CN103981083A - Closed type mixotrophic culture method for microalgae and culture system thereof - Google Patents
Closed type mixotrophic culture method for microalgae and culture system thereof Download PDFInfo
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Abstract
The invention relates to microalgae culture technologies and particularly relates to a closed type mixotrophic culture method for microalgae and a culture system thereof. The method comprises the steps: adding a culture medium, sterile water and a microalgae species into a material supplementing device, and then, conveying into a photo-reactor for constant-speed-flowing culture for 7-10 days under the conditions that the light intensity is 40-100 micromole/m<2>/s, the culture temperature is 25-37 DEG C and the flowing speed is 0.2-0.6m/s; enabling an algae solution to flow into a harvesting separator from the photo-reactor, discharging 70-90% of algae solution (taken as a fresh algae solution) out of the harvesting separator through an algae solution collection opening, refluxing the rest 10-30% of algae solution (taken as the microalgae species) into the material supplementing device from an algae species charging opening through a reflux pump, simultaneously supplementing the culture medium and the sterile water, then, continuing constant-speed-flowing culture, and repeating the above culture steps round by round, thereby completing the closed continuous culture of the microalgae. The culture method disclosed by the invention has the advantages that the continuous production of the microalgae is realized, the yield of the microalgae is increased, and the production cost is reduced.
Description
Technical field
The present invention relates to micro-algae culture technique, be specifically related to closed the holding concurrently of a kind of micro-algae and support culture method and culture systems thereof.
Background technology
Micro-algae (microalgae) is the unicellular algae existing with individual, chain or group form of a large class microcosmic, and size is not from several microns to hundreds of micron etc.Micro-algae is of a great variety, and nearly 200000-800000 kind, wherein has the kind more than 35000 that has recording according to estimates.Micro-algae is divided into the micro-algae of autotrophy, heterotrophic microalgae and holds concurrently and support micro-algae.From the sixties in 20th century, since first Japan start to carry out the extensive both culturing microalgae of commercialization, people can realize micro-algae at multiple fields such as protective foods, medical material, beauty treatment, feeds and commercially produce so far.
The cultivation of micro-algae large-scale autotrophy is mainly adopted in two ways: open microalgae cultivation system and sealed microalgae cultivating system.Open cultivation system is exactly that the outdoor natural sunlight that utilizes carries out both culturing microalgae, has the advantages such as expansion scale ratio is easier to, cost is lower.But open cultivation system is easily subject to the impact of external environment, as intensity of illumination, light application time, temperature and weather; Also be easily subject to the pollution of other algae kinds, bacterium and pathogenic microorganism.Sealed microalgae cultivating system refers to and utilizes substratum in airtight container, to carry out both culturing microalgae.Sealed microalgae cultivating system can be divided into fermentor tank, culture bag, plate smooth biochemical reactor and cast light biochemical reactor; Micro-algae large-scale heterotrophism cultivation is to utilize industrial fermentation method heterotrophism to cultivate micro-algae, can save space, improves output, avoids the pollution of assorted algae and bacterium, and culture condition easily controls, and is the development trend of micro-algae large-scale industrialized production.Part produce oil algae can make its training method transfer heterotrophism to by autotrophy by metabolic regulation, owing to not relying on illumination, does not also consume CO
2, can utilize common fermentor device to cultivate, but will strictly keep sterile state in culturing process.
The double foster cultivation of micro-algae can utilize organism, carbon source, carries out photosynthesis simultaneously.Micro-algae is held concurrently and supports in culturing process, and photoautotrophy and chemoheterotrophy are synchronous and relatively independent processes.Illumination has impact to two pathways metabolisms, but influence degree is different.
Available Microalgae refer to those batch production productions or application prospect, can be by a large amount of kinds of cultivating of biotechnology, in its cell, contained certain or some composition can be utilized by people.At present, Available Microalgae has a few class fronds such as spirulina, chlorella, Dunaliella salina, haematococcus pulvialis.Wherein, the fat content of chlorella is high, can be used as raw material prepared by the renewable energy resources.Chlorella can utilize at sun power and have function to carry out Fast Growth and accumulation grease.(2013) results of study such as Li Tingting show: adopt different training methods, carry out biomass and the lipid production experiment of chlorella.Under the condition of supporting of holding concurrently, can obtain and compare independent light autotrophy or independent heterotrophism, better growth performance, chlorella lipid content also increases considerably.
At present, the cultivation of chlorella large-scale commercial is mainly to carry out in bioreactor, and its floor space is large, the production cycle long, stand density is lower etc., has in a disguised form increased production cost, has limited extensive chlorella cultivation development.Finding a kind of high-density, chlorella cultural method cheaply, is important to promoting chlorella aquaculture industry development pole.
Summary of the invention
The object of this invention is to provide closed the holding concurrently of a kind of micro-algae and support culture method, adopt closed cultured continuously mode, in culturing process, with constant speed, emit the fresh algae liquid of part and process purification, another part is as the cultivation that refluxes of algae kind, supplement corresponding substratum and sterilized water simultaneously and make it to realize the closed cultured continuously of micro-algae, improve the productive rate of micro-algae.
Another object of the present invention is to provide closed the holding concurrently of a kind of micro-algae and supports culture systems, and this system floor space is few, simple in structure, cost is low.
In order to realize above technique effect, the present invention realizes as follows:
Closed the holding concurrently of micro-algae supported a culture systems, it is characterized in that: this culture systems is that feeder, photoreactor, results separator and reflux pump connect successively by pipeline;
The end face of described feeder is provided with substratum charging opening, sterilized water charging opening, initial algae kind charging opening and circulation algae kind charging opening, and the outlet of this feeder is connected with the import of photoreactor;
The outlet of described photoreactor is connected with the import of results separator;
Described results separator is provided with algae liquid and collects mouth and algae kind discharge outlet, and this algae kind discharge outlet is connected with described reflux pump, and the circulation algae kind charging opening on backflow pump outlet and feeder communicates.Preferably, described photoreactor is duct type or flat photoreactor; Described results separator is centrifugal or air-flotation type separator.
Closed the holding concurrently of above-mentioned micro-algae supported culture method, and its step comprises:
A, substratum, sterilized water and initial micro-algae algae kind are added in feeder and form and cultivate mixed solution from substratum charging opening, sterilized water charging opening and initial algae kind charging opening respectively, then be delivered to the mobile 7-10 days of cultivation of constant speed in photoreactor, intensity of illumination is 40-100 μ mol m
-2s
-1, culture temperature is 25-37 ℃, the flow velocity of algae liquid in photoreactor is 0.2-0.6m/s.The concentration of described initial micro-algae algae kind is 50-100mg/L.Preferably, the flow velocity of algae liquid in photoreactor is 0.3m/s.
Algae liquid in B, photoreactor flow in results separator, the algae liquid of 70%-90% in results separator is collected a mouthful discharge as fresh algae liquid from algae liquid, the algae liquid of residue 10%-30% passes through reflux pump as micro-algae algae kind, from circulation algae kind charging opening, be back in feeder, supplemental medium and sterilized water, then flow to the mobile cultivation of constant speed in photoreactor simultaneously; After fresh algae liquid is collected and mouthful to be discharged from algae liquid, and pass through the methods such as centrifugal, the air supporting results of dewatering.Micro-concentration of algae of separating in described results separator is 3-4g/L, and total fat content is 35%-50%.
C, repeating step (A) and step (B).Continuous being cycled to repeat, realizes the closed cultured continuously of micro-algae like this, and the flow velocity of whole double sample culture systems is 0.3m/s.
In feeder (1), adding substratum, to make the nutritive ingredient concentration of the cultivation mixed solution that contains in feeder be 7-12g/L glucose, 1-3g/L KNO
3, 600-650mg/L NaH
2pO
4h
2o, 85-95mg/LNa
2hPO
42H
2o, 240-250mg/L MgSO
47H
2o, 9-10mg/L EDTA, 0.05-0.07mg/LH
3bO
3, 14-15mg/L CaCl
22H
2o, 6-8mg/L FeSO
47H
2o, 0.2-0.3mg/L ZnSO
47H2O, 0.01-0.02mg/L (NH4)
6mo
7o
244H
2o, 0.1-0.2mg/L MnSO
4h
2o, 0.002-0.003mg/LCuSO
45H
2o.Preferably, the nutritive ingredient concentration of the cultivation mixed solution containing in feeder is 9g/L glucose, 1.7g/L KNO
3, 621mg/L NaH
2pO
4h
2o, 89mg/LNa
2hPO
42H
2o, 246.5mg/LMgSO
47H
2o, 9.3mg/L EDTA, 0.061mg/L H
3bO
3, 14.7mg/L CaCl
22H
2o, 6.95mg/LFeSO
47H
2o, 0.287mg/L ZnSO
47H2O, 0.01235mg/L (NH4)
6mo
7o
244H
2o, 0.169mg/LMnSO
4h
2o, 0.00249mg/L CuSO
45H
2o.
Described micro-algae is chlorella.
The invention has the beneficial effects as follows:
1, the present invention adopts closed the holding concurrently of micro-algae to support cultural method, by micro-algae algae kind, substratum and sterilized water be invested according to a certain percentage in culture systems, carry out closed hold concurrently to support cultivate, and the cultured algae liquid of the 10-30% in culture systems is re-used as algae kind, the serialization that training method realized micro-algae that is cycled to repeat is like this produced, and has improved the productive rate of micro-algae.
2, biomass and the total fat content of micro-algae that the present invention's cultivation obtains can reach 3-4g/L and 35%-50% respectively, the biomass of the micro-algae all obtaining higher than the common foster training method of holding concurrently and always fat content.And the maximum biomass dry weight of micro-algae of the present invention's production can reach the more than 1.8 times of heterotrophism cultivation, the more than 5.2 times of light autotrophy cultivation, the yield of biomass that glucose consumption obtains can reach 0.82g/g, and under the same terms, heterotrophism is cultivated only has 0.34g/g.
3, the floor space of the closed foster culture systems of holding concurrently of micro-algae of the present invention's employing is few, simple in structure, cost is low, and the productive rate of micro-algae is high, can realize large-scale industrial production.
Accompanying drawing explanation
Fig. 1 is that closed the holding concurrently of micro-algae of the present invention supported the structural representation of culture systems.
Fig. 2 is the biomass of the chlorella in embodiment 1 and the comparison diagram of total fat content and incubation time.
Wherein, 1 in Fig. 1 is feeder, the 2nd, and photoreactor, the 3rd, results separator, the 4th, reflux pump, the 11st, substratum charging opening, the 12nd, sterilized water charging opening, the 13rd, initial algae kind charging opening, the 14th, circulation algae kind charging opening, the 31st, algae liquid is collected mouth, the 32nd, algae kind discharge outlet.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1
(1), closed the holding concurrently of micro-algae supported the establishment of culture systems:
Feeder (1), photoreactor (2), results separator (3) and reflux pump (4) are connected to form to closed the holding concurrently of micro-algae successively by pipeline and support culture systems.Wherein, photoreactor (1) is duct type photoreactor; Results separators (3) are centrifugal separator.
The end face of described feeder (1) is provided with substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) for supplementing.The outlet of feeder is connected with the import of photoreactor; The outlet of photoreactor (2) is connected with the import of results separators (3); Results separator is provided with algae liquid and collects mouthful (31) and an algae kind discharge outlet (32), 70-90% through cultured micro-algae algae liquid in photoreactor discharges from algae kind discharge outlet (32), and through the centrifugal or air supporting mode results of dewatering, the algae liquid of 10%-30% is discharged from algae kind discharge outlet (32), through reflux pump, from circulation algae kind charging opening (14), squeeze in feeder.The concrete structure schematic diagram of the present embodiment as shown in Figure 1.
(2), closed the holding concurrently of chlorella supported cultural method
(A), the configuration of substratum: configure substratum and carry out high-temperature sterilization processing.
(B), the substratum configuring, sterilized water and micro-algae algae kind are added in feeder (1) from substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) respectively, the nutritive ingredient concentration of the cultivation mixed solution containing in feeder (1) is 9g/L glucose, 1.7g/L KNO
3, 621mg/L NaH
2pO
4h
2o, 89mg/LNa
2hPO
42H
2o, 246.5mg/L MgSO
47H
2o, 9.3mg/LEDTA, 0.061mg/L H
3bO
3, 14.7mg/L CaCl
22H
2o, 6.95mg/L FeSO
47H
2o, 0.287mg/L ZnSO
47H2O, 0.01235mg/L (NH4)
6mo
7o
244H
2o, 0.169mg/LMnSO
4h
2o, 0.00249mg/L CuSO
45H
2o.The concentration of initial micro-algae algae kind is 50-100mg/L, then micro-algae algae kind is delivered to the mobile 7-10 days of cultivation of constant speed in photoreactor (2), and intensity of illumination is 40-100 μ mol m
-2s
-1, culture temperature is 25-37 ℃, flow velocity is 0.3m/s.
(C) the algae liquid in photoreactor (2) flow in results separators (3), the algae liquid of 70%-90% in results separators (3) is collected mouthful (31) as fresh algae liquid from algae liquid and is discharged, residue 10%-30%, the concentration algae liquid that is 3-4g/L as micro-algae algae kind by reflux pump (4), from circulation algae kind charging opening (14), be back in feeder (1), simultaneously supplemental medium and sterilized water, then flow to and in photoreactor (2), carry out constant speed again and flow and cultivate.Subsequently cultured algae liquid is carried out to separation through results separators (3), fractional dose is that the algae liquid of 70%-90% is collected mouthful (31) discharge as fresh algae liquid from algae liquid, after being circulated in feeder (1), cultivates again the algae liquid that the concentration of residue 10%-30% is 3-4g/L, constantly recirculation, realizes the serialization of chlorella and produces.
From photoreactor, carry out sampling analysis, concrete outcome as shown in Figure 2, as can be seen from Figure, total fat content of chlorella constantly increases along with incubation time, can reach more than 45%, biomass cultivate within the 3rd day, to reach maximum be 3.7g/L, next tend towards stability.
Claims (7)
1. closed the holding concurrently of micro-algae supported a culture systems, it is characterized in that: this culture systems is that feeder (1), photoreactor (2), results separator (3) and reflux pump (4) connect successively by pipeline;
The end face of described feeder (1) is provided with substratum charging opening (11), sterilized water charging opening (12), initial algae kind charging opening (13) and circulation algae kind charging opening (14), and the outlet of this feeder (1) is connected with the import of photoreactor (2);
The outlet of described photoreactor (2) is connected with the import of results separators (3);
Described results separator (3) is provided with algae liquid and collects mouthful (31) and an algae kind discharge outlet (32), this algae kind discharge outlet (32) is connected with described reflux pump (4), and the circulation algae kind charging opening (14) on the outlet of reflux pump (4) and feeder (1) communicates.
2. closed the holding concurrently of micro-algae according to claim 1 supported culture systems, it is characterized in that: described photoreactor (2) is duct type or flat photoreactor; Described results separator (3) is centrifugal or air-flotation type separator.
3. closed the holding concurrently of micro-algae supported a culture method, and its step comprises:
A, substratum, sterilized water and initial micro-algae algae kind are added in feeder (1) and form and cultivate mixed solution from substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) respectively, then be delivered to the mobile 7-10 days of cultivation of constant speed in photoreactor (2), intensity of illumination is 40-100 μ mol m
-2s
-1, culture temperature is 25-37 ℃, flow velocity is 0.2-0.6m/s;
Algae liquid in B, photoreactor (2) flow in results separators (3), the algae liquid of 70%-90% in results separators (3) is collected mouthful (31) as fresh algae liquid from algae liquid and is discharged, and through the centrifugal or air supporting mode results of dewatering, the algae liquid of residue 10%-30% passes through reflux pump (4) as micro-algae algae kind, from circulation algae kind charging opening (14), be back in feeder (1), supplemental medium and sterilized water, then flow to the mobile cultivation of constant speed in photoreactor (2) simultaneously;
C, repeating step (A) and step (B).
4. closed the holding concurrently of micro-algae according to claim 3 supported culture method, it is characterized in that: the concentration of the initial micro-algae algae kind in the cultivation mixed solution in described step (A) is 50-100mg/L.
5. closed the holding concurrently of micro-algae according to claim 3 supported culture method, it is characterized in that: the nutritive ingredient concentration of adding the cultivation mixed solution after substratum in described feeder (1) is 7-12g/L glucose, 1-3g/LKNO
3, 600-650mg/L NaH
2pO
4h
2o, 85-95mg/LNa
2hPO
42H
2o, 240-250mg/L MgSO
47H
2o, 9-10mg/L EDTA, 0.05-0.07mg/L H
3bO
3, 14-15mg/L CaCl
22H
2o, 6-8mg/LFeSO
47H
2o, 0.2-0.3mg/L ZnSO
47H2O, 0.01-0.02mg/L (NH4)
6mo
7o
244H
2o, 0.1-0.2mg/L MnSO
4h
2o, 0.002-0.003mg/L CuSO
45H
2o.
6. closed the holding concurrently of chlorella according to claim 3 supported culture method, it is characterized in that: in described step (B), micro-concentration of algae of separating from results separator is 3-4g/L, and total fat content is 35%-50%.
7. closed the holding concurrently of chlorella according to claim 3 supported culture method, it is characterized in that: described micro-algae is chlorella.
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| CN106434284A (en) * | 2016-11-10 | 2017-02-22 | 山东建筑大学 | Modular microalgae culture system with rapid algal species expanding culture device |
| CN109467192A (en) * | 2018-12-10 | 2019-03-15 | 中国石油化工股份有限公司 | Nitric acid waste produces the processing method that waste water is cleaned in microalgae |
| CN113136344A (en) * | 2020-01-19 | 2021-07-20 | 中国石油化工股份有限公司 | Method and system for mixotrophic-autotrophic co-cultivation of photosynthetic microorganisms and method for production of biomass and bioenergy |
| CN113136339A (en) * | 2020-01-19 | 2021-07-20 | 中国石油化工股份有限公司 | Method for mixotrophic-autotrophic continuous culture of photosynthetic microorganisms, culture system and application thereof |
| CN113136339B (en) * | 2020-01-19 | 2023-07-14 | 中国石油化工股份有限公司 | Method for continuously culturing photosynthetic microorganisms by mixotrophic-autotrophic culture, culture system and application thereof |
| CN111500463A (en) * | 2020-04-13 | 2020-08-07 | 青岛旭能生物工程有限责任公司 | Method for continuously culturing chrysophyceae |
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