CN103103129B - Production method for lipid through synchronous mixed culture of microbes - Google Patents
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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 the fat content of microalgae cell simultaneously.
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 there is the features such as renewable, readily biodegradable, storing and transporting security, the capability of antidetonance be good, can be used as the substitute of fossil energy.
Micro-algae is one of oil-containing microorganism, and the nutritive ingredients such as micro-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, have that algae is as chlorella slightly, its body fat acid content can account for 30%～60% of dry weight.Utilize the micro-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 there is huge market potential, and there is outstanding social value.
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 supplement 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 micro-algae industrial production photosynthetic organism reactor assembly " (CN03128138.9) all adopt and in photo-bioreactor system, add a kind of installation method, realize CO
2supply, also can realize certain oxygen simultaneously and resolve effect.Heterotrophic growth process is to utilize organic carbon source for the CO in the alternative autotrophy process of substrate
2carry out the accumulation of micro-algal biomass, Growth of Cells speed, but in cell, oil and fat accumulation level is lower.
Utilize other non-alga microbial and micro-algae to carry out the research of mixed culture also many, such as CN200910038908.0 discloses a kind of method that genus bacillus regulates and controls planktonic microalgae mixed culture, discovery when the method is used genus bacillus to carry out mixed culture to each micro-algae in co-culture system, 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 micro-algae to occur extreme advantage or inferior position, so the method is mainly used in controlling certain micro-algae spreading unchecked in water body.CN200910038910.8 discloses a kind of method that lactobacillus regulates and controls micro-algae mixed culture 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 micro-algaes, make each algae growth balanced, good stability, reaches the object to the denitrification dephosphorization of cultivation waste discharge by the synergy between bacterium, algae.Above-mentioned prior art is by thalline, to control the growth equilibrium of algae in water body, 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 micro-algae of autotrophy to carry out 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, simultaneously less on the fermenting process impact of facultative anaerobic bacteria, realize both and synchronously carry out.
Microbial synchronous mixed culture of the present invention is produced the method for grease, comprises following content:
(1) cultivate facultative anaerobic bacteria seed liquor;
(2) cultivate the micro-algae seed liquor of autotrophy;
(3) the micro-algae seed liquor of facultative anaerobic bacteria seed liquor and autotrophy is mixed, and carries out synchronized mixes cultivation in illuminated bio-reactor, in synchronized mixes culturing process, supplements 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 facultative anaerobic bacteria tunning is generally 1, ammediol and/or organic acid etc., the tunning that different facultative anaerobic bacterias obtains is different, different and different according to the kind of facultative anaerobic bacteria.It is method well known to those skilled in the art that facultative anaerobic bacteria seed liquor is cultivated, and as adopted stirring type bioreactor, adds substratum and facultative anaerobic bacteria, under suitable condition, cultivates.
In the inventive method, the micro-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 micro-algae seed liquor of autotrophy is cultivated, as adopts conventional air lift type illumination bio-reactor to carry out seed liquor cultivation.
In the inventive method, seed culture medium is respectively facultative anaerobic bacteria substratum and micro-algae SE substratum.
In the inventive method, in synchronized mixes culturing process, the initial access volume ratio of facultative anaerobic bacteria seed liquor and micro-algae seed liquor is 1:1～1:10.The initial medium that synchronized mixes is cultivated be take the required basic medium of facultative anaerobic bacteria as main, adds microalgae cell grow required inorganic salt and trace element (as the composition interpolation related substances by SE substratum) etc. simultaneously.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 general illumination bio-reactor that adopts of synchronized mixes cultivation, the general employing of condition that synchronized mixes is cultivated and the condition of the conditional likelihood of facultative anaerobic bacteria fermenting process, as temperature is generally 20 ℃～37 ℃, pH value is generally 6～9, is preferably 6.5～7.5 etc.In 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 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 facultative anaerobic bacteria cell fermentation
2) be self growth carbon source, maintain system osmotic pressure condition (pH value) more stably simultaneously, by supplementing organic carbon source assurance facultative anaerobic bacteria cell, constantly grow, 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 unit fermentation system in single oil-containing micro-algae culturing process, thereby laid a good foundation for the preparation of microbial oil.Simultaneously very little on the fermenting process impact of facultative anaerobic bacteria cell, can obtain required tunning simultaneously.Carbon source and tunning that micro-algae is used fermenting process all have tolerance, do not affect the growth of micro-algae and the accumulation of grease.
Accompanying drawing explanation
Fig. 1 is a kind of concrete technology schematic flow sheet of the present invention.
Wherein: 1-facultative anaerobic bacteria seed, the micro-algae seed of 2-, 3-facultative anaerobic bacteria seed liquor is cultivated reactor, the micro-algae seed liquor of 4-is cultivated reactor, 5-synchronized mixes pond, 6-mixed culture medium and acid-base neutralisation agent, 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 cultivation of facultative anaerobic bacteria seed liquor, adopting illuminated bio-reactor to carry out the micro-algae seed liquor of autotrophy cultivates, in stirring type bioreactor, comprise facultative anaerobic bacteria, fermentation using bacteria substratum, in illuminated bio-reactor, comprise micro-algae algae kind and autotrophy substratum.Before mixed culture, two kinds of microorganisms carry out separately respectively seed liquor cultivation, after seed liquor cultivation is qualified, through synchronized mixes pond two kinds of microorganism seed liquid being carried out to 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 grown for facultative anaerobic bacteria, and the CO that facultative anaerobic bacteria growth produces
2for micro algae growth.It is 1:1～1:10 that synchronized mixes is cultivated the grain weight ratio that two kinds of microorganisms of initial access are facultative anaerobic bacteria and micro-algae, 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 facultative anaerobic bacteria utilization, after facultative thin anaerobic bacterial fermentation, produces more carbonic acid gas, excretes and enters in culture systems, the CO that facultative anaerobic bacteria cell is discharged
2in dissolved state, by the microalgae cell in system, utilized immediately, 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 micro-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, facultative anaerobic bacteria cell is realized Fast Growth under 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 inorganic carbon source content in culture system is increased, and microalgae cell utilizes these inorganic carbon sources to carry out photosynthesis, obtains the growth of microalgae cell self.Simultaneously facultative anaerobic bacteria cell fermentation declines the pH value of system, and along with the growth of microalgae cell, utilizes the CO of dissolving
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, to add acid, alkali to realize the control of system pH by external source.
In present method, pH control acid neutralizing agent is HCl, H
2sO
4, HNO
3etc. conventional mineral acid, alkali neutralizing agent is NaOH, NaHCO
3, KOH, Ca(OH)
2, the conventional mineral alkali such as ammoniacal liquor.
In the inventive method, mixed cultivation process temperature is controlled as 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 nutrient solution volume change, passing into gas volume speed and culture systems dress liquid volume ratio is the unit gas volume that 0.1 vvm ~ 1.0 vvm(unit liquid volume per minute passes 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, according to nutrient solution, become sample analysis of components, the organic carbon source concentration level of detection system, carries out stream by the organic carbon source mother liquor configuring by spending rate and adds at any time.
As shown in Figure 1, facultative anaerobic bacteria seed 1 is cultivated reactor 3 incubation growth through facultative anaerobic bacteria seed liquor and is reached biomass OD=5.0～15.0, and micro-algae seed 2 is cultivated No. 4 incubation growth of reactor through micro-algae seed liquor and reached biomass OD=2.0～10.0.Cultured bacterium seed liquor and microalgae cell seed liquor are carried out to mixed culture in airlift agitation formula bioreactor, after adopting bacterial cell and microalgae cell to be prepared into seed liquor respectively in this mixed cultivation process, synchronously proceed in synchronized mixes pond 5, in suitable ratio, mix, and mixed culture medium and acid-base neutralisation agent 6 are added in mixing pit and are mixed, simultaneously through detection analyze mix after in culture systems pH value regulate pH to 6.5 ~ 7.5 of mixed-culture medium.
In the inventive method, bioreactor used is facultative anaerobic bacteria fermentation culture tank, is also the photosynthetic reactor that micro-algae is cultivated, 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 facultative anaerobic bacteria cell generates
2in culture system, as micro-algae, carry out photosynthetic carbon source, 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%, in process, by anaerobic bacterium Growth of Cells, consume, organic carbon source is constantly added through adding pump, and synchronized mixes culturing process maintains 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 300mL shaking flask, and used medium is anaerobic bacterium glycerin medium, after cultivation 20h, obtains required seed liquor.By in the facultative anaerobic bacteria seed liquor access 10L bio-reactor of cultivating, used medium is mixed culture medium, and reactor is 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 basis, press SE incubation period simultaneously 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 within 5 days, stop afterwards cultivating, 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 maintain content be 15g/L left and right, and cultivating residual glycerol content while finishing 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 (purchased from Inst. of Hydrobiology, Chinese Academy of Sciences's algae kind storehouse 1#) is carried out to the cultivation of light autotrophy in shaking flask, and used medium is SE substratum, cultivates and obtains required seed liquor after 2 days.Cultured chlorella seed liquid is seeded in 10L bioreactor.Reactor is airlift agitation formula, can realize the back-mixing of nutrient solution, and reactor is vitreum, and fluorescent lamp source is set, and automatically controls the switching time of setting light, forms the dark process conversion of light in chlorella culturing process.In reactor, there is temperature to control coil pipe, and pH, O
2and CO
2sensor.10L bioreactor culture condition is with shown in comparative example scheme 1.
Algae kind is Chlorella vulgaris, and the substratum of chlorella culturing process is with the mixed culture medium of scheme 1.
Ventilation adopts 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, cultivates and within 5 days, stops afterwards 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, with comparative example scheme 2, is used substratum separately to carry out single culture Cray Bai Shi pneumobacillus and chlorella in shaking flask, cultivates and obtains required seed liquor.Cultured Cray Bai Shi pneumobacillus seed liquor is seeded in the 10L bioreactor containing mixed culture medium together with chlorella cells seed liquor, reactor is 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 chlorella culturing process.In reactor, there is temperature to control coil pipe, and pH, O
2and CO
2sensor.Mixed culture medium, with the mixed culture medium of scheme 1, supplements organic carbon source by scheme 2 simultaneously.
Synchronized mixes is cultivated ventilation and is adopted 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 and within 5 days, stops afterwards 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 (purchased from Inst. of Hydrobiology, Chinese Academy of Sciences's algae kind storehouse 357#) seed liquor culture condition is with comparative example scheme 2, in shaking flask, use substratum separately to carry out single culture Cray Bai Shi pneumobacillus and grape algae, cultivate and obtain required seed liquor.Cultured Cray Bai Shi pneumobacillus seed liquor is seeded in the 10L bioreactor containing mixed culture medium together with grape frustule seed liquor, reactor is 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.In reactor, there is temperature to control coil pipe, and pH, O
2and CO
2sensor.Mixed culture medium, with the mixed culture medium of scheme 1, supplements organic carbon source by scheme 2 simultaneously.
Synchronized mixes is cultivated ventilation and is adopted 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 and within 5 days, stops afterwards 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 is as following table 1:
The synchronized mixes cultivation results of each embodiment of table 1
| Scheme | Microorganism dry weight | Fat content | Grease harvest yield | Facultative anaerobe fermentation level |
| 1 | 7.02g/L | 2.2% | 0.15g/L | 54.2g/L |
| 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 |
From above-mentioned data, can find out, the inventive method (scheme 3 and 4) has greatly improved the harvest yield of microorganism cells, and fat content is also 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 obtaining has obtained larger raising.In the unit fermentation system that synchronized mixes is cultivated, grease harvest yield is improved significantly.By the fermentation level correlation data of facultative anaerobic bacteria, can find out, in the situation that whether microalgae cell exists, klebsiella carries out anaerobically fermenting, and to produce the level of 1,3-PD substantially unaffected simultaneously.This inventive method is described, when realizing biomass collection and then obtaining bio-oil, can also realizes fermentation using bacteria and produce other target products, thereby improve the utilising efficiency of microorganism.
Claims (10)
1. microbial synchronous mixed culture is produced a method for grease, it is characterized in that comprising following content:
(1) cultivate facultative anaerobic bacteria seed liquor;
(2) cultivate the micro-algae seed liquor of autotrophy;
(3) the micro-algae seed liquor of facultative anaerobic bacteria seed liquor and autotrophy is mixed, and carries out synchronized mixes cultivation in illuminated bio-reactor, in synchronized mixes culturing process, supplements 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, facultative anaerobic bacteria utilize organic carbon source meta-bolites for 1,3-PD and/or organic acid.
3. in accordance with the method for claim 1, it is characterized in that: facultative anaerobic bacteria seed liquor is cultivated and adopted stirring type bioreactor, the micro-algae seed liquor of autotrophy is cultivated and is adopted air lift type illumination bio-reactor.
4. it is characterized in that in accordance with the method for claim 1: the micro-algae of autotrophy comprises chlorella, grape algae, little ring algae or diatom.
5. according to the method described in claim 1,2 or 4, it is characterized in that: in synchronized mixes culturing process, the initial access volume ratio of facultative anaerobic bacteria seed liquor and micro-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, add microalgae cell grow required inorganic salt and trace element simultaneously.
7. according to the method described in claim 1,2 or 6, it is characterized in that: 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 pH value is 6～9.
9. in accordance with the method for claim 1, it is characterized in that: facultative anaerobic bacteria seed liquor is cultured to biomass OD=5.0～15.0, micro-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: it is 1.0% ~ 5.0% that the initial organic carbon source concentration of bacteria culture medium be take organism quality densitometer, and synchronized mixes culturing process maintains organic carbon source concentration 1.0% ~ 2.0%.
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| CN104560749B (en) * | 2013-10-29 | 2017-08-22 | 中国石油化工股份有限公司 | A kind of method of open type fast culture oil-containing microorganism |
| 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 |
| 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 |
| 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 |
| CN108085283B (en) * | 2018-01-19 | 2019-12-06 | 杭州渔森农业技术开发有限公司 | method for culturing high-density algae through symbiosis of bacteria and algae |
| CN110484589B (en) * | 2019-09-25 | 2023-05-05 | 浙江海洋大学 | Method for improving culture conditions to improve oil production capacity of microalgae |
| 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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