after
Take out, with deionized water desalination 1-2 it is small when.
A kind of 2. preparation method of immobilized microalgae according to claim 1, it is characterised in that culture medium in the S1
Raw material includes according to parts by weight：1-2 parts of glucose, 4-6 portions of starch sugars, 0.01-0.03 parts of zinc sulfate, 0.1-0.3 parts of magnesium sulfate,
0.15-0.3 parts of ammonium sulfate, 0.2-0.4 parts of potassium dihydrogen phosphates, 0.1-0.3 parts of sodium nitrate, 9-12 parts of sodium chloride and 240-260 parts
Deionized water.
3. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that the S1 light darks are followed
In ring alternate culture, when the time cultivated under light environment is 8-10 small, intensity of illumination 3500-3800Lux, in dark ring
When the time cultivated under border is 11-13 small, light environment, dark surrounds cycle alternation carry out.
4. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that train microalgae in the S2
The pH value of nutrient solution is adjusted to 5.1-5.4.
5. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that add lemon in the S2
The amount of sour sodium is the 1.0%-2.5% of micro algae culturing liquid weight.
6. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that add lemon in the S2
The amount of sour sodium is the 1.5%-1.7% of micro algae culturing liquid weight.
7. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that add chlorination in the S2
The amount of calcium is the 3%-5% of micro algae culturing liquid weight.
8. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that add chlorination in the S2
The amount of calcium is the 3.8%-4.3% of micro algae culturing liquid weight.
A kind of 9. preparation method of immobilized microalgae according to claim 1, it is characterised in that carrier solution in the S3
In be 2%-3% containing mass fraction sodium alginate and mass fraction be 0.6-0.8% magnesium chloride.
10. the preparation method of a kind of immobilized microalgae according to claim 1, it is characterised in that used in the S3
The amount of carrier solution is 2.3-2.7 times of microalgae liquid weight.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109046290A (en) * | 2018-09-12 | 2018-12-21 | 潘钕 | The preparation and application of fly ash base resin sorbent |
| CN109225164A (en) * | 2018-11-14 | 2019-01-18 | 浙江海洋大学 | A kind of compound microalgae biological adsorption agent and its preparation method and application |
| CN109395700A (en) * | 2018-11-14 | 2019-03-01 | 浙江海洋大学 | A kind of method of cadmium in compound microalgae biological adsorption agent and its adsorbent absorption waste water |
| CN113999839A (en) * | 2021-12-09 | 2022-02-01 | 河北科技大学 | Immobilization method of microalgae and application thereof |
| CN114134043A (en) * | 2021-12-10 | 2022-03-04 | 陕西关天易景科技有限公司 | Preparation method of environment-friendly sewage treatment agent |
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| CN107140742A (en) * | 2017-06-05 | 2017-09-08 | 舟山市普陀兴海养殖优质种苗选育研究所 | A kind of mixed type algae water cleanser and its application |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109046290A (en) * | 2018-09-12 | 2018-12-21 | 潘钕 | The preparation and application of fly ash base resin sorbent |
| CN109225164A (en) * | 2018-11-14 | 2019-01-18 | 浙江海洋大学 | A kind of compound microalgae biological adsorption agent and its preparation method and application |
| CN109395700A (en) * | 2018-11-14 | 2019-03-01 | 浙江海洋大学 | A kind of method of cadmium in compound microalgae biological adsorption agent and its adsorbent absorption waste water |
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| CN114134043A (en) * | 2021-12-10 | 2022-03-04 | 陕西关天易景科技有限公司 | Preparation method of environment-friendly sewage treatment agent |
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