CN114395481A - Method for harvesting microalgae by using fungal floating biological mat - Google Patents
Method for harvesting microalgae by using fungal floating biological mat Download PDFInfo
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- 238000012258 culturing Methods 0.000 claims description 11
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- 241000195493 Cryptophyta Species 0.000 claims description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 235000015278 beef Nutrition 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 5
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- 241000195663 Scenedesmus Species 0.000 claims description 5
- 239000001110 calcium chloride Substances 0.000 claims description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 5
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 5
- 239000000284 extract Substances 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
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- 239000011684 sodium molybdate Substances 0.000 claims description 5
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 5
- 229910021654 trace metal Inorganic materials 0.000 claims description 5
- 241000195634 Dunaliella Species 0.000 claims description 3
- 241000196321 Tetraselmis Species 0.000 claims description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 241000195585 Chlamydomonas Species 0.000 claims description 2
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 2
- 241000233671 Schizochytrium Species 0.000 claims description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 2
- 229910052603 melanterite Inorganic materials 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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Abstract
The invention discloses a method for harvesting microalgae by using a fungus floating biological mat, which utilizes mucor racemosus (mucor racemosus) to adsorb the microalgae so as to achieve the aim of harvesting the microalgae. According to the method, the mucor racemosus can adsorb a large amount of microalgae biomass, and the mucor racemosa and the microalgae can float on the surface of the culture solution together, so that the microalgae can be harvested by using the mucor racemosa floating biological mat, and the harvesting method is efficient and convenient. In addition, in the process of co-growing the mucor racemosa and the microalgae, the vitality of the microalgae can be enhanced, and the growth and the propagation of the microalgae can be promoted, so that the harvesting efficiency of the microalgae can be improved, and the growth of the microalgae can be enhanced.
Description
Technical Field
The invention relates to the technical field of microalgae harvesting, in particular to a method for harvesting microalgae by using a fungal floating biological mat.
Background
Microalgae is a autotrophic microorganism with high photosynthetic efficiency and can produce various metabolites, for example, some kinds of microalgae can accumulate a large amount of grease and fatty acid in vivo in the growth and propagation processes. Microalgae are used in many fields, such as food, medicine, environmental remediation and energy, and are therefore considered to be one of the most promising microorganisms. However, in the harvesting of microalgae, methods such as precipitation, centrifugation, filtration, flocculation, etc. are mainly adopted, wherein the precipitation and filtration take a lot of time, the harvesting efficiency is not high, the centrifugation requires a lot of power resources, the flocculation is the most applied method at present, but it is not easy to find a high-efficiency flocculant for different microalgae and operation conditions. Therefore, it is an urgent problem to find a simple and efficient method for harvesting microalgae.
Disclosure of Invention
In order to solve the above technical problems, the present invention aims to provide a method for harvesting microalgae using a fungal floating bio-mat, so as to solve the problems of complexity and low efficiency of the existing microalgae harvesting methods.
The technical scheme for solving the technical problems is as follows: provided is a method for harvesting microalgae by using a fungal floating bio-mat, wherein the microalgae is harvested by adsorbing the microalgae by using mucor racemosus (mucor racemosus).
The invention has the beneficial effects that: according to the method, the mucor racemosus can adsorb a large amount of microalgae biomass, and the mucor racemosa and the microalgae can float on the surface of the culture solution together, so that the microalgae can be harvested by using the mucor racemosa floating biological mat, and the harvesting method is efficient and convenient. In addition, in the process of co-growing the mucor racemosa and the microalgae, the vitality of the microalgae can be enhanced, and the growth and the propagation of the microalgae can be promoted, so that the method for harvesting the microalgae by using the fungal floating biological mat can enhance the growth and the harvesting efficiency of the microalgae.
On the basis of the technical scheme, the invention can be further improved as follows
Further, the harvesting of microalgae by adsorbing microalgae with mucor racemosus (mucor racemosus) comprises the following steps:
(1) culturing Mucor racemosus in culture solution at room temperature for 5-10 days;
(2) and (2) adding a culture solution containing microalgae into a microalgae-fungi incubator, inoculating the gross mucor cultured in the step (1) into the culture solution, culturing at room temperature, and collecting the microalgae by adsorbing the microalgae by the gross mucor in 8-12 days.
Further, the microalgae is Chlorella (Chlorella Sp.), Cylindrotheca (Cylindrotheca Sp.), Diatom (Diatom), rhombohedral (Nitzschia Sp.), Schizochytrium (Schizochytrium Sp.), Dunaliella (Dunaliella), Scenedesmus (Scenedesmus Sp.), nannochlorococcus (nannochlorooris Sp.), Chlamydomonas Sp, Tetraselmis (Tetraselmis Sp.), or eudomyces (Eudorina Sp.).
Further, the culture solution in the step (1) is a YPD liquid medium.
Further, the YPD liquid culture medium comprises glucose, beef peptone and yeast extract, wherein the concentration of the glucose in the YPD liquid culture medium is 18-22g/L, the concentration of the beef peptone is 8-12g/L, and the concentration of the yeast extract is 4-6 g/L.
Further, the concentration of glucose in the YPD liquid medium was 20g/L, the concentration of beef peptone was 10g/L, and the concentration of yeast extract was 5 g/L.
Further, the specific method for harvesting microalgae in the step (2) comprises the following steps: and (3) supplying culture solution from the feed inlet of the microalgae-fungi incubator and simultaneously harvesting microalgae from the discharge outlet of the microalgae-fungi incubator.
The beneficial effect of adopting the further technical scheme is as follows: the gross mucor and the microalgae can float on the surface of the culture solution together after being adsorbed, so that the gross mucor adsorbing the microalgae can naturally overflow from a discharge hole of the microalgae-fungi incubator while the culture solution is supplied from a feed hole of the microalgae-fungi incubator, and the 'simultaneous supply and harvest' is realized, namely the continuous harvest of the microalgae is realized.
Further, the culture solution in step (2) is BBM liquid culture medium.
Further, BBM liquid medium comprises NaNO3、K2HPO4、KH2PO4、MgSO4·7H2O、CaCl2·2H2O、NaCl、KOH、EDTA、FeSO4·7H2O, Trace metal solution (mother liquor Sigma-Aldrich reagent from Merck), Na2MoO4And H3BO3(ii) a NaNO in BBM liquid Medium3The concentration of (a) is 230-260mg/L, K2HPO4The concentration of the extract is 70-80mg/L, KH2PO4The concentration of (b) is 160-185mg/L, MgSO4·7H2The concentration of O is 70-80mg/L, CaCl2·2H2The concentration of O is 20-30mg/L, the concentration of NaCl is 20-30mg/L, the concentration of KOH is 25-35mg/L, the concentration of EDTA is 40-60mg/L, and FeSO4·7H2The concentration of O is 4.8-5.2mg/L, the concentration of Trace metal solution (mother liquor purchased from Sigma-Aldrich reagent of Merck) is 20-30mg/L, and Na2MoO4The concentration of (A) is 10-100mg/L, H3BO3The concentration of (A) is 10-12 mg/L.
Further, in BBM liquid medium, NaNO3Has a concentration of 250mg/L, K2HPO4The concentration of (A) is 75mg/L, KH2PO4Has a concentration of 175mg/L, MgSO4·7H2The concentration of O is 75mg/L, CaCl2·2H2The concentration of O is 25mg/L, the concentration of NaCl is 25mg/L, the concentration of KOH is 31mg/L, the concentration of EDTA is 50mg/L, FeSO4·7H2The concentration of O is 4.98mg/L, the concentration of Trace metal solution (mother liquor purchased from Sigma-Aldrich reagent of Merck) is 24.45mg/L, Na2MoO4Has a concentration of 50mg/L, H3BO3The concentration of (B) was 11.42 mg/L.
Further, the inoculation density of the mucor racemosus in the step (2) is 1-4 multiplied by 106OTU/mL。
Further, the volume ratio of the microalgae to the culture solution in the step (2) is 1: 50-70.
Further, the volume ratio of the microalgae to the culture solution is 1: 60.
the invention has the following beneficial effects: the use of the mucor racemosus can enhance the growth and harvesting efficiency of the microalgae, and the microalgae can be harvested while the culture solution is supplied, so that the growth force of the microalgae is improved, the harvesting mode of the microalgae is simplified, the continuous harvesting of the microalgae is realized, and the harvested microalgae can be used for energy conversion, such as extraction of lipid, preparation of biodiesel and the like.
Drawings
FIG. 1 is a microalgae-fungi incubator used in the present invention;
FIG. 2 shows the light absorbencies (OD) of the microalgae of example 1 and comparative example 1680)；
FIG. 3 is a diagram of microalgae harvested in example 1 and comparative example 1;
wherein: 1. an air pump; 2. an air pump controller; 3. an aeration disc; 4. a barrel; 5. lighting; 6. a discharge port; 7. and (4) feeding a material inlet.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The device adopted by the invention is a microalgae-fungi incubator, and comprises a cylinder body 4 and a base arranged at the bottom of the cylinder body, as shown in figure 1; a feed inlet 7 is formed in the top end of the cylinder body 4, a discharge outlet 6 is formed in the side wall of the cylinder body, the discharge outlet 6 is close to the feed inlet 7, a culture solution required by microalgae growth can be added from the feed inlet 7, and meanwhile microalgae are automatically harvested from the discharge outlet 6; the outer wall of the cylinder body is provided with light 4 which can provide illumination conditions for the growth of microalgae; be provided with air pump 1 in the base, air pump controller 2 sets up in the base surface, air pump controller 2 and 1 electric connection of air pump, and accessible air pump controller 2 controls the switch of air pump 1 and the flow that the air pump carried oxygen, and the bottom of barrel 4 is provided with aeration dish 3, and aeration dish 3 passes through the oxygen conveyer pipe and is connected with air pump 1, and aeration dish 3 can play the effect of importing barrel 4 with the oxygen dispersion.
Example 1:
a method of harvesting microalgae using a fungal floaters mat, comprising the steps of:
(1) culturing Mucor racemosus in YPD liquid culture medium at room temperature for 8 days; wherein the concentration of glucose in YPD liquid culture medium is 20g/L, the concentration of beef extract peptone is 10g/L, and the concentration of yeast extract is 5 g/L;
(2) adding 3L BBM liquid culture medium (formula of BBM liquid culture medium is shown in Table 1) into a microalgae-fungi culture apparatus, adding 50mL rhombohedral algae, inoculating Mucor racemosus cultured in step (1) into the microalgae-fungi culture apparatus, wherein the inoculation density is 2 × 106OTU/mL (the weight of inoculated total mucor is 8.62g), culturing at room temperature, and harvesting the rhombohedral algae by using the total mucor floating biological mat on the 10 th day, wherein the specific method for harvesting the rhombohedral algae comprises the following steps: and (3) adding the culture solution prepared in the step (2) from a feed inlet 7, and automatically harvesting the rhombohedral algae from a discharge outlet 6.
Example 2:
a method of harvesting microalgae using a fungal floaters mat, comprising the steps of:
(1) culturing Mucor racemosus in YPD liquid culture medium at room temperature for 5 days; wherein the concentration of glucose in YPD liquid culture medium is 22g/L, the concentration of beef extract peptone is 12g/L, and the concentration of yeast extract is 6 g/L;
(2) adding 3LBBM liquid culture medium (formula of BBM liquid culture medium is shown in Table 1) into a microalgae-fungi culture apparatus, adding 60mL of diatom, inoculating the total mucor cultured in step (2) into the microalgae-fungi culture apparatus at an inoculation density of 1 × 106OTU/mL, culturing at room temperature, and harvesting diatom by using a gross mucor planktonic organism pad on the 10 th day, wherein the specific method for harvesting diatom comprises the following steps: and (3) adding the culture solution prepared in the step (3) from a feed port 7, and automatically harvesting diatom from a discharge port 6.
Example 3:
a method of harvesting microalgae using a fungal floaters mat, comprising the steps of:
(1) culturing Mucor racemosus in YPD liquid culture medium at room temperature for 10 days; wherein the concentration of glucose in YPD liquid culture medium is 18g/L, the concentration of beef extract peptone is 8g/L, and the concentration of yeast extract is 4 g/L;
(2) adding 3L BBM liquid culture medium (formula of BBM liquid culture medium is shown in Table 1) into the microalgae-fungi culture apparatus, adding 43mL scenedesmus, inoculating the Mucor racemosus cultured in step (2) into the microalgae-fungi culture apparatus at an inoculation density of 4 × 106OTU/mL, culturing at room temperature, and harvesting Scenedesmus obliquus by using a gross mucor buoyant organism pad on the 10 th day, wherein the specific method for harvesting Scenedesmus obliquus comprises the following steps: and (4) adding the culture solution prepared in the step (3) from a feed inlet 7, and automatically harvesting scenedesmus from a discharge outlet 6.
TABLE 1 BBM liquid Medium component concentrations
Comparative example 1
A method of harvesting microalgae comprising the steps of:
adding 3L BBM liquid medium (the formula of the BBM liquid medium is shown in Table 1) into a microalgae-fungi incubator, adding 50mL rhombohedral algae, and culturing at room temperature, wherein the rhombohedral algae are harvested on the 10 th day, and the specific method for harvesting the rhombohedral algae comprises the following steps: the rhombohedral algae were harvested by centrifugation and filtration.
Test examples
Examples 1-3 the harvesting of microalgae was essentially the same, and therefore example 1 was chosen for the experiments. During the culture of the microalgae of example 1 and comparative example 1, the light absorbance (OD) of the microalgae was measured daily680) The results are shown in FIG. 2, and it can be seen from FIG. 2 that the growth of microalgae can be promoted by using the method for harvesting microalgae using fungal plankton mat according to the present invention; the conditions of the microalgae harvested in example 1 and comparative example 1 are shown in FIG. 3, and it can be seen from FIG. 3 that Mucor racemosusAdsorbing microalgae and floating on the surface of the microalgae-fungi incubator; the microalgae harvested in the example 1 and the microalgae harvested in the comparative example 1 are subjected to dry weight weighing, because the weight of the mucor racemosus inoculated at the beginning in the example 1 is 8.62g, no organic substances required for growth of the mucor racemosa exist in a BBM liquid culture medium, and the weight before and after the mucor racemosa is cultured is kept unchanged, the difference value between the total weight of the mucor racemosa adsorbing the microalgae and the weight of the inoculated mucor racemosa is the weight of the microalgae harvested finally, the result is shown in table 2, and the table 2 shows that the method provided by the invention utilizes the mucor racemosa and the microalgae to be cultured together, and the microalgae are harvested finally through a mucor racemosa floating biological mat, so that the growth of the microalgae can be promoted, and the harvesting efficiency is improved.
Table 2 microalgae dry weights harvested in example 1 and comparative example 1
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A method for harvesting microalgae by using a fungal floating biological mat is characterized in that the microalgae is harvested by adsorbing the microalgae by using mucor racemosus.
2. The method of harvesting microalgae using a fungal bioraft as claimed in claim 1, comprising the steps of:
(1) culturing Mucor racemosus in culture solution at room temperature for 5-10 days;
(2) and (2) adding a culture solution containing microalgae into a microalgae-fungi incubator, inoculating the gross mucor cultured in the step (1) into the culture solution, culturing at room temperature, and collecting the microalgae by adsorbing the microalgae by the gross mucor in 8-12 days.
3. The method of harvesting microalgae using a fungal floating bio-mat according to claim 1 or 2, wherein the microalgae is of the genus chlorella, cylindracea, diatom, rhombohedral algae, schizochytrium, dunaliella, scenedesmus, nannochloropsis, chlamydomonas, tetraselmis, or glomus.
4. The method for harvesting microalgae using fungal plankton mats according to claim 2, wherein the culture fluid in step (1) is a YPD liquid medium.
5. The method of claim 4, wherein the YPD liquid medium comprises glucose, beef peptone and yeast extract, and the concentration of glucose, beef peptone and yeast extract in the YPD liquid medium is 18-22g/L, 8-12g/L and 4-6 g/L.
6. The method for harvesting microalgae using fungal plankton mat according to claim 2, wherein the specific method for harvesting microalgae in step (2) is: and (3) supplying culture solution from the feed inlet of the microalgae-fungi incubator and simultaneously harvesting microalgae from the discharge outlet of the microalgae-fungi incubator.
7. The method for harvesting microalgae using fungal plankton mat according to claim 2 or 6, wherein the culture fluid in step (2) is BBM liquid culture medium.
8. The method of claim 7, wherein the BBM liquid medium comprises NaNO3、K2HPO4、KH2PO4、MgSO4·7H2O、CaCl2·2H2O、NaCl、KOH、EDTA、FeSO4·7H2O、Trace metal solution、Na2MoO4And H3BO3(ii) a NaNO in BBM liquid Medium3The concentration of (a) is 230-260mg/L, K2HPO4The concentration of the extract is 70-80mg/L, KH2PO4The concentration of (b) is 160-185mg/L, MgSO4·7H2The concentration of O is 70-80mg/L, CaCl2·2H2The concentration of O is 20-30mg/L, the concentration of NaCl is 20-30mg/L, the concentration of KOH is 25-35mg/L, the concentration of EDTA is 40-60mg/L, and FeSO4·7H2The concentration of O is 4.8-5.2mg/L, the concentration of Trace metal solution is 20-30mg/L, Na2MoO4The concentration of (A) is 10-100mg/L, H3BO3The concentration of (A) is 10-12 mg/L.
9. The method for harvesting microalgae using fungal bioraft as claimed in claim 2, wherein the inoculation density of Mucor racemosus in step (2) is 1-4 x 106OTU/mL。
10. The method for harvesting microalgae using fungal plankton mat of claim 2, wherein the volume ratio of microalgae to culture fluid in step (2) is 1: 50-70.
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