CN102586116B - Common chlorella as well as culturing method and application thereof - Google Patents
Common chlorella as well as culturing method and application thereof Download PDFInfo
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Abstract
The invention provides a common chlorella, which is chlorella C9-JN2010 with the preservation number of CCTCC NO: M2010373; and the invention further provides a culturing method of the common chlorella. The culturing method comprises the following steps: using secondary outlet water, which is obtained by treating municipal sewage and/or industrial sewage in a sewage treatment plant, as a culture medium; and culturing the chlorella after the chlorella is inoculated, wherein the initial inoculating density is 1.0 multiplied by 107 cfu/ml-6.0 multiplied by 107 cfu/ml, and the culturing condition is that the light cycle is 10h:14h-16h:8h, the light intensity is 5000-15000 lux, the culturing temperature is 20-30 DEG C, the pH value is 6.5-8.5, the CO2 gas throughput is 0.01-0.2 VVm, and the CO2 concentration is 15.0%-30.0% (v/v). The invention further provides an application of the common chlorella in sewage treatment and oil production. The common chlorella provided by the invention has the advantages of strong CO2 tolerance and high production strength; through the adoption of the culturing method provided by the invention, the sewage treatment and the production of the biological energy are combined with other side products (such as proteins, cosmetic raw materials, and animal feeds), so that the production cost is reduced, the waste resources are comprehensively utilized as well as the energy-saving and emission reducing green production is realized; and the culturing method provided by the invention has the advantages of skillful and special design as well as suitability for being popularized and applied on a large scale.
Description
Technical field
The present invention relates to the Alga technology field, more specifically, relate to the chlorella technical field, refer to especially a kind of Chlorella vulgaris and cultural method thereof and application.
Background technology
Energy scarcity, environmental pollution and short-commons are current three big world difficult problems.Global warming is owing to the main greenhouse gas CO in atmosphere
2Increase, with ubiquitous by Eutrophication materials (as, nitrogen and phosphorus etc.) and the water that causes of other pollutent pollute and belong to one of main environmental problem.
Little algae has very 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, production efficiency is high, 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.Little algae has that growth velocity is fast, production intensity is high, the light utilization efficiency advantages of higher, utilizes little algae more and more to receive many-sided concern as the starting material of bioenergy.At present, existing little algae as the bioenergy raw material is mainly CO
2Tolerance is relatively poor, and production intensity is low, and adopts the substratum of high density to cultivate, and production cost is too high, and industrialization is obstructed.
Therefore, need to provide a kind of little algae, its CO
2Tolerance is strong, and production intensity is high, and necessary sewage disposal and produce bioenergy and other byproducts (as, protein, cosmetic material and animal-feed etc.) combine, reduce production costs, can reach the comprehensive utilization of waste resource, realize the green production of energy-saving and emission-reduction.
Summary of the invention
Main purpose of the present invention is exactly the problems and shortcomings for above existence, and a kind of Chlorella vulgaris and cultural method thereof and application are provided, this Chlorella vulgaris CO
2Tolerance is strong, production intensity is high, its cultural method sewage disposal and produce bioenergy and other byproducts (as, protein, cosmetic material and animal-feed etc.) combine, reduce production costs, reach the comprehensive utilization of waste resource, realize the green production of energy-saving and emission-reduction, design ingenious uniqueness, be suitable for large-scale promotion application.
In a first aspect of the present invention, a kind of Chlorella vulgaris is provided, be characterized in, described Chlorella vulgaris is chlorella C9-JN2010, deposit number is: CCTCC NO:M 2010373.
Described Chlorella vulgaris is to separate and pass through artificially breeding to obtain from the seawater of Yellow Sea, called after chlorella C9-JN2010, for green, single spherical frond, wall thin, formal name used at school is categorized as a kind of Chlorella vulgaris (Chlorella vulgaris), it is deposited in Chinese Typical Representative culture collection center (China Center for Type Culture Collection on December 31st, 2010, be called for short CCTCC), the culture presevation registration number is CCTCC NO:M 2010373.
The isolation and screening mode of above-mentioned Chlorella vulgaris is: get the Huanghai Seawater 100mL that contains chlorella, filtrate is put into centrifuge tube, centrifugal 15min under 5000r/min after first removing large-scale particulate matter with filter paper filtering.The evacuation supernatant liquor, after adding base culture base, vibration suspends frustule.Again through the centrifugal supernatant of going.After above-mentioned cleaning frustule step triplicate.After getting 10mL algae suspension first carry out illumination cultivation 5d in the f/2 substratum, sampling 10mL is re-seeded in fresh f/2 substratum again and carries out illumination cultivation.After repetition illumination cultivation 3 times, get the 1mL culture and carry out 10
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6Gradient dilution is got on the flat board that 0.2mL coats the f/2 solid medium, wherein, agar content is 2%, is placed in 25 ℃, and intensity of illumination is 4000lux, periodicity of illumination is 12h: 12h, relative humidity is to cultivate in 75% growth cabinet, and three to surrounding, grows green single algae on solid plate and falls, picking list algae is dropped on and carries out plate streaking on the flat board of f/2 solid medium and separate subsequently, repeatedly after 5 times, obtain pure strain, be Chlorella vulgaris after identifying.Subsequently, the Chlorella vulgaris with above acquisition carries out seed selection again.The take the logarithm chlorella of phase growth is diluted respectively 10,100,1000 and 10000 times and coats on the f/2 solid medium, and at 25 ℃, illumination 4000lux, periodicity of illumination cultivate under the condition of 12h: 12h.After 7d, picking deep green mono-clonal phycomycete falls to being inoculated in liquid medium, selects the 100mL triangular flask, in the 50mL nutrient solution is arranged.Altogether picking 120 mono-clonal phycomycetes fall, with front identical condition under cultivate, shake every day 2 times.After 14d, remove adherent serious and color jaundice, pick out 50 dark green, the not adherent chlorellas of strain algae liquid color.This 50 chlorella is inoculated in respectively new liquid medium, 2 Duplicate Samples, under same culture conditions, rotating speed is 100rpm, with original chlorella vulgaris in contrast, calculates their cell density every day.After 8d, according to its speed of growth and final biomass, pick out 15 chlorellas, the speed of growth and biomass are all greater than contrast, and algae liquid is dark green, without adherent phenomenon.15 chlorellas are inoculated in respectively new liquid medium, use the 250mL triangular flask instead, in-built 150mL nutrient solution arranges 3 Duplicate Samples, and culture condition is the same.With original algae kind in contrast, calculate cell density every day.After cultivating 14d, respectively get 100mL, 4 ℃ centrifugal and remove supernatant, remaining frond is dry in freeze drier, and cold well temperature is-35 ℃, weighs after dry 48h, after this every 1h weighs 1 time, until constant weight.Survey thick grease with gas-chromatography, comprehensive fat content and the speed of growth are picked out the strain of setting out of ultraviolet mutagenesis.Obtain the strain of good chlorella C9-JN2010 algae.
In a second aspect of the present invention, a kind of Chlorella vulgaris cultural method is provided, be characterized in, in photoreactor, secondary effluent after sewage work processes is as substratum take municipal wastewater and/or industrial sewage, inoculate above-mentioned Chlorella vulgaris and cultivate, wherein, Initial seeding density is 1.0 * 10
7Cfu/ml～6.0 * 10
7Cfu/ml, the condition of described cultivation is: periodicity of illumination is 10h: 14h～16h: 8h, and intensity of illumination is 5000lux～15000lux, and culture temperature is 20～30 ℃, and pH is 6.5～8.5, CO
2The gas ventilation amount is 0.01～0.2VVm, described CO
2The CO of gas
2Concentration is 15%～30% (v/v).After cultivating the suitable time, the nutrient solution of centrifugal photoreactor, results Chlorella vulgaris.
Described sewage work can be any sewage work.Preferably, described sewage work is the Sewage Plant of Wuxi curry favour connection Thermal Power Co., Ltd.
Described CO
2Gas can be the CO in any source
2Gas.Preferably, described CO
2Gas is that waste gas and air mixed that factory discharges are formed.Can directly mix, also can to waste gas do certain processing (as, dedusting, sulphur removal etc.) after remix, thereby effectively utilize waste gas, and reduce to pollute.
Described secondary effluent is the second pond water outlet of sewage work's second-stage treatment technique, and in the secondary effluent of different sewage treatment plant, nitrogen phosphorus equal size generally is more or less the same.Usually, in described secondary effluent, total nitrogen is 30～50.0mg/L, and ammonia nitrogen is 10～16.0mg/L, and total phosphorus concentration is 2.0～4.0mg/L, and pH is 6.5～8.0.
Incubation time is definite as required, and when disposing of sewage, preferably, after cultivating 480h, in described photoreactor, frustule concentration reaches 1.0 * 10
8Cfu/ml～5.0 * 10
8Cfu/ml, and the water quality of the described substratum in described photoreactor meets national category-A discharge water standard (total nitrogen concentration is 15.0mg/L, and ammonia nitrogen concentration is 1.0mg/L, and total phosphorus concentration is 0.5mg/L).Namely wherein nitrogen phosphorus equal size meets national category-A discharge water standard, realizes wastewater treatment efficiency.
When adopting Chlorella vulgaris of the present invention to dispose of sewage, preferably, be cultured to that in the described substratum in described photoreactor, total nitrogen concentration is lower than 15.0mg/L, ammonia nitrogen concentration is less than 1.0mg/L, and total phosphorus concentration is lower than 0.5mg/L.
When adopting Chlorella vulgaris accumulation grease of the present invention, preferably, the fat content that is cultured to the described Chlorella vulgaris in described photoreactor reaches 25%～40% (W).
In a third aspect of the present invention, provide the above-mentioned application of Chlorella vulgaris in sewage disposal.For example, be that chlorella C9-JN2010 is as the raw material of bioenergy take above-mentioned Chlorella vulgaris, secondary effluent after sewage work processes is as substratum take municipal wastewater and industrial sewage, by the advanced treatment of chlorella C9-JN2010, wherein, in water, total nitrogen concentration is lower than 15.0mg/L, ammonia nitrogen concentration is less than 1.0mg/L, total phosphorus concentration is lower than 0.5mg/L, and the nitrogen and phosphorus content in sewage has reduced respectively 80.0%～98.0% (W) and 80.0%～99.5% (W), CO
2Concentration reduces by 30.0～75.0% (v/v), and pH is 7.0～8.0.
In a fourth aspect of the present invention, provide the above-mentioned application of Chlorella vulgaris in grease production.For example, the raw material take chlorella C9-JN2010 as bioenergy, the secondary effluent after sewage work processes is as substratum take municipal wastewater and industrial sewage, and by the growth accumulation grease of chlorella, frustule concentration can reach 1.0 * 10
8Cfu/ml～5.0 * 10
8Cfu/ml, fat content 25%～40% (W).
Beneficial effect of the present invention is:
A. compare with other algae kind as the bioenergy raw material, Chlorella vulgaris of the present invention is mainly CO
2Tolerance strong (15%～30% (v/v)), it is strong that dirty ability is fallen, production intensity is high, and can be substratum with municipal wastewater and industrial sewage secondary effluent, do not need to add the nutritive elements such as nitrogen phosphorus, can alleviate the water resources burden and reduce sewage disposal and bioenergy production cost.
B. the Municipal Sewage Advanced Treatment Process that adopts Chlorella vulgaris of the present invention to build is carrying out sewage disposal and CO
2Can obtain bioenergy when reducing discharging, design is unique ingenious, is a kind of green production, reutilization of the sewage and bioenergy production technology cheaply, is suitable for large-scale promotion application.
C. the Municipal Sewage Advanced Treatment Process that adopts Chlorella vulgaris of the present invention to build can be realized nitrogen, phosphorus and CO
2High removal efficient (as, in water, total nitrogen concentration is lower than 15.0mg/L, and ammonia nitrogen concentration is less than 1.0mg/L, and total phosphorus concentration is lower than 0.5mg/L, and the nitrogen and phosphorus content in sewage has reduced respectively 80.0%～98.0% (W) and 80.0%～99.5% (W), C0
2Concentration reduces by 30.0%～75.0% (v/v)), and the content of concentration of algae and grease is high, and (concentration of algae that is respectively in photoreactor reaches 1.0 * 10
8Cfu/ml～5.0 * 10
8Cfu/ml, its fat content are 25%～40% (W)), can reach the double effects that protection of the environment and Biological resources are produced again, realize higher economy and social synthesis's benefit, be suitable for large-scale promotion application.
Description of drawings
Fig. 1 is the opticmicroscope aspect graph of Chlorella vulgaris of the present invention.
Fig. 2 A adopts Chlorella vulgaris cultural method of the present invention to cut apart to cultivate nitrogen, phosphorus and the CO that chlorella C9-JN2010 disposes of sewage
2The removal effect schematic diagram.
Fig. 2 B adopts Chlorella vulgaris cultural method of the present invention to cut apart to cultivate biomass and the interior fatty accumulation effect schematic diagram of born of the same parents that chlorella C9-JN2010 disposes of sewage.
Fig. 3 A is nitrogen, phosphorus and the CO that adopts Chlorella vulgaris cultural method cultured continuously chlorella C9-JN2010 of the present invention to dispose of sewage
2The removal effect schematic diagram.
Fig. 3 B is biomass and the interior fatty accumulation effect schematic diagram of born of the same parents that adopts Chlorella vulgaris cultural method cultured continuously chlorella C9-JN2010 of the present invention to dispose of sewage.
Fig. 4 A is nitrogen, phosphorus and the CO that adopts Chlorella vulgaris cultural method batch culture chlorella C9-JN2010 of the present invention to dispose of sewage
2The removal effect schematic diagram.
Fig. 4 B is biomass and the interior fatty accumulation effect schematic diagram of born of the same parents that adopts Chlorella vulgaris cultural method batch culture chlorella C9-JN2010 of the present invention to dispose of sewage.
Embodiment
In order more clearly to understand technology contents of the present invention, describe in detail especially exemplified by following examples.
In waste water, the transformation efficiency of nitrogen, phosphorus is that clearance is an important indicator weighing this process spent water treatment effect, and method of calculation are as follows:
The transformation efficiency of nitrogen, phosphorus (%)=(C
1-C
2) * 100%/C
1
In formula: C
1The content of initial nitrogen, phosphorus in-wastewater medium, mg;
C
2The content of nitrogen, phosphorus in-processed waste water nutrient solution, mg.
CO in 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 formula: C
1CO in-power plant waste gas
2Volumetric concentration, %;
C
2-CO in waste gas after processing
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 technique, and method of calculation are as follows:
The yield of grease (%)=m
1/ m
0* 100%
In formula: m
0The frond dry weight of-extraction grease, g;
m
1The quality of-thick grease, g.
Embodiment 1
Adopt the secondary effluent of processing through sewage work of fermentation mode that cutting method-two step method is coupled and the municipal wastewater after filtering and impurity removing bacterium or the assorted algae and industrial sewage as substratum, to this inoculation of medium chlorella C9-JN2010, wherein, initially connecing density is 6.0 * 10
7Cfu/ml, periodicity of illumination are 16h: 8h, and intensity of illumination is 15000lux, and culture temperature is 30 ℃, and pH is 7.0, CO
2The CO of gas
2Concentration is 30% (v/v), and air flow is 0.1VVm, and before cutting apart cultivation, after cultivation 48h, the interior concentration of algae of described photoreactor is up to 2.0 * 10
8Cfu/ml, and the nitrogen in described photoreactor, the content of phosphorus carry out fermented liquid when meeting 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 described photoreactor and add fresh municipal wastewater in described photoreactor, the total amount 100% that keeps described substratum, the concentration of algae that continues to be cultured in described photoreactor reaches 5.0 * 10
8Cfu/ml.Remaining described concentrated broth is filled with to have in the long-pending oil tank of sewage substratum that sea salt concentration is 2.5% (W) be the CO of 0.2VVm at air flow
2Concentration is the described CO of 30.0% (v/v)
2Under gas, two stage fermentation carries out oil and fat accumulation 48h, collects afterwards frond and extracts algae oil.
Utilize above-mentioned technique to cultivate chlorella C9-JN2010, nitrogen, phosphorus and CO
2Clearance and oil and fat accumulation relation see shown in Fig. 2 A-2B.
Analyzed as can be known by Fig. 2 A, in 0～48h culturing process, along with incubation time extends, chlorella C9-JN2010 is to nitrogen, phosphorus and CO
2Spending rate accelerate, the fastest at the 48h spending rate, reaches maximum value and be respectively 90.0% (W), 93.0% (W) and 70% (v/v), and frond concentration reaches 2.0 * 10
8Cfu/ml is cut apart subsequently, takes out after wherein 2/3 fermented liquid concentrate, with wherein 1/3 Returning reactor and be filled with salt-free fresh municipal wastewater, keeps substratum total amount 100%, and a part passes into long-pending oil tank Secondary Fermentation and amasss oily 48h in addition.Analyzed as can be known by Fig. 2 B, final concentration of algae reaches 5.0 * 10
8Cfu/ml, fat content are up to 40.0% (W).Later on by above-mentioned cultivation, semicontinuous cultivation, the successive segmentation 10 times again cut apart.
Employing continuously ferment mode and the municipal wastewater after filtering and impurity removing bacterium or the assorted algae and industrial sewage be the secondary effluent processed through sewage work as substratum, to this inoculation of medium chlorella C9-JN2010, wherein, initially connecing density is 3.5 * 10
7Cfu/ml, periodicity of illumination are 14h: 10h, and intensity of illumination is 5000lux, and culture temperature is 25 ℃, and pH is 8.5, CO
2The CO of gas
2Concentration is 25.0% (v/v), and air flow is 0.2VVm, cultured continuously 480h, nitrogen, phosphorus removal effect, CO
2Transformation efficiency (% (v/v)) and oil and fat accumulation relation see shown in Fig. 3 A-3B.
Analyzed by Fig. 3 A and learn, in cultured continuously in earlier stage, along with the time lengthening of cultivating, it is to nitrogen, phosphorus and CO
2Utilization accelerate.The speed of utilizing at 0～120h nitrogen is constantly accelerated, and clearance reaches 80.0% (W) subsequently, and is also unstable at 120～288h, and nitrogen removal efficiency tends towards stability at 80.0% (W) subsequently.The speed of utilizing at 0～96h phosphorus is constantly accelerated, and clearance reaches 80.0% (W) subsequently, and 96～216h is also unstable, and tp removal rate tends towards stability at 80.0% (W) subsequently.At 0～96h CO
2Utilize speed constantly to accelerate, clearance reaches 65.0% (v/v) subsequently, and is also unstable at 96～240h, subsequently CO
2Clearance tends towards stability at 65.0% (v/v).
Analyzed by Fig. 3 B and learn, in cultured continuously in earlier stage, along with the time lengthening of cultivating, the accumulation rate of frustule growth velocity and grease is accelerated.Constantly accelerate in 0～96h frond speed of growth, frond concentration reaches 3.0 * 10
8Cfu/ml, also unstable at 96～240h, frond concentration tends towards stability 3.0 * 10 subsequently
8Cfu/ml.0～192h oil and fat accumulation speed constantly accelerate and fat content up to 25.0% (W), after 192h, the content of frond grease tends towards stability at 25.0% (W).
Embodiment 3
Adopt batch fermentation mode and the municipal wastewater after filtering and impurity removing bacterium or the assorted algae and industrial sewage be through secondary effluent from sewage treatment plant as substratum, to this inoculation of medium chlorella C9-JN2010, wherein, initially connecing density is 1.0 * 10
7Cfu/ml, periodicity of illumination are 10h: 14h, and intensity of illumination is 7500lux, and culture temperature is 20 ℃, and pH is 6.5, CO
2Concentration be 15.0% (v/v), air flow is 0.01VVm, batch culture 480h, nitrogen, phosphorus removal effect, CO
2Transformation efficiency (% (v/v)) and oil and fat accumulation relation see shown in Fig. 4 A-4B.
Analyzed by Fig. 4 A and learn, cultivating early stage, along with the time lengthening of cultivating, it is to nitrogen, phosphorus and CO
2Utilization accelerate.In 0～168h nitrogen utilize speed constantly accelerate and clearance up to 98.0% (W), remain on subsequently this level always in 168～216h, but begin to drop to 91.0% (W) at 240h, remain on afterwards this level always.In 0～168h phosphorus utilize speed constantly accelerate and clearance up to 99.5% (W), remain on subsequently this level always in 168～264h, but slightly descend at 288h, be 98.0% (W), remain on afterwards this level always.CO in 0～168h
2Utilize the continuous quickening of speed and clearance up to 75.0% (v/v), and keep stable in 168～240h, utilize subsequently speed to slow down gradually, final CO
2Clearance only has 30.0% (v/v).
Analyzed by Fig. 4 B and learn, in batch culture in earlier stage, along with the time lengthening of cultivating, the accumulation rate of frustule growth velocity and grease is accelerated.In the batch culture process, reach 1.1 * 10 in the continuous quickening of 0～120h frond growth and frond concentration maximum
8Cfu/ml maintains this level in 120h～168h, frond concentration begins to descend subsequently, is finally 5.2 * 10
7Cfu/ml.The continuous quickening of frond oil and fat accumulation speed and frond fat content up to 30.0% (W), remain on this level subsequently always in 0～168h.
Therefore, of the present invention take municipal wastewater and industrial sewage as substratum and utilize Chlorella vulgaris C9-JN2010 high-density culture technique to carry out sewage disposal, to this inoculation of medium chlorella C9-JN2010, wherein, initially connecing density is 1.0 * 10
7Cfu/ml～6.0 * 10
7Cfu/ml, periodicity of illumination are 10h: 14h～16h: 8h, and intensity of illumination is 5000lux～15000lux, and culture temperature is 20～25 ℃, and pH is 6.5～8.5, CO
2(15.0%～30.0% (v/v)) air flow is 0.01～0.2VVm, after cultivating 480h, frustule concentration is high, and nitrogen, phosphorus equal size in described photoreactor meet national category-A discharge water standard, and (total nitrogen concentration is 15.0mg/L, ammonia nitrogen concentration is 1.0mg/L, and total phosphorus concentration is 0.5mg/L).After advanced treatment by chlorella, in water, total nitrogen concentration is lower than 15.0mg/L, and ammonia nitrogen concentration is less than 1.0mg/L, and total phosphorus concentration is lower than 0.5mg/L, nitrogen and phosphorus content in sewage has reduced respectively 80.0%～98.0% (W) and 80.0%～99.5% (W), CO
2Concentration reduces by 30.0～75.0% (v/v), and pH is 7.0～8.0.By the growth accumulation grease of chlorella, the concentration of algae in final photoreactor reaches 1.0 * 10
8Cfu/ml～5.0 * 10
8Cfu/ml, fat content can reach 25.0%～40.0% (W).Can realize CO
2Reduce discharging and bioenergy production, nitrogen, phosphorus and CO
2High remove efficient, and the content of concentration of algae, grease is high, reaches the double effects of protection of the environment and production Biological resources, easy and simple to handle fast, low, pollution-free, the society of cost and economical synthesis benefit high, be suitable for large-scale promotion application.
In sum, Chlorella vulgaris CO of the present invention
2Tolerance is strong, production intensity is high, its cultural method sewage disposal and produce bioenergy and other byproducts (as, protein, cosmetic material and animal-feed etc.) combine, reduce production costs, reach the comprehensive utilization of waste resource, realize the green production of energy-saving and emission-reduction, design ingenious uniqueness, be suitable for large-scale promotion application.
In this specification sheets, the present invention is described with reference to its specific embodiment.But, still can make various modifications and conversion obviously and 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 (7)
1. a Chlorella vulgaris (Chlorella vulgaris), is characterized in that, described Chlorella vulgaris is chlorella C9-JN2010, and deposit number is: CCTCC NO:M2010373.
2. Chlorella vulgaris cultural method, it is characterized in that, in photoreactor, secondary effluent after sewage work processes is as substratum take municipal wastewater and/or industrial sewage, and in described secondary effluent, total nitrogen is 30.0～50.0mg/L, and ammonia nitrogen is 10.0～16.0mg/L, total phosphorus concentration is 2.0～4.0mg/L, inoculate Chlorella vulgaris according to claim 1 and cultivate, wherein, Initial seeding density is 1.0 * 10
7Cfu/ml～6.0 * 10
7Cfu/ml, the condition of described cultivation is: periodicity of illumination is 10h:14h～16h:8h, and intensity of illumination is 5000lux～15000lux, and culture temperature is 20～30 ℃, and pH is 6.5～8.5, CO
2The gas ventilation amount is 0.01～0.2VVm, described CO
2The CO of gas
2Concentration is 15%～30%v/v.
3. Chlorella vulgaris cultural method according to claim 2, is characterized in that, after cultivating 480h, in described photoreactor, frustule concentration reaches 1.0 * 10
8Cfu/ml～5.0 * 10
8Cfu/ml, and the water quality of the described substratum in described photoreactor meets national category-A discharge water standard: total nitrogen concentration is 15.0mg/L, and ammonia nitrogen concentration is 1.0mg/L, and total phosphorus concentration is 0.5mg/L.
4. Chlorella vulgaris cultural method according to claim 2, is characterized in that, is cultured to that in the described substratum in described photoreactor, total nitrogen concentration is lower than 15.0mg/L, and ammonia nitrogen concentration is less than 1.0mg/L, and total phosphorus concentration is lower than 0.5mg/L.
5. Chlorella vulgaris cultural method according to claim 2, is characterized in that, the fat content that is cultured to the described Chlorella vulgaris in described photoreactor reaches 25.0%～40.0%W/W.
6. the application of Chlorella vulgaris according to claim 1 in sewage disposal.
7. the application of Chlorella vulgaris according to claim 1 in grease production.
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| CN103060308A (en) * | 2013-01-14 | 2013-04-24 | 安徽省农业科学院水产研究所 | Mutagenesis and screening method of high-yield chlorella new germplasm by means of chromosome doubling |
| CN103468578B (en) * | 2013-10-14 | 2015-01-14 | 哈尔滨工业大学 | Chlorella strain for synchronously treating municipal sewage and implementing grease accumulation |
| CN103981083B (en) * | 2014-06-09 | 2016-03-16 | 中国水产科学研究院渔业机械仪器研究所 | The closed mixotrophic cultivation method of a kind of micro-algae |
| CN106399109B (en) * | 2016-11-07 | 2020-08-07 | 辽宁东科电力有限公司 | Chlorella strain for synchronously realizing carbon fixation and denitration by combined treatment of wastewater and waste gas |
| CN106706909B (en) * | 2016-12-26 | 2019-04-19 | 中国科学院水生生物研究所 | A kind of method and device applied to assessment microalgae carbon dioxide tolerance |
| CN107140781A (en) * | 2017-04-05 | 2017-09-08 | 湖北工业大学 | Suitable for the advanced purification process of secondary effluent of municipal sewage plant |
| CN107916226B (en) * | 2017-10-13 | 2020-11-27 | 浙江省农业科学院 | Screening of microalgae capable of efficiently treating livestock and poultry sewage in pig farm and realizing resource utilization |
| CN108546648A (en) * | 2018-05-14 | 2018-09-18 | 江南大学 | A kind of environment-friendly type microalgae culture method |
| CN109234167B (en) * | 2018-10-10 | 2021-01-15 | 中国农业大学 | Chlorella and application thereof in biogas slurry purification |
| 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 |
| CN114015493B (en) * | 2021-09-26 | 2024-02-20 | 上海师范大学 | Method for extracting grease from algae |
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