CN103103129A - Production method for lipid through synchronous mixed culture of microbes - Google Patents
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- CN103103129A CN103103129A CN2011103524541A CN201110352454A CN103103129A CN 103103129 A CN103103129 A CN 103103129A CN 2011103524541 A CN2011103524541 A CN 2011103524541A CN 201110352454 A CN201110352454 A CN 201110352454A CN 103103129 A CN103103129 A CN 103103129A
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Abstract
The invention discloses a production method for lipid through synchronous mixed culture of microbes. The method comprises the following steps: (1) culturing facultative anaerobic bacteria seed liquid; (2) culturing autotrophic microalgae seed liquid; and (3) mixing the facultative anaerobic bacteria seed liquid and the autotrophic microalgae seed liquid and carrying out synchronous mixed culture in an illuminated bioreactor, wherein during synchronous mixed culture, an organic carbon source needed in growth and metabolism of the facultative anaerobic bacteria is supplemented. The method provided by the invention has the advantages of increased harvest yield of autotrophic microalgae, an improved utilization rate of an inorganic carbon source (CO2), increased content of microbial lipid, a simplified culture apparatus and the like, and exerts small influence on the fermentation process of the facultative anaerobic bacteria, thereby realizing synchronous culture of the facultative anaerobic bacteria and the autotrophic microalgae.
Description
Technical field
The invention belongs to biological technical field, relate in particular to a kind of autotrophy oil-producing microalgae that utilizes and obtain high cell harvesting amount, improve simultaneously the fat content of microalgae cell.
Background technology
Along with day by day highlighting and problem of environmental pollution more outstanding of the rising steadily of day by day in short supply, the oil price of petroleum resources, oil product imbalance between supply and demand, developing renewable resources by all kinds of means, it is inevitable to become.From the diesel oil of microbial oil have that energy density is high, sulphur content is low, the performance such as sufficient combustion, oilness are good, also have the characteristics such as renewable, readily biodegradable, storing and transporting security, the capability of antidetonance be good, can be used as the substitute of fossil energy.
Little algae is one of oil-containing microorganism, and the nutritive ingredients such as little algae rich in proteins, polysaccharide, unsaturated fatty acids (as spirulina etc.) can be used for food, medicine and energy aspect; Can accumulate in a large number lipid acid, algae such as chlorella are slightly arranged, its body fat acid content can account for 30%～60% of dry weight.Utilize the little algae of cultivation to accumulate oil resource, become and utilized at present the most popular research field of solar energy development renewable resources.Not only have huge market potential, and have outstanding social value.
The microalgae cell growth pattern divides and is generally two kinds of light autotrophy and heterotrophism carbon sources, and light autotrophy process will consume CO
2, CO
2Effective utilization absorb, be the key of culture effect of realizing ideal, exist simultaneously and replenish CO
2Produce O with photosynthesis
2Desorb, the problem of discharge.Liu Jianguo etc. " microalgae mass cultivate pipeline bioreactor " (CN200410020978.0) and Miao JianRen etc. " a kind of little algae industrial production photosynthetic organism reactor assembly " (CN03128138.9) all adopt add a kind of installation method in photo-bioreactor system, realize CO
2Supply, can realize also that simultaneously certain oxygen resolves effect.The heterotrophic growth process is to utilize organic carbon source to be the CO in the alternative autotrophy process of substrate
2Carry out the accumulation of little algal biomass, the Growth of Cells speed, but in cell, the oil and fat accumulation level is lower.
Utilize other non-alga microbial and little algae to carry out the research of mixed culture also many, a kind of method of genus bacillus regulation and control planktonic microalgae mixed culture is disclosed such as CN200910038908.0, when using genus bacillus to carry out mixed culture to each little algae in co-culture system, the method finds, each micro algae growth is balanced, good stability, can avoid the species diversity of algae phase in co-culture system single, avoid the quantity of certain little algae extreme advantage or inferior position to occur, so the method is mainly used in controlling certain little algae spreading unchecked in water body.CN200910038910.8 discloses a kind of method of lactobacillus little algae mixed culture of regulation and control and synergistic purification cultivation discharge water, the method is by the direct or indirect effect of lactobacillus, regulate and control the quantitative proportion of various little algaes, make each algae growth balanced, good stability reaches the purpose of denitrification dephosphorization to the cultivation waste discharge by the synergy between bacterium, algae.Above-mentioned prior art is to control the growth equilibrium of algae in water body by thalline, is not to promote the growth of algae and the oil and fat accumulation in frustule.
Summary of the invention
For the deficiencies in the prior art, the invention provides a kind of method of utilizing facultative anaerobic bacteria and the little algae of autotrophy to carry out the synchronized mixes cultivation, the inventive method has the autotrophy of raising microalgae harvesting amount, improves inorganic carbon source (CO
2) utilization ratio, improve microbial oil content, simplify the advantages such as culture apparatus, less on the fermenting process impact of facultative anaerobic bacteria simultaneously, realize that both synchronously carry out.
Microbial synchronous mixed culture of the present invention is produced the method for grease, comprises following content:
(1) cultivate the facultative anaerobic bacteria seed liquor;
(2) cultivate the little algae seed liquor of autotrophy;
(3) the little algae seed liquor of facultative anaerobic bacteria seed liquor and autotrophy is mixed, and carries out synchronized mixes and cultivate in illuminated bio-reactor, in the synchronized mixes culturing process, replenishes the required organic carbon source of facultative anaerobic bacteria growth metabolism.
In the inventive method, facultative anaerobic bacteria comprises genus bacillus, fusiform bacilarmature, bifidus bacillus, Bacterium lacticum and klebsiella etc., organic carbon source is glucose, glycerine, fructose, starch, cellulosic hydrolysate etc., the meta-bolites of facultative anaerobic bacteria, be that the facultative anaerobic bacteria tunning is generally 1, ammediol and/or organic acid etc., the tunning that different facultative anaerobic bacterias obtains is different, and is different and different according to the kind of facultative anaerobic bacteria.It is method well known to those skilled in the art that the facultative anaerobic bacteria seed liquor is cultivated, and as adopting stirring type bioreactor, adds substratum and facultative anaerobic bacteria, cultivates under suitable condition.
In the inventive method, the little algae of autotrophy comprises chlorella, grape algae, little ring algae, diatom etc., and it is method well known to those skilled in the art that the little algae seed liquor of autotrophy is cultivated, as adopts conventional air lift type illumination bio-reactor to carry out seed liquor and cultivate.
In the inventive method, seed culture medium is respectively facultative anaerobic bacteria substratum and little algae SE substratum.
In the inventive method, in the synchronized mixes culturing process, the initial access volume ratio of facultative anaerobic bacteria seed liquor and little algae seed liquor is 1:1～1:10.The initial medium that synchronized mixes is cultivated adds the required inorganic salt of microalgae cell growth and trace element (adding related substances as the composition of pressing the SE substratum) etc. simultaneously take the required basic medium of facultative anaerobic bacteria as main.The synchronized mixes culturing process in batches or the required organic carbon source of continuous supplementation facultative anaerobic bacteria fermenting process.
In the inventive method, the illumination bio-reactor is generally adopted in the synchronized mixes cultivation, and the condition of the conditional likelihood of the general employing of the condition that synchronized mixes is cultivated and facultative anaerobic bacteria fermenting process is generally 20 ℃～37 ℃ as temperature, the pH value is generally 6～9, is preferably 6.5～7.5 etc.In the synchronized mixes culturing process, can pass into nitrogen or carbonated gas.
The inventive method utilizes facultative anaerobic bacteria cell and microalgae cell to carry out mixed culture, and the facultative anaerobic bacteria cell is different from microalgae cell growth conditions and required carbon source, complements each other and promotes.Microalgae cell is discarded carbon source (CO after utilizing the facultative anaerobic bacteria cell fermentation
2) be the carbon source of self growing, keep simultaneously system's osmotic pressure condition (pH value) more stably, constantly grow by replenishing organic carbon source assurance facultative anaerobic bacteria cell, by optical condition, microalgae cell is constantly grown, thereby realize the synchronized mixes culturing process of facultative anaerobic bacteria cell and microalgae cell.Select suitable kind facultative anaerobic bacteria cell and suitable kind microalgae cell, by controlling ratio and the condition of facultative anaerobic bacteria cell and microalgae cell, make both form stable co-cultivation system, and set up the dependence of different iuntercellular symbiosis, realized the efficient process of growth of oil-containing micro-algae, and improved the accumulative effect of grease, improved the grease harvest yield in the unit fermentation system in single oil-containing micro-algae culturing process, thereby laid a good foundation for the preparation of microbial oil.Very little on the fermenting process impact of facultative anaerobic bacteria cell simultaneously, can obtain simultaneously required tunning.Little algae all has tolerance to carbon source and the tunning that fermenting process uses, and does not affect the growth of little algae and the accumulation of grease.
Description of drawings
Fig. 1 is a kind of concrete technology schematic flow sheet of the present invention.
Wherein: 1-facultative anaerobic bacteria seed, the little algae seed of 2-, 3-facultative anaerobic bacteria seed liquor is cultivated reactor, the little algae seed liquor of 4-is cultivated reactor, 5-synchronized mixes pond, 6-mixed culture medium and acid-base neutralisation agent, the 7-synchronized mixes is cultivated reactor, the outlet of 8-mixed culture tail gas, 9-mixed culture gas lift inlet mouth.
Embodiment
Microbial synchronous mixed culture of the present invention is produced the method for grease, specifically comprise following content: adopt stirring type bioreactor to carry out the facultative anaerobic bacteria seed liquor and cultivate, adopting illuminated bio-reactor to carry out the little algae seed liquor of autotrophy cultivates, comprise facultative anaerobic bacteria, fermentation using bacteria substratum in stirring type bioreactor, comprise little algae algae kind and autotrophy substratum in illuminated bio-reactor.Before mixed culture, two kinds of microorganisms carry out separately respectively seed liquor and cultivate, seed liquor cultivate qualified after, through the synchronized mixes pond two kinds of microorganism seed liquid being carried out synchronized mixes, and add mixed culture medium nutritive ingredient and acid-base neutralisation agent to be adjusted to the required growth conditions of mixed culture.Wherein the organic carbon source in mixed culture medium is used for the facultative anaerobic bacteria growth, and the CO that the facultative anaerobic bacteria growth produces
2Be used for micro algae growth.The grain weight ratio that it is facultative anaerobic bacteria and little algae that synchronized mixes is cultivated two kinds of microorganisms of initial access is 1:1～1:10, after inoculation mixed cultivation process by normal pH control, the control culturing process such as temperature control and air flow.Wherein organic carbon source most preferably adopts fed-batch mode to carry out during mixed culture, and it is grown by the facultative anaerobic bacteria utilization, produces more carbonic acid gas after facultative thin anaerobic bacterial fermentation, excretes to enter in culture systems the CO that the facultative anaerobic bacteria cell is discharged
2Be in dissolved state, immediately by the microalgae cell utilization in system, as the carbon source supply of microalgae cell growth.
In the inventive method, bioreactor can be the closed reactors such as board-like, tubular type, airlift agitation formula.The nitrogen that can pass into or carbon dioxide containing gas are back-mixing power.Other operational condition of bio-reactor bacterium and little algae culture condition routinely controlled.The determinator of culture system carbonic acid gas and dissolved oxygen content can be set, adjust as required air flow, to obtain good effect.
In the inventive method, the facultative anaerobic bacteria cell is realized Fast Growth under the organic carbon source condition, and facultative anaerobic bacteria cell consumption organic carbon source generates bacterium living beings matter and various meta-bolites, and fermentation simultaneously produces CO
2Gas excretes, and makes inorganic carbon source content increase in culture system, and microalgae cell utilizes these inorganic carbon sources to carry out photosynthesis, obtains the growth of microalgae cell self.Simultaneously the facultative anaerobic bacteria cell fermentation descends the pH value of system, and along with the growth of microalgae cell, utilizes the CO that dissolves
2, make the pH value of system increase, both act on each other, and the pH of regulation system is in subject range.Bioreactor provides pH electrode to detect simultaneously, in order to add acid, alkali to realize the control of the pH of system by external source.
In present method, pH control acid neutralizing agent is HCl, H
2SO
4, HNO
3Deng mineral acid commonly used, the alkali neutralizing agent is NaOH, NaHCO
3, KOH, Ca(OH)
2, mineral alkali that ammoniacal liquor etc. is commonly used.
In the inventive method, the mixed cultivation process temperature is controlled to be inner coil pipe type of heating.
In the inventive method, it is nitrogen that mixed cultivation process passes into gas, pass into gas volume and change with the nutrient solution volume change, passing into the long-pending ratio of gas volume speed and culture systems dress liquid is the unit gas volume that 0.1 vvm ~ 1.0 vvm(unit's liquid volume per minutes pass into).
In the inventive method, the required organic carbon source of facultative anaerobic bacteria cell can be glycerine, glucose, fructose, starch, cellulosic hydrolysate etc., the adding of this organic carbon source adopting stream to add arbitrary way carries out, become the sample analysis of components according to nutrient solution, the organic carbon source concentration level of detection system, carry out stream with the organic carbon source mother liquor that configures by spending rate and add at any time.
As shown in Figure 1, facultative anaerobic bacteria seed 1 is cultivated reactor 3 incubation growth through the facultative anaerobic bacteria seed liquor and is reached biomass OD=5.0～15.0, and little algae seed 2 is cultivated No. 4 incubation growth of reactor through little algae seed liquor and reached biomass OD=2.0～10.0.Cultured bacterium seed liquor and microalgae cell seed liquor are carried out mixed culture in airlift agitation formula bioreactor, synchronously change over to after adopting bacterial cell and microalgae cell to be prepared into seed liquor respectively in this mixed cultivation process in synchronized mixes pond 5, mix in suitable ratio, and mixed culture medium and acid-base neutralisation agent 6 added in mixing pit mix, simultaneously through detection analyze mix after in culture systems the pH value regulate the pH to 6.5 of mixed-culture medium ~ 7.5.
In the inventive method, bioreactor used is facultative anaerobic bacteria fermentation culture tank, is also the photosynthetic reactor that little algae is cultivated, the facultative anaerobic bacteria cell utilizes its suitable substratum, and wherein carbon source is organic carbon source, grows, obtain the growth of biomass, consume oxygen and generate CO
2, the CO that the facultative anaerobic bacteria cell generates
2Carry out photosynthetic carbon source as little algae in culture system, thereby obtain the growth of microalgae cell.
In the inventive method, facultative anaerobic bacteria cell seed liquor and microalgae cell seed liquor inoculation volume ratio are 1:1 ~ 1:10.
In the inventive method, the initial organic carbon source concentration of bacteria culture medium is (with organism quality densitometer, lower same) be 1.0% ~ 5.0%, consume by the anaerobic bacterium Growth of Cells in process, organic carbon source is constantly added through adding pump, and the synchronized mixes culturing process is kept organic carbon source concentration in 1.0% ~ 2.0% scope.
In the inventive method, mixed cultivation process adopts facultative anaerobic bacteria substratum and micro-algae culture medium to combine, and comprises the compositions such as organic carbon source, inorganic salt and trace element, becomes mixed culture medium.Preferred culture condition is: total seed liquor inoculum size (accounts for illumination bio-reactor volume, V/V): 5% ~ 20%; Temperature: 25 ℃ ~ 30 ℃; Air flow: 0.1 vvm ~ 1.0 vvm, mixing speed: 100 rpm ~ 400rpm, time: 24h ~ 120h.Dissolved oxygen content in co-culture system (DO) is generally lower than 1mg/L.
Scheme 1(comparative example)
Klebsiella (Chinese microorganism strain preservation center C GMCC0798) is cultivated in the 300mL shaking flask, and used medium is the anaerobic bacterium glycerin medium, obtains required seed liquor after cultivation 20h.In the facultative anaerobic bacteria seed liquor access 10L bio-reactor of cultivating, used medium is mixed culture medium, and reactor is the airlift agitation formula, can realize the back-mixing of nutrient solution, and reactor is vitreum, has temperature to control coil pipe in reactor, and pH, O
2And CO
2Sensor.
Facultative anaerobic bacteria is Cray Bai Shi pneumobacillus, and its substratum is glycerin medium.
Glycerin medium formula (in every liter):
NH
4Cl 5.35 g, KCl 0.75 g, NaH
2PO
41.38 g, Na
2SO
40.28 g, MgCl
26H
2O 0.26 g, CaCl
2H
2O 0.02 g, yeast extract 1.0 g, glycerine 40 g.
SE culture medium prescription (in every liter):
| NaNO 3 | 0.20g |
| K 2HPO 4 . 3H 2O | 0.07g |
| MgSO 4 . 7H 2O | 0.07g |
| CaCl 2 . 2H 2O | 0.03g |
| KH 2PO 4 | 0.18g |
| NaCl | 0.03 |
| Soil extract (soil extract) | 40mL |
| FeCl 3·6H 2O | 0.01 |
| Fe—EDTA | 1mL |
10L bioreactor culture condition is to adopt mixed culture medium (take glycerin medium as the basis, press simultaneously the SE incubation period and form the suitable material of interpolation): inoculum size (V/V): 10%; Temperature: 30 ℃; Air flow (nitrogen): 0.4 vvm, mixing speed: 200 rpm, time: 120h.
Cultivate and stopped afterwards cultivation in 5 days, collect microorganism cells, survey dry weight and fat content, the harvest yield of the unit's of drawing fermentation system microbial oil.Wherein initial glycerin medium organic carbon source (glycerine) quality is 40.0g/L, and process is added organic carbon source, and to keep content be the 15g/L left and right, cultivates that when finishing, residual glycerol content is 5.0g/L.Utilize after the centrifugal collection thalline of liquid-phase chromatographic analysis fermented liquid primary product 1,3-PD concentration in clear liquid to determine the fermentation level of facultative anaerobe.
Scheme 2(comparative example)
Chlorella (available from Inst. of Hydrobiology, Chinese Academy of Sciences's algae kind storehouse 1#) is carried out the light autotrophy cultivate in shaking flask, used medium is the SE substratum, cultivates and obtains required seed liquor after 2 days.Cultured chlorella seed liquid is seeded in the 10L bioreactor.Reactor is the airlift agitation formula, can realize the back-mixing of nutrient solution, and reactor is vitreum, and the fluorescent lamp source is set, and automatically controls the switching time of setting light, forms the dark process conversion of light in the chlorella culturing process.There is temperature to control coil pipe in reactor, and pH, O
2And CO
2Sensor.10L bioreactor culture condition is with shown in comparative example scheme 1.
The algae kind is Chlorella vulgaris, and the substratum of chlorella culturing process is with the mixed culture medium of scheme 1.
Ventilation adopts the air compressor compressed nitrogen to pass into, intake 4.0L/min.
Nutrient solution (is pressed the ratio-dependent of 12:12 hour every day) under set light dark period, cultivate to stop afterwards in 5 days cultivating, and collects microorganism cells, surveys dry weight and fat content, the grease harvest yield of the unit's of obtaining fermentation system.
Scheme 3(embodiment 1)
Cray Bai Shi pneumobacillus seed liquor cultural method is with comparative example scheme 1, and chlorella seed liquid culture condition is with comparative example scheme 2, uses in shaking flask Cray Bai Shi pneumobacillus and chlorella separately that substratum carries out single culture, cultivates to obtain required seed liquor.Cultured Cray Bai Shi pneumobacillus seed liquor and chlorella cells seed liquor are seeded to together contain in the 10L bioreactor that mixes substratum, reactor is the airlift agitation formula, can realize the back-mixing of nutrient solution, reactor is vitreum, fluorescent light source is set, automatically control the switching time of setting light, form the dark process conversion of light in the chlorella culturing process.There is temperature to control coil pipe in reactor, and pH, O
2And CO
2Sensor.Mixed culture medium replenishes organic carbon source by scheme 2 simultaneously with the mixed culture medium of scheme 1.
Synchronized mixes is cultivated ventilation and is adopted the gas compressor compressed nitrogen to pass into, intake 4.0L/min.Nutrient solution is (in the ratio of 12:12 hour every day) under set light dark period, cultivates to stop afterwards in 5 days cultivating, and collects microorganism cells, surveys dry weight and fat content, the grease harvest yield of the unit's of obtaining fermentation system.Utilize after the centrifugal collection thalline of liquid-phase chromatographic analysis fermented liquid product 1,3-PD concentration in clear liquid to determine the fermentation level of facultative anaerobe.
Scheme 4(embodiment 2)
Cray Bai Shi pneumobacillus seed liquor cultural method is with comparative example scheme 1, grape algae (available from Inst. of Hydrobiology, Chinese Academy of Sciences's algae kind storehouse 357#) seed liquor culture condition is with comparative example scheme 2, use in shaking flask Cray Bai Shi pneumobacillus and grape algae separately that substratum carries out single culture, cultivate to obtain required seed liquor.Cultured Cray Bai Shi pneumobacillus seed liquor and grape frustule seed liquor are seeded to together contain in the 10L bioreactor that mixes substratum, reactor is the airlift agitation formula, can realize the back-mixing of nutrient solution, reactor is vitreum, fluorescent light source is set, automatically control the switching time of setting light, form the dark process conversion of light in grape algae culturing process.There is temperature to control coil pipe in reactor, and pH, O
2And CO
2Sensor.Mixed culture medium replenishes organic carbon source by scheme 2 simultaneously with the mixed culture medium of scheme 1.
Synchronized mixes is cultivated ventilation and is adopted the gas compressor compressed nitrogen to pass into, intake 4.0L/min.Nutrient solution is (in the ratio of 12:12 hour every day) under set light dark period, cultivates to stop afterwards in 5 days cultivating, and collects microorganism cells, surveys dry weight and fat content, the grease harvest yield of the unit's of obtaining fermentation system.Utilize after the centrifugal collection thalline of liquid-phase chromatographic analysis fermented liquid product 1,3-PD concentration in clear liquid to determine the fermentation level of facultative anaerobe.
Above-described embodiment experimental result such as following table 1:
The synchronized mixes cultivation results of each embodiment of table 1
| Scheme | The microorganism dry weight | Fat content | Grease harvest yield | The facultative |
| 1 | 7.02g/L | 2.2% | 0.15g/L | 54.2g/ |
| 2 | 5.21g/L | 7.4% | 0.39g/L | - |
| 3 | 10.40g/L | 32.6% | 3.90g/L | 52.8g/L |
| 4 | 11.06g/L | 33.7% | 3.73g/L | 56.2g/L |
Can find out from above-mentioned data, the inventive method (scheme 3 and 4) has greatly improved the harvest yield of microorganism cells, and fat content also is improved.Therefore adopt the inventive method, under identical culture condition, compare with microorganism single culture method, two kinds of microorganisms are cultivated through synchronized mixes, the grease harvest yield that obtains has obtained larger raising.In the unit fermentation system that synchronized mixes is cultivated, grease harvest yield is improved significantly.Can find out by the fermentation level correlation data of facultative anaerobic bacteria simultaneously, in the situation that whether microalgae cell exist, klebsiella carries out anaerobically fermenting, and to produce the level of 1,3-PD substantially unaffected.This inventive method is described, when realizing biomass collection and then obtaining bio-oil, can also realizes that fermentation using bacteria produces other target products, thereby improve the utilising efficiency of microorganism.
Claims (10)
1. a microbial synchronous mixed culture is produced the method for grease, it is characterized in that comprising following content:
(1) cultivate the facultative anaerobic bacteria seed liquor;
(2) cultivate the little algae seed liquor of autotrophy;
(3) the little algae seed liquor of facultative anaerobic bacteria seed liquor and autotrophy is mixed, and carries out synchronized mixes and cultivate in illuminated bio-reactor, in the synchronized mixes culturing process, replenishes the required organic carbon source of facultative anaerobic bacteria growth metabolism.
2. in accordance with the method for claim 1, it is characterized in that: facultative anaerobic bacteria comprises genus bacillus, fusiform bacilarmature, bifidus bacillus, Bacterium lacticum or klebsiella, organic carbon source is glucose, glycerine, fructose, starch or cellulosic hydrolysate, the organic carbon source meta-bolites that utilizes of facultative anaerobic bacteria is 1,3-PD and/or organic acid.
3. in accordance with the method for claim 1, it is characterized in that: the facultative anaerobic bacteria seed liquor is cultivated and is adopted stirring type bioreactor, and the little algae seed liquor of autotrophy is cultivated and adopted air lift type illumination bio-reactor.
4. it is characterized in that in accordance with the method for claim 1: the little algae of autotrophy comprises chlorella, grape algae, little ring algae or diatom.
5. according to claim 1,2 or 4 described methods, it is characterized in that: in the synchronized mixes culturing process, the initial access volume ratio of facultative anaerobic bacteria seed liquor and little algae seed liquor is 1:1～1:10.
6. in accordance with the method for claim 1, it is characterized in that: the initial medium that synchronized mixes is cultivated adopts the required substratum of facultative anaerobic bacteria, adds simultaneously required inorganic salt and the trace element of microalgae cell growth.
7. according to claim 1,2 or 6 described methods, it is characterized in that: the synchronized mixes culturing process in batches or the required organic carbon source of continuous supplementation facultative anaerobic bacteria fermenting process.
8. in accordance with the method for claim 1, it is characterized in that: the condition that synchronized mixes is cultivated is: temperature is 20 ℃～37 ℃, and the pH value is 6～9.
9. in accordance with the method for claim 1, it is characterized in that: the facultative anaerobic bacteria seed liquor is cultured to biomass OD=5.0～15.0, and little algae seed liquor is cultured to biomass OD=2.0～10.0.
10. in accordance with the method for claim 7, it is characterized in that: the initial organic carbon source concentration of bacteria culture medium is take organism quality densitometer as 1.0% ~ 5.0%, and the synchronized mixes culturing process is kept organic carbon source concentration 1.0% ~ 2.0%.
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| CN104560749A (en) * | 2013-10-29 | 2015-04-29 | 中国石油化工股份有限公司 | Method for open-type fast cultivation of oil-bearing microbes |
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| CN107746809A (en) * | 2017-12-13 | 2018-03-02 | 何红娣 | The method for improving algae bio amount |
| CN107746809B (en) * | 2017-12-13 | 2020-04-24 | 杭州渔森农业技术开发有限公司 | Method for increasing algae biomass |
| CN107841464A (en) * | 2017-12-15 | 2018-03-27 | 何红娣 | A kind of cultural method of algae |
| CN107841464B (en) * | 2017-12-15 | 2020-04-24 | 杭州渔森农业技术开发有限公司 | Algae culture method |
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| CN108004190B (en) * | 2018-01-19 | 2019-12-06 | 杭州渔森农业技术开发有限公司 | Method for increasing chlorella biomass by using bacillus |
| CN110484589A (en) * | 2019-09-25 | 2019-11-22 | 浙江海洋大学 | A method of condition of culture is improved to improve microalgae oil productive capacity |
| CN117303679A (en) * | 2023-11-30 | 2023-12-29 | 山东省水利科学研究院 | Water treatment device for efficiently removing nitrate in underground water |
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