CN107586720A - Microalgae culture method - Google Patents
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Abstract
The invention provides a kind of microalgae culture method, this method includes：Configuration step, the nutrient solution of micro algae growth is configured, contains bicarbonate ion in nutrient solution；Step is cultivated, algae kind is accessed into nutrient solution, adjusts the pH value of nutrient solution to default pH value, and adjusts the bicarbonate ion concentration in nutrient solution and reaches the first preset concentration, carries out frustule cultivation, the first preset concentration is 10 30mmol/L；Grease production step, when frustule yield declines, adjust the bicarbonate ion concentration in nutrient solution and reach the second preset concentration, carry out grease production, the second preset concentration is 0 20mmol/L；Step is harvested, when grease yield declines, harvests the frustule of oil-containing.The present invention can ensure the carbon substrate needed for frustule growth, so that frustule quickly increases, improve the yield of microalgae biomass, and frustule is caused to accumulate more greases under the carbon substrate effect of abundance, the content and yield of microalgae grease are improved, and makes full use of the carbon dioxide in air.
Description
Technical field
The present invention relates to microalgae technical field, in particular to a kind of microalgae culture method.
Background technology
Microalgae is a kind of photoautotroph being grown in water, can absorb CO under luminous energy driving2, convert light energy into
The chemical substance stored into the cell, while discharge oxygen.The cell metabolism of microalgae produces the production such as polysaccharide, protein, grease, pigment
Thing, these metabolites are respectively provided with good DEVELOPMENT PROSPECT in fields such as food, medicine, liquid fuels.
With the fast development of global economy, the demand of the energy is increasingly increased so that main energy sources source, as oil,
The conventional fossil fuels such as coal, are increasingly reduced.Also, conventional fossil fuel burning can discharge substantial amounts of CO2, cause global warming
And seawater acidifying, huge environmental problem is caused, therefore, the bioenergy affine to environment turns into development in recent decades
Focus.Conventional bioenergy technology, such as prepares bio-ethanol and biodiesel, in recent years using corn and soybean, sugarcane
Certain achievement is obtained, but because occupying cultivated land so that scale application is restricted.And microalgae bio-fuel because be not take up arable land,
Productivity per unit is high, can fix the reasons such as industrial waste gas becomes the focus that various countries fall over each other research and development.
At present, the industrialization of microalgae biodiesel is prepared also immature, and maximum challenge is both culturing microalgae, harvests, carries
The high Cost Problems of oil.How microalgae biomass yield, fat content and yield are improved, and reduce production cost to turn into solution
The key of this problem.Prior art is supported more using nutrient defective form induction microalgae cell oil-producing in scale
In growing, this mode is still difficult to stablize the high oil-containing algae powder of output, can not still effectively improve content and the production of microalgae grease
Amount.
The content of the invention
In consideration of it, the present invention proposes a kind of microalgae culture method, it is intended to which microalgae can not be effectively improved by solving prior art
The problem of fat content.
The present invention proposes a kind of microalgae culture method, and this method comprises the following steps：Configuration step, configure micro algae growth
Nutrient solution, contain bicarbonate ion in nutrient solution；Step is cultivated, algae kind is accessed into nutrient solution, adjusts the pH of nutrient solution
Value adjusts the bicarbonate ion concentration in nutrient solution and reaches the first preset concentration to default pH value, carries out frustule cultivation,
Wherein, the first preset concentration is 10-30mmol/L；Grease production step, when frustule yield declines, adjust in nutrient solution
Bicarbonate ion concentration reaches the second preset concentration, carries out grease production, wherein, the second preset concentration is 0-20mmol/L；
Step is harvested, when grease yield declines, harvests the frustule of oil-containing.
Further, in above-mentioned microalgae culture method, in configuration step, added into nutrient solution and generation carbon is decomposed in water
The inorganic salts of sour hydrogen radical ion, so as to contain bicarbonate ion in nutrient solution.
Further, in above-mentioned microalgae culture method, cultivate in step, it is 7~9 to preset pH value.
Further, in above-mentioned microalgae culture method, by into nutrient solution add acid or carbon dioxide so that
The pH value of nutrient solution reaches default pH value.
Further, in above-mentioned microalgae culture method, cultivate in step, when the bicarbonate ion concentration in nutrient solution is small
When the first preset concentration, the inorganic salts or carbon dioxide that generation bicarbonate ion is decomposed in water are replenished into nutrient solution
Gas.
Further, in above-mentioned microalgae culture method, in grease production step, the second preset concentration is 0-15mmol/L.
Further, in above-mentioned microalgae culture method, the first preset concentration is more than or equal to the second preset concentration.
Further, in above-mentioned microalgae culture method, grease production step includes：When frustule yield declines, by algae
Cell is seeded in the nutrient solution of auxotrophy, and adjusts the bicarbonate ion concentration in the nutrient solution of auxotrophy to second
Preset concentration, carry out grease production.
Further, in above-mentioned microalgae culture method, the nutrient solution of auxotrophy be lack one kind in nitrogen phosphate and sulfur or
Several nutrient solutions.
In the present invention, adjusted by accessing algae kind into the nutrient solution containing bicarbonate ion, and by the pH value of nutrient solution
Section is to default pH value, and the bicarbonate ion concentration in nutrient solution is adjusted to the first preset concentration and supported with carrying out frustule
Grow, ensure that the carbon substrate needed for frustule growth so that frustule quickly increases, and is effectively improved microalgae biomass
Yield, when frustule yield declines, the bicarbonate ion concentration in nutrient solution is adjusted to the second preset concentration to carry out
Grease production, so that frustule can accumulate more greases under the carbon substrate effect of abundance, it is effectively improved
The content and yield of microalgae grease, solve the problems, such as that prior art can not effectively improve microalgae grease content；In addition, this method
The carbon dioxide in air can be better profited from, that has not only saved carbon dioxide is passed through cost, additionally it is possible to reduces greenhouse
Effect.
Brief description of the drawings
By reading the detailed description of hereafter preferred embodiment, it is various other the advantages of and benefit it is common for this area
Technical staff will be clear understanding.Accompanying drawing is only used for showing the purpose of preferred embodiment, and is not considered as to the present invention
Limitation.And in whole accompanying drawing, identical part is denoted by the same reference numerals.In the accompanying drawings：
Fig. 1 is the structured flowchart of microalgae culture method provided in an embodiment of the present invention；
Fig. 2 is the experimental result picture of the embodiment one of microalgae culture method provided in an embodiment of the present invention；
Fig. 3 is the experimental result picture of the embodiment two of microalgae culture method provided in an embodiment of the present invention；
Fig. 4 is the experimental result picture of the embodiment three of microalgae culture method provided in an embodiment of the present invention；
Fig. 5 is the experimental result picture of the example IV of microalgae culture method provided in an embodiment of the present invention.
Embodiment
The exemplary embodiment of the disclosure is more fully described below with reference to accompanying drawings.Although the disclosure is shown in accompanying drawing
Exemplary embodiment, it being understood, however, that may be realized in various forms the disclosure without should be by embodiments set forth here
Limited.On the contrary, these embodiments are provided to facilitate a more thoroughly understanding of the present invention, and can be by the scope of the present disclosure
Completely it is communicated to those skilled in the art.It should be noted that in the case where not conflicting, embodiment in the present invention and
Feature in embodiment can be mutually combined.Describe the present invention in detail below with reference to the accompanying drawings and in conjunction with the embodiments.
Referring to Fig. 1, it is the structured flowchart of microalgae culture method provided in an embodiment of the present invention.As illustrated, microalgae is trained
Foster method may include steps of：
Configuration step S1, the nutrient solution of micro algae growth is configured, contains bicarbonate ion in nutrient solution.
Specifically, the existing nutrient solution for both culturing microalgae can be used as initial incubation liquid, to initial incubation liquid
It is middle to add the inorganic salts that bicarbonate ion is resolved into water, so as to contain bicarbonate ion in nutrient solution, so as to
It is configured to the nutrient solution of micro algae growth.Wherein, the inorganic salts that bicarbonate ion can be resolved into water can be bicarbonate
Salt or carbonate.When it is implemented, the addition of the inorganic salts can determine according to actual conditions, the present embodiment to this not
Impose any restrictions.
Step S2 is cultivated, algae kind is accessed into nutrient solution, adjusts the pH value of nutrient solution to default pH value, and adjust nutrient solution
In bicarbonate ion concentration reach the first preset concentration, carry out frustule cultivation, wherein, the first preset concentration is 10-
30mmol/L。
Specifically, algae kind is accessed in the nutrient solution configured into configuration step S1, monitors the pH value of nutrient solution, and will training
The pH value of nutrient solution is adjusted to default pH value, and the default pH value should be suitable for micro algae growth.Monitor nutrient solution in bicarbonate radical from
Sub- concentration, and the bicarbonate ion concentration in nutrient solution is adjusted to the first preset concentration, the first preset concentration and enabled to
Frustule carries out proper splitting, and can preferably absorb the bicarbonate ion in nutrient solution, so that microalgae preferably grows.
When the pH value in nutrient solution is default pH value, also, the bicarbonate ion concentration in nutrient solution is the first preset concentration, algae
Cell is grown in the nutrient solution regulated.
Preferably, it is 7~9 to preset pH value so that alkalescent is presented in nutrient solution, so, bicarbonate radical in nutrient solution from
Son or carbanion can more preferably, fix carbon dioxide more quickly, and then for frustule growth provide it is sufficient
It carbon substrate, can also reduce environmental pollution, keep the stability of nutrient solution.When adjusting the pH value of nutrient solution, can pass through
Acid or carbon dioxide are added into nutrient solution to adjust the pH value of nutrient solution to default pH value, i.e., by the pH value of nutrient solution
Adjust to 7~9.When the pH value of nutrient solution grows with frustule and raise, also, two are passed through in order to adjust the pH value of nutrient solution
During carbonoxide, carbon dioxide can regenerate bicarbonate radical, so not only cause the pH value of nutrient solution to keep default pH value, and
The bicarbonate radical of consumption is absorbed supplemented with frustule.
In order to promote the production of microalgae biomass, the first preset concentration is preferably 10-30mmol/L.For example, this is first pre-
If concentration is 15mmol/L, 20mmol/L, 25mmol/L.When the bicarbonate ion concentration in nutrient solution is default less than first dense
When spending, the inorganic salts or carbon dioxide that generation bicarbonate ion is decomposed in water are replenished into nutrient solution.Preferably,
The inorganic salts are bicarbonate.
When it is implemented, after algae kind is accessed, can be according to the bicarbonate ion concentration in nutrient solution, into nutrient solution
Bicarbonate or carbon dioxide are supplemented, so that the bicarbonate ion concentration in nutrient solution reaches the first preset concentration.
In frustule production process, the bicarbonate ion concentration in nutrient solution is monitored in real time, when bicarbonate ion concentration is less than
During the first preset concentration, bicarbonate or carbon dioxide are supplemented into nutrient solution, so that the bicarbonate radical in nutrient solution
Ion concentration maintains the first preset concentration.Also, when supplementing bicarbonate or carbon dioxide into nutrient solution, need
The pH value in nutrient solution is monitored in real time, by adding acid or carbon dioxide into nutrient solution so that the pH value of nutrient solution
Maintain between 7~9.
When it is implemented, it is necessary to monitor the pH value and bicarbonate radical in nutrient solution in real time during frustule grows
Ion concentration, and when pH value does not meet default pH value or do not meet the first preset concentration in bicarbonate ion concentration,
PH value and bicarbonate ion concentration is constantly adjusted according to actual conditions, the final pH value for causing nutrient solution meets default pH
It is worth and bicarbonate ion concentration meets the first preset concentration, so that frustule is preferably produced.
Grease production step S3, when frustule yield declines, the bicarbonate ion concentration adjusted in nutrient solution reaches
Second preset concentration, grease production is carried out, wherein, the second preset concentration is 0-20mmol/L.
Specifically, the yield of frustule in nutrient solution is monitored in real time, when frustule yield declines, by carbonic acid in nutrient solution
Hydrogen radical ion concentration is adjusted to the second preset concentration, so that frustule carries out grease production.
In order to promote grease production, the second preset concentration is preferably 0-20mmol/L.It is more highly preferred to, the second preset concentration
For 0-15mmol/L.For example, second preset concentration is 0mmol/L, 5mmol/L, 10mmol/L.When the bicarbonate in nutrient solution
When ion concentration is less than the second preset concentration, the inorganic salts that generation bicarbonate ion is decomposed in water are replenished into nutrient solution
Or carbon dioxide.Preferably, the inorganic salts are bicarbonate.
It is more highly preferred to, the first preset concentration is more than or equal to the second preset concentration, to advantageously promote microalgae biomass
Production, and improve the content and yield of microalgae grease.
When it is implemented, when frustule carry out grease production when, can according to the bicarbonate ion concentration in nutrient solution,
Bicarbonate or carbon dioxide are supplemented into nutrient solution, so that the bicarbonate ion concentration in nutrient solution reaches second
Preset concentration.During frustule carries out grease production, the bicarbonate ion concentration in nutrient solution is monitored in real time, works as carbon
When sour hydrogen radical ion concentration is less than the second preset concentration, bicarbonate or carbon dioxide are supplemented into nutrient solution, so that
Bicarbonate ion concentration in nutrient solution maintains the second preset concentration.Also, into nutrient solution supplement bicarbonate or
During person's carbon dioxide, the pH value in nutrient solution is monitored in real time, by acid or carbon dioxide into nutrient solution so that culture
The pH value of liquid is maintained between 7~9.
When it is implemented, frustule carry out grease production during, it is necessary in real time monitor nutrient solution in pH value and
Bicarbonate ion concentration, and do not meet default pH value in pH value or do not meet that second is default dense in bicarbonate ion concentration
When spending, pH value and bicarbonate ion concentration is constantly adjusted all in accordance with actual conditions, the final pH value for causing nutrient solution expires
The default pH value of foot and bicarbonate ion concentration the second preset concentration of satisfaction, so that frustule preferably carries out grease production.
Cultivating principle is：CO2And HCO3 -It can be entered by cell membrane in frustule, the CO in frustule2Or HCO3 -Storage
The form of depositing is HCO3 -.In the presence of carbonic anhydrase (CA), according to equationUnderstand,
HCO3 -It is converted into CO2, CO2Rubisco (i.e. ribulose-1,5-bisphosphate, 5- diphosphonic acid carboxylases/oxygenase) is transported nearby to be fixed, because
This, the HCO in frustule3 -It is more, Rubisco substrate CO2Concentration is higher, the CO fixed2It is more, so as to ensure that
Carbon substrate needed for micro algae growth and oil and fat accumulation is sufficient, so that micro algae growth speed is speeded, and when grease induces
More greases can be accumulated.In addition, bicarbonate radical can influence the cell cycle of frustule and certain side of body is formed to cell metabolism
Compel pressure, so as to induce triacylglycerol accumulation, therefore, extracellular bicarbonate ion concentration is maintained into the second preset concentration to pierce
Swash frustule oil-producing, the CO that frustule is fixed2More flow to the grease production of frustule.
To sum up, in the microalgae cell production phase, it is default dense that the bicarbonate ion concentration in nutrient solution is maintained first
Degree, this way it is ensured that the abundance of carbon substrate, also, various nutrients are also all sufficient in nutrient solution, are advantageous to microalgae
The growth of cell.When microalgae cell yield declines, nutrient in nutrient solution is consumed because of the growth of microalgae cell so that
Nutrient deficiency in nutrient solution, at this moment, the bicarbonate ion concentration in nutrient solution is maintained into the second preset concentration, energy
Enough ensure the essential elements carbon substrate abundance of Synthetic Oil and certain stress pressure is formed to cell metabolism, it is thin to be advantageous to microalgae
The grease production of born of the same parents, improve the yield of microalgae grease.
Contain bicarbonate ion in nutrient solution, also, under weak basic condition, i.e., pH value is 7~9, according to equationThe CO of nutrient solution2Or CO3 2-More with HCO3 -Form is present.One HCO3 -By thin
After after birth enters frustule, it is utilized and is converted into CO2, and an OH is left in frustule-So that the proton on frustule film
Pump is opened, H+The neutral stable state in frustule is maintained into cell, so as to cause to add an OH in nutrient solution-, according to
EquationSo that more CO2It is soluble in water.CO2After being dissolved in water, CO2Can
Entered by cell membrane in frustule, the CO in frustule2Storage form be HCO3 -, HCO3 -It is converted into CO2, CO2Fixed,
And then improve concentration of substrate.Therefore, in above process, the CO being passed through in nutrient solution is not only caused2It can obtain preferably
Utilize, be also easy to capture the CO in air2, so as to save CO2Service cost.
When frustule yield declines, grease production can be continued in original nutrient solution, it is necessary to by bicarbonate
Ion concentration is adjusted to the second preset concentration, also, pH value is adjusted to default pH value.
It is of course also possible to so operate：When frustule yield declines, frustule is seeded to the nutrient solution of auxotrophy
In, and the bicarbonate ion concentration in the nutrient solution of auxotrophy is adjusted to the second preset concentration, and regulation auxotrophy
Medium pH value to default pH value, carry out grease production.
Specifically, when frustule yield declines, by frustule by being taken out in original nutrient solution, renewed vaccination to nutrition
In the nutrient solution of defect, grease production is carried out.Wherein, the nutrient solution of auxotrophy is to lack the one or more in nitrogen phosphate and sulfur
Nutrient solution.
Step S4 is harvested, when grease yield declines, harvests the frustule of oil-containing.
It should be appreciated by those skilled in the art that when it is implemented, microalgae frustule cultivation and grease production during,
Need to maintain other necessity regulations, such as suitable illumination, temperature and the necessary battalion of micro algae growth needed for microalgae normal growth
Form and grade.
As can be seen that in the present embodiment, by accessing algae kind into the nutrient solution containing bicarbonate ion, and will culture
The pH value of liquid is adjusted to default pH value, and the bicarbonate ion concentration in nutrient solution is adjusted to the first preset concentration to enter
Row frustule cultivates, and ensure that the carbon substrate needed for frustule growth so that frustule quickly increases, and is effectively improved micro-
The yield of algal biomass, when frustule yield declines, the bicarbonate ion concentration in nutrient solution is adjusted to second and preset
Concentration, so that frustule can accumulate more greases under the carbon substrate effect of abundance, has to carry out grease production
The content and yield of microalgae grease are improved to effect, solves the problems, such as that prior art can not effectively improve microalgae grease content；
In addition, this method can better profit from the carbon dioxide in air, that has not only saved carbon dioxide is passed through cost, additionally it is possible to
Reduce greenhouse effects.
Microalgae culture method is illustrated below by specific embodiment.
Embodiment one
Control group one and cultivation group one are inoculated in 5cm light path pipe reactions by one plant of exponential phase from separation algae kind
In device, illumination is 150 μm of ol/ ㎡/s, and whole day illumination in 24 hours, temperature is room temperature, and initial incubation liquid is suitable for microalgae
The improvement BG11 nutrient solutions of growth inducing, wherein, the formula of improvement BG11 nutrient solutions sees attached list 1.
Cultivation group one is that 1g/L NaHCO are added into initial incubation liquid3, then bicarbonate ion concentration is about in nutrient solution
For 12mmol/L.Algae kind is accessed into the nutrient solution configured and proceeds by the cultivation of frustule, and is passed through into nutrient solution
Containing 2%CO2Mixing air so that the pH value of nutrient solution is adjusted between 7~8.During frustule cultivates, monitoring training
The ion concentration of bicarbonate radical in nutrient solution, and CO in mixing air is adjusted according to bicarbonate ion concentration2Content so that
Bicarbonate ion concentration is maintained in the range of 10-15mmol/L.After growth four days, frustule yield is begun to decline, and algae is thin
Born of the same parents are seeded in the nutrient solution (seeing attached list 2) of auxotrophy, dilute frustule concentration, while mend into the nutrient solution of auxotrophy
Fill 1g/L NaHCO3, the ion concentration of bicarbonate radical in the nutrient solution of auxotrophy is maintained in the range of 10-20mmol/L,
Carry out grease production.In oil production, the grease yield in the nutrient solution of auxotrophy is monitored, grease produces at the 4th day
Amount has declined, and harvest the frustule of oil-containing.
Control group one directly starts frustule cultivation in initial incubation liquid, without other additions, is passed through and contains in breeding process
2%CO2Mixing air control nutrient solution pH value between 7~8, growth four days after, frustule is seeded to auxotrophy
Nutrient solution (seeing attached list 2) in, dilute frustule concentration, carry out grease induction, after four days harvest oil-containing frustule.This implementation
The experimental result picture of example is referring to Fig. 2.
Wherein, the mark in subordinate list 1 and subordinate list 2 is5Micro- * solution composition referring to subordinate list 3.
From above experimental result, NaHCO is added3Cultivation group one grow the dry weight of four days and be significantly higher than control group
One, and grow four days after content of fatty acid be also significantly greater than control group one.After starting within 4th day grease induction, oil and fat accumulation adds
It hurry up, the content of fatty acid of cultivation group one is finally significantly higher than control group one.Therefore, the grease yield of cultivation group one is compared with control group one
Improve 21%.
Embodiment two
Control group two, cultivation group two and cultivation group three are inoculated in 5cm light by one plant of exponential phase from separation algae kind
In journey tubular reactor, illumination is 150 μm of ol/ ㎡/s, 8h：16h Dark-light cycles, temperature are room temperature, and initial incubation liquid is
Suitable for the improvement BG11 nutrient solutions of the algae growth inducing, wherein, the formula of improvement BG11 nutrient solutions sees attached list 1.
Cultivation group two and cultivation group three add 1.26g/L NaHCO in initial incubation liquid3, then carbonic acid in nutrient solution
Hydrogen radical ion concentration is each about 15mmol/L.Cultivation group two and cultivation group three are to access algae kind simultaneously into the nutrient solution configured
The cultivation of frustule is proceeded by, cultivation group two adds acetic acid into nutrient solution and adjusts the pH value of nutrient solution between 7-8；Support
Addition hydrochloric acid in group three-dimensional nutrient solution is grown to adjust the pH value of nutrient solution between 7-8.During frustule cultivates, monitoring
Whether the ion concentration of bicarbonate radical is maintained in the range of 10-20mmol/L in nutrient solution, if being less than this scope, into nutrient solution
Add NaHCO3To supplement bicarbonate ion.After growth four days, frustule yield is begun to decline, and frustule is seeded into nutrition
In the nutrient solution (seeing attached list 2) of defect, frustule concentration is diluted, while supplements 1.26g/L's into the nutrient solution of auxotrophy
NaHCO3, bicarbonate ion concentration is about 15mmol/L in the nutrient solution of auxotrophy, and is made in the nutrient solution of auxotrophy
The ion concentration of bicarbonate radical is maintained in the range of 10-20mmol/L, carries out grease production.In oil production, monitoring
Grease yield in the nutrient solution of auxotrophy, when grease yield has declined at the 4th day, harvest the frustule of oil-containing.
Control group two is passed through air in initial incubation liquid and directly starts frustule cultivation, and without other additions, whole process utilizes
Hydrochloric acid controls the pH value of nutrient solution between 7~8, after growth four days, by frustule be seeded to auxotrophy nutrient solution (see
Subordinate list 2) in, grease induction is carried out, the frustule of oil-containing is harvested after four days.The experimental result picture of the present embodiment is referring to Fig. 3.
Wherein, the mark in subordinate list 1 and subordinate list 2 is5Micro- * solution composition referring to subordinate list 3.
From above experimental result, the biomass increment of cultivation group two and cultivation group three is all remarkably higher than control group two.
After grease induces four days, the content of fatty acid of cultivation group two and cultivation group three is finally significantly higher than control group two.Cultivation group two
Grease yield improve 2.3 times compared with control group two, the grease yield of cultivation group three improves 1.8 times compared with control group two.
Embodiment three
Control group three, cultivation group four and cultivation group five are inoculated in 0.2 ㎡ by one plant of exponential phase from separation algae kind
In small raceway pond, light path is 10cm, and illumination is 700 μm of ol/ ㎡/s, 16h：8h Dark-light cycles, temperature are room temperature, initially
Nutrient solution is the improvement BG11 culture mediums suitable for micro algae growth induction, wherein, the formula of improvement BG11 nutrient solutions sees attached list 1.
Cultivation group four is that 0.84g/L NaHCO is added in initial incubation liquid3, then bicarbonate ion is dense in nutrient solution
Degree is about 10mmol/L.Cultivation group five is that 1.68g/L NaHCO is added in initial incubation liquid3, then bicarbonate radical in nutrient solution
Ion concentration is about 20mmol/L.Algae kind is accessed into the nutrient solution configured and proceeds by the cultivation of frustule, cultivation group
Four and cultivation group five be passed through into nutrient solution containing 10%CO2Mixing air with by the pH value of nutrient solution adjust to 7~8 it
Between.During frustule cultivates, the ion concentration of bicarbonate radical in nutrient solution is monitored, and makes the bicarbonate radical of cultivation group four
Ion concentration is maintained in the range of 10-15mmol/L, and the bicarbonate ion concentration of cultivation group five maintains 20-25mmol/L models
In enclosing.After growth five days, frustule yield is begun to decline, and harvests whole frustules, and frustule is seeded into auxotrophy
In nutrient solution (seeing attached list 2), frustule concentration is diluted, also, still in 0.2m2Cultivated in small raceway pond, illumination temperature is constant.
Cultivation group four adds 0.84g/L NaHCO in the nutrient solution of auxotrophy3, then bicarbonate ion concentration is about 10mmol/
L；Cultivation group five adds 1.68g/L NaHCO in the nutrient solution of auxotrophy3, then bicarbonate ion concentration be about
20mmol/L, carry out grease production.In oil production, it is ensured that the ion concentration of the bicarbonate radical of cultivation group four maintains
In the range of 10-15mmol/L, the ion concentration of the bicarbonate radical of cultivation group five is maintained in the range of 15-20mmol/L.Monitoring oil
Fat yield, when grease yield has declined, harvest the frustule of oil-containing.
Control group three directly starts frustule cultivation in initial incubation liquid, and without other additions, whole process is passed through 10%CO2's
Mixing air controls the pH value of nutrient solution between 7~8, and after growing five days, frustule is seeded to the culture of auxotrophy
In liquid (seeing attached list 2), grease induction is carried out, the frustule of oil-containing is harvested after five days.The experimental result picture of the present embodiment is referring to figure
4。
Wherein, the mark in subordinate list 1 and subordinate list 2 is5Micro- * solution composition referring to subordinate list 3.
From above experimental result, the yield of biomass and content of fatty acid of cultivation group four and cultivation group five are above pair
It is in be incremented by situation according to group three, and with the ion concentration rise of bicarbonate radical.After grease induces five days, although cultivation group four and cultivation
The yield of biomass of group five has declined with the ion concentration rise of bicarbonate radical, but to increase trend notable for content of fatty acid,
Finally, the grease yield of cultivation group four and cultivation group five is above control group three.Wherein, the grease yield of cultivation group four relatively compares
Group three improves 14.1%, and the grease yield of cultivation group five improves 3.8% compared with control group three.
Example IV
Control group four, cultivation group six and cultivation group seven are inoculated in 5cm light by one plant of exponential phase from separation algae kind
In journey tubular reactor, illumination is 200 μm of ol/ ㎡/s, and illumination in 24 hours, temperature is room temperature, and initial incubation liquid is suitable
In the improvement BG11 nutrient solutions of the algae growth inducing, wherein, the formula of improvement BG11 nutrient solutions sees attached list 1.
Cultivation group six adds 12mmol/L NaHCO in initial incubation liquid3, cultivation group seven adds in initial incubation liquid
25mmol/L NaHCO3.Algae kind is accessed into the nutrient solution configured for cultivation group six and cultivation group seven and to proceed by algae thin
The cultivation of born of the same parents, and be passed through into nutrient solution containing 2%CO2Mixing air so that the pH value of nutrient solution is adjusted between 7~8.
During frustule cultivates, monitor the ion concentration of bicarbonate radical in nutrient solution, and make the bicarbonate radical of cultivation group six from
Sub- concentration is maintained in the range of 10-15mmol/L, and the bicarbonate ion concentration of cultivation group seven maintains 25-30m mol/L models
In enclosing.After growth five days, frustule yield is begun to decline, and the ion of adjustment cultivation group six and the bicarbonate radical in cultivation group seven is dense
Degree so that the ion concentration of the bicarbonate radical of cultivation group six is maintained in the range of 110-15mmol/L, the bicarbonate of cultivation group seven
The ion concentration of root is maintained in the range of 15-20mmol/L.Grease yield is monitored, grease induces two days later, and grease yield declines
It has been declined that, harvest the frustule of oil-containing.
Control group four directly starts frustule cultivation in initial incubation liquid, without other additions, is passed through and contains in breeding process
2%CO2Mixing air control nutrient solution pH value between 7~8, growth four days after, carry out grease induction, harvested after four days
The frustule of oil-containing.The experimental result picture of the present embodiment is referring to Fig. 5.
Wherein, the mark in subordinate list 1 and subordinate list 2 is5Micro- * solution composition referring to subordinate list 3.
From above experimental result, the dry weight change of cultivation group six and cultivation group seven is little with the difference of control group four,
But the content of fatty acid of cultivation group six and cultivation group seven is all remarkably higher than control group four, and increases with the increase of bicarbonate concentration
It is high.The grease yield of cultivation group six improves 43.7% compared with control group four, and the grease yield of cultivation group seven improves compared with control group four
53.5%.
Table 1 improves BG11 culture medium prescriptions
| NaNO3 | 0.3g/L |
| K2HPO4·3H2O | 0.059g/L |
| MgSO4·7H2O | 0.075g/L |
| CaCl2·2H2O | 0.036g/L |
| Citric acid | 0.006g/L |
| FeCl3·6H2O | 0.00315g/L |
| Na2EDTA·2H2O | 0.00436g/L |
| Na2CO3 | 0.02g/L |
| A5Micro- * | 1ml/L |
The formula of the nutrient solution of the auxotrophy of table 2
| CaCl2·2H2O | 0.036g/L |
| Citric acid | 0.006g/L |
| FeCl3·6H2O | 0.00315g/L |
| Na2EDTA·2H2O | 0.00436g/L |
| Na2CO3 | 0.02g/L |
| A5Micro- * | 1ml/L |
The A of table 35The solute constituent of micro- * solution, wherein, solvent is 1000mL deionized waters
| H3BO3 | 2.86g |
| MnCl2·H2O | 1.81g |
| ZnSO4·7H2O | 0.222g |
| CuSO4·5H2O | 0.079g |
| Na2MoO4·2H2O | 0.390g |
| Co(NO3)2·6H2O | 0.0494g |
In summary, the carbon substrate needed for frustule growth can be ensured so that frustule quickly increases, and effectively carries
The high yield of microalgae biomass, and cause frustule to accumulate more greases under the carbon substrate effect of abundance, have
Improve to effect the content and yield of microalgae grease；In addition, this method can better profit from the carbon dioxide in air, not only
That has saved carbon dioxide is passed through cost, additionally it is possible to reduces greenhouse effects.
Obviously, those skilled in the art can carry out the essence of various changes and modification without departing from the present invention to the present invention
God and scope.So, if these modifications and variations of the present invention belong to the scope of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to comprising including these changes and modification.
Claims (9)
1. a kind of microalgae culture method, it is characterised in that comprise the following steps：
Configuration step, the nutrient solution of micro algae growth is configured, contains bicarbonate ion in the nutrient solution；
Step is cultivated, algae kind is accessed into the nutrient solution, adjusts the pH value of the nutrient solution to default pH value, and described in regulation
Bicarbonate ion concentration in nutrient solution reaches the first preset concentration, carries out frustule cultivation, wherein, described first is default dense
Spend for 10-30mmol/L；
Grease production step, when frustule yield declines, adjust the bicarbonate ion concentration in the nutrient solution and reach
Two preset concentrations, grease production is carried out, wherein, second preset concentration is 0-20mmol/L；
Step is harvested, when grease yield declines, harvests the frustule of oil-containing.
2. microalgae culture method according to claim 1, it is characterised in that in the configuration step,
Added into the nutrient solution in water and decompose the inorganic salts of generation bicarbonate ion, so as to contain in the nutrient solution
Bicarbonate ion.
3. microalgae culture method according to claim 1, it is characterised in that in the cultivation step,
The default pH value is 7~9.
4. microalgae culture method according to claim 3, it is characterised in that
By adding acid or carbon dioxide into the nutrient solution so that the pH value of the nutrient solution reaches described default
PH value.
5. microalgae culture method according to claim 1, it is characterised in that in the cultivation step,
When the bicarbonate ion concentration in the nutrient solution is less than first preset concentration, supplemented into the nutrient solution
The inorganic salts or carbon dioxide of generation bicarbonate ion are decomposed in water.
6. microalgae culture method according to claim 1, it is characterised in that in the grease production step,
Second preset concentration is 0-15mmol/L.
7. microalgae culture method according to claim 1, it is characterised in that
First preset concentration is more than or equal to second preset concentration.
8. microalgae culture method according to claim 1, it is characterised in that the grease production step includes：
When the frustule yield declines, the frustule is seeded in the nutrient solution of auxotrophy, and adjust the battalion
The bicarbonate ion concentration in the nutrient solution of defect is supported to second preset concentration, carries out grease production.
9. microalgae culture method according to claim 8, it is characterised in that the nutrient solution of the auxotrophy is to lack
One or more of nutrient solutions in nitrogen phosphate and sulfur.
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| JPH0679A (en) * | 1992-06-18 | 1994-01-11 | Tokyo Electric Power Co Inc:The | Method for controlling cultivation of microalgae |
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2017
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| CN103484372A (en) * | 2013-10-01 | 2014-01-01 | 陈意民 | Culture method for increasing biomass and grease yield of microalgae |
| KR101765833B1 (en) * | 2016-03-31 | 2017-08-08 | 인하대학교 산학협력단 | Cultivation method of microalgae using bicarbonate as carbon source |
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