CN102443542B - High-density culture process of autotrophic oil-producing microalgae - Google Patents
High-density culture process of autotrophic oil-producing microalgae Download PDFInfo
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Abstract
The invention relates to a high-density culture process of autotrophic oil-producing microalgae. A fermentation method combining a partition method and a two-step method is adopted, CO2 waste gas discharged from a thermal power plant is used as a carbon source, municipal sewage after living contaminant or miscellaneous algae filtering removal is used as a culture medium, the autotrophic oil-producing microalgae is cultured in a high-density way for growth, perfectly, sea salt is added into the municipal sewage for simulating sea water so that the permeation pressure is increased, the living contaminant or miscellaneous algae pollution is prevented, and the autotrophic oil-producing microalgae is selected from one kind or several kinds of materials from autotrophic chlorella, nannochloropsis salina, botryococcus braunii, chaetoceros gracile, marine green algae, isochrysis galbana, nitzschia closterium, phaeodactylum tricornutum, dunaliella and prism-shaped algae. When the high-density culture process is adopted, high removal efficiency of nitrogen, phosphorus and CO2 can be realized, in addition, the algae concentration, the grease content and the protein content are high, the double effects of environment protection and biological resources production are reached, the operation is simple, convenient and fast, the cost is low, no pollution is caused, social and economic integrated benefits are high, and the high-density culture process is suitable for large-scale popularization and application.
Description
Technical field
The present invention relates to the recycling economy technical field of comprehensive utilization of waste materials, energy-saving and emission-reduction green production, be particularly related to the recycling economy technical field of the comprehensive utilization of sewage and waste gas, energy-saving and emission-reduction green production, specifically refer to the high-density culture technology of the little algae of a kind of autotrophy produce oil.
Background technology
Energy scarcity, environmental pollution and short-commons are current three big world difficult problems.Little algae has important effect to solving an above-mentioned difficult problem.Little algae is a kind ofly can carry out photoautotrophic microorganism, is in the primary producer in the ecosystem, in whole material cycle effect huge, the production efficiency height, development prospect is tempting.And little algae is rich in grease, protein, carbohydrate, trace element and other biologically active substance, thereby it not only can be used as bait, and is one of main source of food and bioenergy.
The cutting method of the little algae of produce oil autotrophy and the two step method fermentation method that is coupled is a kind of technology that is applied in association area such as high-density culture, is that the semi-continuous culture systems of carrying out of utilizing seeding tank, photoreactor and long-pending oil tank to form realizes.Cultured little algae seed is inoculated in the photoreactor, after cultivating for some time, after being consumed of the nitrogen in the reactor, phosphorus, its content is during near national category-A emission standard, take out the part nutrient solution, wherein a part directly feeds long-pending oil tank, and the frond cell after another part concentrates inserts in the reactor again, add fresh sewage substratum again and cultivate, repeat aforesaid way afterwards always and carry out cutting method every the regular hour and get algae and two stage fermentation accumulation algae oil.Such working method not only provides a kind of semicontinuous cultivation, also increased the stability of culture systems, can under higher growth velocity, stably keep higher cell concn in the reactor, realize two-step fermentation simultaneously, micro algae growth is fast, the density height, thus make the waste water treatment efficiency of unit volume reactor increase substantially, and fat content height (production efficiency height).
Municipal wastewater is a kind of water body of nitrogenous, eutrophication that phosphorus is high, causes environmental pollution easily, can not directly discharge.At present, the method for wastewater treatment has three major types basically, and modern wastewater treatment mainly is divided into physical treatment process, method of chemical treatment and biological treatment 3 classes.1. physical treatment process: separate, reclaim the undissolved waste water treatment process that is the pollutent (comprising oil film and oil droplet) of suspended state in the waste water by physical action, can be divided into gravitational separation process, centrifugal separation and sieving and hold back method etc.Facture based on heat exchange principle also belongs to physical treatment process.2. method of chemical treatment: separate, remove the pollutent that is dissolving, colloidal state in the waste water or the waste water treatment process that is translated into innoxious substance with effect of mass transmitting by chemical reaction.Wherein use the processing unit of effect of mass transmitting both to have chemical action, associated physical action is arranged again, so also can from method of chemical treatment, branch away, become another kind for the treatment of process, be called physico-chemical processes.3. biological treatment: by the metabolism of microorganism, make the organic pollutant that is solution, colloid and fine suspended state in the waste water, be converted into the waste water treatment process of stable, harmless material.
Yet these several classes are removed in the process application and are all existed some defectives.Particularly, physical treatment process mainly reclaims the undissolved pollutent that is suspended state in the waste water, can't remove the pollutent of the little solubility of molecular weight, the energy consumption height, and efficient is low; The method of chemical treatment complex process, the pharmaceutical chemicals cost height of input, and might bring secondary pollution; Biological treatment mainly is converted into organism stable nontoxic inorganics, but the content of nitrogen phosphorus is still very high, produces a large amount of active sludge, causes secondary pollution easily, and equipment has high input the cost height.
CO in thermal power plant's high-concentration waste gas
2Concentration is about 10.0%～20.0% (V/V), directly is discharged in the atmosphere environment is polluted, and produces Greenhouse effect.
The complicated component of sewage, nutrient composition contents such as nitrogen, phosphorus are higher, directly as the easy pollution microbes of micro-algae culture medium (or algae).
Therefore, need provide the method for the above-mentioned refuse of a kind of comprehensive treating process, it can realize nitrogen, phosphorus and CO
2High remove efficient, and concentration of algae, grease and Protein content height, easy and simple to handle fast, low, pollution-free, the society of cost and economical synthesis benefit height.
Summary of the invention
The objective of the invention is to have overcome above-mentioned shortcoming of the prior art, the high-density culture technology of the little algae of a kind of autotrophy produce oil is provided, adopt this technology, can realize nitrogen, phosphorus and CO
2High remove efficient, and concentration of algae, grease and Protein content height, reach the double effects of protection environment and production Biological resources, easy and simple to handle fast, low, pollution-free, the society of cost and economical synthesis benefit height, be suitable for large-scale promotion application.
To achieve these goals, the high-density culture technology of the little algae of autotrophy produce oil of the present invention is characterized in, the fermentation process that adopts cutting method and two step method to be coupled is with the CO of power plant emission
2Waste gas is carbon source, and as substratum, the little algae of the described autotrophy produce oil of high-density culture grows with the municipal wastewater behind filtering and impurity removing bacterium or the assorted algae.
Wherein, the nitrogen content of municipal wastewater is generally 25.0mg/L～60.0mg/L, and phosphorus content is generally 3.0mg/L～15.0mg/L.
Preferably, in described municipal wastewater, add the sea salt simulated seawater, make its osmotic pressure increase the pollution that prevents assorted bacterium or assorted algae.
Preferably, described municipal wastewater is the high municipal wastewater of handling through sewage work's one-level of inorganic nitrogen, phosphorus content.
The little algae of described autotrophy produce oil is the little algae of ocean autotrophy.Preferably, the little algae of described autotrophy produce oil is selected from autotrophy chlorella (Chlorella autotuophica), little plan ball algae (Nannochloropsis salina), Blang's grape algae (BotyococcusBraunii), very thin Chaetoceros (Chaetoceros Gracilis), sea Chlorococcum (Halochlorococcum marmum), Isochrysis galbana (Isaochrysis galbana Parke), Nitzschia closterium minutissima (Nitzschia closterium), Phaeodactylum tricornutum (Phaeodactylum tricornutum Bohlin), one or more of Dunaliella salina (Dunaliella sp.) and prismatic algae (Nitzschia sp.).
More preferably, the little algae of described autotrophy produce oil is autotrophy chlorella, little plan ball algae or Blang's grape algae.
Preferably, the fermentation process that is coupled of described cutting method and two step method is: be 4.0 * 10 with cultured concentration
7Cfu/ml～3.0 * 10
8The little algae of high density autotrophy produce oil of cfu/ml is inoculated in the photoreactor, and inoculum size is 20%～60% (v/v), and culture temperature is 20～35 ℃, and pH is 6.5～8.5, and periodicity of illumination is 10h～16h, and intensity of illumination is 5000lux～15000lux, described CO
2The concentration of waste gas is 10.0%～20.0% (v/v), and air flow is 0.01～0.2VVm, cut apart cultivation before, cultivate after 1～3 day in the described photoreactor concentration of algae up to 1.0 * 10
8Cfu/ml～3.0 * 10
8Cfu/ml, and the nitrogen in the described photoreactor, the content of phosphorus carry out fermented liquid during near national category-A discharge water standard to be cut apart, take out 1/3～2/3 described fermented liquid and concentrate the acquisition concentrated broth, the described concentrated broth of a part is returned in the described photoreactor and in described photoreactor, add fresh municipal wastewater, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in the described photoreactor reaches 1.0 * 10
8Cfu/ml～3.0 * 10
8Cfu/ml.
More preferably, initial concentration of algae is 2.5 * 10 in the described photoreactor in inoculation back
7Cfu/ml～1.0 * 10
8Cfu/ml.
More preferably,, remaining described concentrated broth is charged in the long-pending oil tank of the sewage substratum that adds sea salt at described CO
2Two stage fermentation carries out oil and fat accumulation under the waste gas, collects frond afterwards and extracts algae oil, fat content 25%～40% (weight).
Further, the concentration of described sea salt is 2.0%～3.5% (weight), described CO
2The concentration of waste gas is 10.0%～20.0% (v/v), and air flow is 0.01～0.2VVm, and the time of oil and fat accumulation is 24h～72h.
More preferably, described photoreactor is the partition type fermentation culture system that the recirculation of part frond and frond are collected.
Beneficial effect of the present invention is specific as follows:
1, the present invention adopts the fermentation process that cutting method and two step method are coupled, and utilizes the high municipal wastewater of inorganic nitrogen, phosphorus content of sewage work to make substratum, feeds the high concentration CO that thermal power plant collects again
2Waste gas is carbon source, to remove nitrogen, phosphorus and the CO of the overwhelming majority
2, carry out the high-density cultured continuously and obtain algae oil and protein etc., can realize nitrogen, phosphorus and CO
2High remove efficient, and concentration of algae, grease and Protein content height, reach the double effects of protection environment and production Biological resources, easy and simple to handle fast, low, pollution-free, the society of cost and economical synthesis benefit height, be suitable for large-scale promotion application.
2, the present invention adds the sea salt simulated seawater in municipal wastewater, makes its osmotic pressure increase the pollution that prevents assorted bacterium (or algae), thereby can be with the CO of power plant emission
2Waste gas is carbon source,, cultivates little algae and grows as substratum with the municipal wastewater that adds sea salt, and novel uniqueness is suitable for large-scale promotion application.
3, raw material required for the present invention is waste pollutant entirely, carries out wastewater treatment, CO by comprehensive processing technique
2Reduce discharging and produce comprehensive benefit height such as microalgae grease, production cost is low, and the comprehensive cost of disposing of sewage is about 0.60～1.20 yuan/ton, is at half approximately than the cost of traditional water technology.
4, sewage disposal of the present invention and effects of energy saving and emission reduction are good, and nitrogen, tp removal rate are respectively 80.0%～95.0% (weight) and 85.0%～98.0% (weight), CO in power plant's waste gas in the sewage
2Concentration has reduced more than 55.0～80.0% (v/v), and the discharging that has reduced a large amount of greenhouse gases realizes green production, has good social benefit.
5, the present invention can realize high-density culture and high yield algae oil, and little concentration of algae can reach 1.0 * 10
8Cfu/ml～3.0 * 10
8Cfu/ml, fat content are 25.0%～40.0% (weight), imitate better than traditional fermentation method, obtain high economic benefit.
6, operation is simple in the present invention, and equipment maintenance is convenient, and the large-scale industrial production process is from traditional water treatment process and the CO of power plant
2The reforming equipment investment of collection process is less.
7, suitability of the present invention is wide: be not subjected to the restriction of ambient conditions, algae kind etc., be applicable to the high-density culture of the little algae of multiple ocean autotrophy.
Description of drawings
Fig. 1 be adopt the present invention to cultivate that the autotrophy chlorella is disposed of sewage and little gonidium in fatty accumulation effect synoptic diagram.
Fig. 2 be adopt the present invention to cultivate that little plan ball algae is disposed of sewage and little gonidium in fatty accumulation effect synoptic diagram.
Fig. 3 be adopt the present invention to cultivate that Blang's grape algae is disposed of sewage and little gonidium in fatty accumulation effect synoptic diagram.
Fig. 4 A is cut apart cultivation, cultured continuously and the batch culture nitrogen removal effect synoptic diagram of disposing of sewage.
Fig. 4 B is cut apart cultivation, cultured continuously and the batch culture phosphorus removal effect synoptic diagram of disposing of sewage.
Fig. 4 C is cut apart cultivation, cultured continuously and the batch culture CO that disposes of sewage
2Transformation efficiency effect synoptic diagram.
Fig. 4 D is cut apart cultivation, cultured continuously and the batch culture oil and fat accumulation effect synoptic diagram of disposing of sewage.
Fig. 4 E is cut apart cultivation, cultured continuously and batch culture to dispose of sewage that concentration of algae changes synoptic diagram in the process.
Fig. 5 is the synoptic diagram that concerns that adopts sodium chloride concentration of the present invention (% (weight)) and microbiological contamination rate (%).
Embodiment
In order more to be expressly understood technology contents of the present invention, describe in detail especially exemplified by following examples.
The transformation efficiency of nitrogen, phosphorus is that clearance is an important indicator weighing this process spent water treatment effect in the waste water, and method of calculation are as follows:
The transformation efficiency of nitrogen, phosphorus (%)=(C
1-C
2) * 100%/C
1
In the formula: C
1The content of initial nitrogen, phosphorus in the-wastewater medium, mg;
C
2The content of nitrogen, phosphorus in the-processed waste water nutrient solution, mg.
CO in the waste gas
2Transformation efficiency be CO
2CER is an important indicator weighing this technology waste gas treatment effect, and method of calculation are as follows:
CO
2Transformation efficiency (%)=(C
1-C
2) * 100%/C
1
In the formula: C
1CO in the-power plant waste gas
2Volumetric concentration, %;
C
2-CO in the waste gas after handling
2Volumetric concentration, %.
The content of microalgae grease or the yield of grease are to weigh an important indicator of the overall economic efficiency of this technology, and method of calculation are as follows:
The yield of grease (%)=m
1/ m
0* 100%
In the formula: m
0The frond dry weight of-extraction grease, g;
m
1The quality of-thick grease, g.
Little algae culturing process, it is huge to prevent that assorted bacterium (algae) from polluting the production influence of entire treatment process and microalgae grease, protein etc., is one of key factor that determines the fermentation success or failure, and method of calculation are as follows:
Microbiological contamination rate (%)=N
2/ N
1* 100%
In the formula: N
1-handle batch number of times;
N
2-pollute batch, number of times.
As substratum, be 4.0 * 10 with cultured concentration with the municipal wastewater behind filtering and impurity removing bacterium or the assorted algae (nitrogen content 30.0mg/L, phosphorus content are 5.0mg/L)
7Cfu/ml～3.0 * 10
8The high density autotrophy chlorella (Chlorellaautotuophica) of cfu/ml is inoculated in the photoreactor, and inoculum size is 40% (v/v), and culture temperature is 28 ℃, and pH is 7.0, and periodicity of illumination is 12h, and intensity of illumination is 10000lux, with the CO of power plant emission
2Waste gas (CO
2Concentration is 13.0% (v/v)) for carbon source, air flow is 0.2VVm, cut apart cultivation before, cultivate behind the 48h in the described photoreactor concentration of algae up to 1.5 * 10
8Cfu/ml, and the nitrogen in the described photoreactor, the content of phosphorus carry out fermented liquid during near national category-A discharge water standard to be cut apart, take out 1/3～2/3 described fermented liquid and concentrate the acquisition concentrated broth, the described concentrated broth of a part is returned in the described photoreactor and in described photoreactor, add fresh municipal wastewater, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in the described photoreactor reaches 1.9 * 10
8Cfu/ml.Remaining described concentrated broth charged into to have in the long-pending oil tank of sewage substratum that sea salt concentration is 2.5% (weight) at air flow be the CO of 0.2VVm
2Concentration is the described CO of 13.0% (v/v)
2Two stage fermentation carries out oil and fat accumulation 48h under the waste gas, collects frond afterwards and extracts algae oil.
Utilize above-mentioned technology to cultivate autotrophy chlorella (Chlorella autotuophica), nitrogen, phosphorus and CO
2Clearance and oil and fat accumulation relation see shown in Figure 1.
By Fig. 1 analysis as can be known, in 0～48h culturing process, along with incubation time prolongs, the autotrophy chlorella is to nitrogen, phosphorus and CO
2Spending rate accelerate, the fastest at the 48h spending rate, reaches maximum value and be respectively 90.0% (weight), 93.0% (weight) and 70% (v/v), and frond concentration reaches 1.5 * 10
8Cfu/ml is cut apart subsequently, takes out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keeps substratum total amount 100%, and a part feeds long-pending oil tank Secondary Fermentation and amasss oily 48h in addition.Finally, concentration of algae reaches 1.9 * 10
8Cfu/ml, the content of algae oil are up to 35% (weight).Cut apart cultivation again by above-mentioned later on, semicontinuous cultivation is cut apart 10 times continuously.
As substratum, be 4.0 * 10 with cultured concentration with the municipal wastewater behind filtering and impurity removing bacterium or the assorted algae (nitrogen content 30.0mg/L, phosphorus content are 5.0mg/L)
7Cfu/ml～3.0 * 10
8The little plan ball of the high density of cfu/ml algae (Nannochloropsissalina) is inoculated in the photoreactor, and inoculum size is 20% (v/v), and culture temperature is 20 ℃, and pH is 8.5, and periodicity of illumination is 16h, and intensity of illumination is 15000lux, with the CO of power plant emission
2Waste gas (CO
2Concentration is 20.0% (v/v)) for carbon source, air flow is 0.1VVm, cut apart cultivation before, cultivate behind the 48h in the described photoreactor concentration of algae up to 3.0 * 10
8Cfu/ml, and the nitrogen in the described photoreactor, the content of phosphorus carry out fermented liquid during near national category-A discharge water standard to be cut apart, take out 1/3～2/3 described fermented liquid and concentrate the acquisition concentrated broth, the described concentrated broth of a part is returned in the described photoreactor and in described photoreactor, add fresh municipal wastewater, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in the described photoreactor reaches 2.95 * 10
8Cfu/ml.Remaining described concentrated broth charged into to have in the long-pending oil tank of sewage substratum that sea salt concentration is 2.0% (weight) at air flow be the CO of 0.1VVm
2Concentration is the described CO of 20.0% (v/v)
2Two stage fermentation carries out oil and fat accumulation 48h under the waste gas, collects frond afterwards and extracts algae oil.
Utilize above-mentioned technology to cultivate little plan ball algae (Nannochloropsis salina), nitrogen, phosphorus removal effect, CO
2Transformation efficiency and oil and fat accumulation relation see shown in Figure 2.
By Fig. 2 analysis as can be known, in 0～48h culturing process, along with incubation time prolongs, little plan ball algae is to nitrogen, phosphorus and CO
2Spending rate accelerate, the fastest at the 48h spending rate, reaches maximum value and be respectively 95.0% (weight), 97.0% (weight) and 80% (v/v), and frond concentration reaches 3.0 * 10
8Cfu/ml is cut apart subsequently, takes out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keeps substratum total amount 100%, and a part feeds long-pending oil tank Secondary Fermentation and amasss oily 48h in addition.Finally, concentration of algae reaches 2.95 * 10
8Cfu/ml, the content of algae oil are up to 30% (weight).Cut apart cultivation again by above-mentioned later on, semicontinuous cultivation is cut apart 10 times continuously.
As substratum, be 4.0 * 10 with cultured concentration with the municipal wastewater behind filtering and impurity removing bacterium or the assorted algae (nitrogen content 30.0mg/L, phosphorus content are 5.0mg/L)
7Cfu/ml～3.0 * 10
8High density Blang's grape algae (BotyococcusBraunii) of cfu/ml is inoculated in the photoreactor, and inoculum size is 60% (v/v), and culture temperature is 35 ℃, and pH is 6.5, and periodicity of illumination is 10h, and intensity of illumination is 5000lux, with the CO of power plant emission
2Waste gas (CO
2Concentration is 10.0% (v/v)) for carbon source, air flow is 0.01VVm, cut apart cultivation before, cultivate behind the 48h in the described photoreactor concentration of algae up to 1.0 * 10
8Cfu/ml, and the nitrogen in the described photoreactor, the content of phosphorus carry out fermented liquid during near national category-A discharge water standard to be cut apart, take out 1/3～2/3 described fermented liquid and concentrate the acquisition concentrated broth, the described concentrated broth of a part is returned in the described photoreactor and in described photoreactor, add fresh municipal wastewater, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in the described photoreactor reaches 1.3 * 10
8Cfu/ml.Remaining described concentrated broth charged into to have in the long-pending oil tank of sewage substratum that sea salt concentration is 3.5% (weight) at air flow be the CO of 0.01VVm
2Concentration is the described CO of 10.0% (v/v)
2Two stage fermentation carries out oil and fat accumulation 48h under the waste gas, collects frond afterwards and extracts algae oil.
Utilize above-mentioned technology to cultivate Blang grape algae (Botyococcus Braunii), nitrogen, phosphorus and CO
2Removal effect and oil and fat accumulation relation are seen shown in Figure 3.
By Fig. 3 analysis as can be known, in 0～48h culturing process, along with incubation time prolongs, Blang's grape algae (BotyococcusBraunii) is to nitrogen, phosphorus and CO
2Spending rate accelerate, the fastest at the 48h spending rate, reaches maximum value and be respectively 82.0% (weight), 77.0% (weight) and 65% (v/v), and frond concentration reaches 1.0 * 10
8Cfu/ml is cut apart subsequently, takes out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keeps substratum total amount 100%, and a part feeds long-pending oil tank Secondary Fermentation and amasss oily 48h in addition.Finally, concentration of algae reaches 1.3 * 10
8Cfu/ml, the content of algae oil are up to 40% (weight).Cut apart cultivation again by above-mentioned later on, semicontinuous cultivation is cut apart 10 times continuously.
Adopt batch fermentation, continuously ferment that (nitrogen content is 30.0mg/L to the municipal wastewater of three kinds of training methods of fermentation after identical culture condition is with filtering and impurity removing bacterium or assorted algae that is coupled with cutting method-two step method, phosphorus content is 5.0mg/L) as substratum, be 4.0 * 10 with cultured concentration
7Cfu/ml～3.0 * 10
8The little algae of high density produce oil autotrophy of cfu/ml is inoculated in the photoreactor, and inoculum size is 35% (v/v), and culture temperature is 28 ℃, and pH is 7.0, and periodicity of illumination is 12h, and intensity of illumination is 10000lux, with the CO of power plant emission
2Waste gas (CO
2Concentration is 13.0% (v/v)) be that carbon source, air flow are to cultivate the autotrophy chlorella under the 0.1VVm, cultivate 480h, nitrogen, phosphorus removal effect, CO
2Transformation efficiency (% (v/v)) and oil and fat accumulation relation see shown in Fig. 4 A-4E.
Learnt that by Fig. 4 A analysis along with the time lengthening of cultivating, three kinds of training methods are accelerated the utilization of nitrogen.In the culturing process that cutting method-two step method is coupled, in the culturing process of 0～48h, the utilization of nitrogen speeds up and carries out fermented liquid at 48h and cuts apart, take out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keep substratum total amount 100%, after a part feeds the long-pending oily 48h of long-pending oil tank Secondary Fermentation in addition, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart 10 batches continuously.The clearance of nitrogen is up to 90% (weight).In the culture of continuous cultivation, constantly accelerate in the speed of utilizing of 0～96h nitrogen, clearance reaches 80% (weight) subsequently, and also unstable at 96～288h, nitrogen removal efficiency tends towards stability in 80% (weight) subsequently.In the batch culture process, 0～96h nitrogen utilize speed constantly accelerate and clearance up to 95% (weight), remain on this level subsequently always.
Learnt that by Fig. 4 B analysis along with the time lengthening of cultivating, three kinds of training methods are accelerated the utilization of phosphorus.In the culturing process that cutting method-two step method is coupled, in 0～48h culturing process, the utilization of nitrogen speeds up and carries out fermented liquid at 48h and cuts apart, take out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keep substratum total amount 100%, after a part feeds the long-pending oily 48h of long-pending oil tank Secondary Fermentation in addition, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart 10 batches continuously.The clearance of phosphorus is up to 85% (weight), and clearance is stabilized in 90% (weight) subsequently.In the culture of continuous cultivation, constantly accelerate in the speed of utilizing of 0～72h phosphorus, clearance reaches 80% (weight) subsequently, and 72～216h is also unstable, and tp removal rate tends towards stability in 80% (weight) subsequently.In the batch culture process, 0～120h phosphorus utilize speed constantly accelerate and clearance up to 95% (weight), remain on this level subsequently always.
Learn that by Fig. 4 C analysis along with the time lengthening of cultivating, three kinds of training methods are to CO
2Utilization accelerate.In the culturing process that cutting method-two step method is coupled, in the culturing process of 0～48h, CO
2Utilization speed up and carry out fermented liquid at 48h and cut apart, take out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keep substratum total amount 100%, after a part feeds the long-pending oily 48h of long-pending oil tank Secondary Fermentation in addition, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart 10 batches continuously.CO
2Clearance up to 70% (v/v), clearance is stabilized in 70% (v/v) subsequently.In the culture of continuous cultivation, at 0～72h CO
2Utilize speed constantly to accelerate, clearance reaches 65% (v/v) subsequently, and is also unstable at 72～240h, subsequently CO
2Clearance tends towards stability at 65% (v/v).In the batch culture process, at 0～96h CO
2Utilize the continuous quickening of speed and clearance up to 75% (v/v), keep stable at 96～144h, utilize speed to slow down gradually subsequently, final CO
2Clearance has only 30% (v/v).
Learnt that by Fig. 4 D analysis along with the time lengthening of cultivating, three kinds of training methods are accelerated the utilization of phosphorus.In the culturing process that cutting method-two step method is coupled, in the culturing process of 0～48h, oil and fat accumulation speeds up and carries out fermented liquid at 48h and cuts apart, take out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, keep substratum total amount 100%, after a part feeds the long-pending oily 48h of long-pending oil tank Secondary Fermentation in addition, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart 10 batches continuously.The fat content of frond in the oil and fat accumulation jar is up to 35% (weight), and be basicly stable in 35% (weight) subsequently.In the culture of continuous cultivation, 0～192h oil and fat accumulation speed constantly accelerate and fat content up to 25% (weight), the frond content of oil and grease tends towards stability in 35% (weight) after the 192h.In the batch culture process, 0～144h frond oil and fat accumulation speed constantly accelerate and the frond fat content up to 30% (weight), remain on this level subsequently always.
Learnt that by Fig. 4 E analysis along with the time lengthening of cultivating, three kinds of training methods are accelerated the utilization of nitrogen.In the culturing process that cutting method-two step method is coupled, in the culturing process of 0～48h, the frond speed of growth is accelerated and is carried out fermented liquid at 48h (frond concentration is 1.5 * 10
8Cfu/ml) cut apart, take out after wherein 2/3 fermented liquid concentrates, with wherein 1/3 Returning reactor and charge into salt-free fresh municipal wastewater, maintenance substratum total amount 100%, after a part feeds the long-pending oily 48h of long-pending oil tank Secondary Fermentation in addition, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart 10 batches continuously.Cut apart cultivation back frond concentration and maintain 1.85 * 10
8Cfu/ml.In the culture of continuous cultivation, constantly accelerate in 0～96h frond speed of growth, frond concentration reaches 1.5 * 10
8Cfu/ml, also unstable at 96～240h, frond concentration tends towards stability 1.5 * 10 subsequently
8Cfu/ml.In the batch culture process, reach 1.5 * 10 in the continuous quickening of 0～96h frond growth and frond concentration
8Cfu/ml maintains this level at 96h～144h, and frond concentration begins to descend subsequently, finally is 8.0 * 10
7Cfu/ml.
Little algae culturing process, it is huge to prevent that assorted bacterium (algae) from polluting the production influence of entire treatment process and microalgae grease, protein etc., is one of key factor that determines the fermentation success or failure.The sewage substratum of cultivating the algae kind is not sterilized and is directly added different concentrations of sodium chloride and prevent living contaminants, utilize the technology identical with embodiment 3 to cultivate the autotrophy chlorella respectively, little plan ball algae and three kinds of little algaes of Blang's grape algae, carry out 20 batches operation continuously, the concentration of sodium-chlor (% (weight)) is seen shown in Figure 5 with the relation of microbiological contamination rate (%).
Learnt by Fig. 5 analysis, in the low salt concn scope, increase along with salt concn, the microbiological contamination rate is increasing, and the concentration about 1% is that pollution rate is the highest, may be more suitable for a lot of microbial growths in the physiology salinity range, when the concentration of sodium-chlor is higher than 3% (weight), near seawater, pollution rate almost is zero, and preventive effect is remarkable.
The fermentation process that the present invention utilizes municipal wastewater to adopt cutting method and two step method to be coupled carries out little algae high-density culture, thereby disposes of sewage and produce algae wet goods active substance, and comprise step: at first (concentration is 4.0 * 10 to high density
7Cfu/ml～3.0 * 10
8Cfu/ml) little algae (inoculum size is 20%～60% (v/v)) seed is transferred and is cultivated into the fermentor tank of salt-free sewage substratum, culture temperature is 20～35 ℃, pH6.5～8.5, periodicity of illumination is that (intensity is that 5000lux～15000lux), air flow is 0.01～0.2VVm (waste gas CO of thermal power plant to 10h～16h
2Concentration is 10.0%～20.0% (v/v)), frond concentration reaches 1.0 * 10 in reactor
8Cfu/ml～3.0 * 10
8During cfu/ml, fermented liquid is cut apart, each 1/3～2/3 of fermented liquid that takes out, after it is concentrated, to wherein also in reactor, add fresh salt-free municipal wastewater in a small amount of back light reactor, keep substratum total amount 100%, remaining part charges into the long-pending oil tank middle and high concentration CO that sea salt concentration is the sewage substratum of 2.0%～3.5% (weight)
2(air flow is 0.01～0.2VVm, CO
2Concentration is 10.0%～20.0% (v/v)) descend two stage fermentation to carry out oil and fat accumulation 24h～72h, collect frond afterwards and extract algae oil.Semicontinuous cultivation is cut apart continuously, can reach 8～15 batches.Nitrogen in the sewage and phosphorus and content have reduced by 80.0%～98.0% (weight) and 75.0%～95.0% (weight), CO respectively as a result
2Concentration reduces by 55.0～80.0% (v/v), and final concentration of algae is 1.0 * 10
8Cfu/ml～3.0 * 10
8Cfu/ml, fat content are 25.0%～40.0% (weight).
The fermentation process that utilizes municipal wastewater to adopt cutting method and two step method to be coupled of the present invention carries out the sewage disposal of little algae high-density culture and produces the technological design of algae oil ingenious, ferments with traditional batch and compares this technology nitrogen, phosphorus and CO with continuous fermentation process
2Remove efficient height, concentration of algae, grease and Protein content height, easy and simple to handle fast, cost is low, pollution-free, comprehensive benefit is high, be suitable for large-scale industrialization production, particularly nitrogen, phosphorus and CO
2Removal efficient height is conducive to the processing of sewage, and concentration of algae and fat content height are conducive to the production of biofuel.
To sum up, the high-density culture technology of the little algae of autotrophy produce oil of the present invention can realize nitrogen, phosphorus and CO
2High remove efficient, and concentration of algae, grease and Protein content height, reach the double effects of protection environment and production Biological resources, easy and simple to handle fast, low, pollution-free, the society of cost and economical synthesis benefit height, be suitable for large-scale promotion application.
In this specification sheets, the present invention is described with reference to its certain embodiments.But, still can make various modifications and conversion obviously and can not deviate from the spirit and scope of the present invention.Therefore, specification sheets and accompanying drawing are regarded in an illustrative, rather than a restrictive.
Claims (10)
1. the high-density cultivation method of the little algae of autotrophy produce oil is characterized in that, the fermentation process that adopts cutting method and two step method to be coupled is with the CO of power plant emission
2Waste gas is carbon source, described CO
2The concentration of waste gas is 10.0%～20.0%(v/v), and as substratum, the little algae of the described autotrophy produce oil of high-density culture grows with the municipal wastewater behind filtering and impurity removing bacterium or the assorted algae, and the initial concentration of the little algae of described autotrophy produce oil is 4.0 * 10
7Cfu/ml～3.0 * 10
8Cfu/ml is cultured to the content of the interior nitrogen of described photoreactor, phosphorus near national category-A discharge water standard.
2. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 1 is characterized in that, adds the sea salt simulated seawater in described municipal wastewater, makes its osmotic pressure increase the pollution that prevents assorted bacterium or assorted algae.
3. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 1 is characterized in that, described municipal wastewater is the high municipal wastewater of handling through sewage work's one-level of inorganic nitrogen, phosphorus content.
4. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 1, it is characterized in that the little algae of described autotrophy produce oil is selected from one or more of autotrophy chlorella, little plan ball algae, Blang's grape algae, very thin Chaetoceros, extra large Chlorococcum, Isochrysis galbana, Nitzschia closterium minutissima, Phaeodactylum tricornutum, Dunaliella salina and prismatic algae.
5. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 4 is characterized in that, the little algae of described autotrophy produce oil is autotrophy chlorella, little plan ball algae or Blang's grape algae.
6. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 1 is characterized in that the fermentation process that described cutting method and two step method are coupled is: be 4.0 * 10 with cultured concentration
7Cfu/ml～3.0 * 10
8The little algae of high density autotrophy produce oil of cfu/ml is inoculated in the photoreactor, inoculum size is 20%～60%(v/v), culture temperature is 20～35 ℃, pH is 6.5～8.5, periodicity of illumination is 10h～16h, and intensity of illumination is 5000lux～15000lux, and air flow is 0.01～0.2VVm, before cutting apart cultivation, the interior concentration of algae of described photoreactor is up to 1.0 * 10 behind cultivation 24～72h
8Cfu/ml～3.0 * 10
8Cfu/ml, and the nitrogen in the described photoreactor, the content of phosphorus carry out fermented liquid during near national category-A discharge water standard to be cut apart, take out 1/3～2/3 described fermented liquid and concentrate the acquisition concentrated broth, the described concentrated broth of a part is returned in the described photoreactor and in described photoreactor, add fresh municipal wastewater, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in the described photoreactor reaches 1.0 * 10
8Cfu/ml～3.0 * 10
8Cfu/ml.
7. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 6 is characterized in that, initial concentration of algae is 2.5 * 10 in the described photoreactor in inoculation back
7Cfu/ml～1.0 * 10
8Cfu/ml.
8. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 6 is characterized in that, remaining described concentrated broth is charged in the long-pending oil tank of the sewage substratum that adds sea salt at described CO
2Two stage fermentation carries out oil and fat accumulation under the waste gas, collects frond afterwards and extracts algae oil, fat content 25%～40%(weight).
9. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 8 is characterized in that, the concentration of described sea salt is 2.0%～3.5%(weight), described CO
2The concentration of waste gas is 10.0%～20.0%(v/v), and air flow is 0.01～0.2VVm, and the time of oil and fat accumulation is 24h～72h.
10. the high-density cultivation method of the little algae of autotrophy produce oil according to claim 6 is characterized in that, described photoreactor is the partition type fermentation culture system that the recirculation of part frond and frond are collected.
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| CN103642694A (en) * | 2013-12-13 | 2014-03-19 | 嘉兴泽元生物制品有限责任公司 | High-yield cultivation method of botryococcus |
| CN105331646A (en) * | 2014-08-06 | 2016-02-17 | 中国科学院大连化学物理研究所 | Culture method for improving oil production of microalgae |
| CN104498362A (en) * | 2014-12-26 | 2015-04-08 | 甘肃德福生物科技有限公司 | Biomass cultivation method for chlorella |
| CN105063131B (en) * | 2015-08-27 | 2020-02-14 | 江南大学 | Saccharifying enzyme fermentation process for improving use efficiency of fermentation tank by fermentation liquor division method |
| CN105174476B (en) * | 2015-09-28 | 2017-08-01 | 中国科学院城市环境研究所 | Activated sludge and microalgae coupling particle system and its structure and operation method for wastewater treatment |
| CN109467191A (en) * | 2018-12-10 | 2019-03-15 | 中国石油化工股份有限公司 | The method for handling high nitre waste water using microalgae |
| CN109650555B (en) * | 2019-01-17 | 2021-09-21 | 东北师范大学 | Method for treating phosphorus-containing wastewater by using microalgae |
| CN110387332A (en) * | 2019-07-09 | 2019-10-29 | 天津大学 | It is a kind of to be accumulated using artificial municipal wastewater culture chlorella and extract the research method of protein |
| CN113005041B (en) * | 2021-03-23 | 2022-05-06 | 山东大学 | Diamond algae, culture method thereof and application thereof in super-salt oil production |
| CN114015493B (en) * | 2021-09-26 | 2024-02-20 | 上海师范大学 | Method for extracting grease from algae |
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