CN108384720A - A method of it is flocculated using combination flocculant and harvests chlorella pyrenoidosa - Google Patents
A method of it is flocculated using combination flocculant and harvests chlorella pyrenoidosa Download PDFInfo
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- CN108384720A CN108384720A CN201810413415.XA CN201810413415A CN108384720A CN 108384720 A CN108384720 A CN 108384720A CN 201810413415 A CN201810413415 A CN 201810413415A CN 108384720 A CN108384720 A CN 108384720A
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- chlorella
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- chlorella pyrenoidosa
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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Abstract
The present invention provides a kind of method for the harvesting chlorella pyrenoidosa that flocculates using combination flocculant, belongs to technical field of aquaculture.Party normal direction OD680Combination flocculant of hundred a ten thousandth of addition chlorella volume to a ten thousandth in well-grown bead algae solution that light absorption value is 0.9~1.2, adjust pH to 6~10, it is stirred 5~20 minutes with 20~100rpm speed, it is then allowed to stand 0.5~8 hour, so that chlorella flocculation sediment, discharge supernatant is cultivated for chlorella, collects chlorella concentrate, chlorella is harvested by centrifugation, freeze-drying obtains edible or the chlorella powder as feed.This method reduces flocculant usage by adjusting flocculation system pH, a small amount of polymeric aluminum is added into biological flocculant and shortens flocculation time, and flocculating rate is higher than 90%, and the chlorella of harvest is safe and reliable, it can be used for food and animal feed, can be applied to chlorella industrialization harvest.
Description
Technical field
The present invention relates to technical field of aquaculture, particularly relate to a kind of small using combination flocculant flocculation harvesting pyrenoids
The method of ball algae.
Background technology
Chlorella is to grow most fast animals and plants in the world, can increase by 4 times at 20 hours, this contains uniqueness mainly due to it
Growth factor.Rich in protein, a variety of pigments, B family vitamin, essential amino acid, trace element and some bioactivity generations
Thank to product.With anticancer, radioresistance, anti-infective, anti-oxidant, anti-curing hyperlipemia, anti-treat constipation and a systems such as bone marrow suppression
Row physiological hygiene function.Chlorella is good green nourishing source food, important health products, containing the unique CGF factors, praised
For mankind's 21st century most ultimate food and astronaut's special food, it is mainly used in food, health products, feed, medicine etc.
Field.
Chlorella is one of function microalgae of current large-scale farming, and chlorella diameter is about 3~8 μm, thus filters nothing
Method harvests；Centrifugation is to harvest the best method of chlorella, but since chlorella cultivation concentration is very low, the cost of centrifugation is too
It is high.Membrane filter method harvesting cost is too high, is not suitable for industrial applications.Therefore, the cost of chlorella harvesting usually accounts for chlorella
20~30% or so of production cost are a main causes for causing chlorella industrialization difficult.
Flocculence microalgae recovery chlorella has many advantages, such as that simple and easy to do, energy consumption is relatively low, is that most possible realize industrializes
One of method of harvesting.The key that chlorella is harvested using flocculence is it is ensured that the flocculating settling agent used will not to environment
It causes damages, it is safe and non-toxic, while having the effect of good concentration microalgae cell.Inorganic flocculating agent such as ferric sulfate, aluminum sulfate
Deng flocculation rate is high, flocculating effect is good, but the needs such as sulfate radical in water body, aluminium ion can be caused further to locate after flocculating
Reason, while containing above-mentioned flocculant in the frond harvested, it cannot be directly used to food or animal feed.Although biological flocculant is wadded a quilt with cotton
Solidifying rate and flocculating rate do not have that inorganic flocculating agent is good, but the chlorella for harvesting of flocculating can be directly as animal feed or food
Product, thus be most to be hopeful one of industrial applications flocculant.
Invention content
The present invention is to solve the problems, such as that current chlorella pyrenoidosa harvests harvesting of high cost and difficult, provides a kind of utilize and combines
The method of flocculant flocculation harvesting chlorella pyrenoidosa.
It is as follows that the method comprising the steps of：
(1) chlorella is inoculated into the fresh culture that pH is 6~8, illumination cultivation 3~5 days obtains bead algae solution
OD680It is 0.9~1.2；
(2) into the bead algae solution of the middle gained of step (1) according to chlorella pyrenoidosa volume (mL)：Combination flocculant (g)
=10 000~1 000 000:Combination flocculant is added in 1 ratio；
(3) chlorella pyrenoidosa flocculation system pH to 6~10 in regulating step (2)；
(4) the chlorella pyrenoidosa liquid for obtaining step (3) is stirred 5~20 minutes with 20~100rpm speed, then quiet
.5~8 hour are set to 0, chlorella is made to flocculate；
(5) supernatant after chlorella flocculation in step (4) is taken out, is cultivated for chlorella pyrenoidosa；
(6) the concentration algae solution after chlorella flocculation in step (4) is centrifuged 15~30 at 4 DEG C, 6 000~10 000g
Minute, the chlorella pyrenoidosa freeze-drying of harvest obtains chlorella powder.
Wherein, illumination cultivation condition is in step (1)：OD680It is 6~8 for 0.3~0.5, pH, temperature is 25~35 DEG C,
Light regime：Illumination：Dark=8~14:10~16h, 3 000~5 000lx of illumination.
It is biological flocculant in mass ratio in combination flocculant in step (2)：Food grade polymeric aluminium=100~10000：1.
Wherein, biological flocculant is that carboxymethyl chitosan or carboxymethyl chitosan addition chitosan, inulin and carboxymethyl are fine
Tie up one or more of element.
When biological flocculant is that methyl chitosan adds one or more of chitosan, inulin and carboxymethyl cellulose
When, mass ratio of each component is：Carboxymethyl chitosan：Chitosan：Inulin：Carboxymethyl cellulose：Food grade polymeric aluminium=90~95:
0~10:0~10:0~10：0.01~1.
The above-mentioned technical proposal of the present invention has the beneficial effect that：
In said program, using the biological flocculant and minimal amount of food grade polymeric being mainly made of carboxymethyl chitosan
The combination flocculant of aluminium composition, the chlorella pyrenoidosa for harvesting of flocculating are used directly for animal feed and food, securely and reliably；
It minimal amount of food grade polymeric aluminium is added in combination flocculant can significantly improve biological flocculant and settle slow disadvantage, optimization flocculation
System pH can significantly improve the flocculation rate of chlorella, reduce the dosage of flocculant.This method is simple for process, flocculating effect can
It leans on, can be used for chlorella pyrenoidosa industrialization harvesting.
Description of the drawings
Fig. 1 is the method and process flow chart of the harvesting chlorella pyrenoidosa that flocculated using combination flocculant of the present invention.
Specific implementation mode
To keep the technical problem to be solved in the present invention, technical solution and advantage clearer, below in conjunction with attached drawing and tool
Body embodiment is described in detail.
The problem of present invention harvests of high cost and difficult harvesting for existing chlorella pyrenoidosa provides a kind of utilize and combines
The method of flocculant flocculation harvesting chlorella pyrenoidosa.
As shown in Figure 1, this method generally comprises following steps：
(1) chlorella is inoculated into the fresh culture that pH is 6~8, temperature is 25~35 DEG C, light regime：Illumination：
Dark=8~14:10~16h, illumination are 3 000~5 000lx, and illumination cultivation 3~5 days obtains the OD of bead algae solution680For
0.9~1.2；
(2) into the bead algae solution of the middle gained of step (1) according to chlorella pyrenoidosa volume (mL)：Combination flocculant (g)
=10 000~1 000 000:Combination flocculant is added in 1 ratio；
(3) chlorella pyrenoidosa flocculation system pH to 6~10 in regulating step (2)；
(4) the chlorella pyrenoidosa liquid for obtaining step (3) is stirred 5~20 minutes with 20~100rpm speed, then quiet
.5~8 hour are set to 0, chlorella is made to flocculate；
(5) supernatant after chlorella flocculation in step (4) is taken out, is cultivated for chlorella pyrenoidosa；
(6) the concentration algae solution after chlorella flocculation in step (4) is centrifuged 15~30 at 4 DEG C, 6 000~10 000g
Minute, the chlorella pyrenoidosa freeze-drying of harvest obtains chlorella powder.
It is explained with reference to specific embodiment.
Embodiment 1
(1) chlorella pyrenoidosa used in the present embodiment is that Jiangxi Province's both culturing microalgae is preserved with deep processing project technique center
Algae, culture medium prepared with tap water, and chlorella pyrenoidosa culture medium is the culture medium of engineering center's optimization.
(2) chlorella vulgaris is added in the bead algae culture medium that pH is 6, initial OD680It is adjusted to 0.3, is 25 in temperature
DEG C, light regime be illumination：Dark=12h:12h is cultivated 4 days in the illumination box that intensity of illumination is 3 000lx.
(3) by OD680For 1.0 be placed on 500 milliliters in 300 milliliters of chlorella pyrenoidosa liquid for growing logarithmic phase three
In the bottle of angle, 50 milligrams of combination flocculant (carboxymethyl chitosan is added：Inulin：Food grade polymeric aluminium=950:10:1).
(4) it is stirred under the conditions of rotating speed is 50rpm 5 minutes with mechanical agitator, is then allowed to stand 5 hours, keeps pyrenoids small
Ball algae is flocculated, flocculating rate 92%, and upper water column is added nutriment and cultivated for chlorella pyrenoidosa, lower layer's flocculation pyrenoids
For chlorella in 4 DEG C, centrifugal force to be centrifuged 15 minutes under the conditions of 8 000g, chlorella pyrenoidosa is placed on refrigerator freezing, then freezes
It is dry to obtain chlorella freeze-dried powder.
Embodiment 2
(1) chlorella pyrenoidosa used in the present embodiment is that Jiangxi Province's both culturing microalgae is preserved with deep processing project technique center
Algae, culture medium prepared with tap water, and chlorella pyrenoidosa culture medium is the culture medium of engineering center's optimization.
(2) chlorella vulgaris is added in the chlorella pyrenoidosa culture medium that pH is 7, initial OD680It is adjusted to 0.4, in temperature
Degree is 30 DEG C, light regime is illumination：Dark=16h:8h is cultivated 4 days in the illumination box that intensity of illumination is 4000lx.
(3) by OD680For 1.1 be placed on 500 milliliters in 300 milliliters of chlorella pyrenoidosa liquid for growing logarithmic phase three
In the bottle of angle, 50 milligrams of combination flocculant (carboxymethyl chitosan is added：Inulin：Chitosan：Food grade polymeric aluminium=950:20：
50：5).
(4) it is in rotating speed with mechanical agitator:It is stirred 10 minutes under the conditions of 20rpm, is then allowed to stand 6 hours, makes pyrenoids
Chlorella is flocculated, flocculating rate 95%, and upper water column is added nutriment and cultivated for chlorella pyrenoidosa, lower layer's flocculation albumen
For core chlorella in 4 DEG C, centrifugal force to be centrifuged 15 minutes under the conditions of 8 000g, chlorella pyrenoidosa is placed on refrigerator freezing, then cold
Dry acquisition chlorella pyrenoidosa freeze-dried powder is lyophilized.
Embodiment 3
(1) chlorella pyrenoidosa used in the present embodiment is that Jiangxi Province's both culturing microalgae is preserved with deep processing project technique center
Algae, culture medium prepared with tap water, and chlorella pyrenoidosa culture medium is the culture medium of engineering center's optimization.
(2) chlorella vulgaris is added in the chlorella pyrenoidosa culture medium that pH is 8, initial OD680It is adjusted to 0.35,
Temperature is 30 DEG C, light regime is illumination：Dark=12h:12h is cultivated in the illumination box that intensity of illumination is 5 000lx
3 days.
(3) by OD680For 0.9 be placed on 500 milliliters in 300 milliliters of chlorella pyrenoidosa liquid for growing logarithmic phase three
In the bottle of angle, 50 milligrams of combination flocculant (carboxymethyl chitosan is added：Chitosan：Food grade polymeric aluminium=900：100：0.1).
(4) it is in rotating speed with mechanical agitator:It is stirred 5 minutes under the conditions of 40rpm, is then allowed to stand 8 hours, keeps pyrenoids small
Ball algae is flocculated, flocculating rate 98%, and upper water column is added nutriment and cultivated for chlorella pyrenoidosa, lower layer's flocculation pyrenoids
For chlorella in 4 DEG C, centrifugal force to be centrifuged 15 minutes under the conditions of 8 000g, chlorella pyrenoidosa is placed on refrigerator freezing, then freezes
It is dry to obtain chlorella pyrenoidosa freeze-dried powder.
The above is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, without departing from the principles of the present invention, several improvements and modifications can also be made, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (5)
1. a kind of method for the harvesting chlorella pyrenoidosa that flocculated using combination flocculant, it is characterised in that：Including steps are as follows：
(1) chlorella is inoculated into the fresh culture that pH is 6~8, illumination cultivation 3~5 days obtains the OD of bead algae solution680
It is 0.9~1.2；
(2) into the bead algae solution of the middle gained of step (1) according to chlorella pyrenoidosa volume (mL)：Combination flocculant (g)=10
000~1 000 000:Combination flocculant is added in 1 ratio；
(3) chlorella pyrenoidosa flocculation system pH to 6~10 in regulating step (2)；
(4) the chlorella pyrenoidosa liquid for obtaining step (3) is stirred 5~20 minutes with 20~100rpm speed, is then allowed to stand 0.5
~8 hours, chlorella is made to flocculate；
(5) supernatant after chlorella flocculation in step (4) is taken out, is cultivated for chlorella pyrenoidosa；
(6) the concentration algae solution after chlorella flocculation in step (4) is centrifuged 15~30 minutes at 4 DEG C, 6 000~10 000g,
The chlorella pyrenoidosa freeze-drying of harvest obtains chlorella powder.
2. the method for the harvesting chlorella pyrenoidosa according to claim 1 that flocculated using combination flocculant, it is characterised in that：
Illumination cultivation condition is in the step (1)：OD680It is 6~8 for 0.3~0.5, pH, temperature is 25~35 DEG C, light regime：
Illumination：Dark=8~14:10~16h, 3 000~5 000lx of illumination.
3. the method for the harvesting chlorella pyrenoidosa according to claim 1 that flocculated using combination flocculant, it is characterised in that：
It is biological flocculant in mass ratio in combination flocculant in the step (2)：Food grade polymeric aluminium=100~10 000：1.
4. the method for the harvesting chlorella pyrenoidosa according to claim 3 that flocculated using combination flocculant, it is characterised in that：
The biological flocculant is that carboxymethyl chitosan or carboxymethyl chitosan add in chitosan, inulin and carboxymethyl cellulose
It is one or more of.
5. the method for the harvesting chlorella pyrenoidosa according to claim 4 that flocculated using combination flocculant, it is characterised in that：
In the biological flocculant, mass ratio of each component is：Carboxymethyl chitosan：Chitosan：Inulin：Carboxymethyl cellulose：Food-grade
Polymeric aluminum=90~95:0~10:0~10:0~10：0.01~1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109430651A (en) * | 2018-11-26 | 2019-03-08 | 江苏科技大学 | A kind of chlorella health beverages and preparation method thereof |
| CN113736829A (en) * | 2021-07-22 | 2021-12-03 | 同济大学 | Simple photobiological hydrogen production system and preparation method and application thereof |
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| CN113736829A (en) * | 2021-07-22 | 2021-12-03 | 同济大学 | Simple photobiological hydrogen production system and preparation method and application thereof |
| CN113736829B (en) * | 2021-07-22 | 2023-07-14 | 同济大学 | Simple photobiological hydrogen production system and preparation method and application thereof |
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Application publication date: 20180810 |